roczniki państwowego zakładu higieny - Wydawnictwa NIZP-PZH
Transcription
roczniki państwowego zakładu higieny - Wydawnictwa NIZP-PZH
ISSN 0035-7715 ROCZNIKI PAŃSTWOWEGO ZAKŁADU HIGIENY ANNALS OF THE NATIONAL INSTITUTE OF HYGIENE Quarterly 2014 Volume 65 Number 2 EDITOR and PUBLISHER: NATIONAL INSTITUTE OF PUBLIC HEALTH – NATIONAL INSTITUTE OF HYGIENE Warsaw, Poland ROCZNIKI PAŃSTWOWEGO ZAKŁADU HIGIENY (ANNALS OF THE NATIONAL INSTITUTE OF HYGIENE) Published since 1950 Quarterly, 4 issues in 1 volume per year (No 1 - March, No 2 - June, No 3 - September, No 4 - December) The journal is devoted to research studies on food and water safety, nutrition, environmental hygiene, toxicology and risk assessment, public health and other related areas Available at http://www.pzh.gov.pl/roczniki_pzh/ Edited and published by the National Institute of Public Health - National Institute of Hygiene, Warsaw, Poland Editor-in-Chief Deputy Editors Technical Editor Linguistic Editor Statistical Editor Kazimiera Ćwiek-Ludwicka Sławomir Garboś, Paweł Struciński Piotr Supranowicz Piotr Hołownia Daniel Rabczenko Subject Editors: Kazimierz Karłowski – food safety, Ewa Bulska – food and environmental analysis, Anna Gronowska-Senger – nutrition, Barbara Gworek – environmental hygiene, Jan K. Ludwicki – toxicology and risk assessment, Mirosław J. Wysocki – public health INTERNATIONAL EDITORIAL BOARD Stanisław Berger, Warsaw, Poland Jens Peter Bonde, Copenhagen, Denmark Brian T. Buckley, Piscataway, NJ, USA Krzysztof Chomiczewski, Warsaw, Poland Adrian Covaci, Antwerp, Belgium Małgorzata M. Dobrzyńska, Warsaw, Poland Jerzy Falandysz, Gdansk, Poland Antoni K. Gajewski, Warsaw, Poland Aleksander Giwercman, Malmö, Sweden Muhammad Jamal Haider, Karachi, Pakistan Bo Jönsson, Lund, Sweden Masahide Kawano, Ehime, Japan Grażyna Kostka, Warsaw, Poland Tao Li, Yunnan, China Honggao Liu, Kunming, China Halina Mazur, Warsaw, Poland Julia Melgar Riol, Lugo, Spain Regina Olędzka, Warsaw, Poland Krzysztof Pachocki, Warsaw, Poland Andrea Raab, Aberdeen, Scotland, UK Mark G. Robson, New Brunswick, NJ, USA Martin Rose, York, UK Kenneth S. Sajwan, Savannah, USA Józef Sawicki, Warsaw, Poland Jacques Scheres, Maastricht, The Netherlands Marcello Spanò, Rome, Italy Andrzej Starek, Cracow, Poland Ujang Tinggi, Archerfield Qld, Australia Bogumiła Urbanek-Karłowska, Warsaw, Poland Jesús Olivero Verbel, Cartagena, Colombia Stefan M. Waliszewski, Veracruz, Mexico Bogdan Wojtyniak, Warsaw, Poland Jan Żmudzki, Puławy, Poland Indexed/abstracted in: MEDLINE/Pubmed, Index Copernicus Int., EBSCO, Agro Base, Food Science and Technology Abstracts, Global Health, NISC SA Databases, EMBASE/Excerpta Medica, Polish Medical Bibliography/ Central Medical Library, Polish Ministry of Science and Higher Education (MNiSW), Web of knowledge Abstracts and full text are freely accessible on the journal’s website: http://www.pzh.gov.pl/roczniki_pzh/ The printed version of the journal is an original reference version. Editorial office address: Narodowy Instytut Zdrowia Publicznego - Państwowy Zakład Higieny ul. Chocimska 24, 00-971 Warsaw, Poland Phone: +48 22 54 21 266; Fax +48 22 849 35 13 e-mail: [email protected] © Copyright by the National Institute of Public Health - National Institute of Hygiene, Warsaw, Poland Edition: 515 copies ROCZNIKI PAŃSTWOWEGO ZAKŁADU HIGIENY [ANNALS OF THE NATIONAL INSTITUTE OF HYGIENE] Volume 65 2014 Number 2 CONTENTS REVIEW ARTICLES Flavonoids – food sources and health benefits. A. Kozłowska, D. Szostak-Węgierek .................................................................................................................................. 79 Diacetyl exposure as a pneumotoxic factor: a review. B. Starek-Świechowicz, A. Starek ...................................................................................................................................... 87 ORIGINAL ARTICLES Development and validation of a method for determination of selected polybrominated diphenyl ether congeners in household dust. W. Korcz, P. Struciński, K. Góralczyk, A. Hernik, M. Łyczewska, K. Czaja, M. Matuszak, M. Minorczyk, J. K. Ludwicki ................................................................................................................................................................... 93 Variations of niacin content in saltwater fish and their relation with dietary RDA in Polish subjects grouped by age. M. Majewski, A. Lebiedzińska ......................................................................................................................................... 101 Evaluating adult dietary intakes of nitrate and nitrite in Polish households during 2006-2012. A. Anyżewska, A. Wawrzyniak ......................................................................................................................................... 107 School pupils and university students surveyed for drinking beverages containing caffeine. M. Górnicka, J. Pierzynowska, E. Kaniewska, K. Kossakowska, A. Woźniak ................................................................ 113 The use of vitamin supplements among adults in Warsaw: is there any nutritional benefit? A. Waśkiewicz, E. Sygnowska, G. Broda , Z. Chwojnowska ........................................................................................... 119 Energy and nutritional value of the meals in kindergartens in Niš (Serbia). K. Lazarevic, D. Stojanovic, D. Bogdanović .................................................................................................................. 127 Comparing diabetic with non-diabetic overweight subjects through assessing dietary intakes and key parameters of blood biochemistry and haematology. K. Gajda, A. Sulich, J. Hamułka, A. Białkowska ............................................................................................................ 133 Nutritional values of diets consumed by women suffering unipolar depression. E. Stefańska, A. Wendołowicz, U. Kowzan, B. Konarzewska, A. Szulc, L. Ostrowska .................................................... 139 Awareness of factors affecting osteoporosis obtained from a survey on retired Polish subjects. N. Ciesielczuk, P. Glibowski, J. Szczepanik .................................................................................................................... 147 Responsiveness to the hospital patient needs in Poland. L. Gromulska, P. Goryński, P. Supranowicz, M.J. Wysocki ............................................................................................ 155 Instruction for authors ................................................................................................................................................. 165 Abstracts and full texts: http:// www. pzh.gov.pl/roczniki_pzh/ About the guide This provides comprehensive definitions and explanations to the terminologies used throughout toxicology, ecotoxicology, food safety, risk assessment, public health and other related disciplines. Over 1700 words or phrases explained The definitions given are based on many sources, however the main ones are from European Union legislation, OECD and FAO/ WHO documents together with official releases by the European Food Safety Authority (EFSA). To ensure convenient use for readers, the guide is divided into four parts as follows; o terms in Polish, their definitions and equivalent terms in English o terms in English, their equivalents and definitions in Polish o a list of Polish and English abbreviations o a list of the most important references About the authors The authors are leading experts in Poland employed in the Department of Toxicology and Risk Assessment at the National Institute of Public Health - National Institute of Hygiene in Warsaw. Their remit covers toxicology and risk assessment. They are also members of various working groups at the European Commission, EFSA, OECD, Codex Alimentarius Commission of the FAO/WHO and the Risk Assessment team set up by the Polish Chief Sanitary Inspector as well as other national and international bodies responsible for effecting safety policy and strategy on chemicals, food, water and environmental threats to health. How to place an order? Please apply to: Ms Diana Kowalczyk, Research Library National Institute of Public Health - National Institute of Hygiene 24 Chocimska Street,00-791 Warsaw, Poland Tel: +48 (22) 54 21 264, +48 (22) 54 21 262 e-mail: [email protected] Price: 85 PLN Rocz Panstw Zakl Hig 2014;65(2):79-85 FLAVONOIDS - FOOD SOURCES AND HEALTH BENEFITS Aleksandra Kozłowska1, Dorota Szostak-Węgierek2* 1 Department of Preventive Medicine and Hygiene, Institute of Social Medicine, Medical University of Warsaw, Poland 2 Department of Human Nutrition, Faculty of Health Science, Medical University of Warsaw, Poland ABSTRACT Flavonoids are a group of bioactive compounds that are extensively found in foodstuffs of plant origin. Their regular consumption is associated with reduced risk of a number of chronic diseases, including cancer, cardiovascular disease (CVD) and neurodegenerative disorders. Flavonoids are classified into subgroups based on their chemical structure: flavanones, flavones, flavonols, flavan-3-ols, anthocyanins and isoflavones. Their actions at the molecular level include antioxidant effects, as well the ability to modulate several key enzymatic pathways. The growing body of scientific evidence indicates that flavonoids play a beneficial role in disease prevention, however further clinical and epidemiological trials are greatly needed. Among dietary sources of flavonoids there are fruits, vegetables, nuts, seeds and spices. Consumption of these substances with diet appears to be safe. It seems that a diet rich in flavonoids is beneficial and its promotion is thus justifiable. Key words: flavonoids, cancer, cardiovascular diseases, neurodegenerative disorders STRESZCZENIE Flawonoidy to grupa związków bioaktywnych występujących powszechnie w żywności pochodzenia roślinnego. Aktualne dane literaturowe wskazują, że substancje te, spożywane wraz z dietą człowieka, wykazują działanie ochronne przed wieloma chorobami przewlekłymi, w tym przed niektórymi nowotworami oraz schorzeniami układu sercowo-naczyniowego, a ponadto pozytywnie wpływają na układ nerwowy. W zależności od struktury chemicznej wyróżnia się takie podklasy flawonoidów jak: flawony, flawanony, flawonole, flawanole, antocyjany i izoflawony. Przypuszcza się, że mechanizm działania tych substancji opiera się na ich silnych właściwościach antyoksydacyjnych oraz innych mechanizmach, takich jak zdolność do modulowania licznych szlaków enzymatycznych. W wielu badaniach wykazano ich korzystne działanie w prewencji chorób przewlekłych. Jednakże poznanie dokładnego metabolizmu tych substancji wymaga prowadzenia dalszych badań. Źródłami flawonoidów w diecie człowieka są warzywa, owoce, orzechy i nasiona, a także niektóre przyprawy. Spożywanie tych substancji wraz z dietą człowieka wydaje się być bezpieczne. Uzasadnionym zatem wydaje się promowanie diety bogatej we flawonoidy. Słowa kluczowe: flawonoidy, nowotwory, choroby sercowo-naczyniowe, choroby neurodegeneracyjne INTRODUCTION Flavonoids are a diverse group of plant metabolites with over 10,000 compounds that have been identified until now. However, only very few of them have been investigated in detail [25]. They have several important functions in plants, such as providing protection against harmful UV radiation or plant pigmentation. In addition, they have antioxidant, antiviral and antibacterial properties. They also regulate gene expression and modulate enzymatic action [25]. All naturally occurring flavonoids possess three hydroxyl groups, two of which are on the ring A at positions five and seven, and one is located on the ring B, position three. Biochemical actions of flavonoids depend on the presence and position of various substituent groups, that affect metabolism of each compound. They can be found in free or bound forms: aglycones or β-glycosides [17]. The flavonoid subclasses, based on types of chemical structure, include: flavonols, flavones, flavanones, flavanols, anthocyanins and isoflavones [17, 20]. Table 1 shows some common examples according to this classification. Antioxidant properties of foodstuffs depend not only on polyphenol content, but also on their type. For instance, quercetin and catechin demonstrate the greatest antioxidant properties in vitro [6, 11, 36]. However, their *Corresponding author: Dorota Szostak-Węgierek, Zakład Żywienia Człowieka, Warszawski Uniwersytet Medyczny, ul. Ciołka 27, 01-445 Warszawa, tel. +48 22 8360913, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 80 Nr 2 A. Kozłowska, D. Szostak-Węgierek Table 1. Subclasses of flavonoids; authors’ selection based on [17] Subclass Flavonols Flavones Flavanones Flavanols, Anthocyanins Isoflavones Examples of compounds Quercetin, kaempferol, myricetin Luteolin, apigenin, tangeretin Naringenin, hesperetin Catechin, epicatechin, epigallocatechin, glausan-3-epicatechin, proanthocyanidins Cyanidin, delphinidin, pelargonidin, malvidin Genistein, daidzein human metabolism is incompletely understood. Current studies on biological effects of flavonoids focuse on their absorption mechanisms, metabolism and bioavailability. Thus, in order to elucidate their physiological role, molecular studies are required. The results would enable to evaluate their effectiveness in the treatment and prevention of certain diseases, together with eventual risks arising from their use [18, 33]. FLAVONOIDS CONSUMPTION AND SAFETY At present, consumption of dietary flavonoids is regarded as safe. Nevertheless, it is worth noting that the use of pharmaceutical products that contain high doses of bioactive substances is increasing. Such supplements provide an alternative source of flavonoids to those obtained from the diet. It is of concern that the toxicity of concentrated sources of flavonoids is unknown, together with their interactions with other dietary components or taken medications [12]. Administration of large doses of a single flavonoid may decrease bioavailability of trace elements, vitamins or folic acid. Besides, it may exert an adverse effect on the thyroid function [12]. There is a special concern about possible side effects of taking several flavonoid-containing products at the same time as flavonoid-flavonoid interactions are little known so far [12]. A consumer may be misled that flavonoids are entirely safe because they are so-called ’natural’ products. Uncontrolled use of pharmaceutical preparations containing flavonoids may come out disadvantageous for health. Furthermore, the packaging labels for some dietary supplements have scant information about safety, adverse reactions, interactions, contraindications, and efficacy [7, 12]. It is clear that the molecular mechanisms of action of flavonoids need to be thoroughly understood and intensive research on this problem should be performed. However, it should be emphasised that in the light of recent findings the best and safest source of these substances is a properly balanced diet. DIETARY CONSUMPTION AND SOURCES OF FLAVONOIDS It is estimated that inhabitants of the Western Europe consume on average 100 – 1000 mg flavonoids/day/ person [17, 36]. This was confirmed by the European Prospective Investigation into Cancer and Nutrition (EPIC) study, which showed the median daily intake of flavonoids in Greek and Spaniard subjects equal to 93 mg (n ≥ 28,000) and 126.1 mg (n = 40,683) per Table 2. Content of flavonoids, according to their sub-classes, in chosen foodstuffs (mg/100g foodstuff); authors’ selection based on [2] Flavanones Artichokes Grapefruit juice Orange juice Oranges Limes Lemons Grapefruit Dried oregano 12.51 18.98 18.99 42.57 46.40 49.81 54.50 412.13 Flavonols Apples Cooked brussel sprouts Fresh figs Dried & sweetened cranberries Buckwheat Chicory Morello cherries American bilberries Blackcurrants Cooked asparagus Fresh cranberries Goji berries Red onions Flavones Kohlrabi 1.3 Red grapes 1.3 Lemons 1.9 Chicory 2.85 Celeriac 3.90 Green pepper 4.71 Artichokes 9.69 Fresh parsley 216.15 Dried oregano 1046.46 Dried parsley 4523.25 Anthocyanins Mean Hazel nuts 3.40 5.24 Morello cherries 5.47 Pears 6.91 Black grapes 7.09 8.94 9.41 10.59 11.53 15.16 21.59 31.20 38.34 Red table wine Pecan nuts Strawberries Red bilberries Raspberries Red cabbage Red currants Blackberries American bilberries Black currants Chickpeas Bilberries Rocket lettuce Radish Sorrel Elderberry juice concentrate 69.27 78.09 102.20 Dried parsley 331.24 Fresh capers 493.03 Flavanols Cooked broad 5.96 beans 8.41 Blackberries 8.6 Cocoa, dry powder 9.17 Dark chocolate 11.05 Black tea, brewed 15.99 Green tea, brewed Apple juice Apricots Peaches Apples Red table wine Pecan nuts 108.16 Aronia Elderberry juice concentrate 6.71 7.45 12.18 21.63 23.18 25.02 27.76 40.15 40.63 63.50 75.02 90.64 141.03 154.77 262.49 285.21 349.79 411.4 20.63 42.5 52.73 108.60 115.57 116.15 Nr 2 Flavonoids – food sources and health benefits person respectively [9, 39]. The Greek survey was performed in 1992 – 1996 and thus the results may be underestimated as the database concerning flavonoid levels in foodstuffs was incomplete that time. On the other hand, it is assumed that inhabitants of countries in the Far East, such as Japan, because of high intake of legumes, soy and tea, may consume up to 2 g of flavonoids daily [36]. In contrast, the Polish National Multi-centre Health Survey (WOBASZ) demonstrated that the mean flavonoid intake in the Polish population was 1 g/person/day [41]. Important dietary sources of flavonoids are vegetables, fruits, seeds, some cereals, together with wine, tea and certain spices. Table 2 demonstrates flavonoid content in chosen foodstuffs. It should be noted that the presence of particular flavonoids in vegetables and fruits depends on the crop variety, location and type of cultivation, as well as the specific plant morphological part [13]. Differences in flavonoid contents between varieties of species are usually small, although in a few cases very high amounts have been observed, e.g. in certain berries and tea prepared from leaves of the Quingmao tree [2]. EFFECTS OF FLAVONOIDS ON THE CARDIOVASCULAR SYSTEM The well recognised anti-oxidant properties of flavonoids resulted in the interest about their potential role in prevention of cardiovascular diseases [18]. For example, a recent study clearly showed health benefits of dietary flavonoids as there was a positive association between their intake and reduction of the risk of cardiovascular death in adult Americans [22]. The study demonstrated that both male and female subjects who consume large amounts of flavonoids (the top quintile) had the 18% lower mortality risk of cardiovascular diseases (CVD) compared to those whose intake was in the lowest quintile. Another study [3] demonstrated that high flavonoid consumption (flavones and flavanols) protected against hypertension. Subjects whose intake of these substances was in the top quintile, in comparison with those of the lowest consumption, exhibited the 8% risk reduction of development of this condition [3]. Atherosclerosis is a multifactorial disease. High blood concentration of oxidatively modified low density lipoproteins (ox-LDL) accelerates its development. Other causative factors include blood vessel inflammation and disorders of coagulation [18, 31]. Because of their antioxidant and chelating properties, flavonoids inactivate reactive oxygen species (ROS) and this way counteract plasma LDL oxidation and ameliorate inflammation of the blood vessel endothelium. Furthermore, flavonoids decrease activity of xanthine oxidase, 81 NADPH oxidase, and lipoxygenase ie. the enzymes that increase ROS production. Anti-arteriosclerotic action of flavonoids is related also to the reduction of inflammation in the blood vessel wall through inhibition of the influx of leucocytes. Flavonoids also decrease activity of such enzymes as 15-lipoxygenase (15-LOX) and cyclooxygenase (COX, particularly COX-2). These enzymes participate in formation of, prostaglandins and leukotrienes, substances that mediate inflammation, from arachidonic acid. Decline in their secretion results in reduction of synthesis of prostaglandin PGE2, leukotriene B4 and thromboxane A2, what in turn leads to decrease in inflammation and platelet aggregation. Inhibition of these enzymes results also in protection of LDL against oxidation and regulates capillary pressure back to normal [18]. Beyond of protection of blood vessels against ox-LDL, antiatheromatous action of flavonoids results also from suppression of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA) activity. This enzyme plays a key role in the synthesis of cholesterol in the human body, and thereby influences its plasma levels. Inhibition of its activity lowers intracellular cholesterol concentrations and results in the following increase in expression of LDL receptors. This in turn raises the cellular lipoprotein uptake and removal of cholesterol from the circulation. Hesperetin is a good example of a flavonoid, found in lemons and oranges, which reduces blood cholesterol level in the aforementioned way [18]. Furthermore, a randomised, double-blind study, that included cell culture, demonstrated in subjects with metabolic syndrome that oral administration of 500 mg of hesperetin daily over 3 weeks stimulated endothelial nitric oxide (NO) formation, what was probably related to the decreased activity of proinflammatory cytokines. This study showed that a three week hesperetin supplementation improves endothelial function, reduces inflammation and beneficially affects lipid profile in patients with metabolic syndrome [27]. It was shown that such flavonoids as rutin and its derivatives, along with hesperetin, help seal and reinforce blood vessel walls [18, 23]. These substances, similarly to vitamin C, enhance collagen synthesis and thus make the connective tissue in blood vessels more elastic. Rutin and its derivatives are used as a medication aimed at regulation of capillary permeability and improvement of peripheral circulation [23]. Flavonoids, such as quercetin or rutin, have anti-aggregating properties, and thereby reduce the risk of clot formation near the damaged endothelium [18]. By interaction with platelet integrins, these substances prevent platelets from sticking. They also stimulate NO formation in the vascular endothelium what facilitates vasodilation, and thus plays a key role in regulation of blood pressure [18]. 82 A. Kozłowska, D. Szostak-Węgierek Obesity is an important and independent risk factor for CVD and is strongly associated with dyslipidaemia, insulin resistance and type 2 diabetes [29]. Research on the effects of long-term flavonoid dietary supplementation in obese or normal body mass mice, in comparison with diet without addition of these substances, showed improved lipid profile, decreased insulin resistance and reduced visceral adipose tissue mass. The non-obese mice that consumed flavonoids demonstrated reduced levels of atherogenic cholesterol fractions (non-HDL cholesterol) [29]. These findings confirm the protective effects of flavonoids on the cardiovascular system. EFFECTS OF FLAVONOIDS ON THE NERVOUS SYSTEM Effectiveness of flavonoids in prevention of age-related neurodegenerative diseases has been much investigated in the recent years. It concerns particularly dementia, Parkinson’s and Alzheimer’s diseases. It seems that flavonoids can modulate neuronal function [21, 26, 34, 38]. Diets rich in these substances were shown to beneficially affect maintenance of human cognitive functions, probably through protection of neurons, enhancement of their function and regeneration [38]. Reactive oxygen and nitrogen species are involved in the development of many neurodegenerative diseases, whilst dietary flavonoids have been shown to counteract effectively oxidative neuronal damage. It was demonstrated that use of the extract from the gingobiloba plant, that is rich in flavonoids, may beneficially influence treatment of the age-related dementia and Alzheimer’s disease [1]. Tangeretin, a flavonoid that belongs to the flavone subclass, found mainly in citrus fruits, was shown to provide protection in Parkinson’s disease. Animal models of this condition is based on striatal damage by the neurotoxic substance 6-hydroxydopamine, what in turn leads to damage of the nigrostriatal pathway that connects the substantia nigra with the striatum. The latter is responsible, amongst others, for planning of body movements. Damage of this area underlies Parkinson’s disease. It was shown that tangeretin given to mice passes the blood-brain barrier (BBB) and protects the nigrostriatal pathway against adverse effects of 6-hydroxydopamine [8]. The PAQUID study (Personnes Age’es QUID), published in 2007, convincingly demonstrated that dietary flavonoids in the elderly support their cognitive functions [16]. The 10 years long observation was performed in 1640 subjects aged above 65 years, free from dementia at baseline. The data about flavonoid consumption were obtained by means of a food frequency questionnaire that listed foodstuffs containing these substances. At each visit (four times) every subject underwent cogni- Nr 2 tive tests including Mini-Mental State Examination, Benton’s Visual Retention Test and the ‘Isaacs’ Set Test. Participants whose flavonoid intake was in the two highest quartiles (ie. above 13.6 mg/day) had better cognitive function after 10 years than those who consumed less of these compounds. Moreover, subjects who ingested the least amounts of flavonoids (below 10.38 mg/day), lost on average 2.1 points in the Mini-Mental State Examination scale, while those with the highest consumption (above 17.7 mg/day) lost only 1.2 points. These findings demonstrated that regular consumption of dietary flavonoids exerts beneficial effect on cognitive function maintenance during aging [16]. The multitude of effects resulting from consuming flavonoids, both with foodstuffs and concentrated sources, appears to be related to two parallel processes. The first is regulation of the neuronal signal cascade what results in the inhibition of cell apoptosis that is caused by the action of neurotoxic substances. This promotes neuronal survival and differentiation [34]. Secondly, flavonoids seem to exert beneficial effects on the peripheral and central nervous systems by generation of changes in the cerebral blood flow. This can induce angiogenesis and growth of new nerve cells in the hippocampus. These processes are important for maintenance of neuronal and cognitive brain functions [34]. It seems that regular consumption of foods rich in flavonoids reduces the risk of neurodegenerative diseases and counteracts or delays the onset of age-related cognitive disorders. However, mechanisms of flavonoid action are not entirely clear. The question then arises as to when to use these substances to ensure their optimal effectiveness and which of them produce the strongest protection of the nervous system. Further studies on this wide group of compounds are therefore necessary to provide satisfactory answers to these questions. ANTICANCER ACTION OF FLAVONOIDS Chemoprevention is defined as the use of natural or synthetic substances to inhibit or reverse carcinogenesis [24]. Much attention, in this respect, is focused on flavonoids [4, 5, 10, 14, 19, 28, 35]. Epidemiological and clinical studies suggest that these compounds can prevent cancer through their interaction with various genes and enzymes [4]. It seems that biologically active substances found in foodstuffs may affect such stages of carcinogenesis as initiation, promotion and progression [24]. Many mechanisms of flavonoid action have been discovered. In the initiation and promotion stages, they include: inactivation of the carcinogen, inhibition of cell proliferation, enhancement of DNA repair processes, and reduction of oxidative stress. In the progression phase flavonoids may induce apoptosis, inhibit angio- Nr 2 83 Flavonoids – food sources and health benefits genesis, exhibit antioxidant activity, and also cytotoxic or cytostatic action against cancer cells [4, 19, 24, 40]. Prevention of metabolic activation of procarcinogens is related to flavonoid interaction with phase I enzymes that are responsible for metabolism of various endogenous or exogenous substrates. This results from inhibition of the cytochrome P450 enzymes, such as CYP1A1 and CYP1A2. Flavonoids thus protect against cellular damage arising from the activation of carcinogenic factors. Another mechanism of their action is related to reinforcement of mutagen detoxification through induction of the phase II enzymes, such as glutathione S-transferase (GST) and UDP-glucuronyl transferase (UDP-GT), which detoxify and eliminate carcinogens from the body [4, 15]. The anticancer effects of flavonoids can also be explained by the cell cycle inhibition. There are two classes of regulatory molecules responsible for cell cycle progression: cyclins and cyclin-dependent kinases (CDKs), which are activated under the influence of mitogenic signals within the cell. The uncontrolled activation of CDKs plays a key role in the pathogenesis of cancer. Various types of cancer are linked to excessive CDKs activity through gene mutation. For this reason, much research is increasingly focused on substances that can inhibit or modulate CDKs. These actions may exhibited by such flavonoids as: genistein, quercetin, daidzein, luteolin, kaempferol, apigenin, and epigallocatechin. Current evidence about the anticarcinogenic potential of flavonoids are however still equivocal. Some studies, that were performed in animals or various cell models, indicate that certain flavonoids may inhibit both cancer initiation and progression [10, 30, 37]. However, experiments on rats, conducted to determine the effect of tangeretin and quercetin on the risk of cancer occurrence arising from alphatoxin B1 induction (initiation and promotion of hepatic cancer) showed that whereas tangeretin administrated during tumour initiation reduced the number of precancerous lesions, quercetin did not exhibit such effect [30]. Another study showed that the development of lung cancer in mice exposed to tobacco smoke was arrested by consumption of both black and green teas. The results demonstrated that catechins contained in tea may protect against development of cancer [37]. A further research that tested influence of selected compounds on cultured human liver cells demonstrated that luteolin and apigenin also provided effective protection against cancer development. These flavonoids seem to inhibit CDKs. However, other studies indicate that flavonoids have weaker actions in vivo compared to that in vitro [11, 32]. An investigation on whether quercetin prevents lung cancer in mice showed that this substance, in spite of its strong biological activity, is not absorbed by these animals efficiently enough. The authors however suggest that further work should be focused on making the absorption mechanism of this substance more effective what would probably promote the expected anticancer action [32]. The studies quoted above were performed in animals, and so the conclusions should be extrapolated to humans with caution. Observational studies conducted on various human populations are also equivocal [10, 14, 35]. The Iowa Women’s Health Study investigated the effect of dietary flavonoid consumption on the incidence of cancer of the lung, colon, breast and pancreas in 34,708 post-menopausal women who were observed in 1986 – 2004. Their dietary habits were determined by means of a food frequency questionnaire. Results showed that regular flavonoid consumption significantly reduced the risk of the lung cancer, particularly in the women who had stopped smoking. However, there was no evident effect of flavonoid consumption on the risk of other cancers [5]. Another study, performed in 34,408 women (aged above 45 years), demonstrated no significant link between intake of foods rich in flavonoids and the risk of cancer [35]. Despite of these findings, a meta-analysis of 12 studies showed a reduced risk of breast cancer in women, especially postmenopausal, who consumed large amounts of flavonoids, such as flavonols and flavones [14]. Further studies are therefore required to assess the promising influence of flavonoids on the human body. SUMMARY Flavonoids exhibit manifold effects in protection of the human body. However, the underlying mechanisms are still not fully understood. According to current knowledge, a diet that includes flavonoid containing products should be promoted. Among foods that provide large amounts of these substances there are: citrus fruits, blueberries, blackberries, onions, peppers, a variety of teas, and also oregano and parsley. However, it should be emphesized that toxicity of flavonoids consumed in large doses remains unknown. For this reason, use of their dietary supplements should be considered with caution. The question arises as to when to use these substances to enable their most effective action, and as to which flavonoids are the most beneficial to human health. It is presumed that flavonoids exert stronger effects in vitro than in vivo, and thus it is important to determine their mechanisms of action at the molecular level. Further studies in this area are therefore greatly needed. Acknowledgement This paper was financed by the Warsaw Medical University, Poland 84 A. Kozłowska, D. Szostak-Węgierek Conflict of interest The authors declare no conflict of interest. REFERENCES 1. Bastianetto S., Zheng W.H., Quirion R.: The Ginkgo biloba extract (EGb 761) protects and rescues hippocampal cells against nitric oxide-induced toxicity: involvement of its flavonoid constituents and protein kinase. J Neurochem 2000; 74:2268–2277. 2. Bhagwat S., Haytowits D. B., Holden J. M.: USDA Database for the flavonoid content of selected foods. Nutrient Data Laboratory, Beltsville Human Nutrition Research Center Agricultural Research Service U.S. Departament of Agriculture 2011;1-159. 3. Cassidy A., O’Reilly E.J, Kay C., Samson L., Franz M., Forman J.P., Curhan G., Rimm E.B.: Habitual intake of flavonoid subclasses and incident hypertension in adults. Am J Clin Nutr 2011;93:338-347. 4. Chahar M.K., Sharma N., Dobhal M.P., Joshi Y.C.: Flavonoids: A versatile source of anticancer drugs. Pharmacogn Rev 2011; 5(9):1-12. 5. Cutler G.J., Nettleton J.A., Ross J.A., Harnack L.J., Jacobs D.R., Scrafford C.G., Barraj L.M., Mink P.J., Robien K.: Dietary flavonoid intake and risk of cancer in postmenopausal women: The Iowa Women’s Health Study. Int J Cancer 2008; 123(3):664-671. 6. Czeczot H., Podsiad M.: Antioxidant status of quercetin. Brom Chem Toks 2005;38(4):329-334 (in Polish). 7. Czerwiecki L.: Contemporary view of plant antioxidants role in prevention of civilization diseases. Rocz Panstw Zakl Hig 2009; 60(3):201-206 (in Polish). 8. Datla K.P., Christidou M., Widmer W. W., Rooprai H.K., Dexter D.T.: Tissue distribution and neuroprotective effects of citrus flavonoid tangeretin in a rat model of Parkinson’s disease. NeuroReport 2001; 12:3871–3875. 9. Dilis V., Trichopoulou A.: Antioxidant intakes and food sources in Greek Aduls. J Nutr 2010; 140:1274-1279. 10. Dong H., Lin W., Jing W., Taosheng C.: Flavonoids activate pregnane x receptor-mediated CYP3A4 gene expression by inhibiting cyclin-dependent kinases in HepG2 liver carcinoma cells. BMC Biochemistry 2010; 11:23. 11. Duthie G., Morrice P.: Antioxidant capacity of flavonoids in hepatic microsomes is not reflected by antioxidant effects in vivo. Oxid Med Cell Longev 2012; 2012:1-6 Article ID 165127. 12. Egert S., Rimbach G.: Which sources of flavonoids: complex diets or dietary supplements? Adv Nutr 2011; 2: 8-14. 13. Hallmann E., Rembiałowska E., Szafirowska A., Grudzień K.: Importance of fruits and vegetables from organic production in preventive medicine at the example of peppers from organic farming. Rocz Panstw Zakl Hig 2007; 58(1):77-82 (in Polish). 14. Hui C., Qi X., Qianyong Z., Xiaoli P., Jundong Z., Mantian M.: Flavonoids, flavonoid subclasses and brest cancer risk: a meta-analysis of epidemiologic studies. Plos One 2013;8:e54318. Nr 2 15. Krajka-Kuźniak V.: Induction of phase II enzymes as a strategy in the chemoprevention of cancer and other degenerative diseases. Postępy Hig Med Dosw 2007; 61:627-638 (in Polish). 16. Letenneur L., Proust-Lima C., Gouge A.L., Dartigues J.F., Barberger-Gateau P.: Flavonoid intake and cognitive decline over a 10-year period. Am J Epidemiol 2007; 165:1364-137. 17. Majewska M., Czeczot H.: Flavonoids in prevention and therapy diseases. Ter Leki 2009; 65(5):369-377 (in Polish). 18. Majewska-Wierzbicka M., Czeczot H.: Flavonoids in the prevention and treatment of cardiovascular diseases. Pol Merk Lek 2012; 32:50-54 (in Polish). 19. Majewski G., Lubecka-Pietruszewska K., Kaufman-Szymczak A., Fabianowska-Majewska K.: Anticarcinogenic capabilities of plant polyphenols: flavonoids and stilbene. Pol J Public Health 2012; 122(4):434-439. 20. Małolepsza U., Urbanek H.: Plant flavonoids as biochemical active compounds. Wiad Bot 2000;44(3/4):27-37 (in Polish). 21. Macready A.L., Kennedy O.B., Ellis J.A., Williams C.M., Spencer J.P.E., Butler L.T.: Flavonoids and cognitive function: a review of human randomized controlled trial studies and recommendations for future studies. Genes Nutr 2009; 4:227-242. 22. McCullough M.L, Peterson J.J., Patel R., Jacques P.F., Shah R., Dwyer J.T.: Flavonoid intake and cardiovascular disease mortality in a prospective cohort of US adults. Am J Clin Nutr 2012;95:454-464. 23. Miller E., Malinowska K., Gałęcka E., Mrowicka M., Kędziora J.: Role of flavonoids as antioxidants in human organism. Pol Merk Lek 2008; 24:556-560. 24. Olejnik A., Tomczyk J., Kowalska K., Grajek W.: The role of natural dietary compounds in colorectal cancer chemoprevention. Postępy Hig Med Dosw 2010; 64:175187 (in Polish). 25. Pollastri S., Tattini M.: Flavonols: old compounds for old roles. Ann Bot 2011; 108:1225-1233. 26. Prasain J.K., Carlson S.H., Wyss J.M.: Flavonoids and Age Related Disease: Risk, benefits and critical windows. Maturitas 2010; 66(2):163-171. 27. Rizza S., Muniyappa R., Iantorno M., Kim J.A., Chen H., Pullikotil P., Senese N., Tesauro M., Lauro D., Cardillo C., Quon M.J.: Citrus polyphenol hesperidin stimulates production of nitric oxide in endothelial cells while improving endothelial function and reducing inflammatory markers in patients with metabolic syndrome. J Clin Endocrinol Metab 2011;30(2):182-187. 28. Samuel T., Fadlalla K., Mosley L., Katkoori V., Turner T., Manne U.: Dual-mode interaction between quercetin and DNA-damaging drugs in cancer cells. Anticancer Res 2012; 32(1):61-71. 29. Shabrova E.V., Tarnopolsky O., Singh A.P., Singh A.P., Plutzky J., Vorsa N., Quadro L.: Insights into the molecular mechanism of the anti-atherogenic actions of flavonoids in normal and obese mice. Plos One 2011; 6:e24634. Nr 2 Flavonoids – food sources and health benefits 30. Siess M.H., Le Bon A.M., Canivenc-Lavier M.C., Suschetet M.: Mechanisms involved in the chemoprevention of flavonoids. Biofactors 2000; 12(1-4):193-199. 31. Szostak-Węgierek D.: The role of flavonoids in the prevention of atherosclerosis. Med Metabol 1999;3(2): 28-40 (in Polish). 32. Tan B., Liu Y., Chang K., Lim B.K., Chiu G.N.: Perorally active nanomicellar formulation of quercetin in the treatment of lung cancer. Int J Nanomedicine 2012; 7:651-661. 33. Tarko T., Duda-Chodak A., Zając N.: Digestion and absorption of phenolic compounds assessed by in vitro simulation methods. A review. Rocz Panstw Zakl Hig 2013; 64(2):79-84. 34. Vauzour D., Vafeiadou K., Rodrigues-Mateos A., Rendeiro C., Spencer J.P.E.: The neuroprotective potential of flavonoids: a multiplicity of effects. Genes Nutr 2008; 3:115-126. 35. Wang L., Lee I., Zhang S.M., Blumberg J.B., Buring j.E., Sesso H.D.: Dietary intake of selected flavonols, flavones, and flavonoid-rich foods and risk of cancer in middle-aged and older women. Am J Clin Nutr 2009;89:905-912. 36. Wilczyńska A., Retel M.: Evaluation of polyphenol dietary intake considering participation of honey. Probl Hig Epidemiol 2011; 92(4): 709-712 (in Polish). 85 37. Yang C.S., Chung J.Y., Yang G., Chhabara S.K., Lee M.J.: Tea and tea polyphenols in cancer prevention. J Nutr 2000; 130:472-478. 38. Youdim K. A., Joseph J.A.: A possible emerging role of phytochemicals in improving age-related neurological dysfunctions: a multiplicity of effects. Free Radic Biol Med 2001; 30:583–594. 39. Zamora-Ros R., Andres-Lacueva C., Lamuela-Raventos R., Berenguer T., Jakszyn P., Barricarte A., Ardanaz E., Amiano P., Dorronsoro M., Larranaga N., Martinez C., Sanchez M.J., Navarro C., Chirlaque M.D., Tormo M.j., Quiros J.R., Gonzalez C.A.: Estimation of dietetary sources and flavonoid intake in Spanish adult population (EPIC-Spain). J Am Diet Assoc 2010;110:390-398. 40. Zalega J., Szostak-Węgierek D.: Nutrition in cancer prevention. Part I. Plant polyphenols, carotenoids, dietary fiber. Probl Hig Epidemiol 2013; 94:41-49 (in Polish). 41. Zujko M.E., Witkowska A.M., Waśkiewicz A., Sygnowska E.: Estimation of dietary intake and patterns of polyphenol consumption in Polish adult population. Adv Med Sci 2012; 57(2):375-384. Received: 07.11.2013 Accepted: 12.03.2014 Mrs. President of the Polish Society of Nutritional Sciences Berlin, 20th February 2014 Dear Prof. Dr. hab. Anna Brzozowska: It is my pleasure to inform you that the website of the next 12th Conference of the Federation of European Nutrition Societies (FENS), which will be held in Berlin from the 20th to the 23rd October 2015 with the following title: “Nutrition and health throughout life-cycle – Nutritional sciences for the benefit of European consumers“ is now open. The website address is www.fensberlin2015.org. Please also find attached the Conference announcement. We kindly request you to upload it on the website of your society. From now on, you will receive information via the newsletter the technical secretariat will be sending. Please help us spread the information and forward the newsletters to all partners and professionals in your country. Prof. Dr. Heiner Boeing (Chairman of the Organizing Committee) and Dr. Helmut Oberritter (Secretary of the Organizing Committee), along with other members of the Organizing and Scientific Committees, are preparing a scientific programme which will be of great interest from both an educational and a participative level for all of us. I would like to highlight that Berlin is one of the most attractive European cities with one of the richest cultures. This perfect combination of science and culture will make the FENS conference a memorable event. I would like to thank you in advance for your support and cooperation and I look forward to meeting you in Berlin to exchange ideas and knowledge on the wide field of Nutrition. Yours sincerely, Ascensión Marcos President of FENS Rocz Panstw Zakl Hig 2014;65(2):87-92 DIACETYL EXPOSURE AS A PNEUMOTOXIC FACTOR: A REVIEW Beata Starek-Świechowicz, Andrzej Starek* Chair of Toxicology, Department of Biochemical Toxicology, Jagiellonian University, Medical College, Kraków, Poland ABSTRACT Diacetyl (2,3-butanedione) is a natural ingredient in foodstuffs which is not generally regarded health risk to consumers. Nevertheless, when manufactured for use as a synthetic flavouring/additive in processed foods (e.g. microwave popcorn), it poses a human health threat at the workplace. Its pneumotoxic action consists of inflammation, obstruction and restriction in the distal respiratory tract. One of the factors causing bronchiolitis obliterans is also recognised to be diacetyl. The scientific literature mostly describes human exposure to diacetyl in factory settings where functional disorders and structural changes of the respiratory system have been recorded, particularly bronchiolitis obliterans. Moreover, differential diagnosis shows pathological changes in the distal respiratory tract and in the pneumotoxic actions of diacetyl. Key words: food flavourings, additives, diacetyl, bronchiolitis obliterans STRESZCZENIE Diacetyl (2,3-butandion) jako naturalny składnik żywności nie wydaje się stwarzać zagrożenia dla zdrowia konsumentów. Związek ten będąc syntetycznym dodatkiem do żywności przetworzonej jest czynnikiem szkodliwym dla zdrowia pracowników zatrudnionych przy jego syntezie i stosowaniu w produkcji prażonej kukurydzy do mikrofalówek. Pneumotoksyczne działanie tego związku manifestuje się zmianami zapalnymi, obturacyjnymi i restrykcyjnymi, szczególnie w dystalnych drogach oddechowych. Diacetyl uznano za czynnik etiologiczny zarostowego zapalenia oskrzelików. Na podstawie piśmiennictwa przedstawiono narażenie na diacetyl w warunkach przemysłowych, zaburzenia czynnościowe i zmiany strukturalne w układzie oddechowym u osób narażonych, ze szczególnym uwzględnieniem zarostowego zapalenia oskrzelików. Ponadto zwrócono uwagę na diagnostykę różnicową zmian patologicznych w dystalnych drogach oddechowych oraz na mechanizmy pneumotoksycznego działania diacetylu. Słowa kluczowe: środki aromatyzujące do żywności, diacetyl, zarostowe zapalenie oskrzelików INTRODUCTION Diacetyl (2,3-butanedione; CAS: 431-03-8) is natural ingredient of butter, caramel, beer, coffee, cocoa, honey, vegetable oil, whisky, brandy and some other foodstuffs. It arises during primary milk maturation or in the manufacture of butter or margarine [1, 6] and is synthesised from methyl ethyl ketone and by special fermentation of glucose via methylacetylcarbinol [26, 38]. Diacetyl imparts an aroma/flavour similar to other diketones such as 2,3-pentanodione, 2,3-hexanodione and 2,3-heptanodione used in liquid form, pastes or powders to intensify such food flavour/aroma. It is a foodstuff ingredient of butter flavourings [5]. Synthetic diacetyl is used in the manufacture of popcorn, chips (i.e. french fries), confectionery, dairy products that include cheese, sour cream, mayonnaise, sauces, mari- nades and other processed foods and beverages where it imparts a buttery taste and aroma to foodstuffs [31, 37]. In order to fortify the natural odour of milk, the final concentrations of diacetyl used are 1-3 mg/kg. In microwave popcorn, levels of diacetyl used to range 1-25%, however this has now been decreased or indeed replaced by other substances with similar properties. Most confectionery flavouring contain 1% diacetyl [31]. In USA, France, Belgium, Norway and Sweden diacetyl is permitted as an additive to foodstuffs and in the EU it is manufactured on a large scale in Italy and the UK. The physico-chemical properties of diacetyl are given in Table 1. Industrial scale of microwave popcorn production is multi-stage. Sweet corn seed is stored in silos for a maximum of 2 months followed by sieving, air purification and roasting. By automated means, the product *Corresponding author: Andrzej Starek, Chair of Toxicology, Department of Biochemical Toxicology, Jagiellonian University, Medical College, Poland, Medyczna Street 9, 30-688 Kraków, phone +48 12 62 05 651, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 88 B. Starek-Świechowicz, A. Starek Table 1. Physico-chemical properties of diacetyl [1, 31, 37] Molecular formula Structural formula Appearance Sensory qualities Molecular mass Melting point Boiling point Density Vapour pressure Saturated vapour density Concentration of saturated steam Ignition temperature Auto-ignition temperature Limits of flammability Partition coefficient (log Pow) Solubility C4H6O2 CH3-CO-CO-CH3 Yellowish green liquid Buttery odour, similar to benzoquinone or chlorine, odour threshold in water 4 x 10-3 mg/l, in air 0.323 mg/m3 x 10-3, taste threshold: butter - 1 mg/kg, in milk – 1.4 x 10-2 – 2.9 10-2 mg/kg, in water - 5.4 x 10-3 mg/l 86.09 -1.2°C 88.0°C (1013 hPa) 1.1 (water = 1) 7.6 kPa (25°C) 3.0 (air = 1) 268,500 mg/m3 27.0°C (closed cup) 365.0°C 2.4-13.0% vol. (in air) -1.34 Soluble in water at 200 g/l (15°C), and in benzene, tetrachloromethane, acetone, propylene glycol, glycerol and ethanol. is then packed into polyethylene bags together with flavourings that contain diacetyl; with all these processes taking place in premises where the flavourings are also mixed. Flavourings consist of soya oil, salt, butter flavourings and food colourings which are mixed together at 64 - 66°C. When ready, the liquid form mixture is transferred into storage at temperatures >51°C [22]. Because of the relatively low boiling-point of diacetyl (88°C) but its high vapour pressure (7.6 kPa at 25°C), this compound very readily permeates the air atmosphere at the workplace. Human exposure to diacetyl only ever becomes toxic under industrial conditions. During its synthesis however, which occurs at temperatures ~360°C under enclosed conditions, there is no present of exposure; this only happens when the reactor is opened. Within the Dutch chemicals industry, operators where diacetyl was manufactured were exposed to 1.8 – 351 mg/m3 or 3 – 396 mg/m3 in more specific tasks. In addition to diacetyl, it was found that in such places the workplace air also contained around 0.4 – 29 mg/m3 of acetaldehyde [38]. It should note that emission of diacetyl from liquid or paste forms is more intense than from powders. In the USA microwave popcorn plants, exposure to diacetyl was from below the limits of detection of the analytical method (LOD) to 350.8 mg/m3. The mean arithmetic concentrations ± standard deviation (M ± SD) was 29 ± 66.2 mg/m3 whilst those in the microwave mixing rooms Nr 2 were 135.3 ± 98.8 mg/m3 [22]. In Poland, the average air concentrations of diacetyl found at the manufacture of confectionary were on average 51 mg/m3 [15]. However, the maximum admissible concentration (MAC) value for diacetyl has not yet been established. In the EU, an occupational exposure limit (OEL) for this compound is recommended at level of 0.352 mg/m3 [37], whereas the ACGIH in USA proposes using threshold limit value - time weighted average (TLV-TWA) and threshold limit value - short-term exposure limit (TLV-STEL) values of 0.04 mg/m3 and 0.07 mg/m3, respectively [1]. It has been found that there are over 100 different volatile organic compounds (VOCs) in the microwave area where microwave popcorn is manufactured, and the flavourings are in storage. These mostly included the ketons: diacetyl, butanone, 3-hydroxybutanone (acetoin), 2-nonanone and acetic acid. The average diacetyl concentration reached 125.5 (9.6 – 325.4) mg/m3. For machine operators this value was 5.6 (0.86 - 18.4) mg/m3 and for those packaging 6.8 (1.5-17.7) mg/m3; all other places were below 2.0 mg/m3. In addition, aerosols of salt and oil were found at mean concentrations of 0.13 ± 0.11 mg/m3 [22]. For quality control purposes, the opening of microwave popcorn bags released 780 µg diacetyl/bag into the air as well as other VOCs [36]. Thus, the actual human exposure to flavouring ingredients is in general mixed. Tobacco smoke contains 300-430 µg diacetyl/cigarette [8]. Occupational exposure controls implemented in the USA during 2000-3 led to decrease diacetyl levels at the workplace air. When the flavourings are mixed, peak diacetyl levels were reduced from 462 to 0.97 mg/m3 [34]. In turn, average concentrations of this compound at the places where mixing occurs, machines are operated and the popcorn product is packaged, have been reduced from 205, 9.9 and 2.9 mg/m3 in 2000 to 10.3 mg/m3 or below LOD in 2003, respectively [17]. ACUTE TOXIC EFFECTS Repeated exposures to diacetyl at single instances or at short intervals lead to pronounced irritation to the eyes, respiratory tract and skin. Symptoms include persistent cough, muco-purulent secretion from the respiratory tract, wheezing, dyspnoea/breathlessness, fatigue, mild fever, generalized body aches and skin rash. The substance can also cause central nervous system (CNS) depression, sometimes leading to a loss of consciousness [14, 37]. A case study on a 36 year old never-smoking man with normal lung function and normal serum α-1 antitrypsin activity, but exposed to diacetyl for several hours revealed sore and reddened his eyes, painful eyes and eyelids, together with a sticky conjunctival secretion. Spirometry was normal after 3 months of exposure, but Nr 2 89 Diacetyl exposure as a pneumotoxic factor when repeated 6 months later, showed decease the flow rate of the midportion of the expiratory spirogram (the FEF 25%–75%) attaining 30% of the predicted value, thus indicating altered small-airway function [12]. CHRONIC TOXIC EFFECTS OBLITERATIVE BRONCHIOLITIS Occupational exposure to diacetyl-containing food flavourings has led to respiratory disease including obstruction of the small airways [3, 4, 16, 21-24] frequently coupled with persistent dry cough and breathlessness after exertion [25], together with spirometric changes [4, 16, 18, 22-24, 33, 38] (Table 2). Bronchiolitis obliteranswas found in 5/184 persons exposed to diacetyl during its synthesis or in microwave popcorn manufacture. This exposure was however mixed because the air at the workplace also contained acetoin, acetaldehyde and acetic acid [4, 38]. The incidence of bronchiolitis obliterans, often named constructive bronchiolitis or obliterative bronchiolitis, is a rather rare but irreversible disease of the lungs, where obstruction occurs in the distal air passages [11, 40]. The bronchioles become inflamed, exhibit submucosal fibrosis and fibrous tissue proliferation in capillary adventitia and adjacent interalveolar septa [4]. Centrifugal scarring can lead to more frequent obstruction of the small-airways and then complete blockage. These obstructions arise through excess fibroblast proliferation and accumulation of collagen deposits. A loss of lung tissue elasticity resulting from damage to collagen and elastin fibres, as well as secondary atonia of the lung parenchyma causes the peripheral bronchi to collapse and increases air flow resistance in the bronchiole ends; which explains the symptoms of shortness of breath upon exertion. Clinical symptoms are a dry cough and dyspnoea, particularly during expiration that can either appear progressively or suddenly. Blocked airways prevent the rapid emptying of the distal part of the respiratory tract during expiration [11]. This leads to excessive lung aeration resulting in ‘air-trapping’ pockets which become visible during radiology [4, 38]. The next stage in the progression, is an increase in total lung capacity (TLC) and other the volumes of the lungs, termed hyperinflation. Such changes are permanent and are not reversible by drugs that dilate the bronchi [11]. Factors responsible for these changes can be irritants like chlorine, sulphur dioxide, phosgene or ammonia [20]. Bronchiolitis obliterans is diagnosed by histopathology of a lung biopsy [4]. Whenever respiratory tract blockages are coupled with changes observed by radiology, using high resolution computed tomography (HRCT), then this condition is defined as bronchiolitis obliterans syndrome – BOS [4, 9, 39]. If there are no radiological changes observed then it is recommended Table 2. Epidemiological findings for chronic effects of diacetyl on the respiratory system 3. Subject numbers Diacetyl concentration Study results studied (mg/m3) Flavouring manufacture for foodstuffs Cross-sectional 34 0.11 - 0.80 (PS) Decreased FEV1 or TLV values, lung obstruction. BO, decreased FEV1 or TVC values. Longitudinal 175 3.04 - 404.5 (PS) Increased neutrophils, chronic cough, bronchial 1.83 - 356.9 (AS) asthma. Microwave popcorn manufacturing Case report series 3 Not available Decreased FEV1 or TVC values. 4. Cross-sectional 5. Case report series 6. No. 1. 2. Study type 117 2.0 – 115.5 (PS) 9 6.8 - 115.6 (AS) Cross-sectional 108 0.09 - 26.9 (AS) 7. Cross-sectional 135 8. Cross-sectional 9. Cross-sectional 10. Longitudinal References [16] [38] [39] [33] Lung obstruction, dysopnoea , fatigue, skin irritation. BO, decreased FEV1 or TVC values. [21] [3] 0.09 - 26.9 (AS) Lung obstruction, increased neutrophils-OR: 3.8 (1.3 – 11.5). Increased muscle tone of Chest. 537 0.72 - 4.3 (AS) 0.07 - 3.6 (PS) Decreased FEV1 or FVC values, dyspnoea, chronic cough, wheezing. [18] 3 < LOD Decreased FEV1 or FVC values, lung obstruction. [23] 725 1.25 - 3.08 (PS) Decreased FEV1 or FVC values, lung obstruction. [24] AS – area sampling PS – personal sampling LOD – detection limit BO – bronchiolitis obliterans confirmed by histology and radiology FEV1 – the forced expiratory volume in 1 second FVC – the forced vital capacity [4] [2] 90 B. Starek-Świechowicz, A. Starek that the concept of ‘fixed airways obstruction’ be used. A non-invasive method of evaluation the levels of obstructive changes in the airways is spirometry. Here, a forced expiratory volume in 1 second (FEV1) value below 60% of the predicted value, as well as lowered forced vital capacity (FVC) and the ratio of these measures (FEV1/ FVC) indicate fixed airflow obstruction of the distal respiratory tract [9]. Because this condition is rare, there is a potential for mis-diagnosing it as either bronchial asthma, bronchitis, emphysema or pneumonia. There are a number of differences between bronchiolitis obliteransand other more common obstructive lung diseases such as asthma or chronic obstructive pulmonary disease (COPD). For example, in asthma, the degree of airway obstruction expressed by the FEV1/ FVC ratio is not long lasting and alters from day to day. Furthermore, FEV1 values return to normal when treating asthma with short-term bronchiole dilators. Moreover, COPD nearly always results in decreased diffusion capacity of the lungs for carbon dioxide (CO2) together with excessive reactivity of respiratory tract. These described symptoms are not characteristic features of bronchiolitis obliterans. This condition can be distinguished from fibrotic changes of the lung, such as those in idiopathic pulmonary fibrosis or asbestosis by means of impairment of air flow but not FVC value. Notwithstanding, during the early disease stage, the TLC value is raised however, when fibrotic lung changes occur, this indicator becomes lowered [11]. When screening for early signs of bronchiolitis obliterans, the FEF 25-75% value is recommended coupled with both the diffusing capacity of the lung for carbon monoxide (DLCO) and lung volumes which are highly regarded diagnostic features of this condition [4, 38]. When diagnosing airway inflammation, the bronchoalveolar lavage (BAL) is used. Amongst other things, this procedure provides a profile of inflammatory cells and interleukin concentrations of IL-6 and Il-8; these being mediators of inflammation [11]. Salivary tests can also yield relevant information, where workers exposed to food flavourings show increased levels of neutrophils (>1.63x105 ml-1) with odds ratio (OR) 3.8 (95% CI: 1.311.5) as well as increased IL-8 and eosinophil cationic protein (ECP) concentrations [3]. Bronchiolitis obliterans can indeed lead to death or qualify for a lung transplant. However milder symptoms of obstructed or restricted airways are usually seen in those persons exposed to diacetyl. Studies performed on workers at an USA microwave popcorn factory showed 9/450 persons (aged 27 - 51 years) suffering from bronchiolitis obliterans, of whom only 2 were also confirmed histologically. Five out of these nine workers were employed as mixers of flavourings. Most subjects had never smoked cigarettes and the lengths of employment varied between 1 – 17 years. Those who had worked from 5 Nr 2 months to 9 years experienced coughing, dyspnoea and wheezing. TEV1 values were 14.0 – 66.8% of expected whilst HRCT showed significant bronchial thickening. Once the exposures had stopped then all persons recovered normal lung function within 2 years [4]. TOXICOKINETICS AND MECHANISMS OF PNEUMOTOXIC ACTION Diacetyl is a hydrophilic substance and is readily absorbed by the upper respiratory tract; as observed in rat studies [29]. The pharmacokinetics of diacetyl using a physiologically-based pharmacokinetic (PBPK) modelling, indicates that inhaling diacetyl penetrates more deeply in those persons breathing through their mouths than in rats breathing through their noses. Uptake of this substance by the upper respiratory mucosa was clearly intensified by its metabolism. Mucociliary clearance of diacetyl was dominated by its biotransformation and slow reaction with arginine. The absorption efficiency in rats was greater at lower levels of exposure. At higher exposures, the enzymes metabolizing diacetyl become saturated and hence levels of the parent compound are increased which pass into the distal regions of the respiratory tract. It has been calculated that when rats are exposed to 3.58 mg/ m3 diacetyl concentration, only 2% reach the bronchi; for persons breathing through their noses this value is 8%. However in the latter case, when breathing via the mouth then this amount becomes slightly increased. During mild exercise and whilst breathing through the mouth, 24% of the received diacetyl dose passes into the bronchi. The calculated amount of diacetyl present in the bronchial tissue of an exposed person, at rest, breathing through their nose is 5 times higher than in rats but 7 times higher when breathing through the mouth, but in the latter becomes 20 – 40 times higher during mild exercise [10, 29]. Concentration differences in the distribution of diacetyl within the respiratory tract for humans and rodents provoke differences in the localisation of any pathological changes. Whilst in the former these occur mainly in the distal regions, in rodents they affect the upper respiratory tract. Exposing rats or mice to relatively high concentrations of diacetyl results in fibrinopurulent inflammation or necrotic rhinitis of regions such as nose, larynx, trachea and bronchus together with the loss of microvilli and cilia of ciliated epithelium [13, 28]. Diacetyl, like acetoin and 2,3-butanediol is a metabolite of acetaldehyde. The former two are rapidly reduced in mammalian tissues to 2,3-butanediol which undergoes glucuronidation before being excreted. Acetoin is enzymatically formed through the pyruvate dehydrogenase complex or by a non-enzymatic reaction between ace- Nr 2 91 Diacetyl exposure as a pneumotoxic factor taldehyde and pyruvate in the presence of thiamine. Within mammalian hepatic tissue or homogenates thereof, acetoin and 2-3-butanediol are very slowly oxidized to diacetyl and as a result they accumulate in other tissue e.g. brain [32]. Diacetyl is therefore an endogenous compound with a significant toxicological role. Its reduction is catalysed by nicotinamide nucleotide dependent diacetyl reductase present in mammalian tissue [30, 32], which is regarded as a detoxification mechanism. This reduction is inhibited by butyric acid which is present in foodstuffs as flavourings/additives. It has been demonstrated that the efficiency of absorbing diacetyl alone in the isolated upper respiratory tract of the rat is 36% of the received dose, but in the presence of butyric acid this value then significantly (statistically) decreases to 31%. Thus inhibiting diacetyl metabolism in the upper airways, may increase its transport to the bronchioles, where its toxicological effects are manifested [29]. The mechanism by which bronchiolitis obliterans arises still remains unknown. It is suggested that diacetyl directly damages airway epithelia resulting from an induced inflammatory process and by carbonyl and oxidative stress caused by the generation of reactive dicarbonyl and reactive oxygen species [41]. The presence of two adjacent carbonyl groups in the diacetyl’s carbon chain enhances the reactivity of these groups with protein amino groups. When such products are formed, this leads to excessive cytokine production and chronic states of inflammation. Another source of these products comes from cross-linking with structural proteins like collagen and laminin or they can be the result of sclerosis processes in the lungs, blood vessels or other tissue [27]. Furthermore, this leads to, amongst other effects, inhibition of muscle enolase [35] and pyruvate kinase in erythrocytes [19], key enzymes of glycolysis. Diacetyl also modifies arginine in the inner mitochondrial membrane, impairing its permeability [7]. Such disruption leads to an energy deficit and cell death. It is postulated that the processes of repairing the distal respiratory tract occur during the uncontrolled phases of fibroblast and myoblast proliferation. This then leads to an accumulation of fibroblasts and collagen deposition, as well as scarring that is responsible for the partial or complete obstruction of bronchioles [11]. However, during the formation and development of bronchiolitis obliterans, symptoms of allergic pulmonary inflammation, asthma , diffuse interstitial fibrosis and granuloma can be excluded [11]. CONCLUSIONS 1. Diacetyl, as an ingredient of foodstuff flavourings/ additives, is a pneumotoxic substance under specific workplace conditions. 2. Chronic exposure to this substance, particularly during the manufacture of microwave popcorn, leads to obstructive changes in the distal regions of the respiratory tract. 3. Diacetyl is one of the factors responsible for causing bronchiolitis obliterans. 4. Reducing the occupational exposure to diacetyl in industry seems to be the best method for preventing bronchiolitis obliterans. 5. The presence of diacetyl as a flavouring/additive in foodstuffs does not appear to constitute a health hazard to consumers. Conflict of interest The authors declare no conflict of interest. REFERENCES 1. ACGIH, Threshold Limit Values for Chemical Substances and Physical Agents & Biological Exposure Indices. American Conference of Governmental Industrial Hygienists 2012. 2. Akpinar-Elci M., StempleK.J., Elci O.C., Dweik R.A., Kreiss K., Enright P.L.: Exhaled nitric oxide measurement in workers in a microwave popcorn production plant. Int J Occup Environ Health 2006; 12:106-110. 3. Akpinar-Elci M., Stemple K.J., Enright P.L., Fahy J.V., Bledsoe T.A., Kreiss K., Weissman D.N.: Induced sputum evaluation in microwave popcorn production workers. Chest 2005; 128:991-997. DOI: 10.1378/chest.128.2.991. 4. Akpinar-Elci M., Travis W.D., Lynch D.A., Kreiss K.: Bronchiolitis obliterans syndrome in popcorn production plant workers. Eur Respir J 2004;24:298-302. DOI: 10.1183/09031936.04.00013903. 5. Boylstein R., Piacitelli C., Grote A., Kanwal R., Kullman G., Kreiss K.: Diacetyl emissions and airborne dust from butter flavorings used in microwave popcorn production. J Occup Environ Hyg 2006; 3:530-535. 6. Colley J., Gaunt L.F., Lansdown A.B.G., Grasso P.: Acute and short-term toxicity of diacetyl in rats. Fd Cosmet Toxicol 1969; 7:571-582, quoted [1]. 7. Eriksson O., Fontaine E., Bernardi P.: Chemical modification of arginines by 2,3-butanedione and phenylglyoxal causes closure of the mitochondrial permeability transition pore. J Biol Chem 1998; 273(20):12669-12674. 8. Fujioka K., Shibamoto T.: Determination of toxic carbonyl compounds in cigarette smoke. Environ Toxicol 2006;21:47-54. 9. Galbraith D., Weill D.: Popcorn lung and bronchiolitis obliterans: a critical appraisal. Int Arch Occup Environ Health 2009; 82:407-416. DOI: 10.1007/s00420-008-0337-x. 10. Gloede E., Cichocki J.A., Baldino J.B., Morris J.B.: A validated hybrid computional fluid dynamics-physiologically based pharmacokinetic model for respiratory tract vapor absorption in the human and rat and its application to inhalation dosimetry of diacetyl. Toxicol Sci 2011; 123:231-246, quoted [1]. 92 B. Starek-Świechowicz, A. Starek 11. Harber O., Saechao K., Boomus C.: Diacetyl-induced lung disease. Toxicol Rev 2006;25(49):261-272. 12. Hendrick D.J.: „Popcorn worker’s lung“ in Britain in a man making potato crips flavouring. Thorax 2008; 63:267-268. 13. Hubbs A.F., Goldsmith W.T., Kashon M.L., Frazer D., Mercer R.R., Battelli L.A., Kullman G.J., Schwegler-Berry D., Friend S., Castranova V.: Respiratory toxicologic pathology of inhaled diacetyl in Sprague-Dawley rats. Toxicol Path 2008; 36:330-344. DOI: 10.1177/0192623307312694. 14.IPCS, International Programme on Chemical Safety. International Chemical Safety Cards. ICSC: 1168. 2,3-Butanedione 2007. 15. Jeżewska A.: Personal information, CIOP, 2007 (data not published). 16. Kanwal R., Kullman G.: HETA 2006-0303-3043, Report on severe fixed obstructive lung disease in workers at a flavoring manufacturing plant. Health Hazard Evaluation Report. National Institute for Occupational Safety and Health (NIOSH), Cincinnati, OH 2007, quoted [25]. 17. Kanwal R., Kullman G., Fedon K.B., Kreiss K.: Occupational lung disease risk and exposure to butter-flavoring chemicals after implementation of controls at a microwave popcorn plant. Public Health Rep 2011; 126:480-494. 18. Kanwal R., Kullman G., Piacitelli C., Boylstein R., Sahakian N., Martin S., Fedan K., Kreiss K.: Evaluation of flavorings-related lung disease risk at six microwave popcorn plants. J Occup Environ Med 2006;48(2):149157. DOI: 10.1097/01.jom.0000194152.48728.fb. 19. Kilinc K., Özer N.: Irreversible inactivation of human erythrocyte pyruvate kinase by 2,3-butanedione. Arch Biochem Biophys 1984;230(1):321-326. 20. King T.E., jr.: Bronchiolitis. In: Schwarz M.I., King T.E., Jr. eds. Interstitial Lung Disease. 3rd Edn. Hamilton-London, B.C. Decker, Inc., 1998. 21. Kreiss K., Gomaa A., Kullman G., Fedan K., Simoes E.J., Enright P.L.: Clinical bronchiolitis obliterans in workers at a microwave-popcorn plant. N Eng J Med 2002; 347(5):330-338. 22. Kullman G., Boylstein R., Jones W., Piacitelli C., Pendergrass S., Kreiss K.: Characterization of respiratory exposures at a microwave popcorn plant with cases of bronchiolitis obliterans. J Occup Environ Hyg 2005; 2:169-178. DOI: 10.1080/15459620590923091. 23. Kullman G., Sahakian N.: HETA 2006-0195-3044: Yatsko’s Popcorn, Sand Coulee, Montana. NIOSH Health Hazard Evaluation Report. National Institute of Occupational Safety and Health (NIOSH), Cincinnati, OH 2007, quoted [25]. 24. Lockey J.E., Hilbert T.J., Levin L.P.: Airway obstruction related to diacetyl exposure at microwave popcorn production facilities. Eur Resp J 2009; 34(1):63-71. DOI: 10.1183/09031936.00050808. 25. Maier A., Kohrman-Vincent M., Parker A., Haber L.T.: Evaluation of concentration-response options for diacetyl in support of occupational risk assessment. Reg Toxicol Pharmacol 2010; 58:285-296. DOI: 10.1016/j.yrtph.2010.06.011. 26. Merck, The Merck Index, an Encyclopedia of Chemicals, Drugs, and Biologicals, 14th ed., Merck & Co., Inc. Whitehouse Station, NJ, USA 2006, p. 504. Nr 2 27. Miller A.G., Gerrard J.A.: Assessment of protein function following cross-linking by α-dicarbonyls. Ann N Y Acad Sci 2005; 1043:195-200. 28. Morgan D.L., Flake G.P., Kirby P.J., Palmer S.M.: Respiratory toxicity of diacetyl in C57BI/6 mice. Toxicol Sci 2008; 103(1):169-180.DOI: 10.1093/toxsci/kfn016. 29. Morris J.B., Hubbs A.F.: Inhalation dosimetry of diacetyl and butyric acid, two components of butter flavoring vapors. Toxicol Sci 2009; 108:173-183, quoted [1]. 30. Nakagawa J., Ishikura S., Asami J.: Molecular characterization of mammalian dicarbonyl/L-xylulose reductase and its localization within the kidney. J Biol Chem 2002; 277:17883-17891. 31.NIOSH, Criteria for a Recommended Standard Occupational Exposure to Diacetyl and 2,3-Pentanedione. Department of Health and Human Services. Centers for Disease Control and Prevention. National Institute for Occupational Safety and Health. August 12, 2011. 32. Otsuka M., Mine T., Ohuchi K., Ohmori S.: A detoxication route for acetaldehyde: metobolism of diacetyl, acetoin, and 2,3-butanediol in liver homogenate and perfused liver of rats. J Biochem 1996; 119(2):246-251. 33. Parmet A.J., von Essen S.: Rapidly progressive, fixed airway obstructive disease in popcorn workers: a new occupational pulmonary illness? J Occup Environ Med 2002; 44:216-218. 34. Pendergrass S.M.: Method development for the determination of diacetyl and acetoin at a microwave popcorn plant. Environ Sci Technol 2004; 38(3):858-861. 35. Pietkiewicz J., Wolny M.: Inactivation of enolase from carp (Cyprinus carpio) muscle by 2,3-butanedione. Biochem Int 1991;23(1):69-74. 36. Rosati J.L., Krebs K.A., Liu X.: Emissions from cooking microwave popcorn. Crit Rev Food Sci Nutr 2007;47:701-709. 37.SCOEL, Recommendation from the Scientific Committee on Occupational Exposure Limits for diacelyt. SCOEL/SUM/149 February 2010. 38. van Rooy F.G.B.G.J., Rooyockers J.M., Prokop M., Houba R., Smit L.A.M., Heederik D.J.J.: Bronchiolitis obliterans syndrome in chemical workers producing diacetyl for food flavorings. Am J Respir Crit Care Med 2007;176:498-504. DOI: 10.1164/rccm.200611-16200c. 39. van Rooy F.G.B.G.J., Smit L.A.M., Houba R., Zaat V.A.C., Rooyackers J.M., Heederik D.J.J.: A cross-sectional study of lung function and respiratory symptoms among chemical workers producing diacetyl for food flavourings. Occup Environ Med 2009; 66:105-110. DOI: 10.1136/ oem.2008.039560. 40. Visscher D.W., Myers J.L.: Bronchiolitis. The Pathologist’s perspective. Proc Am Thorac Soc 2006;3:41-47. DOI: 10.1513/pats.200512-124JH. 41. Wondrake G.T., Cervantes-Laurean D., Roberts M.J.: Identification of α-carbonyl scavengers for cellular protection against carbonyl stress. Biochem Pharmacol 2002; 63:361-373. Received: 26.02.2014 Accepted: 11.04.2014 Rocz Panstw Zakl Hig 2014;65(2):93-100 DEVELOPMENT AND VALIDATION OF A METHOD FOR DETERMINATION OF SELECTED POLYBROMINATED DIPHENYL ETHER CONGENERS IN HOUSEHOLD DUST Wojciech Korcz*, Paweł Struciński, Katarzyna Góralczyk, Agnieszka Hernik, Monika Łyczewska, Katarzyna Czaja, Małgorzata Matuszak, Maria Minorczyk, Jan K. Ludwicki Department of Toxicology and Risk Assessment, National Institute of Public Health – National Institute of Hygiene, Warsaw, Poland ABSTRACT Background. Polybrominated diphenyl ethers (PBDEs) belong to group of so-called persistent organic pollutants (POPs). These compounds occur in nearly all elements of the environment, including household dust which constitutes one of a major route for human exposure. Their main adverse effects on human health are associated mainly with endocrine disruption – they interfere with thyroid function exhibit anti-androgenic action. Objectives. To develop and validate analytical method for determination of BDE-47, BDE-99, BDE-153, and BDE-209 congeners in household dust. Material and methods. Household dust was sampled in residences from Warsaw and the surrounding areas. An automated Soxhlet extraction of samples was then performed and PBDE congeners were subsequently measured in cleaned-up extracts by GC-μECD. The identity of quantified compounds was confirmed by GC/MS. Results. Household dust samples were fortified at levels of 2.88, and 28.8 ng g-1 for BDE-47, BDE-999, and BDE-153, and for BDE-209 at levels of 101.2, and 540 ng g-1. Recoveries ranged between 72 – 106%. The relative standard deviations (RSD) were less than 16% for all PBDE congeners analysed. The relative error determined on the basis of multiple analyses of certified reference material ranged from 1.07 – 20.41%. The method’s relative expanded uncertainty varied between 16 – 21%. Conclusion. The presented method was successfully validated and can be used to measure concentrations of BDE-47, BDE-99, BDE-153 and BDE-209 congeners in household dust. Key words. PBDEs, dust, method validation, recovery STRESZCZENIE Wprowadzenie. Polibromowane difenyloetery (PBDE) zaliczane są do trwałych zanieczyszczeń organicznych. Wykrywane są praktycznie we wszystkich elementach środowiska, także w kurzu. Kurz jest istotnym źródłem pobrania polibromowanych difenyloeterów przez człowieka. Szkodliwy wpływ PBDE na zdrowie człowieka wiązany jest głównie z zaburzaniem równowagi układu hormonalnego – zaburzają one m.in. funkcjonowanie hormonów tarczycy oraz działają antyandrogennie. Cel badań. Opracowanie i walidacja metody analitycznej umożliwiającej oznaczanie kongenerów BDE-47, BDE-99, BDE153 i BDE-209 w kurzu domowym. Materiał i metody. Materiał do badań stanowiły próbki kurzu pochodzące z domów osób zamieszkałych w Warszawie i okolicach. PBDE ekstrahowano z kurzu z wykorzystaniem aparatu do automatycznej ekstrakcji Soxhlet. Ekstrakt oczyszczano i poddawano analizie instrumentalnej. Oznaczenia zawartości analizowanych kongenerów PBDE prowadzono na GC-μECD, a tożsamość potwierdzano na GC-MS. Wyniki. Próbki kurzu były wzbogacane na poziomie 2,88 ng g-1 i 28,8 ng g-1 dla BDE-47, BDE-99, BDE-153 oraz 101,2 ng g-1 i 540 ng g-1 dla BDE-209. Odzysk mieścił się w zakresie 72 - 106%. Względne odchylenie standardowe (RSD) było mniejsze niż 16% dla wszystkich analizowanych kongenerów PBDE. Błąd względny wyznaczony na podstawie wielokrotnej analizy certyfikowanego materiału referencyjnego wynosił od 1,07% do 20,41%. Względna niepewność rozszerzona zawierała się w zakresie 16-21%. Wniosek. Metoda została zwalidowana i może być wykorzystywana do oznaczania zawartości kongenerów BDE-47, BDE99, BDE-153 i BDE-209 w próbkach kurzu domowego. Słowa kluczowe: PBDE, kurz, walidacja metody, odzysk *Corresponding author: Wojciech Korcz, Department of Toxicology and Risk Assessment, National Institute of Public Health – National Institute of Hygiene, Chocimska 24, 00-791 Warsaw, Poland, phone: +48 22 5421421, fax: +48 22 8497441, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 94 W. Korcz, P. Struciński, K. Góralczyk et al. INTRODUCTION One of the negative consequences of a modern lifestyle is the plethora of harmful chemical compounds present in the environment. Those environmental contaminants that are of particular concern are ones with long persistence, the ability to readily migrate and ones which are lipophilic. such persistent organic pollutants (POPs) include the aforementioned polybrominated diphenyl ethers (PBDEs) which consist of 209 congeners that differ in the numbers and position of bromine substitutions on the two aromatic rings of the diphenyl ether moiety. All of these congeners have an octanol: water partition coefficient (LogOW) greater than 5, thereby showing their high fat solubility [12, 14]. PBDEs were first used in the 1960s of the previous century as flame retardants. They are commercially available as three mixture types known as penta-BDE, octa-BDE and deca-BDE. The former mainly consists of BDE47, BDE-99 and BDE-100, whilst the middle type is principally composed of BDE-183, BDE-190, BDE-197 and BDE-196, but the latter is practically only made up of BDE-209 (making up 97% of the content) [31]. Since 15th August 2004 there has been a ban in force throughout the EU on penta-BDE and octa-BDE. Furthermore, from 2008 the European Union Court of Justice extended this ban to include deca-BDE used in electric and electronic goods. Some USA states, like California in 2006, banned the use of penta-BDE and octa-BDE products which thus abolished their manufacture from the USA. At the end of 2013 the manufacture of deca-BDE also became banned [4, 9, 40]. Despite this stepwise withdrawal of PBDE and its manufactured flame retardant products, such products will be nevertheless still present on the market for a long time yet, as well as in the immediate environment [32]. It is estimated that the PBDEs as flame retardant component may constitute up to 30% of the plastic casings of computers, televisions, fabrics (automobile seats and air), together with flooring and polyurethane foams (in household furniture, mattresses or car seats) [29]. As non-permanently chemically bonded components of products, they can be readily released into the environment during the operation of electrical devices when heating is generated. A 5 °C rise in temperature has been shown to increase the emission of PBDEs from television casings to the environment from 40 to 70% [37]. Another factor responsible for such PBDE release is by UV irradiation [38]. PBDEs can in fact be found throughout the environment that includes plant and animal tissue [10, 16, 25], together with human specimens [15, 34]. Many published studies demonstrate that a significant source of human exposure to PBDEs are various foodstuffs and Nr 2 dust [5, 18, 27], where those particularly vulnerable to exposure are small children aged 6 months to two years [19, 41]. It has been shown that these compounds are human endocrine disruptors altering thyroid, pituitary and hypothalamic function as well as having neurotoxic effects, that lead to behavioural changes and thought process disorders [3, 17, 20, 21, 28]. The non-respirable fraction of inhaled dust is a heterogeneous mixture of dander, skin, hair, food debris, sand, fragments of fibres from carpets, clothes and cigarette ash etc. [41]. PBDEs in dust are mainly determined by gas chromatographic (GC) methods with various means of detection eg. GC/ MS (mass spectrometric) or GC-ECD (electron capture) [23, 24]. The aim of the study was to develop a simple method for measuring PBDE congeners in dust, serving as a basis to thereby determine human exposures from this source. Through performing a literature review, four congeners were chosen, namely: BDE47 (2,2’,4,4’- tetrabromodiphenyl ether), BDE-99 (2,2’,4,4’,5-pentabromodiphenyl ether, BDE-153 (2,2′,4,4′,5,5′-hexabromodiphenyl ether) and BDE-209, (2,2’,3,3’,4,4’,5,5’,6,6’-decabromodiphenyl ether) [8, 13, 30, 33, 35, 39]. MATERIAL AND METHODS Reagents and Standards Certified standard solutions of PBDEs (ie. BDE-47, BDE-99, BDE-153 and BDE-209) were commercially obtained in 1.2 mL aliquots, each at 50 µg/mL concentrations (in nonane), from Cambridge Isotope Laboratories (Andover, USA). Merck (Darmstadt, Germany) supplied the following; n-hexane and acetone for GC/ECD and GC/FID, dichloromethane (for analysing pesticide residues), n-dodecane (for synthesis), silica gel (60 extra pure 70-230 mesh ASTM; for column chromatography), activated aluminium oxide 90 neutral (also for column chromatography) and florisil. Cellulose extraction thimbles (43 x 123 mm) were bought from Munktell (Bärestein, Germany) whilst certified reference material SRM 2585 (NIST-2585) was provided by the LGC Standards. Test sample material Dust samples obtained from households in Warsaw and the surrounding areas constituted the test material on which the method was developed and validated. These were taken using a vacuum cleaner at each place of residence and, as quickly as possible, were gathered at the laboratory so that a visual segregation of large object could be done to eliminate any plastics, wood, metal or hair and then sieved on 150 µm vibrational steel sieving (Retsch AS 200 basic). Samples were then placed into closed aluminium vessels and stored at -20 °C ready for further use. Nr 2 Determination of polybrominated diphenyl ethers in dust - method validation This was necessary to prevent photolytic debromination of any PBDE occurring in the dust samples [1, 2]. Extraction Before use, Florisil was heated for 2 hours at 130 °C and left in an exsiccator until cooled, followed by deactivation through adding 2% of distilled water. Next, 1 g dust samples were placed into cellulose extraction thimbles to which 3 g of deactivated florisil were added. Automated extraction was then performed in a Soxhlet B-811 (Büchi) extractor by adding a 100 mL mixture of n-hexane:acetone (3:1, v:v). The details of extraction conditions are shown in Table 1. To each extract, 50 µL of n-dodecane (keeper) was added, which due to its high boiling point (200 °C) prevents any analytical losses in later stages of evaporation and changing of solvent [9]. The extract was then evaporated to almost dryness and the residue reconstituted in 2 mL n-hexane. Table 1. Parameters for Soxhlet’s extraction method (Büchi B-811 system) Parameter Soxhlet warm mode: Lower heating level Upper heating level Number of cycles Rinse: Lower heating level Time setting 10 2 30 8 15 min Column clean-up Prior to use, the aluminium oxide and silica gel were heated at 130 °C for 24 hours, after which each were respectively deactivated by adding 6% and 4.5% water. Extracts were then purified on columns containing 10 g silica gel and 5 g aluminium oxide that had been pre-conditioned with 50 mL n-hexane. Samples were then eluted with 75 mL mixture of dichloromethane:n-hexane (1:9, v:v) and collected eluates were evaporated to dryness followed by reconstitution in 1 mL n-hexane. Using glass Pasteur pipettes, samples were transferred into glass amber vials ready for instrumental analysis. Chromatography Concentrations of the chosen PBDEs in the dust samples were measured using a GC with μECD (electron capture detection) instrument; Agilent Technologies 6890N with automated sample injection (Agilent 7863) controlled by Agilent ChemStation. Chromatographic run conditions were as follows; DB-5MS column (30 m x 0,32 mm i.d. and film thickness 0.25 µm. The GC oven temperature ramp programme was 70 ºC (1.7 min) – 30 ºC min-1 – 210 ºC (0 min) – 5 ºC min-1 – 300 ºC (28 min). The PTV injector temperature ramp programme in ‘solvent vent’ mode was 40 ºC (0.2 min) – 700 ºC min-1 – 220 ºC (1 min) – 700 ºC min-1 – 260 ºC (2 min). Detector temperature was 330 °C, sample volume 1 µL with helium as the carrier gas. Retention times of the chosen PBDEs were; BDE-47 – 13.077 min, BDE-99 Sample weight 1 g dust +3 g florisil (2 % water) Extraction acetone : n-hexane (1:3, v:v) 100 mL + 50 µL n-dodecane Evaporation + 2 mL n-hexane + 50 mL n-hexane (column conditioning) 95 Clean-up column 5 g aluminium oxide (6% water), 10 g silica gel (4,5% water) +75 mL mixture of dichloromethane : n-hexane (1:9, v:v) Evaporation + 1 mL n-hexane GC - µECD Figure 1. Diagram of the analytical method for the determination of selected PBDE congeners in the dust samples Figure 1. Diagram of the analytical method for the determination of selected PBDE congeners in the dust samples. 96 W. Korcz, P. Struciński, K. Góralczyk et al. – 15.942 min, BDE-153 – 19.034 min and BDE-209 – 46.026 min The detailed scheme is shown in Figure 1. The identity of quantified PBDE congeners was confirmed by means of gas chromatography coupled with mass spectrometric detector with the exception of BDE-209, because of its thermal instability. The detector was an ion-trap Varian 4000 and run conditions of the GC system, using the same column, were as follows; 70 ºC (1 min); 30 ºC min-1 – 170 ºC, 8 ºC min-1 – 300 ºC (15 min), with temperatures of the detector and injector at 200 °C and 250 °C, a sample volume of 2 µL with helium again being the carrier gas. Characteristic ions of the PBDE congeners were chosen: BDE-47 – 326 and 486 m/z, BDE-99 – 406 and 564 m/z and BDE-153 – 484 m/z. RESULTS AND DISCUSSION Validation of a given analytical method enables the assessment of possibility of an accurate and precise measurement of analyte concentration to be made. The present study was carried out in accordance with the published recommendations [6, 7, 11, 22]. The limits of quantification (LOQ) corresponding to the lowest points on the calibration curve were 1 ng mL-1 for BDE-47, BDE-99 and BDE-153, and 20 ng mL-1 for BDE-209 (equal to 1 and 20 ng per gram of dust). However due to the influence of co-extracting matrix complex components, an approach applied by Król et al. (2012) has been applied, and LOQs were finally estimated at 2 ng g-1 for BDE-47, BDE-99, and 30 ng g-1 for BDE-209. Parameters characterizing the method are shown in Table 2. Recoveries were determined at two PBDE levels as follows; 2.88 ng g-1 and 28.8 ng g-1 for BDE-47, BDE99 and BDE-153 whilst at 101.2 ng g-1 and 540 ng g-1 for BDE-209. For this purpose, a test portion of the dust inserted in the cellulose thimble was spiked with known volume of standard solution containing mixture Nr 2 of PBDEs and was followed by the adopted scheme. The fortification levels were adopted through performing a review of scientific papers describing levels of these compounds in dust [8, 13, 30, 33, 35]. Recoveries for BDE-47, BDE-99 and BDE-153 ranged from 75% – 82%. The method’s relative expanded uncertainty for both fortification levels was estimated to vary between 16 – 21%. In the case of BDE-209, similar recoveries were found at 73 and 72% respectively at levels of 101.2 ng g-1 and 540 ng g-1 with the relative expanded uncertainty in both cases being 18%. In estimating the method’s uncertainty, only the intra-laboratory analytical procedure was taken into account that included the PBDE congeners recoveries. Precision of the method was also estimated and expressed as the repeatability limit (r) of measurement. During the validation, each stage of the method was checked to determine which contributed to the greatest measurement uncertainty; this was found to be the solvent evaporation. The n-dodecane, as a keeper, was checked for its effects on PBDE recoveries during evaporation in the following manner; two sets of five test tubes containing 1 mL of mixture of standards in n-hexane (BDE-47 – 50.6 ng mL-1, BDE-99 – 51.2 ng mL-1, BDE-153 – 51.2 ng mL-1 and BDE-209 – 253 ng mL-1), were prepared with 50 µL of n-dodecane being added to only the first set. Both sets were then evaporated to dryness and then reconstituted with 1 mL n-hexane. As shown in Table 3, n-dodecane reduces recovery losses in the lower-brominated PBDE congeners (e.g. BDE-47) but reduces recoveries in decabromodiphenyl ether. For BDE-47, BDE-99 and BDE-153 congeners an intra-laboratory reproducibility was checked for dust samples (n=6) spiked with 28.8 ng-1 of these compounds. Recoveries of the PBDEs varied from 78 – 95%, with RSDs (relative standard deviation) ranging 11 – 15% that were lower than the 27% RSDR value (relative standard deviation of reproducibility) calculated according to the Horwitz equation [26]. Table 2. Summary of validation parameters for the method Parameters Working range [ng g-1] Fortification level [ng g-1] Average recovery [%] (n=6) SD [ng g-1] RSD [%] Repeatability limit (r) Relative expanded uncertainty [%] Fortification level [ng g-1] Average recovery [%] (n=6) SD [ng g-1] RSD [%] Repeatability limit (r) Relative expanded uncertainty [%] BDE-47 2- 506 106 0.49 15.98 1.36 16 82 3.65 15.38 10.23 20 PBDE congeners BDE-99 BDE-153 2 - 512 2 - 512 2.88 104 95 0.29 0.38 9.93 13.74 0.82 1.05 15 21 28.8 76 74 3.23 2.72 14.8 12.77 9.04 7.60 16 19 BDE-209 30 - 759 101.2 73 8.81 11.97 24.66 18 540 72 35.26 9.10 98.74 18 Nr 2 Determination of polybrominated diphenyl ethers in dust - method validation 97 Table 3. The role of keeper (n-dodecane) addition at the stage of solvent evaporation PBDE congeners Concentation [ng mL-1] BDE-47 BDE-99 BDE-153 BDE-209 50.60 51.20 51.20 253.00 Evaporation with keeper (n=5) Measured concentration SD [ng mL-1] [ng mL-1] 52.12 0.47 55.42 1.26 54.83 1.48 215.02 11.48 The certified reference material SRM 2585 (NIST) was used also for validation which is the sieved dust contained numerous contaminants, including analysed PBDEs [31]. Results of analysis of 6 such reference dust samples are shown in Table 4. Table 4. Results of standard reference material NIST SRM 2585 analyses (n=6) PBDE congener BDE-47 BDE-99 BDE-153 BDE-209 Certified concentration [ng g-1] 497 892 119 2510 Measured concentration ± SD [ng g-1] 491.66 ± 18.17 751,64 ±20.95 94.71 ± 13.46 2746.54 ± 160.63 Relative error a (%) 1.07 15.74 20.41 9.42 calculated as (|average measured concentration– certified concentration| ∕ certified concentration)*100 a Evaporation without keeper (n=5) Measured concentration SD [ng mL-1] [ng mL-1] 25.83 6.65 46.07 3.39 54.47 1.96 251.34 22.70 The results demonstrate the adequacy of the developed method for measuring the defined analytes. It should however be mentioned that PBDE levels in the certified reference material were very high (especially for BDE-99 and BDE-209) and therefore either dilutions are required at the final stages to fall within the working range of calibration curve or smaller samples of dust need to be taken. Indeed the latter option was used, where only 100 mg amounts of dust were sampled with the final result being accordingly adjusted as per 1 g dust. Because PBDEs are vulnerable to debromination by photolysis, it is important to limit their exposure to UV light at each stage of the analytical procedure, particularly those involving organic solvents where this process occurs most readily [36, 38]. The speed of photo-degradation also increases with the number of Figure 2. GC- µECD chromatograms obtained from standard reference material NIST SRM 2585 sample (a) and household dust sample (b). 98 Nr 2 W. Korcz, P. Struciński, K. Góralczyk et al. Figure 3. GC-MS chromatogram of household dust sample obtained in SIS mode. bromine substitutions on the PBDE moiety [1, 2]. By using the ‘warm extraction’ mode with the automated Soxhlet, it was possible to shorten the extraction time by 3 hours as compared to the original procedure [8, 9, 28, 35]. Due to dust being an extremely heterogeneous matrix, the chromatograms of successive samples may differ in the numbers of peaks observed and their intensities. Figure 2 shows chromatograms of a reference SRM 2585 material and a sample of household dust obtained by GC-µECD. Figure 3 demonstrates an example of a chromatogram obtained from a dust sample using GC/MS in the SIS (selected ion storage) mode which increases the sensitivity of detecting specifically chosen ions, and thus enables full identification of test substances. In the presented sample, the presence of BDE-47 and BDE-99 were confirmed, however BDE-153 was absent. CONCLUSIONS 1. This developed and validated method can be used for measuring the concentrations of the selected polybrominated diphenyl ethers congeners: BDE-47, BDE-99, BDE-153, and BDE-209 in dust samples, thus enabling human exposure to these substances to be assessed within household environments or other similar confined spaces, such as those found in automobiles. 2. Recoveries and relative standard deviations are analytically appropriate as are the repeatability and reproducibility within the working conditions used in the study. The method is also robust to changes in the laboratory environment. Acknowledgements The presented study was funded by the National Science Centre, Poland (Grant No. N N404 0881140). Conflict of interest The authors declare no conflict of interest. REFERENCES 1. Ahn M., Filley T.R., Jafvert C.T., Nies L., Hua I., Bezares-Cruz J.: Photodegradation of decabromodiphenyl ether absorbed onto clay minerals, metal oxides, and sediment. Environ Sci Technol 2006;40:215-220. Nr 2 Determination of polybrominated diphenyl ethers in dust - method validation 2. Allen J.G., McClean M.D., Stapleton H.M., Webster T.F.: Critical factors in assessing exposure to PBDEs via house dust. Environ Int 2008;34:1085-1091. 3. Darras V.M.: Endoctrine disrupting polyhalogenated organic pollutants interfere with thyroid hormone signaling in the developing brain. Cerebellum 2008;26-37. 4. Dodson R.E., Perovich L.J., Covaci A., Van den Eede N., Ionas A.C., Dirtu A.C., Brody J.G., Rudel R.A.: After the PBDE phase-out: a broad suite of flame retardants in repeat house dust samples from California. Environ Sci Technol 2012;46:13056-13066. 5. Domingo J.L.: Polybrominated diphenyl ethers in food and human dietary exposure. A review of the recent scientific literature. Food Chem Toxicol 2012;50:238249. 6. EURACHEM / CITAC Guide CG 4 “Quantifying uncertainty in analytical measurement” third edition, 2012. Available from: http://www.eurachem.org (accessed on 4.01.2014). 7. EURACHEM guide “The fitness for purpose of analytical methods. A Laboratory Guide to method validation and related topics” 1998. Available from: http://www. eurachem.org (accessed on 4.01.2014). 8. Fromme H., Körner W., Shahin N., Wanner A., Albrecht M., Boehmer S., Parlar H., Mayer R., Liebl B., Bolte G.: Human exposure to polybrominated diphenyl ethers (PBDE), as evidenced by data from duplicate diet study, indoor air, house dust, and biomonitoring in Germany. Environ Int 2009;35:1125-1135. 9. Gevao B., Al-Bahloul M., Nabi Al-Ghadban A., Al-Omair A., Ali L., Zafar J., Helaleh M.: House dust as a source of human exposure to polybrominated diphenyl ethers in Kuwait. Chemosphere 2006;64: 603–608. 10. Góralczyk K., Hernik A., Czaja K., Struciński P., Korcz W., Snopczyński T., Ludwicki J.K.: Organohalogen compounds – new and old hazards for peoples. Rocz Panstw Zakl Hig 2010;61(2):109-117 (in Polish). 11. Góralczyk K., Hernik A., Struciński P., Czaja K., Ludwicki J.K.: Methods validation and uncertainty of results in the analysis of pesticide residues in food. Rocz Panstw Zakl Hig 2003;54(1):39-48 (in Polish). 12. Góralczyk K., Struciński P., Czaja K., Hernik A., Ludwicki J.K.: Flame retardants – use and hazards for humans. Rocz Panstw Zakl Hig 2002;53,293-305 (in Polish). 13. Harrad S., Ibarra C., Diamond M., Melymuk L., Robson M., Douwes J., Roosens L., Dirtu A.C., Covaci A.: Polybrominated diphenyl ethers in domestic indoor dust from Canada, New Zealand, United Kingdom and United States. Environ Int 2008;34:232-238. 14. Hernik A., Góralczyk K., Czaja K., Struciński P., Korcz W., Ludwicki J.K.: Polybrominated diphenyl ethers (PBDE) – new threats? Rocz Panstw Zakl Hig 2007;58(2):403-415 (in Polish). 15. Hernik A., Góralczyk K., Struciński P., Czaja K., Kucharska A., Korcz W., Snopczyński T., Ludwicki J.K.: Polybrominated diphenyl ethers, polychlorinated biphenyls and organochlorine pesticides in human milk as markers of environmental exposure to these compounds Ann Agric Environ Med 2011;18:113-118. 99 16. Johanson I., Héas-Moisan K., Guiot N., Munschy C, Tronczyński J.: Polybrominated diphenyl ethers in mussels from selected French coastal sites: 1981-2003. Chemosphere 2006;64:296-305. 17. Johnson P., Stapleton H.M., Mukherjee B., Hauser R., Meeker J.D.: Associations between brominated flame retardants in house dust and hormone levels in man. Sci Total Environ 2013;455:177-184. 18. Johnson-Restrepo B., Kannan K.: An assessment of sources and pathways of human exposure to polybrominated diphenyl ethers in the United States. Chemosphere 2009;76:542–548. 19. Jones – Otazo H. A., Clarke J. P., Diamond M. L., Archbold J. A., Ferguson G., Harner T., Richardson G. M., Ryan J. J., Wilford B.: Is House Dust the Missing Exposure Pathway for PBDEs? An Analysis of the Urban Fate and Human Exposure to PBDEs. Environ Sci Technol 2005;39:5121–5130. 20. Karpeta A., Gregoraszczuk E.: Mixture of dominant PBDE congeners (BDE-47, -99, -100 and -209) at levels noted in human blood dramatically enhances progesterone secretion by ovarian follicles. Endocr Regul 2010;44:49-55. 21. Kiciński M., Viaene M.V., Den Hond E., Shoeters G., Covaci A., Dirtu A.C., Nelen V., Bruckers L., Croes K., Sioen I., Baeyens W., Van Larabeke N., Nawrot T.S.: Neurobehavioral function and low-level exposure to brominated flame retardants in adolescents: a cross-sectional study. Environ Health 2012;11:86. 22. Korcz W., Góralczyk K., Czaja K., Struciński P., Hernik A., Snopczyński T., Ludwicki J.K.: The application of statistical methods in chemical experiments. Rocz Panstw Zakl Hig 2008;59(2):117-129 (in Polish). 23. Król S., Zabiegała B., Namieśnik J.: Determination of polybrominated diphenyl ethers in house dust using standard addition method and gas chromatography with electron capture and mass spectrometric detection. J Chromatogr A 2012;1249:201-214. 24. Król S., Zabiegała B., Namieśnik J.: PBDEs in environmental samples: Sampling and analysis. Talanta 2012;93:1-17. 25. Law R.J., Covaci A., Harrad S., Herzke D., Abdallah M.A.-E., Fernie K., Toms L.L., Takigami H.: Levels and trends of PBDEs and HBCDs in the global environment: Status at the end of 2012. Environ Int 2014;65:147-158. 26. Linsinger T.P.J., Josephs R.D.: Limitations of the application of Horwitz equation. Trac-Trend Anal Chem 2006;25(11):1125-1130. 27. Lorber M.: Exposure of Americans to polybrominated diphenyl ethers. J Expo Sci Env Epid 2008;18:2-19. 28. Meeker J.D., Johnson P.I., Camann D., Hauser R.: Polybrominated diphenyl ethers (PBDE) concentration in house dust are related to hormone levels in men. Sci Total Environ 2009;409:3425-9. 29. Rahman F., Langford K.H., Scrimshaw M.D., Lester J.N.: Polybrominated diphenyl ether (PBDE) flame retardants. Sci Total Environ 2001;275:1-17. 30. Roosens L., Cornelis C., D’Holeander W., Bervoets L., Reyndenrs H., Van Campenhout K., Van Den Heuvle R., Neels H., Covaci A.: Exposure of the Flemish population 100 W. Korcz, P. Struciński, K. Góralczyk et al. to brominated flame retardants: Model and risk assess ment. Environ Int 2010;36,368-376. 31. Stapleton H.M., Harner T.,Shoeib M., Keller J.M., Shantz M.M., Leight S.D., Wise S.A.: Determination of polybrominated diphenyl ethers in indoor dust standard reference materials. Anal Bioanal Chem 2006;383:791-800. 32. Stapleton H.M., Klosterhaus S., Keller A., Ferguson P.L., van Bergen S., Cooper E., Webster T.F., Blum A.: Identification of flame retardants in polyurethane foam collected from baby products. Environ Sci Technol 2011; 45:5323-5331. 33. Thuresson K., Björklund J.A., de Wit C.A,: Tri-decabrominated diphenyl ethers and hexabromocyclododecane in indoor air and dust from Stockholm microenvironments 1: Levels and profiles. Sci Total Environ 2012; 414:713721. 34. Toft G., Lenters V., Vermeulen R., Heederik D., Thomsen C., Becher G., Giwercman A., Bizzaro D., Manicardi G.C., Spanò M., Rylander L., Pedersen H.S., Struciński P., Zviezdai V., Bonde J.P.: Exposure to polybrominated diphenyl ethers and male reproductive function in Greenland, Poland and Ukraine. Reprod Toxicol 2014; 43:1-7. 35. Vorkamp K., Thomsen M., Frederiksen M., Pedersen M., Knudsen L.E.: Polybrominated diphenyl ethers (PBDEs) in the indoor environment and associations with prenatal exposure. Environ Int 2011; 37:1-10. Nr 2 36. Wang J., Chen S., Nie X., Tian M., Luo X., Taicheng A., Mai B.: Photolytic degradation of decabromodiphenyl ethane (DBDPE). Chemosphere 2012; 89:844-849. 37. Waye S. K., Anderson A., Corsi R.L., Ezekoye O.A.: Thermal effects on polybrominated diphenyl ethers mass transfer and emission from computer cases. Int J Heat Mass Tran 2013; 64:343-351. 38. Wei H., Zou Y., Li A., Christensen E.R., Rockne K.J.: Photolytic debromination pathway of polybrominated diphenyl ethers in hexane by sunlight. Environ Pollut 2013; 174:194-200. 39. Whitehead T., Metayer C., Buffler P., Rappaport M.: Estimating exposures to indoor contaminants using residential dust. J Expo Sci Env Epid 2011; 21:549-564. 40. Whitehead T.P., Brown F.R., Metayer C., Park J., Does M., Petreas M., Buffler P., Rappaport S.M.: Polybrominated diphenyl ethers in residential dust: Sources of variability. Environ Int 2013; 57-58:11-24. 41. Wilford B.H., Shoeib M., Harner T., Zhu J., Jones K.C.: Polybrominated diphenyl ethers in indoor dust in Ottawa, Canada: Implications for sources and exposure. Environ Sci Technol 2005; 39: 7027-7035. Received: 14.01.2014 Accepted: 04.04.2014 Rocz Panstw Zakl Hig 2014;65(2):101-105 VARIATIONS OF NIACIN CONTENT IN SALTWATER FISH AND THEIR RELATION WITH DIETARY RDA IN POLISH SUBJECTS GROUPED BY AGE Michał Majewski1*, Anna Lebiedzińska2 Department of Pharmacology and Toxicology, Faculty of Medical Sciences, University of Varmia and Masuria, Olsztyn, Poland 2 Chair and Department of Bromatology, Medical University of Gdańsk, Gdańsk, Poland 1 ABSTRACT Introduction. A rich and natural source of readily assimilated dietary protein together with invaluable vitamins and minerals are fish, particularly the saltwater species. The quality of any given foodstuff is determined by its nutritional value, which in turn depends on the food type and methods used for manufacture, processing and storage. Many fish products contain fewer water soluble vitamins than the source foodstuff as a result of using various technologies during food processing, such as smoking or deep freezing, where vitamins are often either degraded or leached out. In the case of niacin it is relatively easy to make good such losses by eating niacin-rich foods or by taking dietary supplements e.g. the essential amino acid L-tryptophan. Objectives. To determine niacin content in sea fish that are commonly available on the Polish market and to assess whether this dietary source is sufficient to satisfy the RDA requirements for various age groups of selected subjects living in Poland. Material and methods. Niacin levels were measured firstly in 10 saltwater fish species together with butterfish and Norwegian salmon that formed a separate group. Altogether, 15 types of fish products were analysed in all. They consisted of smoked fish: whitefish, butterfish, sprat, trout, herring (kippers) and mackerel, and frozen fish: butterfish, Norwegian salmon, sole, grenadier and panga. Each product was measured as ten replicates, thus in total 150 analyses were performed. A microbiologically-based method was used for the niacin determination, with enzyme hydrolysis by 40 mg papain and diastase on a 2 g sample (according to the AOAC procedure) to release the free form from the bioavailable form that is bound to NAD and NADP. Results. The most plentiful sources of niacin were found in smoked fish with the highest amounts in butterfish, after warm temperature smoking, and in mackerel; respectively 9.03 and 8.90 mg/100 g. Such 100 g portions of smoked fish are a good dietary source of niacin, in that for men and women above 19 years of age, they constitute respectively 22% - 56% and 25% - 64% of the RDA (Recommended Daily Allowance). The highest levels of niacin in frozen fish were found in butterfish and Norwegian salmon; respectively 8.05 and 5.75 mg/100 g which in turn represent respectively 10% - 50% and 11% - 56% of the RDA in men and women aged above 19 years. Conclusions. Niacin concentrations varied according to fish species. The richest dietary sources were smoked fish consisting of butterfish, after warm temperature smoking, and mackerel. In frozen fish, butterfish and Norwegian salmon had the highest niacin amounts. A 100 g serving of such sea fish can, to quite a large extent, satisfy the adult RDA. Key words: niacin, nicotinic acid, nicotinamide, fish STRESZCZENIE Wprowadzenie. Ryby zwłaszcza morskie stanowią naturalne źródło łatwo przyswajalnego białka oraz wielu cennych witamin i minerałów. Witamina B3 to grupa związków w skład których wchodzą kwas nikotynowy (niacyna) oraz amid kwasu nikotynowego (nikotynamid). Stosunkowo łatwo uzupełniać niedobory niacyny spożywając regularnie produkty bogate w tą witaminę, jak i białko lub szeroko dostępne na rynku suplementy diety. Cel badań. Celem pracy było oznaczenie zawartości niacyny w łatwo dostępnych na rynku rybach morskich, a także ocena analizowanych ryb jako potencjalnego dobrego źródła niacyny w diecie człowieka (RDA) w różnych grupach wiekowych. Materiał i metody. Oznaczono zawartość niacyny w piętnastu rodzajach ryb słonowodnych. w rybach wędzonych (sieja, ryba maślana, szprot, pstrąg, śledź oraz makrela) i mrożonych (ryba maślana, łosoś norweski, sola, grenadier, panga). Łącznie przebadano 150 produktów rybnych. Niacynę oznaczono metodą mikrobiologiczną według AOAC stosując hydrolizę en*Corresponding author: Michał Majewski, Department of Pharmacology and Toxicology, Faculty of Medical Sciences, University of Varmia and Masuria, Żołnierska Street 14 C, 10-561 Olsztyn, Poland, phone: + 48 89 524 61 88, fax: (89) 524 61 88, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 102 M. Majewski, A. Lebiedzińska Nr 2 zymatyczną za pomocą papainy i diastazy w celu wyodrębnienia witaminy z analizowanych próbek. Metoda enzymatyczna pozwala na wyodrębnienie tylko biologicznie dostępnych form niacyny związanych w NAD i NADP. Wyniki. Najlepszym źródłem niacyny były ryby wędzone, a najwięcej witaminy stwierdzono w wędzonych na ciepło rybie maślanej (9,03 mg/100 g) i makreli (8,90 mg/100 g). Porcja ryby wędzonej (100 g) może być bardzo dobrym źródłem niacyny realizując normy dziennego zapotrzebowania dla kobiet i mężczyzn w wieku powyżej 19 lat, odpowiednio w zakresie wartości od 24% do 64% i od 21% do 56%. W grupie badanych ryb mrożonych najwyższą zawartość niacyny zawierała ryba maślana (7,89 mg/100 g) i łosoś norweski (5,75 mg/100 g). Porcja ryby mrożonej (100 g) pokrywała dzienne zapotrzebowanie na niacynę normy dla kobiet i mężczyzn w wieku powyżej 19 lat, odpowiednio w zakresach od 11% do 56% i od 10% do 49%. Wnioski. Przeprowadzone analizy zawartości niacyny wykazały zróżnicowanie pomiędzy poszczególnymi gatunkami ryb. Wykazano, iż najlepszym źródłem niacyny są ryby wędzone, spośród których najwięcej analizowanej witaminy posiadają ryba maślana wędzona na ciepło oraz makrela. W grupie ryb mrożonych najwyższą zawartość niacyny oznaczono w rybie maślanej oraz w łososiu norweskim. Porcja ryby morskiej (100 g) może być bardzo dobrym źródłem niacyny. Słowa kluczowe: niacyna, kwas nikotynowy, amid kwasu nikotynowego, ryby INTRODUCTION Sea food, especially that consisting of so called ‘dark meat’, provides an excellent source of dietary niacin. Furthermore, the presence of tryptophan, which lends the meat its dark colouration, is a precursor in the biosynthesis of kynurenine, serotonin and NAD; being the biologically active form of niacin (Figure 1). A 60 mg amount of tryptophan is sufficient for generating L-Tryptophan NH2 (TRP) O HO N H O L-Kynurenine NH2 (KYN) O OH NH2 NADPH, FAD, B , B 6 O 2 Quinolinic acid (QUIN) OH O N 1 mg niacin. This reaction pathway requires B group vitamins as enzyme cofactors. Reasons for any niacin deficiencies may be malnourishment, alcoholism, medicines used for treating Parkinson’s Disease or hydrazine derivatives used in treating tuberculosis and inflammation. A diet containing fish affords many nutritional advantages [5, 17]. It should be stressed that fish protein has a high nutritional value and fish also contain long chain polyunsaturated fatty acids EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid), microelements and vitamins [12-14, 18, 19, 21, 22]. A 100 g portion of fish covers half the daily requirement for tryptophan rich protein; tryptophan being a niacin precursor. Some publications suggest that there may be significant differences in vitamin and fat content between farmed fish with those living free. This may also depend on the fish species, age, the season when fished (captured) and the type and availability of feed. Data on these topics are sparse. [16]. The study aims were to determine the niacin content in various species of sea fish, that included those who had been smoked (under warm or cold conditions) or deep frozen in relation to the sources of the human dietary requirement for niacin; RDA [12-14]. The types of fish chosen were ones that were fatty, saltwater species, easily available on the market and frequently consumed in Poland, as determined from previous dietary surveys. OH O PRPP, Mg2+ MATERIAL AND METHODS OH N Niacin (Na) Nicotinic acid mononucleotide (NaMN) NAD - active form of niacin Figure 1. The niacin pathway of tryptophan metabolism Fig. 1. The niacin pathway of tryptophan metabolism. The study material were samples of fish products that had been smoked (under either warm or cold conditions) or deep frozen; Table 2 and Table 3. The fish samples consisted of fillets, flakes, cutlets and whole carcasses. Ten fish species were analysed; whitefish, butterfish, sprat, trout, herring, mackerel, Norwegian salmon, sole, grenadier and panga that amounted to Nr 2 103 Niacin in saltwater fish 19 years by their concordance with amended reference values of nutrition in Poland, supplied by the Polish Institute of Food and Nutrition [8]. Table 1. Accuracy and precision of niacin determination Niacin content in fish (mg/100 g) n 7.89 ±015 10 Spiked Recovery (mg/100 g) (%) 3 6 97.87 103.04 SD (%) 2.89 3.13 Relative error (%) - 2.13 +3.04 RESULTS AND DISCUSSION n - number of samples, SD- standard deviation Niacin concentrations found in the tested fish samples are shown in Tables 2 and 3. In smoked fish, the highest niacin concentrations were found in butterfish (warm treatment) and mackerel at respectively 9.03 and 8.90 mg/100 g. The lowest levels in smoked fish were recorded in whitefish and sprats (whole and flesh), respectively 3.50 and 4.06 – 4.58 mg/100 g (Table 2). In the deep frozen fish, the highest amounts measured were in butterfish (respectively 8.05 and 7.11 mg/100 g in cutlets and fillets) and Norwegian salmon (5.05 8.85 mg/100 g), whereas the lowest levels were in sole, grenadier and panga; respectively 1.96, 1.71 and 1.53 mg/100 g (Table3). The observed differences in niacin concentrations between smoked and deep frozen fish could be explained by water losses incurred in the latter during thawing, as well as the type of treatment (warm or cold) used in smoking. As aforementioned, the thawing loss was 6% due to the discarding of water. Results were subjected to statistical analysis by ANOVA using p ≤ 0.05 as showing significance. For butterfish, the treatment differences between warm and cold smoking was compared and found to be significantly different at p = 0.001. The niacin levels found in the 100 g fish portions are shown in relation to RDA requirements; for smoked fish (Table 2) and deep frozen fish (Table 3) - taking into account the age, gender and physiological status of the human population. Using the mean RDA values for children, a 100 g portion of butterfish (warm smoked) and smoked mackerel, fulfils their RDA by respectively 113 and 111% (Table 2). Niacin reference values for boys are equivalent to those of adult men; the same 15 fish products in total. Each were analysed as 10 replicates. Three samples of fish flesh were obtained after homogenising and mixing each fish product type. Previously, the fish had been thawed at 4°C, after being frozen for over 24 hours under laboratory conditions, then the thawed water was discarded. This being similar to how fish is prepared domestically in the kitchen and on average the discarded water amounted to a 6% loss. Niacin was isolated from the samples (2 g) after enzymatic hydrolysis using papain and diastase (40 mg) according to the AOAC method [1, 15]. A microbiological method [1, 7] was then used to determine niacin using the Lactobacillus plantarum ATCC No. 8014 strain. Niacin is one of the most stable water soluble vitamins in solution and its biological activity is retained following thermal, light, pH or oxidation treatment. Both acid or enzymatic hydrolysis is thus possible for releasing free niacin from its biologically bound form where it can be liberated from coenzymes or through matrix degradation. When performing mineral acid hydrolysis, this process is however non-physiological and may release nicotinic acid which is not normally bio-available. Studies by Ndaw et al. [18] have demonstrated that by replacing acid hydrolysis by enzymes it is possible to isolate niacin liberated from its NAD and NADP bound forms. The precision and accuracy of the method were established, at highly acceptable levels, on samples spiked with known amounts of niacin (Table 1). Results were checked to see if levels were sufficient to satisfy the RDA requirements for adult subjects aged above Table 2. Niacin content in smoked fish according to the RDA for the Polish population Fish type Smoked butterfish (warm) Smoked mackerel Smoked trout Smoked butterfish (cold) Smoked herring Smoked sprats -flesh Smoked sprats -whole Smoked whitefish RDA requirement in 100 g of fish product (%) Niacin content (mg/100 g) X± SD Children 1-9 years 10 9.03± 0.05 10 10 n Men Women 10–18 years ≥ 19 years 10–18 years ≥ 19 years pregnant nursing 112.88 56.44 56.44 64.50 64.50 50.17 53.12 8.90 ± 0.09 5.65± 0.04 111.25 70.63 55.63 35.31 55.63 35.31 63.57 40.36 63.57 40.36 49.44 31.39 52.35 33.24 10 5.54± 0.13 69.25 34.63 34.63 39.57 39.57 30.78 32.59 10 10 10 10 4.99± 0.20 4.58± 0.28 4.06± 0.10 3.50± 0.21 62.38 57.25 50.75 43.75 31.19 28.63 25.38 21.88 31.19 28.63 25.38 21.88 35.64 32.71 29.00 25.00 35.64 32.71 29.00 25.00 27.72 25.44 22.56 19.44 29.35 26.94 23.88 20.59 n - number of samples; X- average; SD- standard deviation 104 Nr 2 M. Majewski, A. Lebiedzińska Table 3. Niacin content in frozen fish according to the RDA for the Polish population Fish type Butterfish (cutlet) Butterfish (fillet) Norwegian salmon steak, tail Norwegian salmon cutlet Sole, fillet Grenadier, fillet Panga, fillet n* 10 10 10 10 10 10 10 Niacin content (mg/100 g) X± SD 8.05 ± 0.15 7.11 ± 0.15 5.85 ± 0.12 5.05 ± 0,15 1.96 ± 0.08 1.71 ± 0.11 1.53 ± 0.10 Children 1-9 years 100.63 88.88 73.13 63.13 24.50 21.38 19.13 RDA requirement in 100 g of fish product (%) Men Women 10–18 10– 18 10–18 10–18 10–18 years years years years years 50.31 50.31 57.50 57.50 44.72 44.44 44.44 50.79 50.79 39.50 36.56 36.56 41.79 41.79 32.50 31.56 31.56 36.07 36.07 28.06 12.25 12.25 14.00 14.00 10.89 10.69 10.69 12.21 12.21 9.50 9.56 9.56 10.93 10.93 8.50 nursing 47.35 41.82 34.41 29.71 11.53 10.06 9.00 n– number of samples; X- average; SD- standard deviation applying to girls and adult women. Here, it was found that the RDA was satisfied by respectively 56% and 64% in men and women for both warm treated smoked butterfish and smoked mackerel; these being at the highest levels. Those fish showing the lowest RDA fulfilment were whitefish at 44% RDA in children and 22% and 25% respectively for men and women (Table 3). In the frozen fish, sole grenadier and panga least satisfied the RDA where respectively they supplied 24%, 21% and 19% in children. For men the corresponding results were 12%, 11% and 10% whilst 14%, 12% and 11% for women. A butterfish portion (100 g cutlet) best satisfied the RDA in children and in men and women; respectively 50% and 57%. There were almost 1.6 and 1.3 fold higher niacin levels in butterfish smoked respectively under warm and cold conditions compared to deep frozen butterfish. It is suggested that this arose from technological losses incurred during processing and the water loss during thawing. As a component of two vital coenzymes NAD+ and NADP+ in electron transport, niacin takes part in oxidation/reduction reactions catalysed by dehydrogenases [22]. It is vital for normal nervous system function where it protects against oxidative stress and takes part in the syntheses of the sex hormones: cortisol, thyroxin and insulin [6, 9]. Dietary niacin deficiency in children leads to many functional disorders, leading to the development of diet-related diseases, developmental and mental dysfunction [7]. As a nicotinic acid, niacin increases plasma HDL-cholesterol, whilst at the same time decreases fatty acids that induce arteriosclerosis, such as triglycerides, VLDL-cholesterol, LDL-cholesterol and Lipoprotein A [2]. Furthermore, a high dose of niacin can reduce inflammation [8]. Current nutritional recommendations clearly indicate that fish should be eaten 2 – 3 times weekly and that the dietary presence of ‘oily fish’ and certain ‘fruits of the sea’ is beneficial to the health of those at risk of cardiovascular disease, in pregnant women and the elderly [4, 7, 10, 11, 17, 20]. Due to their high nutritional value, fish should be consumed much more than is currently the case in Poland, where in fact fish consumption is falling. CONCLUSIONS 1. The study demonstrated wide variations of niacin content for different fish species. The highest levels were found in warm smoked butterfish or smoked mackerel whilst those levels highest in frozen fish were butterfish and Norwegian salmon. 2. A 100 g portion of smoked fish can be an important dietary source of niacin, satisfying the RDA by 22 - 56% in men and 25 - 64% in women. 3. In frozen fish, a 100 g portion satisfies the niacin RDA by 10 - 50% in men and 11 – 57% in women. Conflict of interests The authors declare no conflict of interest. REFERENCES 1. AOAC. Niacin and Niacinamide (Nicotinic Acid and Nicotinamide) in Vitamin Preparations. 2003 Maryland. http://www.eoma.aoac.org/methods/info.asp?ID=14717. 2. Backes J.M., Padley R.J., Moriarty P.M.: Important considerations for treatment with dietary supplement versus prescription niacin products. Postgrad Med 2011;123(2):70-83. 3. Balasubramanyam A., Coraza I., Smith E.O., Scott L.W., Patel P. et al: Combination of niacin and fenofibrate with lifestyle changes improves dyslipidemia and hypoadiponectinemia in HIV patients on antiretroviral therapy: results of “heart positive,” a randomized, controlled trial. J Clin Endocrinol Metab 2011;96(7):2236-2247. 4. Bassan M.: A case for immediate-release niacin. Heart Lung 2012;41(1):95-98. 5. Goede J., Verschuren W.M., Boer J.M., Kromhout D., Geleijnse J.M.: Gender-specific associations of marine n-3 fatty acids and fish consumption with 10-year incidence of stroke. PLoS One 2012;7(4):1-14. Nr 2 Niacin in saltwater fish 6. Hamoud S., Kaplan M., Meilin E., Hassan A., Torgovicky R. et al: Niacin administration significantly reduces oxidative stress in patients with hypercholesterolemia and low levels of high-density lipoprotein cholesterol. Am J Med Sci 2013;345(3):195-199. 7. Jarosz M.: Nutrition standards for the Polish population – revision. Warsaw, IŻŻ, 2012 (in Polish). 8. Kapoor A., Thiemermann C.: Niacin as a novel therapy for septic shock? Crit Care Med 2011;39(2):410-411. 9. Kirkland J.B.: Niacin requirements for genomic stability. Mutat Res 2012;733(1–2):14–20. 10. Kołodziejczyk M.: Consumption of fish and fishery products in Poland – analysis of benefits and risks. Rocz Panstw Zakl Hig 2007;58(1): 287-293. 11. Lavigne P., Karas R.: The Current State of Niacin in Cardiovascular Disease Prevention. J Am Coll Cardiol 2013;61(4):440-446. 12. Lebiedzinska A.: Fish and shellfish as a source of vitamins B – own results in a view of literature data. Polish J Environ Stud 2006;15(2):1322-1327. 13. Lebiedzińska A., Majewski M., Szefer P.: Butterfish as a source of niacin. Rocz Panstw Zak. Hig 2008;59(2):197201 (in Polish). 14. Lebiedzińska A., Majewski M., Szefer P.: Niacin content in canned tuna fish. Bromat Chem Toksykol 2008;1:2933 (in Polish). 15. Ndaw S., Bergaentzle M., Hasselmann C.: Enzymatic extraction procedure for liquid chromatographic determi- 105 nation of niacin in foodstuffs. Food Chem 2002;78:129– 134. 16. Nettleton J.A., Exler J.: Nutrition in wild and farmed fish and shellfish. J Food Sci 1992;57(2):257-260. 17. Oudin A., Wennberg M.: Fish consumption and ischemic stroke in southern Sweden. Nutr J 2011;10:109. 18. Polak-Juszczak L.: Mineral elements content in smoked fish. Rocz Panstw Zakl Hig 2008;59(2):187-196. 19. Regulska-Ilow B., Ilow R., Konikowska K., Kawicka A., Różańska D., Bochińska A.: Fatty acid profile of the fat in selected smoked marine fish. Rocz Panstw Zakl Hig 2013;64(4):299-307. 20. Robinson J.G.: What is the role of advanced lipoprotein analysis in practice? J Am Coll Cardiol 2012;60(25):2607-2615. 21.WHO. Global strategy on diet, physical activity and health. Fifty-seven world health assembly, Agenda item. 6.12.2004. http://www.who.int/dietphysicalactivity/strategy/ eb11344/strategy_english_web.pdf 22. Zając M.: Vitamins and microelements. Poznan, Kontekst, 2000 (in Polish). Received: 04.11.2013 Accepted: 16.03.2014 Rocz Panstw Zakl Hig 2014;65(2):107-111 EVALUATING ADULT DIETARY INTAKES OF NITRATE AND NITRITE IN POLISH HOUSEHOLDS DURING 2006-2012 Anna Anyżewska*, Agata Wawrzyniak Chair of Nutritional Assessment, Department of Human Nutrition, Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Sciences, Warsaw, Poland ABSTRACT Introduction. Nitrates and nitrites commonly occur throughout nature as well as in foodstuffs. Their excess consumption can however pose health risks, for example, arising from methaemoglobinaemia or from the formation of N-nitrosamines. Objectives. To determine whether the levels of domestic nitrate and nitrite consumption are safe in Polish households during 2006-2012. Material and methods. Appropriate consumption data was obtained from the Central Statistical Office in Poland (GUS), whilst nitrate and nitrite intakes were estimated from nationally available data on foodstuff content taken from the literature. Results. Mean nitrate and nitrite intakes were respectively 147 mg NaNO3 and 3.26 mg NaNO2 /per person/day, corresponding to 41% and 45% of the ADI (acceptable daily intake). Statistically significant differences in intakes were observed between types of households, with the highest seen in those of retired subjects; however the ADIs were not exceeded. Conclusions. Domestic intakes of nitrates and nitrites were found to be at safe levels; nevertheless control over their intake should be maintained because of potentially adverse health threats. Key words: nitrates, nitrites, intake, households STRESZCZENIE Wprowadzenie. Azotany(V) i (III) występują zarówno w przyrodzie jak i w żywności. Nadmierne ich spożycie może powodować zagrożenie zdrowia, np. methemoglobinemię lub może przyczynić się do powstawania N-nitrozoamin. Cel badań. Celem badań było oszacowanie pobrania azotanów(V) i azotanów(III) z żywością w gospodarstwach domowych w Polsce w latach 2006-2012. Materiał i metody. Oszacowanie pobrania azotanów(V) i azotanów(III) w gospodarstwach domowych wykonano na podstawie danych o spożyciu żywności Głównego Urzędu Statystycznego (GUS) oraz zebranych krajowych danych z piśmiennictwa dotyczących zawartości tych związków w produktach spożywczych. Wyniki. Średnie pobranie azotanów(V) i azotanów(III) w latach 2006-2012 wynosiło 147 mg NaNO3/os/dobę (41% ADI) i 3,26 mg NaNO2/os/dobę (45% ADI). Pobranie azotanów(V) i azotanów(III) różniło się istotnie statystycznie w badanych typach gospodarstw domowych. Największe średnie pobranie zarówno azotanów(V) jak i azotanów(III) zaobserwowano w gospodarstwach emerytów, jednak wartości ADI nie zostały przekroczone. Wnioski. Średnie pobranie azotanów(V) i azotanów(III) w gospodarstwach domowych w latach 2006-2012 kształtowało się na bezpiecznym poziomie, niemniej jednak należy kontrolować pobranie tych związków z dietą ze względu na ryzyko możliwych negatywnych skutków zdrowotnych. Słowa kluczowe: azotany (V), azotany (III), pobranie, gospodarstwa domowe INTRODUCTION Both nitrates and nitrites are widespread and naturally occurring ions, mainly arising from organic decomposition of nitrogenous substances. They are found also in mineral salts as well as water. In the latter, their presence is due to the run-off from industry or agriculturally used fertilisers and constitutes the main environmental source [10]. Nitrates can also be present in foodstuffs, depending on the product type, resulting from the technological method of manufacture; for e.g. in the making of cured meats or from using fertiliser for *Corresponding author: Anna Anyżewska, Chair of Nutritional Assessment, Department of Human Nutrition, Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Sciences (SGGW), Nowoursynowska Street 159c, 02-776 Warsaw, Poland, phone +48 22 59 37 122, fax +48 22 59 37 129, [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 108 Nr 2 A. Anyżewska, A. Wawrzyniak plant cultivation. Nitrates have been shown to be decidedly less toxic than nitrites, where the latter arises from nitrate reduction and may cause methaemoglobinaemia [14]. In addition, nitrites can enzymatically react with primary (I), secondary (II) and tertiary (III) amines (via nitro-reductase), together with amino acids, amides, indoles and phenylamines forming N-nitrosoamine products that are well recognised to be carcinogens [9]. The study aims were to estimate nitrate and nitrite intakes from foodstuffs in adults living in defined household groups throughout Poland during 2006-2012. These were then related to ADI values. MATERIAL AND METHODS The study was conducted using foodstuff consumption data from 2006-2012, within Polish household budgets, as made available by the Central Statistical Office in Poland. The following household group categories were selected; manual and non-manual workers, farmers/farm labourers, those self-employed, retired persons and pensioners. Using national data, mainly from the last seven years, the average nitrate and nitrite contents in foodstuffs was obtained from which consumption and intakes were calculated for each of the defined groups. Technological food losses and meal leftovers were not taken into account. In order to relate the findings to ADI for each household type, the following nitrate/nitrite values established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) were used; respectively 5.0 mg NaNO3 and 0.1 mg NaNO2 /kg body mass /24 hours [5], adopting the average persons adult body mass in Poland [11]. The structure of nitrate and nitrite intakes could be presented as well as their sources the total intakes were taken as being 100%. Statistical analyses was performed by the ‘Statistica 10’ computer programme using ANOVA, taking P<0.05 as the critical value for significance. RESULTS The mean nitrate intake during 2006-2012 was 147 mg NaNO3/person/24hours, which decreased by 8% throughout this time (Table 1). Statistically significant differences between household groups were noted (p<0.05), with pensioners having the highest nitrate intakes (198 mg NaNO3/person/24hours). Lower values (by respectively 7% and 9%) were observed for farmers/ farm labourers and retired persons. All other groups Table 1. Dietary nitrate intakes in households during 2006-2012 Type of household Non-manual Self-employed Years Total Manual workers Farmers workers workers mg NaNO3/per person/day 2006 154 132 131 195 134 2007 151 130 131 186 131 2008 149 131 130 187 130 2009 149 130 128 189 130 2010 144 127 124 182 124 2011 142 124 124 172 119 2012 141 125 121 175 121 x 147 128a 127a 184b 127a ±SD 5 3 4 8 5 * mean± standard deviation; results flagged with identical letters did not differ significantly Retired persons Pensioners 206 203 198 198 194 193 191 198c 5 185 185 181 180 175 180 170 179b 5 Table 2. Dietary nitrate intakes in households during 2006-2012 compared with ADI values in adults Type of household Years Total Manual workers Non-manual workers 2006 2007 2008 2009 2010 2011 2012 x ±SD 43 42 41 41 40 39 39 41 1 36 36 36 36 35 34 34 35a 1 36 36 36 35 34 34 33 35a 1 Farmers % of adult ADI 54 51 52 52 50 48 48 51b 2 Self-employed workers Retired persons Pensioners 37 36 36 36 34 33 34 35a 1 57 56 55 55 54 53 53 55c 1 51 51 50 50 48 50 47 49b 1 Nr 2 had substantially lower intakes by 35-36%. None of the household groups exceeded the nitrate ADI, which on average were found to be 41% of this value (Table 2). The highest of the ADI (55%) was in the retired persons group, whilst the lowest (35% ADI) was seen for both the manual and non-manual workers group and those self employed; differences being significant. It was found that vegetables and their processed products were the main foodstuff sources for nitrate (88%); Figure 1. kohlrabi, courgettes, peas, sweet corn as well as root and tuber vegetables. Potatoes constituted 1/4th of the nitrate source and beetroots and cabbage were 15% (Figure 2). Fig. 2. Fig. 1. 109 Nitrates and nitrites intake in Polish households in 2006-2012 Foodstuff sources of dietary nitrates and nitrites in households (%) It was also found, that out of the vegetables listed above, 1/3 of the nitrate source came from the ‘other vegetables’ category that included lettuce, leafy and stem vegetables, cauliflower types, pumpkins, peppers, Selected vegetables sources of dietary nitrates in households (%) Nitrite intakes were significantly different between the selected household groups (p<0.05); Table 3. Likewise as for nitrates, intakes of nitrites were highest in the retired persons group at 3.92 mg NaNO2/person/24hours, whilst the lowest were in the pensioners and farmers group (by 4% and 5% respectively and differences being statistically significant). In all the other household Table 3. Dietary nitrite intakes in households during 2006-2012 Years Total Manual wokers 2006 2007 2008 2009 2010 2011 2012 x ±SD 3.23 3.20 3.18 3.36 3.31 3.31 3.21 3.26 0.07 3.02 3.04 3.02 3.20 3.17 3.17 3.08 3.10a 0.08 Type of household Non-manual Self-employed Farmers Retired persons workers workers mg NaNO2/per person/day 2.87 3.77 2.88 3.82 2.86 3.74 2.81 3.79 2.85 3.72 2.80 3.80 2.97 3.82 2.93 4.07 2.92 3.76 2.86 4.03 2.92 3.76 2.86 4.03 2.85 3.45 2.82 3.89 2.89b 3.72c 2.85b 3.92d 0.04 0.12 0.05 0.12 Pensioners 3.59 3,61 3.70 3.89 3.87 3.87 3.84 3.77c 0.13 Table 4. Dietary nitrite intakes in households during 2006-12 compared with ADI values in adults Type of household Years Total Manual workers 2006 2007 2008 2009 2010 2011 2012 x ±SD 45 44 44 46 46 46 44 45 1 42 42 42 44 44 44 42 43a 1 Non-manual workers 40 39 39 41 40 40 39 40b 1 Farmers % of adult ADI 52 52 51 53 52 52 48 51c 2 Self-employed Retired persons workers 40 39 39 40 39 39 39 39b 1 53 52 52 56 56 56 54 54d 2 Pensioners 50 50 51 54 53 53 53 52c 2 110 groups (i.e. manual, non-manual workers and the self employed) nitrite intakes were respectively 21%, 26% and 27% lower than those for retired persons. The intake of nitrites in all groups did not exceed the ADI and varied between 39% of this value for the self employed to 54% in pensioners; average 45% (Table 4). Over three quarters of the nitrite foodstuff source was meat and its processed products (Figure 1), including cold meats and other processed meat products (cold poultry meat, offal, tinned meat, delicatessen products and other culinary specialities, e.g. meat in aspic); Figure 3. High quality cold meats and sausages made up 1/5th of the nitrite intake source. Fig. 3. Nr 2 A. Anyżewska, A. Wawrzyniak Meat and meat product sources of dietary nitrites in households (%) DISCUSSION The amount of nitrate and nitrite intakes depends not only on the original content in foodstuffs, but also by the method of cooking used and the proportion of source foods consumed within a given diet [7]. Other studies have shown very wide variations in nitrate intakes, as for instance between New Zealand and Japan; 72 vs 1545 mg NaNO3/person/24hours, respectively representing 20% and 500% of the ADI. For nitrite intakes this correspondingly ranged from 0.84 mg NaNO2/ person/24hours in New Zealand to 1.6 mg in Korea; respectively 14 and 38% of the ADI ) [2, 12, 13]. In Europe, the ranges for nitrate intakes were between 215 and 626 mg NaNO3/person/24hours (respectively 71 and 205% of the ADI), whilst for nitrites from 0.29 to 1.14 mg NaNO2/person/24hours; respectively 5 and 20% of the ADI [4]. Analogous results from Poland, during 2006-12, were 132 to 190 mg NaNO3/person/24hours and 3.0 to 3.5 mg NaNO2/person/24hours [15]. The average nitrate intakes within these years were 6% less compared to previous studies, whereas those for nitrites were 3% higher. Moreover, the current study has demonstrated twice higher nitrate and nitrite foodstuff intakes in households compared to those observed in students aged 21 – 24 years [16]. Within Polish households, the nitrate and nitrite intakes have not changed over the years and are maintained at safe levels of around half the ADI. It should however be stressed that certain population groups, especially children and the elderly, are more vulnerable to the effects of nitrates/nitrites and their reactant products. In this respect vegetarians are also a susceptible group, as their main dietary foodstuffs are by definition vegetables, which constitute a rich source of these nitrates/nitrites, compared to those adopting traditional diets. It is thereby estimated that nitrate intakes are three times higher in vegetarians [3, 9, 14]. Excessive nitrite intakes may adversely impact health such as in causing methaemoglobinaemia. Nevertheless, both they and their products also produce beneficial effects on the human body such as on the cardiovascular system, lowering blood pressure and decreasing erythrocyte adhesion and aggregation [4, 12, 17]. Eating vegetables rich in nitrite also decreases the oxygen demand during sub-maximal work whilst consuming leafy vegetables lowers the risk of diabetes in women [1, 6]. CONCLUSIONS 1. Nitrate and nitrite intakes, during 2006-2012, for adults living in various types of households were at appropriate levels of 127 – 198 mg NaNO3 and 2.85 – 3.92 mg NaNO2 /person/24 hours. 2. ADI values for both nitrates and nitrites were not exceeded in any of the studied types households; the mean observed intakes were 41 – 45% of the ADI values. 3. The main dietary source of nitrates was vegetables and their products (88%), whilst for nitrites these consisted of cold and processed meats; both at 77%. 4. Observed nitrate and nitrite intakes were at levels safe for health however their dietary intakes should nevertheless be monitored because of the adverse health effects arising when such levels are exceeded. Acknowledgement This study was financed by the Department of Human Nutrition, Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Sciences (SGGW), Poland Conflict of interest The authors declare no conflict of interest. Nr 2 Nitrates and nitrites intake in Polish households in 2006-2012 REFERENCES 1. Bazzano, L.A., Li, T.Y., Joshipura, K.J., HU, F.B.: Intake of fruit, vegetables, and fruit juices and risk of diabetes in women. Diabetes Care 2008; 31:1311–1317. 2. Chung, S.Y., Kim, J.S., Kim, M., Hong, M.K., Lee, J.O., Kim, C.M., Song, I.S.: Survey of nitrate and nitrite contents of vegetables grown in Korea. Fd Addit Contam 2003; 20:621–628. 3. Du S., Zhang Y., Lin X.: Accumulation of nitrate in vegetables and its possible implications to human health. Agric Sc China 2007;6(10):1246-1255. 4. EFSA Nitrate in vegetables. Scientific opinion of the panel on contaminants in the food chain. EFSA J 2008; 689:1-79, [cited 2011 Feb 10]. Available from: http:// www.efsa.europa.eu/EFSA/Scientific_Opinion/contam_ej_689_nitrate_en.pdf. 5. Food and Agriculture Organization/World Health Organisation. Fifty-ninth report of the Joint FAO/WHO Experts Committee on Food Additives. Techn Rep Ser 913, Geneva 2002. 6. Larsen F.J., Weitzberg E., Lungberg J.O., Ekblom B.: Effects of dietary nitrate on oxygen cost during exercise. Acta Physiol 2007;191(1):59-66. 7. Leszczyńska T., Filipiak-Florkiewicz A., Cieślik E., Sikora E., Pisulewski P.: Effects of some processing methods on nitrate and nitrite changes in cruciferous vegetables. J Food Compos Anal 2009; 22:315-321. 8. Mitek M., Anyżewska A., Wawrzyniak A.: Estimated dietary intakes of nitrates in vegetarians compared to a traditional diet in Poland and acceptable daily intakes; is there a risk? Rocz Panstw Zakl Hig 2013;64(2):105-109. 9. Nowak A., Libudzisz Z.: Carcinogens in human gastrointestinal tract. Zyw Nauk Technol Ja 2008; 4(59): 9-25 (in Polish). 111 10. Piotrowski K. (red.): Selected inorganic compounds: nitrate and nitrite In: Basics of Toxicology, Publisher WNT, Warsaw 2006, 207-208 (in Polish). 11. Report on research: Body mass and overweight the Polish population. Available from: http://www.estymator.com. pl/WYNIKI/WAGA_I_NADWAGA_POLAKOW__komunikat.pdf (7.10.2013r.). 12. Sobko, T., Marcus, C., Govoni, M., Kamiya S.: Dietary nitrate in Japanese traditional foods lowers diastolic blood pressure in healthy volunteers. Nitric Oxide 2010; 22:136–140. 13. Thomson, B.M., Nokes, C.J., Cressey P.J.: Intake and risk assessment of nitrate and nitrite from New Zealand foods and drinking water. Fd Addit Contam 2007; 24:113–121. 14. Traczyk I.: Nitrates and nitrites – occurrence and impact on the human body. Żywn Żyw Prawo Zdr 2000; 1:81-89 (in Polish). 15. Wawrzyniak A., Hamułka J., Pająk M.: Evaluation of nitrites and nitrates food intake in Polish households in years 1996-2005. Rocz Panstw Zakl Hig 2008; 59(1):918 (in Polish). 16. Wawrzyniak A., Hamułka J., Pankowska I.: Evaluation of nitrites and nitrates food intake in the students’ group. Rocz Panstw Zakl Hig 2010; 61(4):367-372 (in Polish). 17. Webb A.J., Patel N., Loukogeorgaktis S., Okorie M., Aboud Z., Misra S., Rashid R., Miall P., Deanfield J., Benjamin N., MacAlister R., Hobbs A.J., Ahluvalia A.: Acute blood pressure lowering, vasoprotective, and antiplatelet properties of dietary nitrate via bioconversion to nitrite. Hypertension 2008; 51(3):784-790. Received:14.10.2013 Accepted:19.02.2014 Rocz Panstw Zakl Hig 2014;65(2):113-117 SCHOOL PUPILS AND UNIVERSITY STUDENTS SURVEYED FOR DRINKING BEVERAGES CONTAINING CAFFEINE Magdalena Górnicka*, Jolanta Pierzynowska, Ewelina Kaniewska, Katarzyna Kossakowska, Agnieszka Woźniak Chair of Nutritional Assessment, Department of Human Nutrition, Faculty of Human Nutrition and Consumer Sciences, University of Life Sciences, Warsaw, Poland ABSTRACT Background. Caffeine is a commonly found ingredient in many beverages. Its main dietary source is coffee, cola drinks and in recent years, energy drinks. Objectives. To compare the consumption of drinks containing caffeine (coffee, colas and energy drinks) and the reasons and circumstances under which they were drunk by middle school (junior high school) pupils and university students. Material and methods. Surveyed subjects were 90 middle school pupils from Warsaw and Kutno together with 100 students attending the Warsaw University of Life Sciences (SGGW). A questionnaire, designed by the authors, was used to determine the amounts, frequency and the reasons or circumstances in which coffee, colas and energy drinks were consumed. Statistics used, consisted of the Mann-Whitney U and Chi-square (χ2) tests, with significance taken as α ≤ 0.05. Results. Cola drinks were found to be the most popularly consumed beverages containing caffeine; 97% pupils and 93% students. Coffee was however drunk twice less by pupils compared to students, whilst similar amounts of energy drinks were consumed by both groups; respectively 48% and 53%. Gender differences were observed for the energy drinks with young men drinking the most. Coffee and energy drink consumption also rose with age by respectively 39% and 57%. The mean caffeine intake in pupils and students were respectively estimated to be 141 and 163 mg/day(d). The reasons why these beverages were drunk varied, from drinking coffee to keeping awake and drinking cola because of its good taste. Pupils also drank energy drinks due to its taste but students because of improved mental performance and in staying awake. Conclusions. Drinking caffeine containing drinks by adolescents can be very variable and comes from many different sources. Thus, its intakes may be very high and so require monitoring, particularly for the youngest. Further observational studies are needed to assess the consumption of energy drinks in relation to physical activity. Key words: caffeine, intake, school pupils/children, students STRESZCZENIE Wprowadzenie. Kofeina jest składnikiem wielu spożywanych napojów. Jej głównym źródłem w diecie jest kawa, napoje typu cola, a w ostatnich latach dodatkowo napoje energetyzujące. Cel badań. Celem badania było porównanie ilości i uwarunkowań spożywania napojów zawierających kofeinę (kawy, napojów typu cola i napojów energetyzujących) przez młodzież gimnazjalną i akademicką. Materiał i metody. Badaniami objęto 90 uczniów gimnazjum (Warszawa i Kutno) i 100 studentów ze Szkoły Głównej Gospodarstwa Wiejskiego w Warszawie. Badania zostały przeprowadzone na podstawie autorskiego kwestionariusza, zawierającego pytania dotyczące ilości, częstotliwości i uwarunkowań spożywania kawy, napojów energetyzujących i typu cola. Uzyskane dane poddano analizie statystycznej za pomocą testu U Manna-Whiteney’a oraz testu Chi2, przyjmując poziom istotności α ≤ 0,05. Wyniki. Powszechnie spożywanym napojem zawierającym kofeinę wśród badanej grupy były napoje typu cola, które spożywało 97% gimnazjalistów i 93% studentów. Spożywanie kawy deklarowało prawie dwukrotnie mniej gimnazjalistów w porównaniu ze studentami, natomiast napoje energetyzujące spożywał podobny odsetek badanych w obydwu grupach (48% gimnazjalistów i 53% studentów). Spożywanie napojów energetyzujących zależało od płci, młodzież męska deklarowała częstsze ich spożywanie. Z wiekiem wzrastało spożycie kawy (o 39%) i napojów energetyzujących (o 57%). Średnie spożycie kofeiny oszacowano w grupie gimnazjalnej na poziomie 141 mg/d, a w grupie studentów na poziomie 163 mg/d. Uwarunkowania spożywania napojów zawierających kofeinę różniły się dla poszczególnych ich rodzajów: kawę spożywano *Corresponding author: Magdalena Górnicka, Chair of Nutritional Assessment, Department of Human Nutrition, Faculty of Human Nutrition and Consumer Sciences, University of Life Sciences, Nowoursynowska 159c, 02-776 Warsaw, phone +48 22 5937122, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 114 M. Górnicka, J. Pierzynowska, E. Kaniewska et al. Nr 2 głównie dla zwalczenia senności, napoje typu cola ze względu na smak. Gimnazjaliści spożywali napoje energetyzujące ze względu na smak, a studenci celem poprawy sprawności umysłowej i zwalczenia senności. Wnioski. Spożywanie napojów zawierających kofeinę wśród młodzieży, z uwagi na fakt, iż zawartość w nich kofeiny może być znacznie zróżnicowana, a łączne dostarczanie jej z różnymi produktami, może powodować znacznie wyższe jej pobranie, wymaga monitorowania, zwłaszcza w młodszych grupach wiekowych. Dalszych badań wymaga zaobserwowana zależność spożywania napojów energetyzujących w związku z wysiłkiem fizycznym. Słowa kluczowe: kofeina, spożycie, gimnazjaliści, studenci INTRODUCTION blood glucose through stimulating adrenal hormones that may lead to type II diabetes [5]. Sweetened fizzy Caffeine is an alkaloid, naturally occurring in coffee drinks, such as colas or energy drinks, contribute tobeans, tea leaves, cola nuts, Yerba-mate leaves, cocoa wards the development of overweight and obesity due beans and guarana seeds. Its main source of intake is to their high sugars’ content [2]. A study by Temple [18] has stressed that caffeine, through drinking infusions of coffee, tea or cocoa. An like sugar, may activate the dopaminergic-reward extract from cola nuts is used in the many cola drinks, however synthetic caffeine is added to energy drinks system and thence lead to addiction. Their joint con[19]. Caffeine is also added to some foodstuffs and me- sumption in foodstuffs and beverages synergistically dicines. Modest intakes of caffeine (i.e. 200 – 300 mg) increases dopamine release and as a result their effects are beneficial to health through being a stimulant of the become potentiated, which, in the long term, during central nervous system (CNS), muscle activity, heart and a critical stages of individual’s development are not kidneys. It also increases mental processes/performance clearly understood. During childhood and adolescence, and decreases fatigue and tension in smooth muscles the brain undergoes intensive development, especially of the vasculature [4]. A safe level of daily caffeine those centres responsible for performance, planning and intake is regarded as one not exceeding 400 mg [23]. emotional control, where frequent caffeine consumption Excessive daily intakes above 400 mg may however by these groups may have adverse health impacts. Additionally, energy drinks can contain ingredients cause agitation, sleep disturbances, anxiety, irritability, such as guarana, taurine, inositol, group B vitamins, nervousness [8, 21], as well as insulin resistance [6]. A glucuronolactone and others which enhance the action regular, long-term and excessive caffeine intake may lead to an addiction and adverse health consequences of caffeine [5]. Because of their composition, energy [10]. A toxic dose of caffeine is difficult to precisely drinks should not be given to children nor adolescents define as the literature reports wide variation between below 16 years, however in many countries, including Poland, they are readily available [5, 8]. Nevertheless, 0.5 to 1.5 g for a healthy individual [23]. Children and adolescents widely consume many drinks are generally popular in these age groups. In caffeine containing foodstuff products. Because the the USA it is estimated that children aged 2 – 5 years effects of caffeine on their development and health consume 16 mg/d caffeine, those aged 6 – 11 years 26 are relatively unknown, surveillance of caffeine intake mg/d and 59 – 80 mg/d for ages above 11 years. As levels becomes necessary. Due to the child’s nervous pointed out by Wierzejska [23], these surveys were system being in growth and developmental stages, it is undertaken in the 1990s of the previous century, based supposed that the effects of caffeine are different when on food interviews, when foodstuff products had much compared to adults. In addition, as children are buil- less caffeine. For Polish children, there is a dearth of ding up their bone mass, drinking caffeine containing data on this topic. For this reason, the presented study beverages when coupled to any calcium deficiencies is therefore focused on comparing the amounts and reasons/circumstances for consuming drinks containing will decrease bone mass density [19, 23]. Only in Canada, have maximum daily intakes of caffeine (coffee, colas and energy drinks) in middle caffeine been established for children aged below 12 school pupils and university students. years at < 2.5 mg/kg body mass/d [23]. Other recommendations for children aged 4 – 12 years state safe MATERIAL AND METHODS doses of 45 – 85 mg/d [20] and for adolescents up to 18 years at < 100 mg/d [16]. Children and adolescents most The study was conducted on 90 pupils attending often consume caffeine with sugar as from colas and middle school at Warsaw and Kutno and 100 students energy drinks. This is detrimental as it very likely leads of the Warsaw University of Life Sciences in 2011-12, to having a predilection for sweet tasting foodstuffs. which had been preceded by a pilot study. A proprietary Furthermore, another effect of caffeine is to increase Nr 2 questionnaire was used to assess the consumption of coffee, colas and energy drinks together with questions on age, gender, height, body mass and the amounts, circumstances/reasons why these drinks had been consumed – Table 3. The relevant features of the subject groups are shown in Table 1. The mean intakes of caffeine for both groups were estimated from taking the average contents of caffeine in coffee, colas and sweet drinks as respectively being; 60 mg/100 ml, 11 mg/100 ml and 32 mg/100 ml. To determine what effect age and gender has, the quantitative data were analysed by the Mann-Whitney U-test whilst the χ2 test was used to analyse the qualitative data using the SPSS statistical software package. Significance was taken as α=0.05. RESULTS Both groups had similar gender proportions. The mean ages of middle school pupils was 15 years and 23 years for university students. The BMI was at normal levels in both groups and did not differ significantly (Table 1). Table 1. Characteristics of the study groups Group pupils n=90 students n=100 115 Consumption of beverages containing caffeine by school pupils and students Gender F M Age (years ) Body mass (kg) Height (cm) BMI (kg/m2) 46 44 15 ± 1 62 ± 9 173 ± 15 21 ± 3 50 50 23 ± 2 68 ± 11 173 ± 13 23 ± 2 F – females, M – males The consumption of cola was found to be particularly popular in pupils and students, however drinking coffee was significantly (p=0.02) more popular with students than pupils (88% vs 51%). There were no age differences in consuming colas and energy drinks. Differences between genders were observed, in that males more frequently (p=0.01) drank energy drinks than females; this being true in both groups (respectively p=0.03 and p=0.001). Females from middle school however, drank more coffee (p=0.02); Table 2. The highest numbers of drinks consumed containing caffeine were colas (average of 3 litres per week), where pupils drank 33% more than students; Table 3. The older student subjects significantly drank more energy drinks than pupils (by 57%; p=0.04). Female students drank more colas compared to males (p=0.01), whilst male pupils drank more energy drinks than females (p=0.02). The mean caffeine intakes were 141 mg/person/d (pupils) and 163 mg/person/d (students). Significantly higher mean caffeine intakes were observed in males; p=0.04, Table 3. The main source of caffeine were coffee and colas, however energy drinks only contributed 10 – 20% of the caffeine consumption. The reasons for consuming caffeine containing beverages varied according to category; Table 4. Coffee was mostly drunk to prevent sleepiness and in students to improve mental alertness (92%) and well-being (83%). Meantime, colas were drank because they tasted good and quenched thirst. Pupils drank energy drinks mainly due to them being tasty (65%) and 51% did so to increase physical efficiency. Students however drank energy drinks to improve their mental process function ie. performance (68%) and for keeping awake (65%); Table 4. Most sub- Table 2. Consumption of caffeine-containing beverages (%) in studied groups Beverages Coffee Colas Energy drinks 1 Subjects pupils n=90 51 97 48 students n=100 88 93 53 p1 0.02 NS NS F n = 96 74 94 34 M p1 n= 94 67 NS 96 NS 67 0.01 F n=46 63 98 36 Pupils M n=44 38 96 59 p1 0,02 NS 0.03 F n=50 84 90 32 Students M p1 n=50 92 NS 95 NS 74 0.001 Chi2 test results, F– females, M – males, NS – statistically insignificant differences Table 3. Average intake of caffeine-containing beverages (ml/week) and estimated average caffeine intake (mg/d) in groups Subjects Beverage Pupils Students p1 Pupils F M p1 F M Students p1 F M p1 N = 46 N = 88 N= 74 N= 63 N=29 N=17 N=42 N=46 752±323 1044±415 NS 912±405 884±336 NS 844±350 660±399 NS 980±360 1108±470 NS N = 87 N = 93 N=90 N=90 N=45 N=42 N=45 N=48 Colas 3666±1184 2750±1024 NS 2494±990 3922±1108 NS 3394±1074 3938±1054 NS 1594±894 3906±1157 0.01 Energy N = 43 N = 53 N=33 N=63 N=17 N=26 N=16 N=37 419±192 656±103 0.04 384±202 692±252 NS 255±199 584±171 0,02 512±125 800±158 NS drinks Caffeine 141±57 163±61 NS 134±48 170±59 0.04 137±58 146±62 NS 132±49 193±67 0.03 Coffee 1 Mann-Whitney U test results, F – Females, M – Males, NS – Statistically insignificant differences 116 Nr 2 M. Górnicka, J. Pierzynowska, E. Kaniewska et al. Table 4. Determinants of caffeine-containing drinks intake (% responses) for middle school pupils and university student subjects Reasons and circumstances for drinks’ intake* Reasons Keeping awake Taste Thirst quenching Improved mood Improved mental alertness Increased physical efficiency Circumstances During studying On social occasions During/after physical exertion Coffee Colas Energy drinks Pupils Students n=43 n=53 Pupils n=46 Students n=88 Pupils n=87 Students n=93 74 60 11 28 5 5 85 78 2 82 92 15 7 85 53 39 10 4 2 90 65 20 20 20 10 65 23 33 33 51 65 24 19 26 68 25 55 40 11 89 75 7 31 65 14 30 78 59 23 30 60 50 18 17 *Several replies could be given (values do not add up to 100) jects also declared that the circumstances for drinking coffee (i.e. caffeine containing beverage) was during studying and social occasions (58%). Colas were drank most often socially but in students also after physical exertion (59%). Pupils drank energy drinks mostly to do with physical exercise (60%), whilst 50% students did so during study. DISCUSSION Caffeine containing products are consumed at all ages, resulting from their availability, popularity and stimulating effect, and are widely enjoyed even by younger population groups. Children and adolescents are however most vulnerable to the adverse effects of caffeine because of their still developing metabolism and nervous system [19]. The risk of untoward effects on a developing individual increases when coupled to raised caffeine intakes from various sources [18, 20, 23]. The current study demonstrates that drinking caffeine containing beverages is widespread irrespective of age and does not differ between the two age groups studied. Studies by Wierzbicka et al. [22] on women, Bartosiuk et al. [3] on female students and Semeniuk [17] on students indicated that energy drinks are becoming increasingly popular. According to Kopacz et al. [12], over half of the student subjects consumed energy drinks, with consumption significantly rising during examination periods. The presented study however showed that cola was the most popular. Significant gender differences were observed in especially energy drink consumption. These findings agree with studies by Attele and Cakir [1] and Wanat and Woźniak-Holecka [19] which reported that males drank more energy drinks than females. Caffeine intakes rose with age, particularly from coffee and energy drinks, consistent with the aforementioned Wanat and Woźniak-Holecka [19] study on high school pupils and university students that found that the latter drank more coffee than the former. In children and adolescents, caffeine intake should not exceed 100 mg/d [16], but in the middle school pupils this value was 141 mg which did not differ with the students (i.e. adults). These estimations were somewhat lower than results reported by Wanat and Woźniak-Holecka [19] or Wierzbicka et al. [22]; at respectively 196 – 241 mg/d and 251 mg/d, but the caffeine sources were not accounted for. Furthermore, these estimations were given as mean caffeine intakes, where the ranges may have shown wide variations. The amount of caffeine in coffee depend on the coffee type and method of preparing the drink. For energy drinks there are no legal regulations and established limits, which results in wildly fluctuating caffeine levels in many varieties of products where added caffeine is ever increasing [23]. The main grounds for consuming caffeine in drinks was to improve well-being so that sleepiness could be prevented and that intellectual or physical performance be enhanced. These were likewise found in other studies [1, 11, 14, 17]. Energy drinks were drunk more due to physical exercise in pupils which concurs with studies by Łagowska et al. [13] and Malinauskas et al. [14], which showed that they are mostly drunk for increasing physical and mental efficiency. Similarly, Bajerska et al. [2] found that adolescents engaged in sport are twice more likely to drink energy drinks compared to their peers undertaking lower levels of physical activity. This requires further studies. Energy drinks can cause much harm and even lead to caffeine poisoning. They are perceived by young consumers as being drinks that have a cool image and intensive advertising campaigns are launched to popularise these products as increasing physical-mental endurance and efficiency without having any ill effects on health [5]. The problem is that there are no restrictions on the sale of these products to Nr 2 Consumption of beverages containing caffeine by school pupils and students children and adolescents who have lower tolerances to caffeine [15]. An 2013 EFSA (European Food Standards Agency), report indicated that 68% teenagers (aged 10 – 18 years) consume energy drinks, of whom 12% do so at rates of 7 litres per month. In conclusion, the study has demonstrated that the popular consumption of such beverages may lead to an excess caffeine intake in middle school pupils which does not differ from their older student counterparts i.e. in effect adults. CONCLUSIONS 1. For adolescents, the consumption of beverages containing caffeine may lead to excessive intakes because of the wide variations in product content. This requires monitoring, particularly for the more vulnerable, younger age groups. 2. Further studies are needed to assess the observed relation between energy drinks and undertaken physical activity. Conflict of interest The authors declare no conflict of interest. REFERENCES 1. Attele S., Cakir B.: Energy-drink consumption in college students and associated factors. Nutrition J 2011;27:316322. 2. Bajerska J., Woźniewicz M., Jeszka J., Wierzejska E.: Częstość spożycia napojów energetyzujących, a aktywność fizyczna i występowania nadwagi i otyłości wśród młodzieży licealnej. Żyw Nauka Technol Ja 2009;4:211217. 3. Bartosiuk E., Markiewicz-Żukowska R., Puścion A., Mystkowska K.: Ocena spożycia żywności typu ‘fast food’ oraz napojów energetyzujących i alkoholu wśród grupy studentek Uniwersytetu Medycznego w Białymstoku. Bromat Chem Toksykol 2012; 3: 766-770. 4. Białas M., Łuczak H., Przygoński K.: Zawartość kofeiny w wybranych napojach kawowych w proszku. Bromat Chem Toksykol 2009;3:426-430. 5. Cichocki M.: Napoje energetyzujące – współczesne zagrożenia zdrowotne dzieci i młodzieży. Przeg Lek 2012;69:854-860. 6. Dworzański W., Opielak G., Burdan F.: Niepożądane działania kofeiny. Pol Merk Lek 2009;161:357-361. 7. EFSA. Energy drinks report. www. efsa.eurpoa.eu/en/ press/news/130306.htm (10.09.2013) 8. Greenberg J.A., Boozer C., Geliebter A.: Coffee, diabetes and weight control. Am J Clin Nutr 2006;84:682-693. 117 9. Gunja N., Brown J.: Energy drinks: health risks and toxicity. Med J Aust 2012;196:46-51. 10. Jarosz M., Wierzejska R., Mojska H., Świderska K., Siuba M.: Zawartość kofeiny w produktach spożywczych. Bromat Chem Toksykol 2009;3:776-781. 11. Kopacz A., Wawrzyniak A., Hamułka J., Górnicka M.: Studies on the determinants of energy in drinks intake by students. Rocz Panstw Zakl Hig 2012;63(4):491-497 (in Polish). 12. Kopacz A., Wawrzyniak A., Hamułka J., Górnicka M.: Evaluation of energy drink intake in selected student groups. Rocz Panstw Zakl Hig 2013;64(1):49-53 (in Polish). 13. Łagowska K., Woźniewicz M., Jeszka J., Posłuszny M.: Ocena częstotliwości spożycia produktów, potraw i napojów o wysokiej wartości energetycznej przez młodzież szkolną o różnym poziomie aktywności fizycznej. Zeszyty Nauk Wielkopol Wyż Szk Turyst Zarządz w Poznaniu 2011;6:91-99. 14. Malinauskas B.M., Aeby V.G., Overton R.F., CarpenterAeby T., Barber- Heidal K.: A survey of energy drink consumption patterns among college students. Nutrition J 2007;6:35-38. 15. Reissig C. J., Strain E. C., Griffiths R. R.: Caffeinated energy drinks – a growing problem. Drug Alcohol Depend 2009;99:1-10. 16. Seifert S.M., Schaechter J.L., Hershorin E.R., Lipshultz S.E.: Health effects of energy drinks on children, adolescents, and young adults. Pediatrics 2011;127:511-528. 17. Semeniuk W.: Spożywanie napojów energetyzujących wśród studentów Uniwersytetu Przyrodniczego w Lublinie. Probl Hig Epidemiol 2011;92:936-939. 18. Temple J.L.: Caffeine use in children: what we know, what we have left to learn, and why we should worry. Neurosci Behav Rev 2009;33:793-806. 19. Wanat G., Woźniak-Holecka J.: Ocena konsumpcji produktów zawierających kofeinę wśród młodzieży akademickiej i licealnej. Probl Hig Epidemiol 2011;92:695699. 20. Warzak W.J., Evans S., Floress M.T., Gross A.C., Stoolman S.: Caffeine consumption in young children. J Pediatr 2011;158:508-509. 21. Whalen D.J., Silk J.S., Semel M., Forbes E.E., Ryan N.D., Axelson D.A., Birmaher B., Dahl R.E.: Caffeine consumption, sleep and affects in the natural environments of depressed youth and healthy control. J Pediat Psychol 2008;33:358-367. 22. Wierzbicka E., Gałkowska K., Brzozowska A.: Ocena spożycia kofeiny z całodzienną racją pokarmową w wybranej grupie dorosłych kobiet. Probl Hig Epidemiol 2010;91:564-571. 23. Wierzejska R.: Caffeine – common ingredients in a diet and its influence on human health. Rocz Panstw Zakl Hig 2012;63(2):141-147 (in Polish). Received: 20.10.2013 Accepted: 06.02.2014 Rocz Panstw Zakl Hig 2014;65(2):119-126 THE USE OF VITAMIN SUPPLEMENTS AMONG ADULTS IN WARSAW: IS THERE ANY NUTRITIONAL BENEFIT? Anna Waśkiewicz 1*, Elżbieta Sygnowska 1, Grażyna Broda 1 , Zofia Chwojnowska 2 Department of CVD Epidemiology, Prevention and Health Promotion, Institute of Cardiology, Warsaw, Poland Independent Unit of Nutritional Epidemiology and Dietary Recommended Intakes, National Food and Nutrition Institute, Warsaw, Poland 1 2 ABSTRACT Background. The use of dietary supplements is widespread and can contribute substantially to total nutrient intake. However, it also generates some potential risks in the case of unreasonable and excessive use of such products. Objective. To estimate the prevalence of supplementation and the vitamin supplement contribution to total intake among Warsaw population aged 20-74 years. Material and methods. Nutrient intake and supplement use were studied in a representative sample of Warsaw population in years 2011/12 (486 men and 421 women) and in 2001 (658 and 671 respectively). The vitamin levels were analyzed in reference to the Recommended Dietary Allowance (RDA) and the tolerable upper intake level (UL). Results. In the years 2011/12 the use of dietary supplements (vitamins and minerals) was reported by 31% men and 40% women. Vitamin intake from food showed the deficiency of vitamins D, B1 and folates and adequate intake of vitamins A, C, E, B2, B6, B12. Supplementing with vitamins D and B1 as well as folic acid contributed to better RDA fulfillment. Supplementing with vitamins A, C, E, B2, B6 and B12 was not justified because these vitamins were taken in sufficient amounts with food. In 1.3%-14.9% supplement users, the total intake of vitamins A, C, E and B6 exceeded the UL. The prevalence of supplementation of vitamins A, C and E did not change between 2001 and 2011/12, but the total intake of vitamin A in both sexes and vitamins C, E in women was significantly higher in 2001. Conclusions. The use of dietary supplements in Warsaw population was widespread and in case of some vitamins- unreasonable. Key words: vitamin supplements, vitamin intake, adult population, recommended dietary allowances, tolerable upper intake level STRESZCZENIE Wprowadzenie. Przyjmowanie suplementów diety jest popularne i może stanowić istotne źródło witamin i składników mineralnych. Jednocześnie niekontrolowane ich pobranie może stwarzać niebezpieczeństwo nadmiernego spożycia. Cel badań. Ustalenie rozpowszechnienia i zasadności stosowania suplementacji wśród mieszkańców Warszawy w wieku 20-74 lat. Materiał i metody. Sposób żywienia oraz przyjmowanie suplementów oceniono w reprezentatywnej próbie populacji Warszawy w roku 2011/12 (u 486 mężczyzn i 421 kobiet) oraz w roku 2001 (u odpowiednio 658 i 671 osób). Pobranie witamin analizowano w odniesieniu do zalecanego dziennego spożycia (RDA) oraz górnych bezpiecznych poziomów spożycia (UL). Wyniki. W latach 2011/2012 suplementy witaminowo-mineralne przyjmowało 31% mężczyzn i 40% kobiet. Spożycie witamin z żywnością było niedoborowe w przypadku witamin D, B1 i folianów oraz zgodne z zaleceniami dla witamin A, C, E, B2, B6, B12. Suplementacja witaminami D i B1 oraz kwasem foliowym przyczyniła się do lepszej realizacji RDA. Natomiast uzupełnianie diety witaminami A, C, E, B2, B6 oraz B12 nie miało uzasadnienia, ze względu na wystarczające ich spożycie z żywnością. W przypadku 1,3%-14,9% osób stosujących suplementy witamin A, C, E, i B6 notowano przekroczenie poziomów UL. Częstość przyjmowania suplementów witamin A, C i E była podobna w latach 2001 i 2011/12, ale sumaryczne pobranie witaminy A u obu płci oraz C, E u kobiet było istotnie wyższe w roku 2001. Wnioski. Wzbogacanie diety suplementami przez mieszkańców Warszawy było szeroko rozpowszechnione, a w przypadku niektórych witamin nieuzasadnione. Słowa kluczowe: suplementy witamin, spożycie witamin, dorosła populacja, zalecane spożycie, górny bezpieczny poziom spożycia *Corresponding author: Anna Waśkiewicz, Department of CVD Epidemiology, Prevention and Health Promotion, Cardinal Stefan Wyszyński Institute of Cardiology, Warsaw, Poland, Alpejska 42 04-628 Warsaw, Tel.: 48 22 8156556; Fax: 48 22 8125586, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 120 A.Waśkiewicz, E. Sygnowska, G. Broda et al. INTRODUCTION If a human organism is to function correctly, it also needs vitamins, most of which it cannot synthesize on its own. Some of them, particularly antioxidants, i.e. vitamins A, C, E and B group (including folates), play an important role in the prevention of chronic diseases, including cardiovascular diseases [10, 13, 14]. It must be stressed that vitamin intake should be supplied by a healthy diet, not by using supplements. Yet it turns out insufficient in numerous situations, such as using low energy diets or such stimulants as tobacco or alcohol, as well as in women in childbearing age. On the one hand, dietary supplements can play a significant role in lowering the risk of the vitamin deficiency; on the other hand, their uncontrolled consumption may result in crossing the thresholds of the tolerable upper intake level [6]. Since the dietary supplements have become popular, they should be considered as a source of vitamins and dietary minerals while evaluating the dietary patterns [22, 23]. It should be emphasised that no current studies regarding the supplementation that would include a representative group of all adult individuals in central Poland have been conducted recently. The results of our project based on standardised methods allowed the evaluation of the discussed issues in the population of Warsaw. The aim of the study was to estimate the patterns of supplementation and to evaluate the vitamin supplement contribution to the total intake among Warsaw adult population. MATERIAL AND METHODS Subjects and study design The material for the analysis comprised the data from the European Health Examination Survey–Joint Action (Polish part) - EHES-JA and from the Warsaw Health Survey – WAW-KARD, which was a continuation of the EHES project [7, 16]. The objective of both projects, performed in 2011/12, was to assess the health condition of Warsaw’s inhabitants in terms of risks leading to the development of cardiovascular and some other chronic diseases. The study included a representative randomized sample of the whole of Warsaw’s population aged 20 years and above – there were 1081 people examined. The operator of randomization was the PESEL system (PESEL – Universal Electronic System for Registration of the Population). The sample randomization scheme was a one-step scheme – a simple sample stratified in terms of sex and place of residence (department of Warsaw). In accordance with Nr 2 the international recommendations on epidemiological studies, a profile of classic risk factors for developing chronic diseases was assessed in all subjects, based on questionnaire, laboratory, anthropometric, blood pressure measurement findings and on subjects’ dietary habits. Dietary patterns and supplement intake were assessed using the 24-hour recall method, in which respondent provides all the products, food and beverages consumed within 24-h before recall. Due to its advantages (low costs, possibility to standardize, a short time of interviewing and no impact on dietary habits) 24-h recall method is commonly used in epidemiological studies. Portion sizes of food consumed was determined based on the album with photographs of more than 200 foodstuffs prepared specifically for this type of research by the National Food and Nutrition Institute (NFNI) (Instytut Żywności i Żywienia). Subjects were asked if they had taken any form of dietary supplement on the recall day and the supplement type, name brand, and dose were recorded. The vitamin intake in the diet was calculated based on the amount of food consumed, with the use of “Polish Food Composition Tables”, including vitamin losses arising during the technological processes of food preparation [15]. The amount of vitamin derived from supplementation was estimated using the NFNI 4D Diet (IŻŻ Dieta 4D) software, that includes a database nutrient pharmaceutical formulations in 1231 supplements, available on the Polish market. The 2001-year data were obtained from the Pol-MONICA bis study, which covered a representative sample of the right-bank Warsaw’s population aged 20-74 years – 679 men and 691 women. Details regarding study were published previously [28]. Their dietary patterns and supplement intake (only vitamins A, C and E, calcium and magnesium) were assessed in the same way as it was done in the EHES and WAW-KARD projects. The analyses included the data on 486 men and 421 women from the EHES and WAW-KARD studies, and 658 men and 671 women from the Pol-MONICA study; who were aged 20-74 years, and whose dietary data were reliable.The vitamin levels were analyzed in reference to the Recommended Dietary Allowance (RDA) [13] and the tolerable upper intake level (UL) [6]. Statistical methods The statistical analyses were performed with the Statistical Analysis System (SAS) 9.2 program using an analysis of covariance (GLM-procedure) and chi2 test (FREQ-procedure) to compare mean values or prevalence of the analyzed factors. The methods of descriptive statistics were employed, the percentage of subjects taking supplements and the mean vitamin intake from food and from supplements were calculated. Nr 2 121 Vitamin supplements use among adults in Warsaw RESULTS Table 1. Prevalence of vitamin/minerals supplementation use in adult Warsaw population in years 2011/12 (%) Dietary supplementation prevalence Adult Warsaw’s inhabitants supplemented their diets with vitamins and minerals quite prevalently. It was more popular among women (40%) than among men (31%). The decision to take such supplementation was most popular among single. The supplementation prevalence was influenced by the subjects’ educational and income status – those with higher education supplemented their diet more often than those with primary education (1.6 times men and around 4.8 times women). The highest supplementation intake was noted among people of the highest income (Table 1). Doses of supplemental vitamins (among supplement users of a selected nutrients) In supplement users, the mean vitamin intake only from this source (excluding food) exceeded the RDA (except for folates). Depending on the vitamin, the actual intake ranged from 130% to 440% of the RDA. Using vitamin doses higher than UL was reported by 1.3%-4,5% of respondents in the case of vitamins A, E and B6 (Table 2). Men 31.3 Women 40.0 31.1 28.5 (ns)* 31.9 40.0 37.4 (ns)* 48.2 21.6 27.2 35.5 9.9 30.3 47.7 29.3 34.3 34.2 45.9 19.5 18.8 35.1 38.2 23.6 33.6 41.1 45.1 Supplement users Age 20-40 years 40-60 years 60-74 years Education ** primary secondary university Marital status ** married single Net income per capita in the family/month** <1000 PLN 1001-2000 PLN 2001-3000 PLN >3000 PLN * - comparison of prevalence of supplementation use between age groups (test chi2) **- value standardized for age structure in Warsaw population for 30 June 2011 Vitamin intake in daily food ration The analysis of the vitamin intake from food, both in the group of supplement users and that of nonusers, showed the deficiency of vitamins D, B1 and folates. The Table 2. Vitamin intake from supplements (among supplement users of a selected nutrient) in relation to Recommended Dietary Allowance (RDA) and the tolerable upper intake level (UL) in Warsaw population in years 2011/12 Vitamins Suplement users number (percentage) Vitamin A (μg) Vitamin C (mg) Vitamin E (mg) 54 (12.4%) 73 (16.9%) 65 (15.6%) Vitamin D (μg) 37 (9.0%) Vitamin B1 (mg) Vitamin B2 (mg) 50 (12.4%) 50 (12.4%) Vitamin B6 (mg) 80 (18.0%) Vitamin B12 (μg) Folate (μg) 30 (7.2%) 31 (7.9%) Vitamin A (μg) Vitamin C (mg) Vitamin E (mg) 67 (16.0%) 83 (19.4%) 70 (15.8%) Vitamin D (μg) 58 (12.7%) Vitamin B1 (mg) Vitamin B2 (mg) 50 (12.2%) 52 (12.6%) Vitamin B6 (mg) 88 (20.5%) RDA Men 900 90 10 5-15 (5)1 1.3 1.3 1.3-1.7 (1.5)1 2.4 400 Women 700 75 8 5-15 (5)1 1.1 1.1 1.3-1.5 (1.4)1 2.4 400 Mean intake % RDA UL Subjects with intakes exceeding UL (%) 130 133 236 3000 1000 300 3.7 0 1.5 197 50 0 265 217 - - 281 25 1.3 330 74 10002 0 137 166 348 3000 1000 300 4.5 0 2.9 151 50 0 137 153 - - 440 25 3.4 10002 0 Vitamin B12 (μg) 24 (5.8%) 193 Folate (μg) 28 (8.0%) 73 1 - values in brackets assumed as RDA 2 - UL for folic acid ref. only to folic acid supplements (without folate in food) 122 Nr 2 A.Waśkiewicz, E. Sygnowska, G. Broda et al. Table 3. Vitamin intake from food and from supplements in daily diets among supplement nonusers and users in Warsaw population in years 2011/12 Supplement nonusers Average intake % RDA from food Vitamins Vitamin A (μg) Vitamin C (mg) Vitamin E (mg) Vitamin D (μg) Vitamin B1 (mg) Vitamin B2 (mg) Vitamin B6 (mg) Vitamin B12 (μg) Folate (μg) 1063±1784 86.4±78.5 11.2±6.6 4.1±5.4 1.27±0.62 1.54±0.76 1.79±0.78 4.08±6.17 252±126 118 96 112 82 98 118 119 170 63 Vitamin A (μg) Vitamin C (mg) Vitamin E (mg) Vitamin D (μg) Vitamin B1 (mg) Vitamin B2 (mg) Vitamin B6 (mg) Vitamin B12 (μg) Folate (μg) 1013±1634 90.2±92.1 8.4±5.3 3.0±4.3 0.88±0.41 1.30±0.62 1.42±0.63 3.56±6.31 225±124 144 150 105 60 80 118 101 148 56 from food Men 1335±1573 105.1±98.6 12.8±6.3 4.0±5.8 1.29±0.62 1.70±0.80 2.02±0.91 4.81±9.34 328±213 Women 1224±1918 73.0±55.8 8.5±4.9 2.0±1.6 0.80±0.31 1.29±0.75 1.36±0.47 3.87±2.68 197±64 intake of other vitamins was within the RDA (except for vitamin C in a group of men who did not use supplementation) (Table 3). Supplementation effectiveness Supplementing the diet with vitamins D, B1 and folic acid was justifiable because it eliminated the deficiency of these nutrients in the diet. In the case of other vitamins, i.e. A, C, E, B2, B6, B12, their supplementation was not necessary because their Supplement users Average intake from supplements total % RDA >UL 1170±1788 119.8±98.6 23.5±53.6 9.8±6.8 3.44±4.25 2.82±4.20 4.21±3.81 7.92±17.9 297±233 2505±2263 224.9±151.6 36.3±54.1 13.8±8.2 4.73±4.22 4.52±4.15 6.23±3.74 12.73±19.9 625±311 278 250 363 276 363 348 415 530 156 8.3 0 1.5 0 1.3 0 1234±2259 124.5±166.0 34.7±89.1 7.6±6.9 1.78±1.25 1.99±1.62 6.60±12.86 4.64±5.38 293±172 2458±3114 197.5±174.0 43.2±90.0 9.6±7.4 2.58±1.26 3.28±1.63 7.96±12.88 8.51±6.42 490±189 351 263 540 192 235 300 569 354 122 14.9 2.4 2.9 0 3.4 0 mean intake with food was sufficient to meet the RDA. The total intake (from food and from supplements) of vitamins mentioned above exceeded RDA within 250%570%. With vitamins A, E, B6 in both sexes and vitamin C in women, UL levels were exceeded (Table 3). Supplementation in 2011/12 in comparison to that in 2001 The prevalence of supplementation with vitamins A, C and E did not change between 2001 and 2011/12. (Figure 1). However, in the years 2011/12 the realization % 25 men women 19,4 20 16,1 17,1 16,9 16,0 16,0 15,6 15,4 15,8 15 10,9 12,6 12,4 10 5 0 vitamin A vitamin C vitamin E vitamin A 2001 vitamin C 2011/12 Figure 1. Prevalence of selected vitamin supplements used in 2001 and 2011/12 Figure 1. Prevalence of selected vitamin supplements used in 2001 and 2011/12 vitamin E nonparametric test Wilcoxon Nr 2 123 Vitamin supplements use among adults in Warsaw % RDA 1100 men 1000 women ** 997 ** 900 844 800 700 600 500 400 300 540 418 ** 405 363 277 351 297 340 ** 263 250 200 100 0 vitamin A vitamin C vitamin E vitamin A 2001 2011/12 vitamin C vitamin E nonparametric test Wilcoxon ** p < 0.001 Figure 2. Total vitamin in relation to among supplement users of a selected 2001 and 2011/12 Figure 2. Total vitamin intakeintake in relation toRDAs RDAs among supplement usersnutrient of a inselected nutrient in 2001 and 2011/12 of RDA for vitamin A in both sexes and vitamins C, E in women was significantly lower than in 2001 (Figure 2). In 2001, the rate of adherence to recommendation for vitamins A and E in women was very high, and reached 844% and 997%, respectively. DISCUSSION The results of the present study show that enriching a nutritional ration with dietary supplements was more popular among the inhabitants of Warsaw (31% - men and 40% - women) than in the population of the whole of Poland. The findings of the study performed by NFNI in Polish households showed that supplementation was used by 20% of all people [25]; in the WOBASZ study, which included a representative randomized sample of the Polish population, dietary supplements were consumed by 4.6% of men and 11.3% of women [24]. Among the elderly Warsaw dwellers with cardiovascular diseases, vitamin and mineral supplementation was declared by 66.3% [26]. In other countries the frequency of supplementation was very varied, e.g. in the US representative group of adults it was 54% [2] and in the group of German women – 40%, men – 33% [21]. The studies carried out by Flynn et al [8] on minerals and vitamins taken with food and supplements by adults in selected European countries showed that the percentage of people taking supplementation was as follows: Finland 32% men and 58% women, Germany respectively 38% and 48%; Ireland 16%, 31%; the Netherlands 21%, 33%; Spain 8%, 10%; UK 29% and 40%. The frequency of supplementation is dependent on many factors, including socioeconomic ones. Both in our study and in other projects, supplementing the diet was more prevalent among women, elderly, single and those of higher socioeconomic status [2]. Of note is the fact that the discrepancy of methods applied to evaluate the supplementation, especially the period covered by the study, creates serious difficulties for making direct comparisons. Our study included only the people who took supplements during the day preceding the test; in other studies the questions about supplementation referred to the period ranging from one day to one year. Additionally, some studies qualified only vitamins and minerals as supplements, others also included herbal supplements. The precondition of effective supplementation is taking such amounts of particular vitamins from pharmaceuticals that their deficiency is leveled, preferably to the values recommended for daily intake in the diet. The safe zone for nutrients intake lies between the recommended value and the tolerable upper intake level (UL). The threat arises when the total vitamin intake, both from food and supplements, exceeds UL. In Warsaw’s population, the amount of vitamins from supplementation (except for folic acid) covered over 100% (130% – 440% range) of their daily recommendation; in the case of vitamins A, E and B6, UL was exceeded (in 1.3% - 4.5% of respondents). An analysis of dietary supplements examined by the Polish National Food and Nutrition Institute showed that the daily vitamin doses in particular preparations (except for niacin) did not exceed UL [22]. It means that at least part of 124 A.Waśkiewicz, E. Sygnowska, G. Broda et al. Warsaw’s inhabitants took a few preparations at a time or more than one daily dose. If only the vitamins taken with food were included in the analysis, Warsaw’s inhabitants, both those taking supplementation and those who did not, suffered from significant deficiency of vitamin D, folates and, to a lesser extent, vitamin B1. Many authors point out the fact of universally present subclinical vitamin D deficiency both in Poland and in other countries [8, 17, 18]. The groups particularly prone to insufficient vitamin D intake are vegans and people who eliminate any dairy from their diet, postmenopausal women, in whom low estrogen concentration is associated with bone mass loss, and also elderly people [13]. A similar problem was observed with folates, which are taken in insufficient amounts both in Poland [12, 27] and in other European countries [5, 8]. Mandatory folic acid fortification can be effective as shown by examples from the US [1] and Northern Ireland [11]. Especially women in childbearing age should have their diet supplemented with appropriate amounts of folic acid in order to diminish the likelihood of neural tube developmental defects and other neurological malformations in the child. Unfortunately, most recent studies have confirmed low intake of this nutrient even in this age group and by pregnant women [4, 9]. Among Warsaw’s inhabitants, the supplementation with the vitamins mentioned was effective because it prevents their deficiency. In the case of other vitamins analyzed, i.e. A, E, B2, B6, B12 and C, in women regardless of supplementing, their mean intake with food was sufficient to meet the RDA. Supplementing the diet with these vitamins was not justified. There are reports that excessive vitamin taking is not beneficial, and UL is not a recommended level which should be reached when nutrition is correct. In Warsaw’s adult population, UL was exceeded mainly in the case of vitamin A (in 8.3% of men and 14.9% of women taking vitamin A supplementation) and to a smaller extent vitamins E and B6 in both sexes and vitamin C in women. High doses of fat-soluble vitamins are particularly worrying – they tend to cumulate in tissue. Vitamin A and β-carotene are mentioned in the EU as those whose excessive intake is risky and which tend to exceed UL [6]. Also in the US, the percentage of people who exceed UL for vitamin A is estimated at 10% – 15% [19]. There are reports in literature warning against excessive vitamin E intake as it is associated with the risk for peroxidative process stimulation [20]. Furthermore, there is no unequivocal scientific evidence that dietary supplementation is justified in cardiovascular prevention, except possibly fish oil and niacin [10, 14]. A meta-analysis of 68 randomized studies did not show any beneficial effect of supplements containing antioxidants (A, E, C, β-carotene and selenium) on Nr 2 mortality rates; in the case of β-carotene and vitamins A and E, the effect may be quite opposite [3]. Although there are no established UL values for vitamins B1, B2 and B12, their intake in the Warsaw population using supplementation was high (235% – 530% of RDA). Yet according to the latest knowledge based on data on the consumption in the EU countries, the risk resulting from the excessive intake of the group B vitamins mentioned above is believed to be nonexistent [6]. It must be also added that the methodology applied in this study did not allow us to differentiate whether excessive intake of some vitamins by Warsaw’s inhabitants was short- or long-term, which might prove significant in assessing how much their health was affected. In summary, it seems that the analysis of benefits and risks arising from the use of dietary supplements by the inhabitants of Warsaw is an important issue. On the one hand, as in the case of vitamins D, B1 and folates, it can contribute to lowering the risk of the deficiency of these nutrients in the diet. On the other, as in the case of vitamins A, C, E, B2 and B12, their sufficient intake with food along with high doses from supplements may not be beneficial for the consumers’ health. Referring to the range of dietary supplementation by Warsaw’s inhabitants in the years 2011/12 vs 2001, it should be noted that the share of people taking vitamins A, C and E was similar even though range of dietary supplements in Poland greatly expanded (in 2003 there were available 557 of them, in 2004 – 1187, and in 2005 as many as 1285) [23]. A positive phenomenon was a significant, over two-fold, drop in the intake of vitamins A and E in the group of women who used supplementation of these vitamins. In the case of vitamin A, the realization of RDA dropped from 844% to 351%; with vitamin E from 997% to 540%. Such a high intake of these vitamins in 2001 might have resulted from their aggressive advertising pointing to their role both in the prevention of chronic diseases and beneficial function in dermatology and cosmetology. CONCLUSIONS 1. Dietary supplementation with vitamins and minerals is very prevalent among Warsaw’s inhabitants, more so among women, unmarried, and of higher socioeconomic status. 2. Supplementing the diet with vitamins D and B1 as well as folic acid contributed to better fulfilling nutritional targets. Supplementing with vitamins A, C, E, B2, B6 and B12 was not justified because these vitamins were taken in sufficient amounts with food. 3. The prevalence of supplementation of vitamins A, C and E did not change between 2001 and 2011/12, but the total intake of vitamin A in both sexes and Nr 2 Vitamin supplements use among adults in Warsaw vitamins C, E in women was significantly higher in 2001. Acknowledgements This study was supported by the Institute of Cardiology grant 2.11/I/13. Conflict of interest The authors declare no conflict of interest. REFERENCES 1. Bailey R.L., Dodd K.W., Gahche J.J., Dwyer J.T., McDowell M.A., Yetley E.A., Sempos Ch.A., Burt V.L., Radimer K.L., Picciano M.F.: Total folate and folic acid intake from foods and dietary supplements in the United States: 2003–2006. Am J Clin Nutr 2010;91:231-237. 2. Bailey R.L., Gahche J.J., Lentino C.V., Dwyer J.T., Engel J.S., Thomas P.R., Betz J.M., Sempos Ch.H., Picciano M.F.: Dietary supplement use in the United States, 20032006. J Nutr 2011;41(2):261-266. 3. Bjelakovic G., Nikolova D., Gluud L.L., Simonetti R.G., Gluud Ch.: Mortality in randomized trials of antioxidant supplements for primary and secondary prevention: systematic review and meta-analysis. JAMA 2007;297:842857. 4. Bojar I., Owoc A., Humeniuk E., Wierzba W., Fronczak A.: Inappropriate consumption of vitamins and minerals by pregnant women in Poland. Ann Agric Environ Med 2012;19:263-266. 5. Dhonukshe-Rutten R.A., de Vries J.H., de Bree A., van der Put N., van Staveren W.A., de Groot, L.C.: Dietary intake and status of folate and vitamin B12 and their association with homocysteine and cardiovascular disease in European populations. Eur J Clin Nutr 2009;63:18-30. 6. European Commission. Health & Consumer Protection Directorate-General. Orientation paper on the setting of maximum and minimum amounts for vitamins and minerals in foodstuffs. July 2007. Available from www. ehpm.org/Food-Supplement-Directive.aspx. 7. European Health Examination Survey. Available from www.ehes.inf (13.03.2013). 8. Flynn A., Hirvonen T., Mensink G.B., Ocke M.C., Serra-Majem L., Stoś K., Szponar L., Tetens I., Turrini A., Fletcher R., Wildemann T.: Intake of selected nutrients from foods, from fortification and from supplements in various European countries. Food Nutr Res 2009;53(supl.):1-51. 9. Hamułka J., Wawrzyniak A., Piątkowska D., Górnicka M.: Evaluation of iron, vitamin B12 and folate intake in the selected group of women at childbearing age. Rocz Panstw Zakl Hig 2011;62:263-270 (in Polish). 10. Hill A.M., Fleming J.A., Kris-Etherton P.M.: The role of diet and nutritional supplements in preventing and treating cardiovascular disease. Curr Opin Cardiol 2009;24:433-441. 11. Hoey L., McNulty H., Askin N., Dunne A., Ward M., Pentieva K., Strain J.J., Molloy A.M., Flynn C.A., Scott J.M.: Effect of a voluntary food fortification policy on 125 folate, related B vitamin status, and homocysteine in healthy adults. Am J Clin Nutr 2007;86:1405-1413. 12. Ilow R., Regulska-Ilow B., Różańska D., Zatońska K., Dehghan M., Zhang X., Szuba A., Vatten L., Janik-Koncewicz K., Mańczuk M., Zatoński W.: Evaluation of mineral and vitamin intake in the diet of a sample of Polish population – baseline assessment from the prospective cohort ‘PONS’ study. Ann Agric Environ Med 2011;18:235-240. 13. Jarosz M. (ed.). The Polish dietary standards- amendments. IŻŻ, Warsaw 2012 (in Polish). 14. Kris-Etherton P.M., Lichtenstein A.H., Howard B.V., Steinberg D., Witztum J.L.: Antioxidant vitamin supplements and cardiovascular disease. Circulation 2004;110:637641. 15. Kunachowicz H., Nadolna I., Przygoda B., Iwanow K.: Food composition tables. PZWL, Warsaw 2005 (in Polish). 16. Kuulasmaa K., Tolonen H., Koponen P., Kilpeläinen K., Avdicová M., Broda G., Calleja N., Dias C., Gösswald A., Kubinova R., Mindell J., Männistö S., Palmieri L., Tell G., Trichopoulou A., Verschuren M.: An overview of the European Health Examination Survey Pilot Joint Action. Arch Public Health 2012;70:1-5. doi:10.1186/07787367-70-20. 17. Lebiedzińska A., Rypina M., Czaja J., Petrykowska K., Szefer P.: Analysis of vitamin D content in daily food rations of Polish adults. Bromat Chem Toksykol 2010;43:255-259 (in Polish). 18. Lee J.H., O’Keefe J.H., Bell D., Hensrud D.D., Holick M.F.: Vitamin D deficiency an important, common, and easily treatable cardiovascular risk factor? J Am Coll Cardiol 2008;52:1949-1956. 19. Murphy S.P., White K.K., Park S.Y., Sharma S.: Multivitamin-multimineral supplements’ effect on total nutrient intake. Am J Clin Nutr 2007;85(suppl.):280-284. 20. Person P., Lewis S.A., Britton J., Young I.S., Fogarty A.: The pro-oxidant activity of high-dose vitamin E supplements in vivo. Biodrugs 2006;20:271-273. 21. Reinert A., Rohrmann S., Becker N., Linseisen J.: Lifestyle and diet in people using dietary supplements. A German cohort study. Eur J Nutr 2007;46:165-173. 22. Stoś K., Krygier B., Głowala A., Jarosz M.: The composition of selected food supplements on the basis of actual requirements. Bromat Chem Toksykol 2011;44:596-603 (in Polish). 23. Stoś K., Szponar L., Bogusz W., Wierzejska R., Głowała A.: Food supplements in Poland – health and legislative aspects. Ann Nutr Metab 2007;51(suppl. 1);402. 24. Sygnowska E., Waśkiewicz A.: Evaluation of prevalence and magnitude of vitamins and minerals supplementation in Polish population. Rocz Panstw Zakl Hig 2009;60:167170 (in Polish). 25. Szponar L., Stoś K., Ołtarzewski M.: Food supplements the possibilities of their use for some diseases prevention in Poland. Żyw Człow Metab 2004;31(supl.):462-471 (in Polish). 26. Tokarz A., Stawarska A., Kolczewska M.: Nutritional habits and supplementation of elderly people with car- 126 A.Waśkiewicz, E. Sygnowska, G. Broda et al. diovascular diseases from Warsaw. Rocz Panstw Zakl Hig 2008;59:467-472 (in Polish). 27. Waśkiewicz A., Sygnowska E., Broda G.: Dietary intake of vitamins B6, B12 and folate in relation to homocysteine serum concentration in the adult Polish population - WOBASZ Project. Kardiol Pol 2010;68(3):275-282. Nr 2 28. Waśkiewicz A.: Nutrition quality of daily ford ration of the Warsaw inhabitants In years 1993-2001. Warsaw Pol-MONICA bis Project. Rocz Panstw Zakl Hig 2003;54:197-205 (in Polish). Received: 28.10.2013 Accepted: 03.03.2014 Rocz Panstw Zakl Hig 2014;65(2):127-131 ENERGY AND NUTRITIONAL VALUE OF THE MEALS IN KINDERGARTENS IN NIŠ (SERBIA) Konstansa Lazarevic 1,2*, Dusica Stojanovic 2,3, Dragan Bogdanović 1,2 State University of Novi Pazar, Serbia 2 Public Health Institute, Niš, Serbia 3 School of Medicine, University of Niš, Serbia 1 ABSTRACT Background. It is well known that high-energy diet, rich in fat and carbohydrates, increases the risk of obesity. Preschool age is an important period to acquire the eating habits continued later in adulthood. Therefore, evaluation of child nutrition in kindergartens is especially important in the prevention of future obesity. Objectives. To determine the energy value and energy density of meals consumed by children in kindergartens in Niš (Serbia), including the different types of food, in respect to a probable risk of obesity. Material and methods. The study had been conducted in the years 1998-2012. Three-hundred samples of the meals were gathered and analysed, and the amount of selected food groups used to prepare the meals in kindergartens was calculated (weight, protein, fat and carbohydrate content) in the accredited laboratory of the Public Health Institute in Niš according to the ISO 17025 recommendation. Results. The mean energy value of meals was 978.9 kcal (range: 810 – 1144 kcal). The energy density was low (mean: 1.02 kcal/g, range: 0.92 – 1.42 kcal/g) and decreased over the years, what would imply a reduction in the risk of obesity. The intake of same high-energy food products, such as fats and oils as well as sweets (13,9% and 7,3%, respectively) was higher compared to low-energy foods (fruits – 5.2% and vegetables – 10.8%). Conclusions. The results of our study indicate that children in kindergarten in Niš, in general, were properly nourished in total energy content. The energy value and energy density of the meals consumed did not pose a risk of developing obesity. However, the distribution of food groups differentiated by the energy density level was unfavourable; the deficit of low-energy foods was observed. Planning the child nutrition in kindergartens, with laboratory control of meals, may be an effective strategy in adequate energy intake and prevention of obesity. Providing the higher amount of low-energy foods (fruits and vegetables) in meals in kindergartens is recommended. Key words: energy intake, diet, children, kindergarten, Serbia STRESZCZENIE Wprowadzenie. Wiadomo, że wysokoenergetyczna dieta, bogata w tłuszcz i węglowodany, zwiększa ryzyko otyłości. Wiek przedszkolny jest ważnym okresem nabywania nawyków odżywiania się kontynuowanych później w wieku dorosłym. Dlatego też ocena żywienia dzieci w przedszkolu jest szczególnie ważna w zapobieganiu przyszłej otyłości. Cel badań. Określenie wartości energetycznej i gęstości energii posiłków spożywanych przez dzieci w przedszkolach w Niš (Serbia), z uwzględnieniem różnych typów żywności, w odniesieniu do potencjalnego ryzyka otyłości. Materiał i metody. Badania prowadzono w latach 1998-2012. Zgromadzono i przeanalizowano 300 próbek posiłków. Obliczono ilość wybranych grup żywności użytej do przygotowania posiłków w przedszkolach (zawartość białka, tłuszczu i węglowodanów). Analizę wykonano w laboratorium Instytutu Zdrowia Publicznego w Niš, akredytowanym zgodnie z normą ISO 17025. Wyniki. Średnia wartość posiłków wynosiła 978,9 kcal (zakres: 810 – 1144 kcal). Gęstość energii była niska (średnia: 1.02 kcal/g, zakres: 0.92 – 1.42 kcal/g) i obniżała się w miarę upływu lat, co mogłoby pociągać za sobą zmniejszenie ryzyka otyłości. Spożycie niektórych produktów żywności takich, jak tłuszcze i oleje, jak również słodycze (odpowiednio: 13,9% i 7,3%) było wyższe w porównaniu z żywnością niskoenergetyczną (owoce – 5,2% i warzywa – 10,8%). Wnioski. Wyniki naszych badań wskazują, że dzieci w przedszkolach w Niš, ogólnie rzecz biorąc, żywione były prawidłowo w zakresie całkowitej zawartości energii. Wartość energetyczna i gęstość energii spożywanych posiłków nie stwarzała ryzyka rozwinięcia się otyłości, Jednakże, rozkład grup żywności różniących się poziomem gęstości energii był niekorzystny; zaobserwowano niedobór żywności niskoenergetycznej. Planowanie żywienia dzieci, z laboratoryjną kontrolą posiłków, może być efektywną strategią odpowiedniego spożycia energii i zapobiegania otyłości. Zalecono dostarczanie większej ilości żywności niskoenergetycznej (owoce i warzywa). Słowa kluczowe: spożycie energii, odżywianie się, dzieci, przedszkole, Serbia *Corresponding author: Konstansa Lazarevic, Public Health Institute, Dr Zorana Djindjica 50, 18-000 Niš, Serbia, phone: +38 1182333587, fax: +38118225974, e-mail: [email protected] or [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 128 Nr 2 K. Lazarevic, D. Stojanovic, D. Bogdanović INTRODUCTION The up to date results of a number of the studies indicate that the prevalence of obesity in preschoolers is the high [8, 17], but little attention is paid to the role of diet in obesity prevention in preschool children [18]. Larson’s at al review of 42 international studies on state nutritional policy in childhood suggested that promoting healthy eating as well as physical activity in child care settings are considered to less extent [19]. Therefore, the kindergarten intervention studies are needed to help in prevention of obesity in preschool children [16, 24, 25], The evidences were provided that a reduction in the meal energy density significantly decreases the energy intake in preschool children [3, 20-22], however, it should be remembered that children choose the energy-dense foods that were able to give them pleasant feelings of fullness [15]. Duffey and Papkin reported that probably reason for increasing the energy intake are: energy density of meals, portion size and number of eating/drinking occasions [10]. All these components of the diet may be successfully controlled in kindergartens. The aim of the study was to examine the value of kindergartens meals, measured by energy value, energy density, and distribution of low- and high-energy food groups, whether they may affect the development of obesity in children. MATERIAL AND METHODS The study had been conducted in the years 19982012 in kindergartens in Niš, Serbia. The nutrition of children, who reside in kindergartens, is planned by a nutritionist, physician and nurse, and consists of three meals: breakfast, lunch and snack. Accordingly to the Serbian Book of Regulations (SBR), the kindergarten meals must provided at least 90% (1600 kcal) of the daily energy requirements of children, if they spend 12 hours in kindergarten [5]. The macronutrient contents in the energy intake should have the following distribution: protein 10-15%, carbohydrates 50-60% and fats 25-30%. The material for analysis was collected as follows: four time per year during five random days the one sample of each meal ingredients was collected from serving on the dinning room table in front of a child, and 300 samples (20 annually) of kindergarten meals were gathered. The collected samples of meals in duplicate were transported to the accredited laboratory of the Institute of Public Health in Niš. The ingredients (i.e. milk, tea, bread, cooked food, salads, fruits, justice, etc.) were weighed separately and the level of moisture, protein, carbohydrates, fat and ash was determined [1]. The analyses were done in accordance to the ISO 17025 recommendations. Descriptive statistics (mean, standard deviation), linear trends of energy density (defined as energy value in kilocalories (kcal) divided by weight in grams (g), and percentage distribution of food groups was calculated using the Microsoft Excel software. RESULTS Mean energy value (kcal), weight (g) and energy density (g) are shown in Table 1. The mean content of energy was 978.9 kcal (range: 810 – 1144 kcal), mean weigh of meals – 991.5 g (range: 823 – 1153.4 g), and energy density – 1.02 kcal/g (range: 0.92 – 1.42 kcal/g). Table 1. Mean energy intake, weight of food intake and energy density of kindergarten meals in Niš in the 1998-2012 period Recommended values Meal energy (kcal) 978.9± 121.8 810-1144 1600 Meal weight (g) 991.5± 95.3 823-1153.4 Energy density 1.02± 0.13 0.92 – 1.42 (kcal/g) Variable Mean ± SD Min-max Table 2 shows the macronutrient contents (protein, fat and carbohydrates) in the analysed meals. The share of macronutrients, protein (14.7%), fat (30.6%) and carbohydrates (54.7%), in the total energy intake were in accordance with the national recommendations. Table 2. Macronutrients (protein, fats and carbohydrate) contents of kindergarten meals in Niš in the 19982012 period Macronutrients Mean ± SD (g) Energy from macronutrients (kcal) Protein Fats Carbohydrates 35.0 ± 4.8 32.2± 6.3 130.6± 14.3 143.5 299.5 535.5 % of total energy intake (kcal) 14.7 30.6 54.7 Recommended % of total energy intake (kcal) 10-15 25-30 55-60 Figure 1 shows that the linear trends of the mean energy density of the meals in kindergartens decreased significantly in the 1998-2012 period. It would imply a reduction in obesity in childhood due to improper diet. Nevertheless, the greater contribution in energy density of child meals, unfortunately, was noted for the high-energy foods, such as fats, oil and sweets, compared to those of low-energy, i.e. fruits and vegetables (Figure 2). Nr 2 1,5 y = -0,013x + 27,124 R2 = 0,2023 1,4 energy density (cal/g) 129 Energy density of meals in kindergartens in NIS, Serbia 1,3 1,2 1,1 1 0,9 1995 2000 2005 2010 2015 year Fig. 1. Trends of mean energy density of kindergarten meals (kcal/g) in Niš in the 1998-2012 period DISSCUSSION Many countries have regulations concerning the recommended level of energy intake in child nutrition, but the agreement between the energy value of child nourishment in kindergartens and the national and world recommendations has rarely been the object of research. Our study confirmed that feeding of kindergarten children in Niš, in general, was proper in total energy content. The energy value of meals did not exceed the level recommended by SBR, and energy density was low and decreased over the years. The maximum energy values presented in the table 1 were lower than the recommended values and were adequate to the time spent by children in the kindergartens, usually shorter than 12 hours. Compared to our results, the Brazilian children in day-care centers consumed meals of lower energy value than required [14]. The children meals had the energy value higher than required due to fat and protein in Poland [13], and higher due to fat in kindergartens of six cities in China [28]. In the 1998 – 1993 period, in 10 out of 24 kindergartens in Zagreb (Croatia), at least one of the analysed parameters of meals (energy value, protein, fat, carbohydrate content) did not meet the national recommendations [6]. Regarding the macronutrients in the child nutrition, the Dietary Reference Intake (DRI) recommends the diet of children aged over 4 years covering: protein 5-20%, carbohydrates 45-65% and fat 30-40% [12]. It should be noted that the SBR recommendations propose a much lower percentage of fat (25 – 30%), and this regulation, established 20 years ago, needs to be changed [5]. The results of our study showed that proportion of selected macronutrients (protein, fat, carbohydrates) in meals of children in kindergarten in Niš met the criteria for both DRI and SBR recommendations. The proper selection of children diet with taking into account the calorie contents of different food products is the subject of a numerous studies. Our investigations found the unfavourable structure of food products differentiated by the level of the energy density in the meals consumed by the kindergarten children in Niš, inconsistent with the Food Guide Pyramid, were fruit and vegetables present the important part of children diet [29]. The distribution of high- and low-energy foods in nutrition of preschool children in care centers varied from country to country, and even between the regions in the same country. The results of the study of 40 child-care centers in New York City indicated that it is necessary to improve the dietary intake of vegetables and foods rich in vitamin E, which was not provided to children in sufficient quantity by preparing meals [11]. In contrast, the children from 20 child-care centers in North Carolina consumed the recommended amount of low-energy foods (whole grains, fruits and vegetables), but also excessive amount of saturated fat and added sugar [2]. Compared with other regions of the world, the Scandinavian children attending daycare centers seem to have the most balanced diet in terms of high- and low-energy foods [23]. 0,2% Other 13,6% Fats and oils 7,3% Sweets 15% Milk and products 5,2% Fruit 10,8% Vegetables 13,9% Meats and products 34% Cereals and products 0 5 10 15 20 25 30 35 Fig. 2. Distribution (%) of food groups in kindergarten meals in Niš in the 1998-2012 period. 130 Nr 2 K. Lazarevic, D. Stojanovic, D. Bogdanović Children in kindergartens formed dietary behaviours developing the preference for certain types of food. The result of the study conducted in Mexico among children aged 3-4 years reported that, in general, children preferred high-energy foods, but those of public daycares were more likely to prefer healthy food of low-energy [7]. Preventing the unhealthy eating habits in preschool children is very important, because a minimum of 400 g fruits and vegetables per day is recommended for protection against the chronic diseases, such as cardiovascular diseases, cancer, diabetes and obesity [29, 30]. The American Dietetic Association obligated the staff of child care settings to promote healthy eating habits in children [26]. The present study has same limitations. The research focused on the children’s diet only in kindergartens. However, it is necessary to know the influence of children’s diet at home in terms of energy and macronutrient intake on their future habits. The children in Brazil received proportionally more energy, proteins and lipids in their meals at home than in the kindergarten [4]. The study conducted in Texas found that the child nourishing at home did not compensate the energy intake due to a low amount of grain and vegetable consuming in the care centers [27]. The role of parents in forming in their children the habits of proper nutrition is essential, but the healthy diet of preschoolers in kindergarten is also important. CONCLUSIONS The findings of our long-term investigations allow us to recognise the trends and current state of nutrition quality of children in kindergartens in Niš with regards to the adequacy of energy intake. In particular, the study showed that: 1. Children in kindergartens were properly nourished in the total energy intake. The mean energy value of meals did not exceed the level statutory recommended. The energy density of meals was low and decreased over the years, what would imply a reduction of the risk of obesity. 2. The distribution of food groups differentiated by the energy density level was unfavourable. The deficit of low-energy foods was observed. Planning the child nutrition in kindergartens, with laboratory control of meals, may be an effective strategy in adequate energy intake. Providing the higher amount of low-energy foods (fruits and vegetables) in kindergarten meals is recommended. Conflict of interest The authors declare no conflict of interest REFERENCES 1. Association of Official Analytical Chemists. Official methods of analysis, 15th Ed. Arlington: Association of Official Analytical Chemists, 1990. 2. Ball SC., Benjamin SE., Ward DS.: Dietary intakes in North Carolina child-care centers: are children meeting current recommendations? J Am Diet Assoc 2008;108(4):718-721. 3. Barlow S.E.: Expert committee recommendations regarding the prevention, assessment, and treatment of child and adolescent overweight and obesity: summary report. Pediatrics 2007;120 (Suppl 4):S164–92. 4. Bernardi J.R., Cezaro C.D., Fisberg R.M., Fisberg M., Vitolo M.R.: Estimation of energy and macronutrient intake at home and in the kindergarten programs in preschool children. J Pediatr (Rio J) 2010;86(1):59-64. 5. Book of regulation on norms of children’s diet in institutions for children, 1994. Official gazette 50/94. 6. Bosnir J., Puntarić D., Tomasić A., Capuder Z.: Caloric and nutritive value of kindergarten meals in Zagreb from 1988 to 1993. Lijec Vjesn 1996;118(10):229-234. 7. De Lira-Garcia C., Bacardi-Gascon M., Jimenez-Cruz A.: Preferences of healthy and unhealthy foods among 3 to 4 year old children in Mexico. Asia Pac J Clin Nutr 2012;21(1):57-63. 8. De Onis M., Blossner M., Borghi E.: Global prevalence and trends of overweight and obesity among preschool children. Am J Clin Nutr 2010; 92(5):1257-1264. 9. DGAC. Report of the DGAC on the Dietary Guidelines for Americans; 2010 (cited 1 February 2011). Available from: www.dietaryguidelines.gov. 10. Duffey K.J., Popkin B.M.: Causes of increased energy intake among children in the u.s., 1977-2010. Am J Prev Med 2013;44(2):e1-8. 11. Erinosho T., Dixon LB., Young C., Brotman LM., Hayman L.L.: Nutrition practices and children’s dietary intakes at 40 child-care centers in New York City. J Am Diet Assoc 2011;111(9):1391-1397. 12. Food and Nutrition Board, Institute of Medicine, National Academy of Sciences. Dietary Reference Intakes for energy, carbohydrate, fiber, fat, fatty acids, cholesterol, protein, and amino acids. Washington, DC: National Academies Press, 2005. 13. Frackiewicz J., Ring-Andrzejczuk K., Gronowska-Senger A.: Energy and selected nutrients content in pre-school children diet of Warsaw district. Rocz Panstw Zakl Hig 2011;62(2):181-185 (in Polish). 14. Gomes RC., da Costa T.H., Schmitz Bde A.: Dietary assessment of pre-school children from Federal District Brazil. Arch Latinoam Nutr 2010;60(2):168-174. 15. Johnson S.L., McPhee L., Birch L.L.: Conditioned preferences: young children prefer flavors associated with high dietary fat. Physiol Behav 1991;50:1245–1251. 16. Jouret B., Ahluwalia N., Dupuy M., Cristini C., NègrePages L., Grandjean H., et al. Prevention of overweight in preschool children: results of kindergarten-based interventions. Int J Obes (Lond) 2009;33(10):1075-1083. Nr 2 Energy density of meals in kindergartens in NIS, Serbia 17. Kosti R.I., Panagiotakos D.B.: The epidemic of obesity in children and adolescents in the world. Cent Eur J Public Health 2006;14(4):151-159. 18. Kuhl E.S., Clifford L.M., Stark L.J. Obesity in preschoolers: behavioural correlates and directions for treatment. Obesity (Silver Spring) 2012;20(1):23-29. 19. Larson N., Ward D.S., Neelon S.B., Story M.: What role can child-care settings play in obesity prevention? A review of the evidence and call for research efforts. J Am Diet Assoc 2011;111(9):1343-1362. 20. Leahy K.E., Birch L.L., Fisher J.O., Rolls B.J.: Reductions in entree energy density increase children’s vegetable intake and reduce energy intake. Obesity (Silver Spring) 2008;16:1559–1565 21. Leahy K.E., Birch L.L., Rolls B.J.: Reducing the energy density of multiple meals decreases the energy intake of preschool-age children. Am J Clin Nutr 2008;88:1459– 1468. 22. Leahy K.E., Birch L.L., Rolls B.J.: Reducing the energy density of an entree decreases children’s energy intake at lunch. J Am Diet Assoc 2008;108:41–8. 23. Lehtisalo J., Erkkola M., Tapanainen H., Kronberg-Kippilä C., Veijola R., Knip M., Virtanen S.M: Food consumption and nutrient intake in day care and at home in 3-year-old Finnish children. Public Health Nutr 2010;13(6A): 957-96. 131 24. Manios Y., Grammatikaki E., Androutsos O., Chinapaw M.J., Gibson E.L., Buijs G., et al. A systematic approach for the development of a kindergarten-based intervention for the prevention of obesity in preschool age children: the ToyBox-study. Obes Rev 2012;(suppl 1):3-12. doi: 10.1111/j.1467-789X.2011.00974.x 25. Mikkelsen B.E.: Policies to promote on physical activity and healthy eating in kindergartens from theory to practice. Int J Pediatr Obes 2011;(suppl 2):8-11. 26. Neelon S.B., Briley M.E.: American Dietetic Association. Position of the American Dietetic Association: benchmarks for nutrition in child care. J Am Diet Assoc 2011;111(4):607-615 27. Padget A., Briley M.E.: Dietary intakes at child-care centers in central Texas fail to meet Food Guide Pyramid recommendations. J Am Diet Assoc 2005;105(5):790793. 28. Yin S., Su Y, Liu Q., Zhang M.: Dietary status of preschool children from day-care kindergartens in six cites of China. Wei Sheng Yan Jiu 2002;31(5):375-378. 29.World Health Organisation. European Region. CINDI dietary guide. Copenhagen: WHO, Europe, 2000. 30. World Health Organisation. Joint WHO/FAO Expert Consultation on Diet, Nutrition, and the Prevention of Chronic Diseases. Geneva, Switzerland: World Health Organization, 2003:160. Received: 20.09.2013 Accepted: 05.02.2014 Rocz Panstw Zakl Hig 2014;65(2):133-138 COMPARING DIABETIC WITH NON-DIABETIC OVERWEIGHT SUBJECTS THROUGH ASSESSING DIETARY INTAKES AND KEY PARAMETERS OF BLOOD BIOCHEMISTRY AND HAEMATOLOGY Karolina Gajda*, Agnieszka Sulich, Jadwiga Hamułka, Agnieszka Białkowska Chair of Nutritional Assessment, Department of Human Nutrition, Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Sciences (SGGW), Warsaw, Poland ABSTRACT Introduction. An important way of preventing type 2diabetes is by adopting a proper diet by which means appropriate control over blood glycaemia and lipids can be achieved. Objectives. To assess selected biochemical and haematological markers in overweight subjects or those suffering from type 2 diabetes in relation to their estimated dietary intake. Material and methods. The study was conducted in 2012 on n=86 overweight or obese subjects living in Warsaw or its environs, of whom n=43 had type 2 diabetes. Dietary intakes were compared between non-diabetics (control group) and diabetics (test group) by 3 day records, whilst the relevant blood biochemistry and haematology results were obtained from medical records; with patient consent. Results. Diabetic subjects had significantly higher serum glucose and CRP levels than controls, respectively; 190 vs 98 mg/ dl and 1.4 vs 1.1 mg/dl. Lipid profiles were however more significantly abnormal in controls, compared to diabetics with respectively; total cholesterol 220 vs 194 mg/dl, LDL-cholesterol 131 vs 107 mg/dl and triglycerides 206 vs 157 mg/dl. There were no significant differences in HDL-cholesterol; respectively 55 vs 51 mg/dl. In the diabetics, calorific intakes from carbohydrates, especially sugars, were significantly lower than controls i.e. 9% vs 13%. The proportional share of calories derived from dietary fats did not differ between groups, nevertheless a positive correlation was observed between dietary fat content with blood cholesterol concentrations in diabetics. Conclusions. Disorders of carbohydrate metabolism were confirmed in both overweight and diabetic (type 2) subjects. In addition, both groups demonstrated untoward lipid profiles that correlated with their improper nutrition. Key words: overweight, obesity, type 2 diabetes, nutrition, lipid profile, C-reactive protein (CRP), adults STRESZCZENIE Wprowadzenie. Prawidłowe żywienie, którego celem jest wyrównanie glikemii oraz profilu lipidowego odgrywa zasadniczą rolę w profilaktyce cukrzycy typu 2. Cel pracy. Ocena wybranych wskaźników biochemicznych krwi (glukoza, lipidogram, białko CRP) u osób z nadmierną masą ciała oraz cukrzycą typu 2 w aspekcie ich sposobu żywienia. Materiał i metody. Badanie przeprowadzono w 2012 roku, wśród 86 mieszkańców Warszawy i okolic, z nadwaga i otyłością, w tym u 43 osób z cukrzycą. Do oceny sposobu żywienia wykorzystano metodę trzydniowego bieżącego notowania. Dane dotyczące wskaźników biochemicznych krwi, za zgodą badanych uzyskano z ich kart zdrowia. Wyniki. U pacjentów z cukrzycą odnotowano istotnie wyższe średnie stężenie glukozy w surowicy krwi (190 vs. 98 mg/dl), jak również wyższe stężenie białka CRP (1,4 vs. 1,1 mg/dl). Biorąc pod uwagę wskaźniki gospodarki lipidowej stwierdzono większe nieprawidłowości w grupie kontrolnej (cholesterol ogółem 220 vs. 194 mg/dl; cholesterol LDL 131 vs. 107 mg/dl; triacyloglicerole 206 vs. 157 mg/dl). Stężenie cholesterolu frakcji HDL nie różniło się istotnie w obydwu grupach (55 vs. 51 mg/dl). Spożycie energii z węglowodanów, zwłaszcza prostych u chorych na cukrzycę było istotnie statystycznie niższe niż w grupie kontrolnej (9 vs. 13%). Procentowy udział tłuszczu w dostarczeniu energii nie różnił się znacząco pomiędzy grupami, przy czym w grupie z cukrzycą odnotowano dodatnią korelację pomiędzy ilością tłuszczu w diecie, a stężeniem cholesterolu we krwi. Wnioski. Uzyskane wyniki potwierdzają występowanie zaburzeń gospodarki węglowodanowej u pacjentów z nadmierną masą ciała oraz cukrzycą typu 2. Ponadto w badanych grupach zaobserwowano niekorzystny profil lipidowy korelujący z ich nieprawidłowym sposobem żywienia. Słowa kluczowe: nadwaga, otyłość, cukrzyca typu 2, sposób żywienia, lipidogram, białko C-reaktywne (CRP), osoby dorosłe * Corresponding author: Karolina Gajda, Chair of Nutritional Assessment, Department of Human Nutrition, Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Sciences (SGGW), Nowoursynowska Street 159c, 02-776 Warsaw, Poland, tel. +48 22 59 37 122, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 134 Nr 2 K. Gajda, A. Sulich, J. Hamułka et al. INTRODUCTION Having an excess body mass (overweight) and type 2 diabetes constitutes a serious health problem in both Poland and worldwide. According to a World Health Organisation (WHO) report, 1 billion people are now overweight (BMI 25-29.9 kg/m2) and 300 million are obese (BMI>30 kg/m2). It was also predicted that in 2015 such levels will rise to 1.5 billion overweight and 700 million obese [27]. The WOBASZ studies conducted in Poland during 2003-5, on subjects aged 20-74 years, demonstrated that respectively 40.4% and 27.9% of men and women were overweight and 20.6% and 20.2% were obese [2]. In somewhat likewise fashion, for type 2 diabetes, there were 285 million people, aged 20-79 years, with this disease worldwide in 2012, whereas in 2030, this figure is expected to rise to 439 million [19]. Studies have demonstrated a close relationship between excess body mass (by the amount of adipose tissue) with a significantly higher risk of suffering from type 2 diabetes [5, 17, 29]. The WHO recognises that obesity, particularly the abdominal variety, accounts for around 80% of type 2 diabetes incidence, with such cases ever increasing [28]. Obese females and males have respectively, an almost 30 and 40 fold risk of developing type 2 diabetes when compared to persons with a normal body mass [5]. An excess of adipose tissue in the body is responsible for a series of metabolic disorders (both endocrinological and immunological) that give rise to type 2 diabetes, hypertension and hyperlipidaemia and in turn lead to accelerated development of arteriosclerosis and increased risk of cardiovascular disease [17]. In recent years, the involvement inflammatory factors has been stressed in the pathogenesis and development of many disease complications that include diabetes [6]. Adopting a proper diet plays a key role in the prevention and treatment of these diseases, especially in the choices made in consuming certain nutrients [8, 14, 22, 26]. The study aim was to assess the significance of selected biochemical markers (ie. lucose, lipid profiles, CRP and haematological parameters) in overweight subjects and those with type 2 diabetes in relation to their diets. MATERIAL AND METHODS The study was conducted in 2012 on 86 adults with excessive body weight (mean age 51 ±14 years) of whom 43 had type 2 diabetes and which constituted the separate test group. Subjects came from Warsaw and the surrounding areas. Dietary intake was assessed by a three day dietary record which covered two working days and one that was work-free. Most of the consumed sizes of foodstuff dishes and meal portions were defined from a photographic album [23] especially designed for such purposes. The daily calorific value of the dietary intake, together with consumption of protein, total carbohydrates (including sugars) and total fat (fatty acids and cholesterol) was estimated by the ‘Diet 5’ computer programme based on ‘Foodstuff composition and nutritional value tables’ [11]. The obtained data were adjusted for nutritional losses incurred during food processing and then compared to reference standards and recommendations [10]. In order to assess the dietary composition of basic components, their proportional (%) share of supplied calories were calculated. For protein and total carbohydrate, this respectively amounted to 10-15% and 50-70% (with sugars being not greater than 10%). The amount of dietary calories obtained from total fat was taken as the reference value of 20-35%, whilst the intakes of saturated fatty acids were assumed to be as low as possible, given that a diet is nutritionally adequate. An acceptable dietary intake value for cholesterol was taken as not being higher than 300 mg/day, whereas for dietary fibre this was taken as being above 25 g/day [10]. Subjects were surveyed by questionnaire to obtain both socio-demographic data (i.e. gender, age, place of residence, self-assessment of health) and anthropometric parameters (height, body mass); the latter two being additionally confirmed during control visits. Blood analysis data were, with subjects’ consent, recorded from their medical records. These consisted of measured concentrations of biochemical markers (glucose, total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides and CRP) and haematological parameters haemoglobin-HGB, hematocrit-HTC, erythrocytes-RBC and leukocytes-WBC). The biochemistry analyses were performed at the Warsaw ALAB medical laboratories. The lipid results were compared to those recommended by the Polish Diabetes Association; PDA [20], whereas the others were assessed according to reference values used at the aforementioned ALAB laboratories. Statistical analyses were performed using the ‘Statistica ver. 10 software’. Data normality was evaluated by the Shapiro-Wilk test, whereas the Mann-Whitney U test determined the significance of differences between study groups for each studied parameter. The strength of any associations were calculated by the Spearman rank correlation coefficient. Significance levels were taken as a p≤0.05 throughout. RESULTS All subjects were overweight, with a high Body Mass Index (BMI) ranging 25-45 kg/m2 (mean 31 ± 6 kg/m2) of whom 50% had type 2 diabetes. Both groups Table 1. Energy values and nutrient composition for the daily diets of studied subjects Overweight subjects Diabetics Non-diabetics p3 n=43 n=43 Energy (kcal) 1947 ± 2761 969 – 25752 2222 ± 387 1376 –2898 0.003 Total protein (g) 84.8 ± 11.2 56.5 – 107.0 84.9 ± 12.6 56.4 – 115.0 NS Total carbohydrate (g) 294 ± 48 133 – 407 333 ± 64 188 – 460 0.0001 Sugars (g) 43.2 ± 12.6 6.5 – 80.3 70.6 ± 24.6 6.6 – 119.4 <0.0001 Dietary fibre (g) 35.0 ± 7.0 19.0 – 48.0 25.0 ± 5.0 12.6 – 33.8 <0.0001 Total fats (g) 62.6 ± 9.2 31.7 – 79.8 70.5 ± 18.5 32.2 – 120.0 0.06 SFA (g) 28.6 ± 5.8 8.6 – 37.0 30.9 ± 7.9 11.3 – 44.6 0.03 MUFA (g) 22.1 ± 4.3 13.8 – 34.3 26.6 ± 9.0 9.3 – 54.5 0.004 PUFA (g) 7.1 ± 2.4 5.0 – 16.4 8.2 ± 3.3 4.1 – 21.7 0.02 283.5 ± 63.0 168.4 – 491.5 297.7 ± 100.0 135.0 – 590.0 NS Cholesterol (mg) 1 Mean±SD; 2min - max; Mann-Whitney-U test results; significant statistically significant differences, p≤ 0.05; NS –not significant differences, p > 0.05; SFA – saturated fatty acids; MUFA monounsaturated fatty acids; PUFA - polyunsaturated fatty acids. Targeted measurement of blood glucose is the most important means used for determining diabetes and evidence of glycaemic control. The diabetic group showed a twice higher fasting glucose compared to controls; 190 vs 98 mg/dl (Table 2). This high glucose concentration was positively and significantly correlated with LDL-cholesterol (r=0.34, p≤0.05), whilst the other lipid markers showed just a positive correlation 60 diabetics * non-diabetics 50 40 % 30 * 20 * 10 A PU F UF A M SF A Fa ts s Su ga r bo hy dr at e te in s 0 Ca r (ie. non-diabetics and diabetics) had similar BMIs. Those in the diabetic group were aged higher than controls; (56 ± 14 vs. 46 ± 13 years), however other socio-demographic features were much the same. In terms of dietary calorific value and nutritional content, there were more irregularities observed for the controls than in diabetics (Table 1, Figure 1). The latter group showed a significantly lower dietary intakes of calories (by 12%), total carbohydrates (by 12%) and total fats (by 11%); (p≤0.05). Cholesterol intakes varied widely from 135-590 mg/day, but with no significant differences between the groups. Mean dietary protein intakes were also similar in both groups at 85 ± 12 g/day. The diabetics, however, on average consumed significantly more dietary fibre (by 40%) than controls. Consumption 135 Nutrition and biochemical parameters in people with excess body weight and type 2 diabetes Pr o Nr 2 *Mann-Whitney-U test results; statistically significant differences, p ≤ 0.05; SFA – saturated fatty acids; MUFA – monounsaturated fatty acids; PUFA – polyunsaturated fatty acids Figure 1.Proportions of calories derived from selected nutrients in the daily diets of studied subjects alone. Both groups had higher than reference values for CRP; this being a marker of inflammation (acute phase response). Mean CRP levels in diabetics were 27% higher than controls and positively correlated with glucose concentration (r=0.29, p≤0.05), leukocyte count (r=0.32, p≤0.05) and the proportional daily share of dietary saturated fatty acids (r=0.23, p≤0.05). Compared to PDA reference values, all subjects demonstrated somewhat high total cholesterol, LDL-cholesterol and triglycerides; respectively 207 mg/dl, 119 mg/dl and 181 mg/dl. Significantly higher levels of these lipids were observed in those non-diabetics with the higher BMIs. HDL-cholesterol concentrations were however similar in both groups and lay within the reference value range. There was much individual variation seen in the lipid parameters irrespective of diabetic status; the greatest being for triglycerides (41-702 mg/dl) and LDL-cholesterol (46-236 mg/dl). Those with diabetes showed respectively a 35%, 44% and 56% agreement with PDA reference levels for total cholesterol, LDL-cholesterol and triglycerides as well as respectively 72% and 50% for men and women in the case of HDL-cholesterol. More normal lipid profiles were seen in both the overweight and diabetic group for those subjects eating healthier diets, particularly in terms of calories, intakes of total fats, fatty acids and fibre compared to the overweight, non-diabetic subjects. Blood morphology results conformed to reference values, however in diabetic women, the erythrocyte count, haemoglobin concentrations and haematocrit were significantly higher than in control women. All other haematological parameters were similar. There was a positive relationship between glucose concentration with intakes of dietary macro-components and cholesterol but a negative association with fibre; but were not statistically significant. Furthermore, in all subjects, positive correlations were found between dietary characteristics, ie. intakes of carbohydrates, 136 Nr 2 K. Gajda, A. Sulich, J. Hamułka et al. Table 2. Results of selected biochemical parameters in blood for studied subjects Analyte Glucose (mg/dl) CRP (mg/dl) Total cholesterol (mg/dl) LDL-Cholesterol (mg/dl) HDL-Cholesterol (mg/dl) Triglycerides (mg/dl) Leukocytes (10³/μl) Erythrocytes (10³/μl) Haemoglobin (g/dl) Hematocrit (%) Platelets (10³/μl) Reference value <110.0 <0.5 <175.0 < 100.0 W* > 50.0 M* > 40.0 < 150.0 4.0-10.0 W* 3.7-5.1 M* 4.1-6.2 W* 12.0-16.0 M* 14.0-18.0 W* 37.0-47.0 M* 40.0-54.0 150.0-450.0 Overweight subjects Diabetics Non-diabetics n=43 n=43 189.7 ± 112.81 98.0 ± 13.1 87.0 – 519.02 51.0 – 129.0 1.4 ± 3.5 1.1 ± 3.3 0.0 – 21.4 0.0 – 21.4 194.2 ± 44.9 219.6 ± 46.6 83.0 – 324.0 91.0 – 333.0 107.0 ± 36.1 130.8 ± 36.5 46.0 – 236.0 58.0 – 236.0 54.9 ± 18.7 50.8 ± 13.1 23.0 – 88.0 33.0 – 85.0 55.5 ± 17.0 50.85 ± 16.9 29.0 – 90.0 29.0 – 99.0 156.8 ± 109.6 205.8 ± 107.1 41.0 – 702.0 55.0 – 702.0 8.3 ± 2.9 8.1 ± 2.4 2.2 – 15.6 4.2 – 13.7 4.0 ± 0.6 4.6 ± 0.4 2.2 – 5.0 3.9 – 5.1 4.3 ± 0.7 5.1 ± 2.2 2.4 – 5.3 3.4 – 15.5 12.1 ± 2.2 13.6 ± 1.5 6.9 – 15.9 9.1 – 15.2 13.3 ± 2.2 13.8 ± 1.8 7.8 – 16.9 10.7 – 17.5 35.7 ± 5.9 40.1 ± 3.5 20.2 – 45.0 29.5 – 43. 38.9 ± 6.1 40.4 ± 5.1 23.7 – 49.9 31.1 – 51.8 280.7 ± 88.6 278.3 ± 79.5 103.0 – 474.0 126.0 – 480.0 p3 <0.001 NS 0.01 <0.001 NS NS <0.001 NS 0.001 NS 0.02 NS 0.002 NS NS Mean ±SD; 2min-max, W* - Women, M* - Men; 3 Mann-Whitney-U test results, statistically significant differences, p ≤ 0.05; NS – statistically not significant differences, p > 0.05 1 sugars, saturated fatty acids and LDL-cholesterol together with the effect of sugar intake on the increase in cholesterol. Such results indicate a relationship between the prevalent dietary habits and in achieving normal levels of lipids during treatment. DISCUSSION As defined by the American Diabetes Association [1], diabetes is a metabolic disease of varied aetiology, demonstrating hyperglycaemia resulting from disorders of insulin secretion or its action or a combination of both. The literature shows that this condition is chronic and is caused by disorders of carbohydrate metabolism, where eating an improper diet leads to glycaemic abnormalities and a disruption of the blood lipid profile [22]. Type 2 diabetes risk factors include genetic disorders and environmental factors, where in the latter, an improper diet, high dietary intakes of calories, saturated fat and cholesterol are important as well as the link to overweight and obesity [14, 21, 25]. In addition, the pathogenesis of type 2 diabetes also includes the role of adiponectin proteins (secreted by white adipose tissue) which improves glucose tolerance. Serum adiponectin levels depend on the BMI and also whether type 2 diabetes is present, where in such cases its levels are lower [15]. Subjects all had excess body mass, which was likewise observed in a study by Włodarek and Głąbska [26], who showed that diabetics above the age of 40 years have, in 87% cases, excess body mass. Studies by Pisarczyk-Wiza et al. [17] and Zielke and Reguła [29] demonstrated that obesity is an important risk factor for acquiring type 2 diabetes, whose effect becomes more pronounced with increasing age. A key factor in the pathogenesis of this condition is insulin resistance that depends on dietary habits which, if improper, leads to an abnormal lipid metabolism. In accordance with PDA guidelines [20], a given diet should not deviate from basic dietary recommendations for healthy people. Nelson et al. [13] found that the majority of diabetics, especially those overweight and obese, do not adopt healthy/appropriate diets. Studies Nr 2 Nutrition and biochemical parameters in people with excess body weight and type 2 diabetes by Mędrela-Kuder [12] on type 2 diabetics have shown that the commonest failings in diets are a lack of eating regular meals, snacking between meals, eating sweets and using inappropriate cooking methods. The presented study found structural shortcomings in dietary habits, particularly as demonstrated by an increase in the share of calories derived from carbohydrates (including sugars) and fats. Fibre intake is important to diabetics because of its beneficial effects in lowering glycaemia and improving the blood lipid profile [8]. Indeed, a high fibre intake was observed in the current study for type 2 diabetics, together with improved dietary habits which may have led to the blood lipid profiles approaching normality - relative to the non-diabetics. This may be reflected in the subject’s conscious decision to eat smaller meals or dishes and the need to keep to dietary recommendations during adopting any dietary therapy for diabetics. Another contributing factor could be that this condition develops over a long period without symptoms, so that un-diagnosed diabetic persons seek treatment only when apparent complications arise. Both type 2 diabetes and obesity can occur independently, nevertheless an excess of body mass will significantly increase the risk of diabetes, and the incidence of hospitalisations [7, 21]. A twofold higher glucose concentration was noted in the presented study for overweight diabetics compared to controls ie. in 84% diabetics and only 16% controls. Similar results were seen in a study by Tripathy et al. [24] on diabetics with average fasting glucose levels of 171 mg/dl compared to normal healthy subjects of 73 mg/dl. Disorders in lipid metabolism are a recognised factor for the incidence of type 2 diabetes. Under non-physiological conditions of high glucose concentration, changes in the lipid profile occur, that are most frequently manifested by high triglyceride levels, low HDL-cholesterol and a normal –moderately high LDL-cholesterol [3]. Moreover, obesity and an excess of abdominal adipose tissue are linked with changes to lipoprotein structure depending on the genes coding for cholesteryl ester transfer protein (CETP). It has been shown that a lack of, or a disorder in its function are factors that affect HDL levels and the effectiveness of reverse cholesterol transport [16]. A study by Fagot-Campagna et al. [4] found that 97% of diabetics had at least one abnormal feature in their lipid profile. Inflammation is recognised to play an important role in type 2 diabetic pathogenesis. In overweight and diabetic persons, hyperglycaemia and adipose tissue are factors that induce chronic inflammatory reactions that appear, amongst others, as an increase in acute phase reaction proteins (such as CRP), which in turn affects atherosclerosis development [6, 18]. The presented study has demonstrated CRP levels higher than the reference value in most subjects studied, with the dia- 137 betics being the most high. A study by Pisaczyk-Wiza et al. [17] has likewise found that obese and diabetic subjects had increased CRP concentrations which had been used as a sensitive marker of inflammation. This relationship was also observed in a Mexican study that a protein marker of inflammation is a significant factor affecting the development of type 2 diabetes and metabolic disorders in women [9]. CONCLUSIONS 1. An abnormal lipid profile in the studied subjects was related to shortcomings in their dietary habits, particularly in the intakes of carbohydrates (including sugars), as well as dietary fibre and fatty acids. 2. Raised serum CRP concentrations in those overweight persons suffering from type 2 diabetes may indicate an inflammatory state arising from long-term hyperglycaemia. 3. Obtained findings illustrate the need for nutritional education and for preventative studies on overweight subjects to reduce the risk of complications resulting from obesity and other conditions so accompanying. Acknowledgement The study was performed as a scientific project financed by the by Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Science (SGGW), Warsaw, Poland Conflict of interest The authors declare no conflict of interest. REFERENCES 1. American Diabetes Association: Diagnosis and classification of diabetes mellitus, Diabetes Care 2010;3(1):62-69. 2. Biela U., Pająk A., Kaczmarczyk-Chałas K., Głuszek J., Tendera M., Waśkiewicz A., Kurjata P., Wyrzykowski P.: The prevalence of overweight and obesity in women and men. The results of the WOBASZ. Kardiol Pol 2005;63(6):632-635 (in Polish). 3. Chapman M.: Metabolic syndrome and type 2 diabetes: lipid and physiological consequences. Diabetes Vasc Dis Res 2007;4(3):5-8. 4. Fagot-Campagna A., Rolka D.B., Beckles G.L.A.: Prevalence of lipid abnormalities, awareness, and treatment in US adults with diabetes. Diabetes 2000;49(1):78-79. 5. Field A.E., Coakley E.H., Must A., Spadano J.L., Laird N., Dietz W.H., Rimm E., Colditz G.A.: Impact of overweight on the risk of developing common chronic diseases during a 10-year period. Arch Intern Med 2001;161(13):1581-1586. 6. Fronczyk A., Majkowska L.: C-reactive protein in diabetes. Diabetol Dośw Klin 2004;4(4):255-259 (in Polish). 138 K. Gajda, A. Sulich, J. Hamułka et al. 7. Gajewska M., Goryński P., Wysocki J.M.: Obesity and type 2 diabetes as main causes of hospitalization in Polish hospitals in 2008. Probl Hig Epidemiol 2011;92(1):132136 (in Polish). 8. Giacco R., Parillo M., Rivellese A.A., Lasorella G., Giacco A., D’Episcopo L., Riccardi G.: Long-term dietary treatment with increased amounts of fiberrich low glycemic index natural foods improves blood glucose control and reduces the number of hypoglycemic events in diabetic patients. Diabetes Care 2000;23:1461-1466. 9. Han T., Sattar N., Williams K., Gonzalez-Villalpando C., Lean M., Haffner S.: Prospective study of C-reactive protein in relations to the development of diabetes and metabolic syndrome in the Mexico City Diabetes Study. Diabetes Care 2002;25:2016-2021. 10. Jarosz M. (red.): Nutritional standards for the Polish population - revision. Warsaw, IŻŻ, 2012 (in Polish). 11. Kunachowicz H., Nadolna I., Przygoda B., Iwanow K.: Component tables and nutritive value of food. Warsaw, PZWL, 2005 (in Polish). 12. Mędrela-Kuder E.: Proper diet in type 2 diabetes as a means of patients’ rehabilitation. Rocz Panstw Zakl Hig 2011;62(2):219-223 (in Polish). 13. Nelson K.M., Reiber G., Boyko E.J.: Diet and exercise among adults with type 2 diabetes: findings from the third national health and nutrition examination survey (NHANES III). Diabetes Care 2002;25:1722–1728. 14. Nishida C., Uany R., Kumanyika S., Shetty P.: The joint WHO/FAO expert consultation on diet, nutrition and the prevention of chronic diseases: process, product and policy implications. Public Health Nutr 2004;7:245-250. 15. Ostrowska L., Fiedorczuk J., Adamska E.: Effect of diet and other factors on serum adiponectin concentrations in patients with type 2 diabetes. Rocz Panstw Zakl Hig 2013; 64(1):61-66. 16. Pachocka M.L., Włodarczyk M., Nowicka G., Kłosiewicz-Latoszek L.: CETP gene TaqIB polymorphism and plasma lipids in patients with overweight and obesity. Rocz Panstw Zakl Hig 2012; 63(2):149-154 (in Polish). 17. Pisarczyk-Wiza D., Zozulińska D., Majchrzak A., Sobieska M., Wiktorowicz K., Wierusz-Wysocka B.: Assessment of selected acute phase proteins in obese type 2 diabetic patients. Diabetol Dośw Klin 2002;2(6):455–460 (in Polish). Nr 2 18. Pradhan A.D., Ridker P.M.: Do atherosclerosis and type 2 diabetes share a common inflammatory basis?. Eur Heart J 2002;23:831–834. 19. Shaw J.E., Sicree R.A., Zimmet P Z.: Global estimates of the prevalence of diabetes for 2010 and 2030. Diabetes Res Clin Pract 2010;87(1):4-14. 20. Stanowisko Polskiego Towarzystwa Diabetologicznego: Clinical recommendations for the management of patients with diabetes, 2011. Diabet Prakt 2011;12;suppl A (in Polish). 21. Supranowicz P., Wysocki M.J., Car J., Dębska A., Gębska-Kuczorwska A., Gromulska L.: The risk of overweight and obesity in chronic diseases among Warsaw inhabitants measured by self-reported method. Rocz Panstw Zakl Hig 2013;64(3):197-204. 22. Szewczyk A., Białek A., Kukielczak A., Czech N., Kokot T., Muc‑Wierzgoń M., Nowakowska-Zajdel E., Klakla K.: Evaluation of eating habits in a group of people with type 1 and 2 diabetes. Probl Hig Epidemiol 2011;92(2):267271 (in Polish). 23. Szponar L., Wolnicka K., Rychlik E.: Photo album of products and foods. Warsaw, IŻŻ, 2000 (in Polish). 24. Tripathy S., Sumathi S., Bhupal Raj G.: Minerals nutritional status of type 2 diabetic subject. Int J Diabetes Dev Ctries 2004;24(1):27-28. 25. Weinstein A.R., Sesso H.D., Lee I.M., Cook N.R., Manson J.E., Buring J.E., Gaziano J.M.: Relationship of physical activity vs body mass index with type 2 diabetes in women. JAMA 2004;292(10):1188-1194. 26. Włodarek D., Głąbska D.: The dietary habits of people with type 2 diabetes. Diabet Prakt 2010;11(1):17-23 (in Polish). 27.World Health Organization. Obesity and overweight. Fact. Sheet. No. 311. September 2006. http://www. who.int/mediacentre/factsheets/JSs311/en/index.html (13.01.2009) 28. WHO: World Health Statistics. World Health Organization, Geneva, 2012. 29. Zielke M., Reguła J.: Diet and physical activity, and anthropometrical parameters in patients with type 2 diabetes. Żyw Człow Metab 2007;34:1131-1137 (in Polish). Received: 04.10.2013 Accepted: 05.03.2014 Rocz Panstw Zakl Hig 2014;65(2):139-145 NUTRITIONAL VALUES OF DIETS CONSUMED BY WOMEN SUFFERING UNIPOLAR DEPRESSION Ewa Stefańska1*, Agnieszka Wendołowicz1, Urszula Kowzan2, Beata Konarzewska2, Agata Szulc2, Lucyna Ostrowska1 Department of Dietetics and Clinical Nutrition, Faculty of Health Sciences, Medical University, Bialystok, Poland 2 Department of Psychiatry, Faculty of Medicine, Medical University, Bialystok, Poland 1 ABSTRACT Background. Previous studies have shown that patients suffering from depression are more likely to adversely change their eating habits (eg. through increases in appetite, comfort eating and compulsive eating), which may result in an abnormal nutritional status. Objectives. To evaluate selected dietary habits, such as the number and type of meals consumed during a normal day and comparing dietary calorific values and nutritional content between women suffering unipolar depression to those without this condition. Material and methods. Subjects were a group of 110 women aged 18-65 years consisting of a test group of 55 women undergoing treatment for unipolar depression at the Department of Psychiatry, Medical University of Bialystok and a control group of 55 women, without depression, attending an Obesity and Diet Related Treatment Centre. A study questionnaire was used to determine their eating habits along with other relevant data. The 24-hour diet recall method was used to obtain quantitative data collected on 3 weekdays and 1 weekend day; results being averaged. The calorific values and nutrient content of selected components, according to mealtimes, were evaluated using the Diet 5.0 computer programme. Results. Those patients with depression showed that the 3 meals/day model dominated whilst the 4 meals/day model was predominant in the control group. The most frequently missed meals for both groups were afternoon tea and the mid-morning meal. Abnormalities in the calorific intake and nutritional contents from various meals were observed in women suffering depression. Conclusions. It seems appropriate to recommend that those women especially suffering from depression should consult with dieticians about their changing dietary habits, particularly for achieving the proper calorific and nutritional values/ balance from their meals. Key words: women, depression, nutritional value of meals STRESZCZENIE Wprowadzenie. Jak wykazały badania u pacjentów chorujących na depresję częściej występują zmiany zachowań żywieniowych (m. in. wzrost apetytu, pocieszanie się jedzeniem, kompulsywne jedzenie), które mogą być przyczyną ich nieprawidłowego stanu odżywienia. Cel. Celem pracy była ocena wybranych nawyków żywieniowych, w tym liczby i rodzaju zwyczajowo spożywanych posiłków w ciągu dnia oraz porównanie wartości energetycznej i odżywczej posiłków w jadłospisach kobiet z depresją jednobiegunową oraz kobiet nie chorujących na depresję. Materiał i metody. Badania przeprowadzono w grupie 110 kobiet w wieku 18-65 lat. Grupę badaną stanowiło 55 kobiet leczonych z powodu depresji jednobiegunowej w Klinice Psychiatrii Uniwersytetu Medycznego w Białymstoku. Grupa kontrolna obejmowała 55 kobiet, u których nie rozpoznano depresji. W badaniach wykorzystano kwestionariusz ankiety zawierający m.in. pytania dotyczące nawyków żywieniowych. Ocenę ilościową dziennych racji pokarmowych dokonano przy użyciu 24-godz. wywiadu żywieniowego zebranego z 3 dni powszednich i 1 dnia weekendowego (wyniki uśredniono). Ocenie poddano (z uwzględnieniem podziału na posiłki) wartość energetyczną oraz zawartość wybranych składników odżywczych wykorzystując program komputerowy Dieta 5.0. Wyniki. Wykazano, iż w grupie pacjentek z depresją dominował model 3 posiłkowy, a w grupie kontrolnej 4 posiłkowy. Najczęściej opuszczanym posiłkiem w obu grupach był podwieczorek i II śniadanie. Wykazano zaburzenia proporcji w dostarczaniu energii z poszczególnych posiłków jak i nieprawidłowości w zakresie ich wartości odżywczej, zwłaszcza w grupie kobiet z depresją. Wnioski. Wydaje się słusznym zalecenie badanym kobietom, zwłaszcza chorującym na depresję konsultacje z dietetykiem nad zmianą nawyków żywieniowych, a w szczególności nad prawidłowym komponowaniem pod względem wartości energetycznej i odżywczej wybranych posiłków. Słowa kluczowe: kobiety, depresja, wartość odżywcza posiłków *Corresponding author: Ewa Stefańska, Department of Dietetics and Clinical Nutrition, Faculty of Health Sciences Medical University, Mieszka I 4b, 15-054 Bialystok, Poland, phone/fax : +48 85 732 82 44, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 140 Nr 2 E. Stefańska, A. Wendołowicz, U. Kowzan et al. INTRODUCTION In recent years the incidence of depression has been increasing, leading to many health complications and thus constituting a serious public health problem [17]. According to World Health Organisation (WHO), unipolar depression was the leading cause of mental illness in the nineties of the previous century and perhaps, together with cardiovascular diseases (CVDs), they may become the most common illnesses in the world by 2020 [6]. Research has shown that patients suffering from depression are more likely to alter their eating behaviour with changes in appetite and food preferences, including avoiding certain foodstuffs and food groups, eating too rarely or by eating high-calorie snacks in between main meals [9, 16, 21, 22, 23]. Monitoring the diet of patients suffering from depression can help to indicate irregularities, which would thereby permit an appropriate adjustment to be made through the development of both healthy eating habits and educational programs. Currently there are no published studies on assessing the nutritional values of foodstuffs consumed by people suffering from unipolar depression. The study was therefore aimed at evaluating and comparing selected dietary habits in women with or without unipolar depression. These included the number and type of meals normally consumed during the day and their dietary calorific and nutritional values. MATERIALS AND METHODS The survey group consisted of 110 women aged 18-65 years. Of these, the test group was 55 women treated for depression at the Department of Psychiatry, Medical University of Bialystok, whilst the control group were 55 women patients who were first reported at the Obesity Treatment Centre and Diet-Related Diseases and were not on a reducing diet. The study took place between September and November 2012. The test group included patients diagnosed with recurrent major depressive disorder (according to ICD-10), lasting up to 5 years, where their current episode of depression did not last longer than a month [24]. Furthermore, their current treatment for depression consisted of one antidepressant from the following; paroxetine, sertraline, venlafaxine, citalopram, mirtazapine, escitalopram and a sedative drug used as needed. The current course of the disease was assessed based on data from interviews and available documentation. Depressive symptoms were measured using the Hamilton Depression Rating Scale (version 17-point), and Beck’s self-esteem scale [3, 10]. Patients participating in the study were informed of its purpose and methodology. Each patient expressed written consent for such consultations. The study was approved by the Bioethics Committee of UMB No. RI-002/325/2011. A dietary questionnaire developed in-house at the Department of Dietetics and Clinical Nutrition, Medical University of Bialystok was used to collect data. The questionnaire contained, amongst others, questions about dietary habits, including the number of commonly consumed meals and any eating in-between. The daily diets were quantitatively analysed using the 24-hour diet recall, including 3 weekdays and 1 weekend day; results being averaged. Patients did not take any additional vitamin and mineral supplements. The daily diet (including mealtimes) were evaluated by their calorific value and nutritional content of selected nutrients, where in the latter case this was performed using the Diet 5.0 computer programme as developed by the Institute of Food and Nutrition in Warsaw, taking into account nutrient losses during the cooking process (Diet 5.0 package for planning and current assessment of individual nutrition, Institute of Food and Nutrition license contract no. HBBxtpINI). In assessing the intake of the selected nutrients, recommended nutrition standards for the Polish population were used as reference [14]. The proportion of calories derived from each meal were thence related to the recommended total calorific intake according to Hasik and Gawęcki [11]. Statistical analysis consisted of calculating averages, standard deviations and percentages with results being evaluated using StatSoft STATISTICA 10.0, by the χ2 and t-test for independent variables. Significance was adopted at the p <0.05 level. RESULTS The subjects groups’ characteristics are presented Table 1. There were no significant differences between the average age (45.8 years in women with depression vs. 41.1 years in the control group), nor correspondingly body height (162.8 cm vs. 162.1 cm), body weight (70.7 kg vs. 69.2 kg) and the average body mass index (26.8 kg/m2 vs. 27.0 kg/m2). The average points score on the Hamilton depression scale for the depressed subjects was 14.0 and 25.4 on the Beck scale. Mean disease duration for 50% of the women was 4 years, for 17% 2-3 years, and for 33% it was less than 1 year. In this group, episodes of depression were on average 2.7. A single episode for depression was reported in 33% of women, 2-4 episodes in 39%, more than 4 episodes occurred in 28% of patients. The average points score on the Hamilton depression scale for the control group was 3.0 and 6.5 on the Beck scale. In the group of depressed women 11% received paroxetine, 26% sertraline Nr 2 141 The nutritional value of meals of women with unipolar depression Table 1. General characteristics of subject groups Variables Age (years) Body height (cm) Body weight (kg) Body mass index (kg/m2) Underweight (%) Normal weight (%) Overweight (%) Obese (%) Waist circumference (cm) Marital status (%) Married Single Education (%) Primary Vocational Secondary University Number of previous depression episodes HAM-D score Beck score Depressed subjects (n=55) 45.8 (12.2) 162.8 (5.4) 70.7 (14.5) 26.8 (5.3) 3.6 45.4 32.7 18.3 89.4 (9.0)** Control group (n=55) 41.1 (13.1) 162.1 (5.2) 69.2 (10.6) 27.0 (5.3) 0.0 47.3 40.0 12.7 83.7 (9.8) 63.6*** 36.4 40.0 60.0 14.5 22.0 54.5 9.0 12.7 12.7 38.2 36.4 2.7(1.7) - 14.0 (7.3) 25.4 (12.8) 3.0 (1.7) 6.5 (3.2) Values for categorical variables are mean, (SD) or percentages of subjects. For continuous variables, the independent t test was used; for categorical variables, chi-square test was used *:<0.05, **:p<0.01, ***:p<0.001 and venlafaxine, 9% citalopram, 12% mirtazapine and 16% escitalopram. Waist circumference (WC) was however statistically higher in the depressed women’s group than controls (89.4 vs. 83.7 cm) and likewise there were significantly more married women in the depressed group than controls. No significant differences in levels of education were observed between groups, however the test group tended to have more women with primary, vocational and secondary education but less with higher education as compared with controls. The number of meals, mealtimes and meal types in both study groups are shown in Table 2, where there were no significant differences in the aforementioned between the test and control groups. Nevertheless, more depressed women (36.4%) preferred eating the usual 3 meals a day whereas those without depression preferred to eat 4 meals daily (43.6%). It was also noted that about 7% of the test women 4% of controls consumed only 2 or fewer meals per day. Eating an optimal 5 meals a day was observed in 31% of women for both compared groups. Nearly 90% of all the women studied consumed three main meals, ie. breakfast, lunch and dinner. The least frequent consumed meals in the two treatment groups were: afternoon tea (consumed by 56% of women with depression and 45% of women in the control group) and the mid-morning meal (consumed by 56% of women with depression and 69% of women Table 2. Details of subjects’ dietary habits selected for assessment Variables Number of meals ≤2 3 4 ≥5 Type of meals Breakfast Mid-morning meal Lunch Afternoon tea Dinner Additional eating inbetween meals Type of additional snacks Sweets Fast food Sandwiches Fruit Vegetables Depressed subjects n=55 n % Control group n=55 n % 4 20 14 17 7.3 36.4 25.4 30.9 2 12 24 17 3.6 21.8 43.6 31.0 51 31 55 31 50 48 92.7 56.3 100.0 56.4 90.9 87.3 49 38 50 25 49 41 89.0 69.0 90.9 45.4 89.0 74.5 28 1 11 8 0 58.3 2.0 23.0 16.7 0.0 17 2 10 12 0 41.4 4.9 24.4 29.3 0.0 without depression). Another detrimental aspect of the observed eating behaviour was the custom of snacking in-between meals as declared by 87% of women with depression and 74% of women in the control group (differences not significant); the main foodstuffs being eaten were mostly sweets. The average calorific values and content of selected nutrients in the daily diets are reported in Table 3. The diets of the depressed had both higher calorific values and nutritional content. Statistically significant differences were observed between respectively the test group and controls for the total daily calorie intakes (1660.1 kcal/day vs. 1431.6 kcal/day), total fat (59.6 g/day vs. 42.7 g/day), saturated fatty acids (SFA) (25.2 g/day vs. 16.9 g/day) and monounsaturated fatty acids (MUFA) (22.8 g/day vs. 15.7 g/day). Moreover, higher dietary contents were respectively seen in women with depression compared to controls of total protein (65.7 g/day vs. 65.2 g/day), polyunsaturated fatty acids (6.3 g/day vs. 5.9 g/day), cholesterol (248.7 mg/day vs. 216.1 mg/ day) and total carbohydrates (230.8 g/day and 212.5 g/ day), although these differences were not statistically significant. The average daily intakes of dietary fibre were too low from normal in both groups of 16.7g/day (test) and 22.7g/day (controls), however these differences were insignificant. The percentage of total calories from dietary protein in women with depression was lower at 15.8% than the 18.2% in the controls; differences being significant. In contrast, the percentage of dietary calories derived from total fat, saturated fatty acids and monounsaturates was significantly higher in women with depression compared with women in the 142 Nr 2 E. Stefańska, A. Wendołowicz, U. Kowzan et al. Table 3. Average calorific values and content of selected nutrients in subjects’ daily diets Variables Calories (kcal/day) Total protein (g/day) Animal protein (g/day) Total fat (g/day) SFA (g/day) MUFA (g/day) PUFA (g/day) Cholesterol (mg/day) Carbohydrates (g/day) Fibre (g/day) Protein (% E) Total Fat (% E) SFA (% E) MUFA (% E) PUFA (% E) Carbohydrates (% E) Depressed subjects (n=55) % of Mean (SD) Standard 1660.1 79.0 (611.5)* 65.7 109.5 (26.2) 41.1 137.0 (19.6) 59.6 85.0 (27.2)*** 25.2 108.1 (12.9)*** 22.8 81.4 (11.5)*** 6.3 35.3 (4.3) 248.7 82.9 (196.2) 230.8 75.8 (88.5) 16.7 55.7 (7.4) 15.8 (3.0)*** 31.3 (7.8)*** 13.7 (2.0)*** 12.4 (2.4)*** 3.4 (1.4) 52.9 (7.7) Control group (n=55) Mean % of (SD) Standard 1431.6 68.2 (410.6) 65.2 108.7 (21.9) 43.5 145.0 (20.8) 42.7 61.0 (18.1) 16.9 72.5 (9.6) 15.7 56.1 (7.4) 5.9 31.5 (3.4) 216.1 72.0 (156.1) 212.5 69.8 (68.9) 22,7 75.7 (27.7) 18.2 (5.2) 26.8 (7.2) 10.6 (3.0) 10.0 (2.0) 3.7 (1.4) 55.0 (8.1) SD-standard deviation %E -percentage of calories For continuous variables the independent t-test was used *:<0.05, **:p<0.01, ***:p<0.001 control group and respectively amounted to 31.3%, 13.7%, 12.4% vs. 26.8%, 10.6%, 10.0%. The percentage of calories derived from dietary polyunsaturated fatty acids was however similar in both groups; being 3.4% in women with depression and 3.7% in the control group. Likewise, there was no significant differences between the test and control groups in the percentage of calories derived from carbohydrate intake; respectively 52.9% vs. 55%. A breakdown of the calorific content for each meal declared to be consumed by both subject groups, along with recommended reference values are presented in Figure 1. This demonstrated that in the depressed women, the calorific content of breakfast and afternoon teas is too low but too high for lunch when compared to recommendations. A significant proportion of calories however derived from snacking in between meals for both the depressed women (11%) and controls (7%). In fact in the former group, these additional calories exceeded those derived from the mid-morning meal and afternoon tea and were mainly provided by carbohydrates and dietary cholesterol (Table 4). The nutritional values of breakfast eaten by both subject groups was similar, however in the depressed women’s group, the intake derived from of total fat and saturated fatty acids was higher than controls. In the former group, the daily diet had significantly less fibre than controls. Significantly lower dietary calorific values and the intakes of total protein and carbohydrates were also seen for the mid-morning meal in women with Table 4. Calorific and nutritional values of meals consumed by subjects Variables Calories (kcal) Total protein (g) Animal protein (g) Total fat (g) SFA (g) MUFA (g) PUFA (g) Cholesterol (mg) Carbohydrates (g) Fibre (g) Calories (kcal) Total protein (g) Animal protein (g) Total fat (g) SFA (g) MUFA (g) PUFA (g) Cholesterol (mg) Carbohydrates (g) Fibre (g) Calories (kcal) Total protein (g) Animal protein (g) Total fat (g) SFA (g) MUFA (g) PUFA (g) Cholesterol (mg) Carbohydrates (g) Fibre (g) Calories (kcal) Total protein (g) Animal protein (g) Total fat (g) SFA (g) MUFA (g) PUFA (g) Cholesterol (mg) Carbohydrates (g) Fibre (g) Depressed subjects (n=55) Mean (SD) Breakfast 321.6 (158.3) 11.9 (7.1) 8.1 (6.0) 14.1 (10.4)** 7.2 (5.0)*** 5.0 (4.3) 1.2 (0.9) 45.1 (47.8) 38.3 (17.3) 2.4 (1.8)*** Mid-morning meal 137.2 (179.9)** 5.0 (7.2)* 3.3 (5.4) 5.2 (8.4) 2.5 (4.3) 1,9 (3.2) 0.4 (0.7) 15.7 (24.1) 18.6 (24.1)* 1.4 (2.4) Lunch 577.1 (184.8)*** 25.5 (8.9) 15.6 (7.7) 14.7 (9.3)** 5.0 (3.4)*** 6.7 (4.7)** 1.8 (1.7) 71.9 (33.1) 91.6 (34.5)*** 7.7 (3.1) Afternoon tea 109.5 (159.7) 3.6 (6.3) 1.6 (3.9) 2.9 (5.6) 0.9 (2.0) 0.9 (1.8) 0.4 (0.6) 9.9 (7.7) 18.5 (28.0) 1.5 (1.8) Control group (n=55) Mean (SD) 291.8 (126.4) 13.1 (6.5) 8.3 (5.0) 10.4 (6.2) 4.4 ( 3.4) 3.9 (2.4) 1.3 (0.9) 48.0 (86.7) 40.8 (19.0) 3.9 (2.4) 211.1 (203.9) 8.5 (10.3) 5.9 (8.3) 6.4 (6.9) 2.8 (3.3) 2,3(2.9) 0.8 (1.9) 17.2 (18.6) 31.6 (34.3) 2.2 (3.0) 427.8 (186.2) 26.6 (13.1) 18.9 (12.1) 10.0 (8.7) 3.2 (2.8) 4.1 (4.2) 1.6 (1.7) 88.4 (100.9) 62.9 (34.2) 6.9 (4.0) 132.0 (154.3) 3.0 (4.9) 1.6 (3.9) 3.3 (6.4) 1.7 (3.4) 1.2 (2.5) 0.4 (0.7) 8.2 (7.2) 22.2 (24.9) 1.3 (1.5) Nr 2 The nutritional value of meals of women with unipolar depression Dinner Calories (kcal) 339.3 (203.8)* 264.5 (184.3) Total protein (g) 13.8 (9.6) 11.9 (10.0) Animal protein (g) 9.7 (8.4) 7.9 (8.8) Total fat (g) 14.7 (12.6)** 8.2 (9.0) SFA (g) 6.5 (6.0) 3.5 (4.0) MUFA (g) 4.9 (4.2)** 2.8 (3.4) PUFA(g) 2.0 (0.2) 1.4 (1.3) Cholesterol (mg) 85.5 (38.4) 47.4 (37.5) Carbohydrates (g) 40.4 (25.8) 38.3(27.6) Fibre (g) 2.5 (1.7) 3.3 (2.8) Snacks in-between meals Calories (kcal) 177.2 (201.0)* 104.4 (108.5) Total protein (g) 4.8 (7.2)* 2.2 (3.6) Animal protein (g) 2.8 (5.9)* 0.9 (2.2) Total fat (g) 7.8 (10.7)* 3.5 (1.1) SFA (g) 2.9 (1.1) 1.6 (1.7) MUFA (g) 3.1 (4.8)* 1.3 (2.6) PUFA (g) 0.9 (1.9) 0.4 (0.9) Cholesterol (mg) 20.6 (13.4)* 6.9 (7.8) Carbohydrates (g) 22.8 (16.3) 16.7 (15.0) Fibre (g) 1.3 (1.9) 1.0 (1.6) SD-standard deviation For continuous variables the independent t-test was used *:<0.05, **:p<0.01, ***:p<0.001 depression compared to controls. In contrast, significantly higher dietary calories at lunchtime, in the depressed women’s group, were observed to be derived from total fat, saturated fatty acids, monounsaturated fatty acids or total carbohydrates compared to the controls. There were no significant differences between the two subject groups in dietary calorific intake nor nutritional content for the afternoon tea. At dinnertime, the women with depression demonstrated had significantly higher dietary calorific values and intakes of total fat and monounsaturated fatty acids compared to controls. Furthermore, the depressed women’s group more frequently adopted the adverse habit of snacking between meals, which provided significantly more calories, total protein, (including animal protein), total fat, monounsaturated fat and dietary cholesterol in their diets compared to the women without depression. 143 DISCUSSION Studies have reported that the usual diets of persons suffering from depression may differ from the principles of rational nutrition [9, 23]. The presented study has shown no significant differences in both the number of meals normally eaten and their type between the two groups of subject women. However the test group ie. depressed women, tended to have more afternoon tea and snacks in-between meals. Whilst the presence of the former in a daily diet is beneficial, attention should be drawn to their doubtful nutritional value, in that these are mainly carbohydrate meals, with small amounts of protein. In addition, the diets of women with depression demonstrated that the nutritional value of in-between meal snacking exceeds that of nutritional foodstuffs consumed during afternoon tea and mid-morning meals, which is also not consistent with the principles of proper nutrition. The current study showed that the most popular snacks were sweets eaten by almost 50% of subjects from the two compared groups. Jeffery et al. [15] reported that depressive symptoms were significantly and positively correlated between the consumption of high-calorie sugary snacks with a high-calorie consumption but negatively correlated with non-sweet food. Other studies have shown that the most common women’s meals skipped were the morning ones and that other mealtimes varied according to the afternoon, evening and night-time hours of one day [19]. Friedrich demonstrated that from the modelling of modern women’s lives, then this shows a lower consumption of breakfast before going to work. Reasons determined were a decreased appetite early in the morning and not enough time for preparing breakfast [7]. According to some research, having the total calorific intake from the daily diet spread over 5 meals is most appropriate for human body health. This 5-meal/ day model should include: breakfast providing 25%, Figure 1. Energy content (%) breakdown per meal determined in both subject groups along with recommended reference values 144 E. Stefańska, A. Wendołowicz, U. Kowzan et al. mid-morning meal -10%, lunch -30%, afternoon tea -15% and dinner -20% of the total calorific daily food intake [11]. The presented results however differ from these cited recommendations and it should be stressed that the habit of snacking in-between meals provided 11% of the diet’s total calorific daily intake for women with depression and 7% in the control group. An even greater amount of calories coming from in-between snacks, compared to the current study, was reported by Carels et al. and Zizza et al. [4, 25], which respectively gave 15% and 22% values of the total calorific intake of the daily diet. Moreover, these were mostly derived from carbohydrate-fat products, a finding also confirmed by the presented study. It was found that, on average, the calorific value of diets in women with depression was significantly higher when derived from total fat, saturated fatty acids and monounsaturated fatty acids compared to controls. Other studies by Grossniklaus et al. and Konttinen et al. respectively however, demonstrated higher calorific values of 1899.8 kcal/day and 2224.0 kcal/day) [9, 16], but lower values were found in studies by Jacka et al., Murakami et al., Park et al., respectively; 1642.0 kcal/d; 1575.0 kcal/d; 1524.5 kcal/d [13, 20, 21]. The presented study found that dietary protein intake from depressed women were insignificantly different to controls and was similar to the values obtained in other studies [5, 9, 21]. However, our study demonstrated an abnormal ratio between the intakes of animal and vegetable derived protein, with a predominance of the former. Dietary fat contents were significantly higher in women with depression. Higher dietary fat intakes in depressed women of 72.0 g - 90.0 g/day than those obtained in the present study were observed in other studies [5, 9]. Further studies on depressed women found similar levels of dietary cholesterol to the current study as follows; 217.7 mg/day in Grossniklaus et al. [9] and 268.8 mg/day in Parks et al. [21]. Total dietary carbohydrate was found to be lower in the presented study compared to others [2, 5, 9], with generally low total carbohydrate intake in both groups together with low dietary fibre. Even lower fibre intakes (ie. than the <16 g/day of the current study), has been reported [9, 21]. Studies have demonstrated that this situation may arise from insufficient food intake in depressed patients from those foodstuffs rich in fibre such as raw fruit and vegetables, whole-grain cereals or dried leguminous seeds [1, 2, 5, 9, 15, 16, 23, 25]. Furthermore, a prospective cohort GAZEL study has shown that the intake of selected groups of foodstuffs such as fruit, vegetables and fish were associated with a reduced risk of subsequent episodes of depression [18]. Studies have shown that the diets of those suffering depression often have an inappropriately excessive composition of fatty acids, especially SFA [23]. Howe- Nr 2 ver, a proper dietary intake of fatty acids, particularly unsaturated fatty acids of omega-3-acid, EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) may have beneficial effects for treating patients suffering from depression. A study by Lakhan found that a high consumption of fish, as a source of unsaturated fatty acids, correlates with fewer patients suffering from mental disorders within the population [17]. Here, it was suggested that a 1 – 2 g daily intake of omega-3 was sufficient for healthy people, whilst consuming 9.6 g appears to be safe and adequate for patients with mental disorders [17]. The antidepressant effect of EPA may be due to its conversion to prostaglandins, leukotrienes and other compounds required for normal brain function. Another theory suggests that EPA and DHA affect signal transduction in brain cells by activating peroxisomal proliferator-activated receptors (PPARs), inhibiting G-proteins and protein kinase C, as well as calcium, sodium, and potassium ion channels [17]. It has been demonstrated that depression caused by monoamine deficiency (acting as a pathophysiological substrate) leads to decreased levels of neurotransmitters serotonin, norepinephrine and dopamine in the central nervous system. Synthesis of serotonin or 5-hydroxytryptamine (5 HT) has been found to be dependent on the availability of the dietary precursor serotonin-tryptophan [12]. High-carbohydrate and low-protein meals may affect mood by increasing the synthesis of this 5-hydroxytryptamine as compared to low carbohydrate and high protein meals [8]. Much research has indicated that patients feel more calm, relaxed after a snack rich in carbohydrates compared to protein-rich and low-carbohydrate meals. This may therefore also explain why the studied subjects took frequent recourse to those foodstuffs providing carbohydrates [8]. CONCLUSIONS 1. There were statistically insignificant differences between the two groups of subjects in both the number and types of meals throughout the day, as well how many snacks were eaten in-between meals, although this unhealthy habit tended to be more common in the depressed women. 2. Despite both subject groups having afternoon tea, the doubtful nutritional value of this actual meal rendered it unhealthy, especially for the depressed women. 3. The composition of diets for those women suffering from depression was incompatible with dietary recommendations regarding calorific intakes. 4. Compared to controls, the diets of women with depression had significantly higher calorific values and contents of total fat, SFA and MUFA as well as Nr 2 The nutritional value of meals of women with unipolar depression the percentage of calories derived from total fat, SFA and MUFA. 5. It is recommended that women suffering from depression should consult with dieticians for improving their dietary habits, particularly for achieving the correct calorific and nutritional values/balance for their meals. Acknowledgement This study was performed as a project of the Faculty of Health Sciences, Medical University, Bialystok (No. 123-16595P). Conflict of interest The authors declare no conflict of interest. REFERENCES 1. Akbaraly T.N., Sabia S., Shipley M.J., Batty G.D., Kivimaki M.: Adherence to healthy dietary guidelines and future depressive symptoms: evidence for sex differentials in the Whitehall II study. Am. J. Clin. Nutr. 2013;97(2):419427. 2. Appelhans B.M., Whited M.C., Schneider K.L., Ma Y., Oleski J.L., Merriam P.A. Waring M.E., Olendzki B.C., Mann D.M., Ockene I.S., Pagoto S.L.: Depression severity, diet quality, and physical activity in women with obesity and depression. J. Acad. Nutr. Diet. 2012;112(5):693-698. 3. Beck A.T., Ward C.H., Mendelson M., Mock J., Erbaugh J.: An inventory for measuring depression. Arch. Gen. Psychiatry.1961;4:53-63. 4. Carels R.A., Young K.M., Coit C., Clayton A.M., Spencer A., Wagner M.: Skipping meals and alcohol consumption. The regulation of energy intake and expenditure among weight loss participants. Apettite. 2008;51(3):538-545. 5. Davison K.M., Kaplan B.J.: Food intake and blood cholesterol levels of community-based adults with mood disorders. BMC Psychiatry. 2012, 12, DOI 10.1186/1471244X-12-10. 6. Eby G.A., Eby K.L.: Rapid recovery from major depression using magnesium treatment. Med. Hypotheses. 2006;67(2):362-370. 7. Friedrich M.: Health-oriented nutritrional education as the factor influencing changes in feeding habits. P. 2: Evaluation of the impact of feeding education on the change of the feeding way of obese, working women age 45-52. Żyw. Człow. 1998;25(3):261-274 (in Polish). 8. Gibson E.L.: Emotional influences on food choice: Sensory, physiological and psychological pathways. Physiology & Behavior. 2006;89(1):53-61. 9. Grossniklaus D.A., Dunbar S.B., Tohill B.C., Gary R., Higgins M.K., Frediani J.: Psychological factors are important correlates of dietary pattern in overweight adults. J. Cardiovasc. Nurs. 2010;25(6):450-460. 145 10. Hamilton M.: A rating scale for depression. J. Neurol. Neurosurg. Psychiatry. 1960; 23:56-62. 11. Hasik J., Gawęcki J.: Żywienie człowieka zdrowego i chorego. Wydawnictwo PWN, Warszawa 2008. 12. Hasler G.: Pathophysiology of depression: do we have any solid evidence of interest to clinicians? World Psychiatry. 2010;9(3):155-161. 13. Jacka F.N., Maes M., Pasko J.A., Williams L.J., Berk M.: Nutrient intakes and the common mental disorders in women. J. Affect. Disord. 2012;141(1):79-85. 14. Jarosz M.: Normy żywienia dla populacji polskiej-nowelizacja. Wydawnictwo IŻŻ, Warszawa 2012. 15. Jeffery J.W., Linde J.A., Simon G.E., Ludman E.J., Rohde P., Ichikawa L.E., Finch E.A.: Reported food choices in older women in relation to body mass index and depressive symptoms. Apettite. 2009;52(1):238-240. 16. Konttinen H., Mӓnnistӧ S., Sarlio-Lӓhtenkorva S., Silventoinen K., Haukkala A:. Emotional eating, depressive symptoms and self-reported food consumption. A population study. Appetite. 2010;54(3):473-479. 17. Lakhan S.E., Vieira K.F.: Nutritional therapies for mental disorders. Review Nutr. J. 2008;7:1-8. 18. Le Port A., Gueguen A., Kesse-Guyot E., Melchior M., Lemogne C., Czernichow H., Nabi H., Goldberg M., Zins M.: Association between dietary patterns and depressive symptoms over time: a 10-year follow-up study of the GAZEL cohort. PLoS One. 2012;7(12): e51593 doi 10.1371/journal.pone 0051593. 19. Mędrela-Kuder E.: Some nutritional habits of a group of overweight and obese women. Rocz Państw Zakl Hig 2005;56(4):371-377 (in Polish). 20. Murakami K., Mizoue T., Sasaki S., Ohta M., Sato M., Matsushita Y., Mishiman M.: Dietary intake of folate, other B vitamins, and ω-3 polyunsaturated fatty acids in relation to depressive symptoms in Japanese adults. Nutrition 2008;24(2):140-147. 21. Park J.Y., You J.S., Chang K.J.: Dietary taurine intake, nutrients intake, dietary habits and life stress by depression in Korean female college students: a case-control study. J. Biomed Sci. 2010;17:S40-S44. 22. Peet M.: International variations in the outcome of schizophrenia and the prevalence of depression in relation to national dietary practices: an ecological analysis. B. J. Psych. 2004;184:404-408. 23. Sanhueza C., Ryan L., Foxcroft D.R.: Diet and the risk of unipolar depression in adults: systematic review of cohort studies. J Hum Nutr Diet. 2013;26(1):56-70. 24. World Health Organisation. International statistical classification of diseases and health-related problems. 10th rev. Geneva, WHO, 1992. 25. Zizza C.A., Tayie F.A., Lino M.: Benefits of snacking in older Americans. J. Am. Diet. Assoc. 2007;107(5):800806. Received: 17.10.2013 Accepted: 19.03.2014 Rocz Panstw Zakl Hig 2014;65(2):147-153 AWARENESS OF FACTORS AFFECTING OSTEOPOROSIS OBTAINED FROM A SURVEY ON RETIRED POLISH SUBJECTS Natalia Ciesielczuk, Paweł Glibowski*, Jolanta Szczepanik Faculty of Food Science and Biotechnology, University of Life Sciences in Lublin, Poland ABSTRACT Background. Osteoporosis is a growing problem facing modern society and currently poses one of the most serious health challenges. It is a progressive skeletal disease characterised by low bone mineral density whose development depends on multiple factors. These principally include increasing age, nutrition, physical activity, endocrine changes, lifestyles, general health condition and taken drugs. Objectives. To assess how much subjects (aged >50 years) are aware of the dangers in contracting osteoporosis along with the effects that lifestyle and dairy product consumption can have on this condition developing. Material and methods. A questionnaire, designed in-house, was used to survey a group of 150 randomly selected individuals aged above 50 years. This was performed in June 2012. Questions were on socio-demographics, milk and dairy product consumption, physical activity as well as assessing knowledge about osteoporosis that included issues such as its incidence, prevention and morbidity. Results. Osteoporosis was established in 60.7% subjects of whom 69.2% were women. Only 9.3% never consumed any dairy products. Physical activity of some kind was undertaken by 77.3% subjects. Within the last year, 38.0% reported having fractures of whom 46.0% had osteoporosis and 25% were healthy. Respondents were aware that consuming dairy products is beneficial to bone health and this awareness was higher among those with osteoporosis. Physical activity was also recognised to be important in preventing this condition. Conclusions. Subjects suffering from osteoporosis had twice as many more fractures than healthy subjects. Key words: osteoporosis, milk, dairy products, calcium STRESZCZENIE Wprowadzenie. Osteoporoza jest coraz większym problemem współczesnego społeczeństwa. Jest to choroba szkieletu, która charakteryzuje się niską gęstością mineralną kości. Jest ona bardzo poważnym wyzwaniem w obecnych czasach, ponieważ na jej rozwój wpływa wiek a także odżywianie, aktywność fizyczna, zaburzenia hormonalne, styl życia, ogólna kondycja zdrowotna a także przyjmowane leki. Cel badań. Celem badań była ocena świadomości zagrożenia osteoporozą oraz wpływu stylu życia i spożywania przetworów mlecznych na jej występowanie u osób w wieku powyżej 50 roku życia. Materiał i metody. Badania zostały przeprowadzone na podstawie ankiety własnego autorstwa w czerwcu 2012 roku. Badaną grupą było 150 losowo wybranych osób w wieku od 50 lat wzwyż. Ankieta zawierała pytania dotyczące sytuacji socjo-demograficznej ankietowanych oraz wiedzy i zagadnień dotyczących osteoporozy, przyczyn jej występowania, zapobiegania, zachorowalności a także spożywania mleka i przetworów mlecznych oraz aktywności fizycznej. Wyniki. 60,7% badanych miało osteoporozę, a 69,2% chorych stanowiły kobiety. Tylko 9,3% badanych nie spożywała produktów mleczarskich. 77,3% respondentów stosowało jakiś rodzaj aktywności fizycznej. 38,0% deklarowało, że w ciągu ostatniego roku wystąpiło u nich złamanie, przy czym 46% wśród nich miało osteoporozę i 25% wśród zdrowych. Uczestniczący w badaniach byli świadomi, że spożywanie produktów mlecznych ma korzystny wpływ na kościec, jednak większą świadomość miały osoby z osteoporozą. Aktywność fizyczna była częstą praktyką wśród respondentów, wskazywana jako ważna część profilaktyki osteoporozy. Wnioski. Nadal niewystarczająca wiedza na temat osteoporozy u osób powyżej 50 roku życia i powinna być poszerzana, co mogłoby wpłynąć na zmniejszenie liczby złamań u takich osób, które są niemal dwukrotnie częstsze niż u osób zdrowych. Słowa kluczowe: osteoporoza, mleko, przetwory mleczne, wapń, *Corresponding author: Paweł Glibowski, University of Life Sciences in Lublin, Faculty of Food Science and Biotechnology, Department of Milk and Hydrocolloids Technology, 8 Skromna Street, 20-704 Lublin, Poland, phone: +48 81 462 33 49, fax +48 81 462 33 54, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 148 Nr 2 N. Ciesielczuk, P. Glibowski, J. Szczepanik INTRODUCTION Healthy and rational nutrition is essential for the proper development of human physical and mental fitness [12]. Inadequate nutrition may lead to a deterioration of health and emergence of lifestyle-related diseases [26]. Due to an aging population, osteoporosis has become an increasing health problem [22]. The World Health Organisation (WHO) defines osteoporosis as ‘an epidemic of the XXI century’, which along with obesity, diabetes and cardiovascular disease, is a serious and growing clinical problem associated with the process of aging. Because developing this disease depends on many factors like ageing (along with the increase in bones loss density and strength), nutrition, physical activity, endocrine changes, lifestyles, general health condition, as well as taken drugs, it nowadays poses one of the most serious health challenges [8, 33]. According to the WHO definition, osteoporosis is a systemic skeletal disease, characterised by low bone mineral density and abnormal bone micro-architecture, which in consequence leads to fragility and increased susceptibility to fractures [24]. It is a serious disease that develops slowly over many years and it is often diagnosed too late. Due to current medical advances, a reduction in the risk and incidence of fractures has been demonstrated as well enabling an increase in bone mineral density. The restoration of normal body weight is also important [25]. The main factors affecting maximum bone mass and strength are genetic predisposition and dietary habits acquired from childhood and adolescence. Nutrients supplied regularly and in sufficient quantities (especially calcium and protein), together with physical activity, contribute in achieving a maximum bone mass [3]. The main risk factor for human osteopaenia (a condition where bone mineral density is lower than normal) and osteoporosis is Vitamin D deficiency. This deficiency is the cause of increased secretion of parathyroid hormone (PTH) and reduced calcium absorption from the gastrointestinal tract, which contributes to a reduction of bone mass [16]. The value of the peak bone mass achieved, is dependent on genetic determinants, nutrition, physical activity and environmental pollution (for e.g. lead adversely affects the bioavailability of nutrients that includes calcium) [29]. Osteoporosis is a disease affecting both sexes, however due to the late start of losing bone mass and a milder course with no sudden changes in hormonal activity, osteoporosis develops less often in men than in women [17]. Diagnosis of osteoporosis proposed by the WHO is based on the measurement of bone mineral density [9]. In Poland, there have been a few studies on osteoporosis conducted on the elderly [10, 28], however to the best of our knowledge, there are no studies on Polish men and women (aged above 50 years) regarding their knowledge of osteoporosis. For this reason, the study aim was to investigate awareness about osteoporosis among retired subjects aged above 50 years and analysing their lifestyle and dairy products consumption. MATERIAL AND METHODS The study was based on an in-house designed questionnaire, and carried out in June 2012. The target group was 150 randomly selected individuals aged above 50 years. Subjects were divided into gender. The questionnaire asked respondents about socio-demographics and their knowledge on osteoporosis issues that included its occurrence, prevention, morbidity as well as their consumption of milk and dairy products and any undertaken physical activity. The questionnaire consisted of 23 questions, including 3 multiple choice ones and 20 that were single choice. RESULTS Subjects consisted of 92 women and 58 men. Socio-demographic details are presented in Table1. Table 1. Socio-demographics of studied subjects Characteristic Age range Location Education Financial situation Variants 50-60 years 60-70 years 70-80 years >80 years Village City; to 20 thousand inhabitants City; 20-50 thousand inhabitants City; > 50 thousand inhabitants Primary Vocational Secondary Higher Bad Average Good Very good Not know Percentage of total 30.0 40.0 20.7 9.3 44.0 29.3 20.0 6.7 21.2 17.3 33.3 26.0 14.7 22.7 38.0 23.3 1.3 Of the 150 subjects surveyed, 91 (60.7%) had osteoporosis, among which 22.0% had been diagnosed with osteoporosis at the age of 50-60, 11.3% at 40-50 years, 9.3% at 60-70 years and 8.0% at up to 30 years as well as 3.3% of people over 80 years, 70-80 years and 30-40 years. In most cases, a genetic influence could be probably discerned, as more than 50% subjects declared Nr 2 149 Awareness of osteoporosis in the retirement age Table 2. Surveyed incidence of osteoporosis types broken down according to gender and age Type of osteoporosis Idiopathic Involutional Senile Post-menopausal Derivative < 30 years 10 W 1W 1M - 30-40 years 1W 1M 1W 1W 1M Ages at which the disease was diagnosed 40-50 years 50-60 years 60-70 years 1W 1W 1M 2W 4W 3W 4M 7M 6W 16 W 2W 3W 4W 3W 3M 3M 3M 70-80 years 2W 1W 2M Over 80 years 1W 2M W- woman, M- man 15.3% in those with senility and 7.3% were stricken by idiopathic-osteoporosis. The lowest morbidity among respondents (4.0%) concerns involutional-osteoporosis. Milk and dairy products had been consumed by 80.0% of respondents in their youth. Almost 11% never consumed any dairy products and a little over 9.0% said they couldn’t remember. At the present time, 45.3% of respondents declared consuming dairy products several times a week, 30.0% every day, 12.0% once a week, 33,9% 3.3% several times a month and 9.3% never at all. A 35% significantly higher consumption of dairy products was 30% recorded for osteoporotic individuals compared to the 25% group as a whole. It was found that 31.0% of patients 20% 14,5% 14,5% consume milk and dairy products daily, 56.0% several 13,0% 15% times weekly, 4.4% once weekly and 1.1% several 8,1% 8,1% 6,5% 10% times a month. However only 4.0% (4/91 osteoporotic 1,6% 5% subjects) did not consume dairy products, which inc0% luded a single subject in the osteoporotic group that suffered fracture (2%). This, therefore indicates a high awareness in the benefits of consuming such kinds of food. Among those respondents who did not get sick, Figure 1.Close relatives of the respondents who were dia- 18.0% (11/59) never ate dairy products, whilst 20.0% gure 1. Close relatives of the respondents who were diagnosed gnosed with osteoporosis (%) with osteoporosis (%) of healthy subjects (3/15) who had fractures never consumed milk nor its products. The most important dairy that osteoporosis had been diagnosed in some of their family members (Figure 1). In addition, 6 osteoporotic subjects (6.6%) were diagnosed with the primary type of this disease. Osteoporosis is a disease that affects people of all ages and both women and men. In our study, 69.2% of those suffering were women. The incidence of the osteoporosis types, according to gender and age, is shown in Table 2. Subjects were found to well recognise and define osteoporosis, where more than 90% gave correct definitions and furthermore, 37.0% were able to provide reasons for its occurrence. In this latter group, 25 subjects (44.7%) reported low bone mineral density as a reason, 18 (32.1%) believed it to be calcium and vitamin D deficiency, 12 (21.5%) indicated an abnormal bone mass, and only one respondent (0.7%) pointed to age. When given the opportunity to select up to three answers, as to the basic factors influencing osteoporosis development, respondents mostly selected old age, a genetic predisposition and menopause (Figure 2). Subjects were aware of the fact that osteoporosis develops more often in women (73.3%), than men (19.3%), or in children (7.3%). The main type of osteoporosis found was the postmenopausal variety affecting more than 17% of the female subjects. Secondary osteoporosis was observed in 16.7% respondents, 23,4% 25,0% 20,0% 20,0% 19,3% 16,3% 15,0% 10,0% 5,0% 8,9% 4,9% 7,3% 0,0% Figure 2. Factors affecting the development of osteoporosis in the opinion of respondents (%) Figure 2. Factors affecting the development of osteoporosis in the opinion of respondents (%) 150 products found to be consumed were milk, curds, and cottage cheese (Figure 3). 30,0% Nr 2 N. Ciesielczuk, P. Glibowski, J. Szczepanik 25,8% 24,5% 25,0% 20,0% 15,1% 15,0% 11,9% 6,9% 10,0% 3,3% 5,0% 5,7% 2,5% 3,4% 0,0% Figure 3. Types of dairy products consumed by respondents (%) gure 3. Types of dairy products consumed by respondents (%) Amongst those that knew about calcium, 55.9% believed that such requirements should be 1200 mg/day, and 44.1% - 1500 mg/day. Over 49% of subjects indicated vitamin D to be a nutrient that supports the absorption of calcium from the gastrointestinal tract with the others being ignorant of this fact. Calcium deficiencies were made up for by 54.0% taking supplements, whilst the remainder did not use these type of products. The majority of respondents derived information on osteoporosis from their GP (General Practitioner) and the Internet. Another important source was also from the family, radio and television (Figure 5). 35,0% 30,0% 25,0% 20,0% 15,0% 10,0% 5,0% 0,0% 30,2% 25,9% Physical activity was considered to be important in 16,0% preventing osteoporosis by 81.0% of subjects, whereas 11,7% 6,9% 16.0% said otherwise and less than 3.0% answered that 5,5% 4,9% they didn’t know. The respondents were found to be 0,0% fairly active physically, where 26.0% said they took walks, 24.7% did exercises, 18.7% went swimming, 16.7% preferred other types of activities and 14.0% said they were passive. Regarding being aware of the importance of physical activity, then 28.0% said they Figure 5.Information sources for respondents concerning undertook this daily, 26.0% a few times weekly Figure 5.and Information sources for respondents concerning osteoporosis (%) osteoporosis (%) 23.3% a few times a month; unfortunately 22.7% were not involved in any sport/exercises at all. 52.0% of respondents were medically examined on Fractures are an inevitable consequence of osteopooccurrence of osteoporosis; the others did not particirosis. The reduced strength of bone tissue due to the pate in such examination. Furthermore, access to such lower mineral density results in decreased resistance to damage as compared to those persons with normal bone examinations is sufficient for less than 23% of responmineral density. It was found that 38.0% subjects had dents, while others declare that it is not sufficient. It was fractures in the last year, where the wrist, forearm and found that 31% (15/59) of the subject group suffering hip bone were broken most frequently (Figure 4). Of from osteoporosis that did not have fractures, didn’t the 91 osteoporotic subjects, 42 (46.0%) had fractures practice sport while 27% (13/49) of these did so daily. over the last year, whilst 15/59 (25%) healthy subjects However, 27.0% (12/44) of healthy persons who didn’t have fractures within the last year were observed not suffered fractures. practice any sports at all, but 34.0% (15/44) practiced it every day. It is worth noting that the results for these 40,0% 35,4% two groups (i.e. osteoporotic and healthy) are very si35,0% milar. In those osteoporotic subjects suffering fractures, 30,0% only 10.0% (4/42) didn’t practice any sport at all and 25,0% likewise in healthy subjects who suffered fracture, only 20,0% 18,5% 20,0% 9.0% (4/44) abstained from any sport. 15,0% 9,2% 10,0% 6,1% 4,6% 5,0% 0,0% Wrist Hip bone Femur Spine Forearm Leg 6,1% Arm Figure 4. Fractures occurred to the surveyed within a year (%) Figure 4. Fractures occurred to the surveyed within a year (%) More than 47.0% of respondents answered that they knew what helps to prevent osteoporosis, but 63.0% didn’t know the importance of daily calcium requirements. DISCUSSION Milk and dairy products are necessary foodstuffs in the daily diet of every person. They provide many essential nutrients, of which calcium is the most important. Nowicka and Panczenko-Kresowska [23] indicate that the calcium deficiency caused by low intakes of milk, give rise to bone abnormalities. According to Coudray [5], insufficient intakes of milk and dairy products by Nr 2 Awareness of osteoporosis in the retirement age children and adolescents leads to a failure in achieving peak bone mass, and in the case of adults, effects the bone tissue remodelling disorder and fosters the loss of bone mass. Furthermore, van Staveren et al. [32] has indicated that an adequate dietary supply of this group of food products helps in preventing osteoporosis. Besides dairy products, drinking water can also be quite an important source of calcium in the diet as reported by Madej et al. [20], with a 15 % of the daily calcium intake derived from drinking water. The Polish WOBASZ study (Multicenter National Study of Population’s Health Status) from 2002-2005 showed that current consumption of milk and dairy products is surprisingly low. Similar results of such a decline were obtained by Bouamra-Mechemache et al. [2]. The presented survey on the randomly selected subjects however, do not confirm this as demonstrated by the quite high consumption of milk and dairy products. Furthermore, the frequency in consuming this type of food products were also satisfactory. The reason for this is probably the dietary habits acquired in childhood. Fischer et al. [11] showed that milk and dairy products are perceived as a major group of foods needed for the growth and development of children and adolescents. A large assortment and wide availability of milk and dairy products on the market allows consumers to choose those which match their preferences. According to Henning et al. [14], development of the dairy industry has had a positive influence on the variety and quality of such available products. Kozłowska-Wojeciechowska [19] showed that 51% men meet the daily demand for milk and dairy products but only 41% women do so. This difference is not so apparent in the current study, as both, male and female subjects were found to regularly consume dairy products. Our study showed that respondents know how important calcium and vitamin D are. This quite high awareness of the recommended calcium intake could be due to the high predominance of osteoporosis in the surveyed group. MarcinkowskaSuchowierska and Sawicka [22] reported that the daily requirement for calcium for older people is 1200 mg/ day, which is consistent with Tang et al. [30], who proved that a calcium intake of 1200 mg/day alone or in combination with vitamin D, reduces the risk of osteoporotic fractures. Marcinkowska-Suchowierska et al. [22] also recommends calcium intake levels of 1200 mg/day and 800-1000 IU/day of vitamin D. Such calcium recommendations (1200 mg/day) for women and men above 50 and 65 years old respectively, can be found in the latest Nutritional Standards established for the Polish population [33]. However, the adequate intake for vitamin D (cholecalciferol) for the same group is 15 µg/day (600 IU) [15]. Subjects recognised that an older age, genetic predisposition, stimulants, drug abuse, poor socio-econom- 151 ic and environmental pollution are factors influencing the development of osteoporosis. Indeed, according to Tanriover et al. [31] and Handa et al. [13], the main factors affecting the development of osteoporosis are age and gender. Center and Eisman [4] point out that genetic factors and environmental conditions also have an impact on this illness. Eastell [9] agrees with the aforementioned factors, but also considers the impact of drugs and other diseases. The incidence of osteoporosis (over 60 %) in the presented study is quite striking and it probably results from the relatively small group of respondents taken. Some estimations report that in Poland, osteoporosis affects 30% of women and 8% of men above 50 years age. According to Koduganti et al. [18], women are more prone to osteoporosis, but men also suffer. The National Osteoporosis Foundation estimates that in people over 50 years age, the risk of bone fractures due to osteoporosis increases up to 50% in women and 25% in men. Zdziemborska et al. [34] reported that osteoporosis occurs in one third of women aged 60-70 years and two thirds of women aged 80 years and above. The results of our study also agree with these data, as over 73% indicated that women suffer from osteoporosis more often. In addition, respondents identified the age of above 50 years as being the threshold when osteoporosis occurs. Czerwinski and Kumorek [6] consider that the incidence of osteoporotic fractures varies across populations, not only in relation to age, gender and race, but also to the region of the world and socio-economic conditions. Data on fractures are alarming, because every year the numbers are increasing. Worldwide, osteoporosis causes more than 8.9 million fractures annually. In Europe, there were 3,119,000 osteoporotic fractures in people aged over 50 years in 2000. Moreover, it is estimated that in 2050 there will be 4.5 million hip fractures, while in 2000 there were only 1.6 million [27]. Badurski et al. [1] suggests that the main goal of treatment is to prevent the risk of osteoporotic fractures. It should include reducing the impact of risk factors for fractures and improve bone density with the use of drug therapy. According to the respondents, osteoporosis tests are necessary and desirable, but equal access to them is not sufficient because of the still small scale of publicising the consequences of this disease. CONCLUSIONS 1. Respondents were well aware that consuming dairy products has a beneficial effect on bone health; with this awareness being higher among persons afflicted with osteoporosis. 152 N. Ciesielczuk, P. Glibowski, J. Szczepanik 2. Physical activity is indicated as a preventative measure against osteoporosis and was often adopted by respondents. 3. The incidence of fractures in people older than 50 years suffering from osteoporosis was almost twice as frequent as in healthy people. 4. It appears that common knowledge and education of people over 50 years old regarding osteoporosis is still insufficient. Conflict of interests The authors declare no conflict of interests. REFERENCES 1. Badurski J., Czerwiński E., Marcinowska-Suchowierska E.: Osteoporosis – fracture risk estimation Status Quo Arte Anno 2007/2008: Review of positions of World Health Organisation (WHO), European Medicine Agency (EMEA), European Society Of Clinical And Economic Aspects Of Osteoporosis And Osteoartritis (ESEAO), International Osteoporosis Foundation (IOF), Polish Foundation Of Osteoporosis (PFO) and Polish Osteoartrology Society (PTOA). Post N Med 2008;6:335-359 (in Polish). 2. Bouamra-Mechemache Z., Réquillart V., Soregaroli C., Trévisiol A. : Demand for dairy products in the EU. Food Policy 2008;33(6):644-656. 3. Caroli A., Poli A., Ricotta D., Banfi G., Cocchi D.: Invited review:. J Dairy Sci 2011; 94, 11, 4: 5249-5262. 4. Center J., Eisman J.: The epidemiology and pathogenesis of osteoporosis. Bailliere’s Clinic Endocrinol Metabol 1997;1:23-62. 5. Coudray B.: The contribution of dairy products to micronutrients intakes in France. J Am Coll Nutr 2011;30(Supl. 1):410-414. 6. Czerwiński E., Kumorek A.: Falls, vitamin D and fractures, Post N Med 2011;3:226-231 (in Polish). 7. Dervis E.: Oral implications of osteoporosis. Oral Surg Oral Med Oral Pathol 2005;100:349-356. 8. Dobrzańska A., Tymolewska-Niebuda B., Lesińska K.: The role of nutrition in prevention and treatment of osteoporosis in children and youth. Nowa Pediatr 2001;1:3338 (in Polish). 9. Eastell R.: Investigating suspected osteoporosis. Women’s Health Medicine 2006;3(4):161-164. 10. Filip R. S., Zagorski J.: Osteoporosis risk factors in rural and urban women from the Lublin Region of Poland. Ann Agric Environ Med 2005;12(1):21-26. 11. Fischer W. J., Schiller B., Tritscher A. M., Stadle R. H.: Contaminants of Milk and DairyProducts/Environmental Contaminants. Encyclopedia of Dietary Sciences (Second Edition), 2011. 12. Głodek E., Gil M., Rudy M., Pawlos M.: Assessment of frequency of consumption of selected sources of dietary fibre by students. Rocz Panstw Zakl Hig 2011;62(4):409412 (in Polish). Nr 2 13. Handa R., Kalla A. A., Maalouf G.: Osteoporosis in developing countries. Best Pract Res Cl Rh 2008;22(4):693708. 14. Henning D.R., Baer R.J., Hassan A.N., Dave R.: Major advances in concentrated and dry milk products, cheese, and milk fat-based spreads. J Dairy Sci 2006;89(4):11791188. 15. Jarosz M., Stoś K., Walkiewicz A., Stolińska H., Wolańska D., Gielecińska I., Kłys W., Przygoda B., Iwanow K.: Vitamins. In: Jarosz M. ed. Nutritional standards for the Polish population – amendment. Food and Nutrition Institute (IŻŻ), Warsaw 2012 (in Polish). 16. Jasik A., Tałałaj M., Paczyńska M., Walicka M., Wąsowski M., Marcinkowska-Suchowierska E.: Vitamin D and osteoporosis, Post N Med 2008;1:8-13 (in Polish). 17. Kita K., Lewandowski B., Klimiuk P. A., Sierakowski S., Domysławska I., Wołczyński S., Małyszko J.: Osteoporosis in men. Nowa Med 2004;3 (in Polish). 18. Koduganti R.R., Gorthi C., Reddy P.V., Sandeep N.: Osteoporosis: A risk factor for periodontitis. J Indian Soc Periodontol 2009;13(2):90-96. 19. Kozłowska-Wojciechowska M.: Council for the Promotion of Healthy Nutrition: Milk and its preserves – necessary products in good health. The newest reports about consumption of milk and dairy preserves and their effect on human health. Warszawa 2007 (in Polish). 20. Madej D., Kałuża J., Antonik A., Brzozowska A., Roszkowski W.: Calcium, magnesium, iron and zinc in drinking water and status biomarkers of these minerals among elder people from Warsaw region. Rocz Panstw Zakl Hig 2011;62(2):159-168 (in Polish). 21. Marcinkowska-Suchowierska E., Czerwiński E., Badurski J., Walicka M., Tałałaj M.: Oateoporosis – diagnostic and treatment in elderly patients. Post Nauk Med 2011;5:410423 (in Polish). 22. Marcinkowska-Suchowierska E., Sawicka A.: Calcium and Vitamin D in prevention of osteoporotic fractures. Post N Med 2012;3:273-279 (in Polish). 23. Nowicka G., Panczenko-Kresowska B.: Coronary heart disease and osteoporosis: factors related to development of both diseases. Prz Lek 2007;64:153-158 (in Polish). 24. Ostrowska B.: Postural stability in women with osteopenia and osteoporosis. Wrocław, Wydawnictwo AWF, 2009 (in Polish). 25. Ravn P., Bidstrup M., Wasnich R. D., Davis J. W., McClung M. R., Balske A., Coupland C., Opinder Sahota O., Amarjot Kaur M., Marianne Daley M., Giovanni Cizza G.: Alendronate and estrogen - progestin in the long term prevention of bone loss: four year results from the early postmenopausal intervention cohort study: a randomized controlled trial, Ann Intern Med 1999;131:935-942. 26. Seidler T., Szczuko M.: Nutrition mode evaluation among university of agriculture students in Szczecin in 2006. Part I. Consumption of selected nutrients and nourishment state. Rocz Panstw Zakl Hig 2009;60(1):59-64 (in Polish). 27. Ström O., Borgström F., Kanis J.A. Compston J. Cooper C. McCloskey E.V., Jönsson B.: Osteoporosis: burden, health care provision and opportunities in the EU. Arch Nr 2 Awareness of osteoporosis in the retirement age Osteoporos International Osteoporosis Foundation and National Osteoporosis Foundation. Springer, 2011. 28. Szałek E.: Teaching menopausal and post-menopausal patients about osteoporosis. Farm Współcz 2013;6:51-56 (in Polish). 29. Śmigiel-Papińska D.: The nutritional role in children and youth from pollutant areas exposed in aspect of osteoporosis prevention. Med Rodz 2002;1:42-45 (in Polish). 30. Tang B.M., Eslick G.D., Nowson C., Smith C., Bensoussan A.: Use of calcium or calcium in combination with witamin D supplementation to prevent fractures of bone in people aged 50 years and older: a meta-analysis. Lancet 2007;370:657-66. 31. Tanriover M.D., Oz S.G., Tanriover A., Kilicarslan A., Turkmen E., Gusen G.S., Saracbasi O., Tokgozoglu M., Sozen T.: Hip fractures in a developing country: Oste- 153 oporosis frequency, predisposing factors and treatment costs. Arch Gerontoland Geriat 2010;50:3: e13-e18. 32. van Staveren W.A., Steijns J.M., de Groot L.C.: Dairy products as essential contributors of (micro-) nutrients in reference food patterns: an outline for elderly people. J Am Coll Nutr 2008,27:6747S-6754S. 33. Wojtasik A., Jarosz M., Stosik K.: Minerals. In: Jarosz M. ed. Nutritional standards for the Polish population – amendment. Food and Nutrition Institute (IŻŻ), Warsaw 2012 (in Polish). 34. Zdziemborska A., Deszczyńska K., Fidecki M.: Osteoporosis – Bisphosphonate – Dentist. Part II. Nowa Stomatol 2012; 1: 15-18. Received: 23.10.2013 Accepted: 05.04.2014 Rocz Panstw Zakl Hig 2014;65(2):155-164 RESPONSIVENESS TO THE HOSPITAL PATIENT NEEDS IN POLAND Lucyna Gromulska1*, Paweł Goryński2, Piotr Supranowicz1, Mirosław Jan Wysocki1 Department of Health Promotion and Postgraduate Education, National Institute of Public Health – National Institute of Hygiene, Warsaw, Poland 1 ABSTRACT Background. The health system responsiveness, defined as non-medical aspect of treatment relating to the protection of the patients’ legitimate rights, is the intrinsic goal of the WHO strategy for 21st century. Objective. To describe the patients’ opinions on treatment they received in hospital, namely: admission to hospital, the role of patient in hospital treatment, course of treatment, medical workforce attitude, hospital environment, contact with family and friends, and the efficacy of hospital treatment in respect to responsiveness to patient’s needs and expectations (dignity, autonomy, confidentiality, communication, prompt attention, social support, basic amenities and choice of provider). Material and methods. The data were collected in 2012 from 998 former patients of the randomly selected 73 hospital in Poland. Results. Dignity: Over 80% of patients experienced kindness, empathy, care and gentleness, and over 90% of them had the sense of security in hospital, met with friendliness during the admission to hospital and never encountered inappropriate comments from medical staff. Autonomy: About 80% of patients accepted the active role of patients in hospital, they perceived they had influence on procedures related to hospitalization and course of treatment, and they felt medical staff responded to their requests and concerns. Over 90 % of them had opportunity to communicate their concerns to medical staff and to discuss the course of treatment. On the other hand, the explanation of the reason for the refusal to meet their requests was given to only 23 % of the patients interested. Confidentiality: 70-80 % of patients declared the respect for privacy and confidentiality during collecting the health information and during medical examinations, and were not examined in presence of other people. Nevertheless, only 23% of patients examined so were asked of their consent. Communication: About 90% of patients declared they trusted their physician, received from him explanation regarding the course of treatment and information about further treatment after discharge from hospital, but physicians devoted the time and attention to only 70% of them. Prompt attention: Over 90% of patients perceived simplicity of the formalities of admission to hospital, and short waiting for treatment and additional tests in hospital (but only 50% received explanation of reason if they waited long). Nevertheless, 10% of them % of them perceived they waited for admission to hospital too long, and over 20% for admission to a ward as long. Social support: The unlimited direct and phone contact with family and friends was declared by 96% of patients. Basic amenities: The high percentage of patients assessed positively the marking in hospital (97%) and cleanliness of linen (89%), followed by the general indoor appearance room in which patient stayed, lack of noise (70-80%), hospital meals, furniture (60-70%), availability of personal hygienic articles (50-60%), cleanliness of hospital room, toilet, showers and bathtubs, and availability of soap (40-50%). Choice of provider: Only 41% of patients declared that they had influence on choice of the hospital. Conclusion. Responsiveness of Polish hospital patient needs is similar to that of the OECD countries of the lowest health system responsiveness. Compared to the Central European countries, the responsiveness in Polish hospitals is lower than that of Czech Republic and only slightly higher of those of Slovenia, Slovakia and Hungary. Key words: responsiveness, patient rights, hospital STRESZCZENIE Wprowadzenie. Wrażliwość systemu opieki zdrowotnej na potrzeby pacjenta, definiowana jako niemedyczny aspekt leczenia odnoszący się do ochrony praw należnych pacjentom, stanowi samoistny cel strategii Światowej Organizacji Zdrowia na 21-szy wiek. Cel badań. Zebranie i przedstawienie opinii pacjentów o leczeniu, które zapewniono im w szpitalu, mianowicie: przyjęcie do szpitala, rola pacjenta w czasie leczenia szpitalnego, przebieg leczenia, postawa personelu medycznego, środowisko szpitalne, kontakt z rodziną i znajomymi oraz skuteczność leczenia szpitalnego, w odniesieniu do wrażliwości na potrzeby i oczekiwania pacjenta (godność, autonomia, poufność, komunikacja, niezwłoczna pomoc, wsparcie społeczne i wybór szpitala). *Corresponding author: Lucyna Gromulska, Department of Health Promotion and Postgraduate Education, National Institute of Public Health – National Institute of Hygiene, Chocimska Street 24, 00-791 Warsaw, Poland, phone +48 22 54 21 203, fax +48 22 54 21 375, e-mail: [email protected] © Copyright 2013 by the National Institute of Public Health - National Institute of Hygiene 156 L. Gromulska, P. Goryński, P. Supranowicz et al. Nr 2 Materiał i metody. Dane zebrano od 998 byłych pacjentów z losowo wybranych 73 szpitali w Polsce. Wyniki. Szacunek: Ponad 80% pacjentów dostrzegało życzliwość, współczucie, troskę i delikatność, a ponad 90% miało poczucie bezpieczeństwa, spotkało się z uprzejmością podczas przyjęcia do szpitala i nie spotkało się z niewłaściwymi uwagami ze strony personelu medycznego. Autonomia: Około 80% pacjentów akceptowało aktywną rolę pacjenta w szpitalu, postrzegało, że mają wpływ na działania związane z pobytem w szpitalu i przebiegiem leczenia i reagowanie przez personel medyczny na ich prośby i wątpliwości. Ponad 90 % miało możliwość przekazywania personelowi medycznemu swoich wątpliwości i omawiać przebieg leczenia z lekarzem. Z drugiej strony, wyjaśnienie powodów odmowy spełnienia ich próśb przekazało tylko 23% zainteresowanych pacjentów. Poufność: Chociaż 70-80% pacjentów deklarowało respektowanie prywatności i poufności w czasie zbierania informacji o zdrowiu i w czasie badań medycznych, a także nie byli oni badani w obecności innych osób, jednakże o zgodę proszono tylko 23% pacjentów badanych w ten sposób. Komunikacja: Prawie 90% pacjentów odczuwało zaufanie do lekarza, otrzymywało od niego wyjaśnienia o przebiegu leczenia i informacje o dalszym leczeniu po wypisaniu ze szpitala, ale lekarz poświęcał swój czas i uwagę tylko 70% z nich. Szybka pomoc: Chociaż ponad 90% pacjentów postrzegało łatwość załatwiania formalności związanych z przyjęciem do szpitala i czas czekania na zabiegi i dodatkowe badania postrzegało jako krótki (ale jeśli długo czekali, to tylko 50% otrzymywało wyjaśnienia o przyczynie), jednakże prawie 10% czekających na przyjęcie do szpitala i ponad 20% czekających na przyjęcie na oddział postrzegało czas oczekiwania jako długi. Wsparcie społeczne: Brak ograniczeń w kontaktowaniu się z rodziną i znajomymi poprzez wizyty i rozmowy deklarowało 96% pacjentów. Podstawowe udogodnienia: Wysoki odsetek pacjentów ocenił pozytywnie oznakowania w szpitalu (97%) i czystość pościeli (89%), a w następnej kolejności: wygląd wnętrza szpitala, salę w której przebywa pacjent, brak hałasu (70-80%), posiłki, mebli (60-70%), zapewnienie papieru toaletowego, ręczników papierowych i suszarek do rąk (50-60%), czystość sali szpitalnej, toalet, pryszniców i wanien oraz dostępność mydła (4050%). Wybór usługodawcy: 41% pacjentów zadeklarowało, że mieli możliwość wyboru szpitala. Wnioski. Wrażliwość na potrzeby pacjentów szpitalnych w Polsce jest podobna do notowanej w państwach OECD o najniższej wrażliwości systemu zdrowia. W porównaniu do państw Europy Środkowej wrażliwość jest niższa niż w Republice Czeskiej i tylko nieco wyższa niż w Słowenii, na Słowacji i na Węgrzech. Słowa kluczowe: wrażliwość na potrzeby pacjenta, prawa pacjenta, szpital INTRODUCTION The concept of responsiveness being, in addition to health outcomes and fair financial contribution, the intrinsic goal of the health system performance assessment was formulated at the beginning of 21st century in WHO strategy aimed at improving health quality and equity [6]. Health system responsiveness is defined as non-medical aspect of treatment relating to the protection of the patients’ legitimate needs and expectations in the way guaranteed to him/her by the human rights and patient rights in particular. It consists of eight domains. Dignity refers to respectful treatment by health care staff, the right to ask questions and provide information during consultations and treatment, and privacy during examination and treatment. Autonomy means the right of an individual to be informed about his/her disease and alternative treatment options, to be consulted about treatment, and to express the informed consent in the context of testing and treatment. Confidentiality involves conducting the consultations with the patients in a manner that protects their privacy and safeguards the confidentiality of information provided by the patient, information relating to an individual’s illness in particular, except in cases where such information needs to be given to a health care provider, or where explicit consent has been gained. Communication refers to clarity of information, careful listening to the patient’s questions and explaining things to be understood. Prompt attention means that patients should be entitled to rapid care in emergency, and they should be entitled to care within reasonable time even in non-emergency health problems or surgery, so waiting lists should not cover long periods. Quality of basic amenities relates to clean surroundings, regular procedures of cleaning and maintenance of hospital buildings, adequate furniture, sufficient ventilation, clean water, toilets and linen, and healthy food. Access to social support during hospitalization should allow for regular visits by relatives and friends and enable religious practices that do not prove an obstacle to hospital or hurt the sensibilities of other patients. Choice of care provider means being able to freely choose a physician and an institution to provide health care [8]. Responsiveness research from the perspective of patients is broadly similar to that of patient’s satisfaction, however they differ in their approach; the latter puts emphasis on increasing the efficacy of medical treatment, whereas the interest of this first mainly relates to ethical issues of treatment [1]. Since 2011, the analysis of factors influencing the opinions of treatment in Polish hospital granted by the Ministry of Science and Higher Education has been carried out in the Department of Health Promotion and Postgraduate Education of the National Institute of Public Health – National Institute of Hygiene in Warsaw (Poland). Nr 2 Responsiveness to hospital patient needs in Poland The aim of present paper was to describe the patients’ opinions on treatment they received in hospital, namely, admission to hospital, the role of patient in hospital cure, course of treatment, medical staff attitude, hospital environment, contact with family and friends, and the efficacy of hospital treatment, in respect with responsiveness to patient’s legitimate needs and expectations (dignity, autonomy, confidentiality, communication, prompt attention, social support, basic amenities and choice of provider). MATERIAL AND METHODS Data collection The hospitals where respondents were recruited, were randomly chosen from the register of Polish hospitals, and 73 public hospitals, proportionally to the number of patients hospitalised in the provinces (voivodeships), were qualified. The study was conducted among the patients of the internal medicine wards after obtaining the permission of the patients themselves and the hospital directors. Two thousand nine hundred and twenty patients being at hospital from April to September 2012 had agreed to participate in the study and provided the contact details. The data were collected from 1000 former patients in December 2012, i.e. after 3 – 9 Table 1. Sample characteristics Demographic factors Total Gender male female Aged 18-29 30-44 45-64 65-79 80 and more Education elementary vocational secondary post-secondary/incomplete higher higher Occupational activity employed unemployed seeking work unemployed not seeking work pensioners Marital status married/in permanent cohabitation divorced widowed single Place of residence town village n 998 % 100 451 547 45.2 54.8 56 129 466 300 47 5.7 12.9 46.6 30.0 4.7 209 258 310 58 153 21.1 26.1 31.4 5.9 15.5 287 39 63 606 28.8 3.9 6.3 60.9 647 54 186 108 65.0 5.4 18.6 10.8 581 417 58.2 41.8 157 months after discharge from hospital, and 998 correctly completed questionnaires were used for analysis. The sample characteristics is presented in Table 1. Questionnaire The developed questionnaire was based on the WHO responsiveness definition and modified to suit the Polish health system conditions. The questions were grouped into seven themes: admission to hospital, the role of patient in hospital care, treatment course, medical staff, hospital environment, contact with family and friends, and the efficacy of hospital treatment. The admission to hospital covered: health status at the time of admission to hospital, procedure of admission, choice of hospital, actual and perceived waiting time for admission to hospital, actual and perceived time of waiting in hospital to be admittted on a ward, simplicity of arranging the formalities of admission to hospital, staff attitude to the patient and to accompanying persons during admission. The role of patient in hospital covered: general opinion concerning the active role of the patient in the hospital, patient’s influence on a course of treatment, discussing and agreeing a course of treatment with the patients, opportunity to communicate the concerns to medical staff, reporting the requests and concerns by medical staff and explanation of the reason for the refusal of fulfilling the request. Course of treatment covered: sense of security during the stay in hospital, respect for privacy and confidentiality when collecting health information, respect for privacy and confidentiality during of medical examinations, presence of unauthorized people during medical examinations or patient-doctor conversations, patient’s consent to the presence of other people during medical examinations or patient-doctor conversations, inappropriate comments from the hospital staff, waiting for treatment or additional tests, explanation of the reason of the long waiting for treatment or additional tests, lack of gentleness of the medical staff during treatment and wearing rubber gloves by the medical staff during treatments. Medical staff assessment covered: kindness of the hospital staff referring to the patients, empathy and care of medical staff towards the patients, confidence to the physician attending, time and attention devoted to patients by the physician, explanation of the course of treatment given by physician, giving the information to the patient about further treatment after discharge from hospital, medical staff assistance in daily activity, quick help from nurses and assessment of the appearance of the medical staff. Hospital environment covered: the interior of the hospital, marking applied in hospital, room in which patient resides, hospital furniture, cleanliness of hospital room, linen, toilet, showers and bathtubs, availability of toilet paper, soap, paper towels and hand dryers, noise and hospital meals. Contact with family and friends cove- 158 Nr 2 L. Gromulska, P. Goryński, P. Supranowicz et al. red: regulation of hospital visits, phone contact with family and friends and nuisance of guest visits to other patients. The efficacy of hospital treatment covered: actual and perceived length of stay in hospital, perceived improvement in health status after hospital treatment and recommending the hospital to family and friends. The usefulness of the questionnaire was validated in the pilot study on 25 patients. Statistical analysis The SPSS program was applied for establishing the database and statistical analysis. According to the WHO recommendations for responsiveness measuring [10], the ordinal variables (except those of two categories) were converted into three-categorised (positive, moderate, negative response), and the prevalence of positive responses (percentages) was set up as a measure of responsiveness in respect to the item creating the domain (i.e. dignity, autonomy, etc.). Responsiveness of a domain was calculated as the mean of percentages of positive responses to the items forming given domain, and the total responsiveness was the mean of percentages of positive responses to all items. RESULTS Admission to hospital The referred planned admissions to hospital were more frequent than those due to emergency or sudden deterioration in health (Table 2). Only two of five patients declared the possibility to choose a hospital. Two-thirds of the patients waited for admission to hospital no longer than 7 days, nevertheless, over 10% had to wait more than 30 days, in that, almost 10% as long as 90 days or more. Also two-third of the patients defined the waiting time as short, while almost 10% perceived that they waited long. Only every fourth patient waited at hospital for admission to a ward up to 15 minutes, and the same proportion of them waited 16-30 minutes, while almost 30% of the patients had to wait one hour, in that, almost 7% as long as four hours and longer. Almost half of the patients described the waiting time as short, however, it was long for every one in five patient, as expected. The vast majority of the patients perceived the arranging the formalities of admission to hospital as simple, and assessed positively the hospital staff attitude to patients and accompanying persons during admission to hospital. The role of patient in hospital Most of the patients recognised the need for the active role of patients in the hospital, while every tenth patient was of the opposite opinion (Table 3). Majority of them experienced the influence on the procedures Table 2. Admission to hospital Factors examined* Health status at the time of admission to hospital walking unaided moving with the help of other person or walker not walking and conscious unconscious Procedure of admission emergency, sudden deterioration in health planned admission to hospital with referral transfer from another hospital Hospital selection (CP) self-choice impossibility of self-choice Waiting time for admission to hospital up to 7 days 7-30 days 30-90 days >90 days Perceived waiting time for admission to hospital (PA) short or did not wait at all middling long Waiting time at the hospital to admit on a ward up to 15 min. 16-30 min. 31-60 min. 61-120 min. 121-240 min. >240 min. Perceived waiting time to admit on a ward (PA) short middling long Simplicity of arranging the formalities of admission to hospital (PA) yes no Staff attitude to the patient during admission to hospital (D) positive negative Staff attitude to the accompanying persons during admission to hospital (D) positive negative n % 733 164 79 22 73.4 16.4 7.9 2.2 433 523 20 43.5 52.5 2.0 200 285 41.2 58.8 180 80 25 11 61.9 26.3 8.2 3.6 318 130 46 64.4 26.3 9.3 231 219 170 113 85 59 26.3 25.0 19.3 13.0 9.7 6.7 318 130 46 49.3 28.0 22.7 880 69 92.7 7.3 933 19 98.2 2.0 655 17 97.5 2.5 * Domains of responsiveness in parentheses: D – dignity, A – autonomy, Cy – confidentiality, Cn – communication, PA – prompt attention, SS – social support, BA – basic amenities, CP – choice of provider. relating to stay in hospital and a course of treatment, however, the opponents were frequent in the latter. The vast majority of the patients discussed and agreed a course of treatment with medical staff and had opportunity to communicate their concerns to them. Every fourth of the patients reported requests and concerns and almost 80% of them received satisfactory reaction from medical staff. Nevertheless, only every fifth was given an explanation for the refusal of their request. Nr 2 159 Responsiveness to hospital patient needs in Poland Table 3. The role of patient in hospital treatment Factors examined* Opinion concerning the active role of the patients in the hospital (A) yes sometimes no Patient’s influence on the procedures related to staying in hospital (A) often sometimes rarely or never Patient’s influence on a course of treatment (A) yes sometimes no Discussing and agreeing a course of treatment with the patients (A) yes no Opportunity to communicate the concerns to medical staff (A) yes rarely or never Reporting the requests and concerns by patients yes no Responding to the patient’s requests and concerns by medical staff (A) satisfactory incomplete getting rid Explanation of the reason for the refusal of fulfilling the request (A) always sometimes never n % 753 115 85 79.0 12.1 8.9 346 92 454 81.2 13.3 5.5 734 101 104 78.2 10.8 11.0 943 40 95.9 4.1 867 79 91.6 8.4 274 724 27.5 72.5 218 31 25 79.6 11.3 9.1 13 22 21 23.2 39.3 37.5 * Domains of responsiveness in parentheses: D – dignity, A – autonomy, Cy – confidentiality, Cn – communication, PA – prompt attention, SS – social support, BA – basic amenities, CP – choice of provider. Treatment course The vast majority of the patients experienced the sense of security during the stay in hospital, while those who felt insecure were 2% (Table 4). Also, the most of the patients experienced respect for privacy and confidentiality when collecting health information and in time of medical examination, nevertheless, every sixth and every eighth patient, respectively, was treated with little respect. One-third of the patients were examined in the presence of unauthorised people, and only every fourth of them were asked of their consent to such presence. Although only 4% of the patients encountered inappropriate comments from the hospital staff. The vast majority of the patients waited shortly for treatment or additional tests, but the explanation of the reason of waiting was given to only the half of those who waited long. Most of the patients were always treated with gentleness , while almost 3% perceived the lack of gentleness of the medical staff during treatment. Wearing the rubber gloves by medical staff during treatments is mandatory, therefore, it is worrying that almost 5% of the patients reported that the gloves were not always worn. Table 4. Course of treatment Factors examined* Sense of security during the staying in hospital (D) yes sometimes no Respect for privacy and confidentiality when collecting health information (Cy) yes sometimes no Respect for privacy and confidentiality in the time of medical examinations (Cy) yes sometimes no Medical examinations or health talks in the presence of other people (Cy) no yes Consent on the presence of other people during medical examinations or health talks (Cy) yes no Inappropriate comments from the hospital staff (D) was not was Waiting for treatment or additional tests (PA) short long Explanation of the reason for the long wait for treatments or additional tests (PA) given not given Lack of delicacy from the medical staff during treatment (D) never sometimes often Wearing rubber gloves by the medical staff during treatments always sometimes never n % 953 24 20 95.6 2.4 2.0 569 70 140 72.3 9.9 17.8 748 55 11 81.8 6.0 12.2 679 298 69.5 30.5 218 745 22.6 77.4 957 40 96.0 4.0 914 63 93.5 6.5 31 32 49.2 50.8 881 72 26 90.0 7.3 2.7 935 41 5 95.3 4.2 0.5 * Domains of responsiveness in parentheses: D – dignity, A – autonomy, Cy – confidentiality, Cn – communication, PA – prompt attention, SS – social support, BA – basic amenities, CP – choice of provider. Medical staff The majority of patients experienced kindness, sympathy and care from medical staff, and only few of them (5,5% and 2,7%) were treated otherwise. (Table 5). The attending physician was mostly described positively in answers to the questions about confidence, explanation the course of treatment, provision of the information about further treatment after discharge from hospital and, to a lesser extent, time and attention 160 Nr 2 L. Gromulska, P. Goryński, P. Supranowicz et al. devoted to patients. Nevertheless, one in twenty patients did not confide in their physician, and one in ten patients was not given enough attention explanation of the treatment, nor was he informed about the future treatment. Medical staff assisted in daily activity three-quarters of patients, but one in thirty patients who needed assistance did not received it. Two-thirds of patients always received the prompt help, however, every tenth of the patients experienced it rarely or never. The appearance (neat, clean) of the medical staff was positively assessed by the majority of patients. Table 5. Medical staff Factors examined* Kindness of the hospital staff referring to the patients (D) always gentle mainly rarely or never Sympathy and care to the patients from medical staff (D) very careful moderately little Confidence to the physician attending (Cn) yes to a limited extent no Time and attention devoted by the physician to patients (Cn) always paid attention to the patients mostly rarely or never Explanation by the physician agreeing the course of treatment (Cn) yes no Providing by physician the information about further treatment after discharge (Cn) yes no Medical staff assistance in daily activity (PA) yes no not need help Quick help from the nurses/orderlies if need (PA) always often rarely or never Assessment of appearance of the medical staff (PA) always neat and tidy mostly rarely or never n % 810 133 55 81.2 13.3 5.5 839 121 27 85.0 12.3 2.7 894 47 54 89.8 4.7 5.5 703 171 118 70.9 17.2 11.9 884 110 88.9 11.1 913 80 91.9 8.1 753 31 212 75.6 3.1 21.3 556 251 74 63.1 28.5 8.4 921 60 14 92.6 6.2 1.4 * Domains of responsiveness in parentheses: D – dignity, A – autonomy, Cy – confidentiality, Cn – communication, PA – prompt attention, SS – social support, BA – basic amenities, CP – choice of provider. Hospital environment The high percentage of patients were satisfied with: the marking in hospital (97%) and cleanliness of linen (89%), the interior of the hospital and patients’ room, lack of noise (70-80%), hospital meals, furniture (6070%), availability of toilet paper and paper towels or hand dryers (50-60%), cleanliness of hospital room, toilet, showers and bathtubs, and soap (40-50%) available (Table 6). Table 6. Hospital environment Factors examined* The interior of the hospital (BA) positive moderate negative Marking applied in hospital (BA) adequate inadequate Patients’ room (BA) spacious middling narrow Hospital furniture (BA) positive moderate negative Cleanliness of hospital room (BA) definitely clean acceptably dirty Cleanliness of linen (BA) changed according to the patient needs not changes Cleanliness of toilet (BA) definitely clean acceptably dirty Availability of toilet paper (BA) always sometimes never has own paper Availability of soap (BA) always sometimes never has own soap Availability of paper towels or hand dryers (BA) always sometimes never Cleanliness of showers and bathtubs (BA) definitely clean acceptably dirty Noise nuisance (BA) no sometimes yes Hospital meals (BA) positive moderately negative n % 742 173 83 74.3 17.3 8.4 955 28 97.1 2.9 740 181 77 74.1 18.1 7.8 649 212 137 65.0 21.2 13.8 487 492 17 48.9 49.4 1.7 84 98 89.6 10.4 466 450 67 47.4 45.8 6.8 572 164 152 99 58.0 16.6 15.4 10.0 446 146 223 171 45.2 14.8 22.6 17.4 488 184 276 51.5 19.4 29.1 403 498 58 42.0 51.9 6,1 709 168 121 71.1 16.8 12.1 565 247 24 67.6 29.5 2.9 * Domains of responsiveness in parentheses: D – dignity, A – autonomy, Cy – confidentiality, Cn – communication, PA – prompt attention, SS – social support, BA – basic amenities, CP – choice of provider. Nr 2 161 Responsiveness to hospital patient needs in Poland Contact with family and friends The vast majority of patients declared they had opportunity to contact family and friends in person, or by phone (Table 7). Only every twelfth patient reported the nuisance of guest visits to other patients. Table 7. Contact with family and friends Factors examined* Regulation of hospital visits (SS) unlimited limited Phone contact with family and friends (SS) possible impossible Nuisance of guest visits to other patients no yes n % 945 39 96.0 4.0 877 37 96.0 4.0 920 77 92.3 7.7 * Domains of responsiveness in parentheses: D – dignity, A – autonomy, Cy – confidentiality, Cn – communication, PA – prompt attention, SS – social support, BA – basic amenities, CP – choice of provider. The efficacy of hospital treatment Over the half of patients stayed in hospital fot 2-7 days, and almost half of them stayed in hospital longer (Table 8). Those of one-day stay were few. From the patient perspective, the length of staying in hospital was adequate to their health needs, for the most of them, while every eighth patient perceived it as too short and every fifteenth as too long. The three-quarters of patients perceived improvement in their health due to hospital treatment, whereas every fifteenth did not note a positive result. The majority of patients would recommend the hospital where they were treated to family and friends. The dissatisfied patients were almost 8%. Table 8. The efficacy of hospital treatment Factors examined Length of staying in hospital 1 day 2-7 days more than 7 days Perceived length of staying in hospital adequate too short too long Perceived improvement in health status after hospital treatment yes hard to say no Recommending the hospital to family and friends yes sometimes no n % 13 540 438 1.3 54.5 44.2 799 118 57 82.0 12.1 5.9 778 154 66 78.0 15.4 6.6 848 74 76 85.0 7.4 7.6 Responsiveness The unweighted means of the responsiveness domains calculated by summing the percentages of positive responses divided by the number of items of the domain were shown in Table 9. The social support presented the highest mean prevalence, followed by dignity, communication, autonomy, prompt attention, basic amenities, confidentiality and choice of provider. The mean of over 80% indicates the provision of patients legitimate needs and expectations (social support, dignity and communication), while that under 70% shows no respect for patient’s rights (choice of provider, confidentiality and basic amenities). Table 9. Unweighted means of the responsiveness domains. Domains Dignity Autonomy Confidentiality Communication Prompt attention Social support Basic amenities Choice of provider Total no. of items 7 7 4 4 5 2 13 1 mean of positive responses (%) 92 76 62 85 70 96 64 41 43 73 range (%) 81 – 98 23 – 96 23 – 82 71 – 92 49 – 94 96 42 – 97 41 23 – 98 DISCUSSION Dignity The WHO ranking of patients’ perceived importance of responsiveness domains was developed using the data from the international study on 117 549 participants from 65 countries. Dignity got the second rating of the importance and its mean was 14.8 points (in Poland – third rating and 13.7 points). The highest rating was noted in Egypt, Lebanon and Chile, while the lowest in Luxemburg, France and New Zealand [10]. The study on the health system responsiveness from the perspective of patients conducted on the sample of 27 521 inpatients from 16 OECD countries showed that the average prevalence of positive responses concerning dignity was 86% (range 61-97%). The higher responsiveness (>90%) was reported in Sweden, United States, United Kingdom, Canada, Luxemburg, France and New Zealand, while the lower (<70%) in Greece and Portugal [9]. The average percentage of positive responses related to dignity in the study conducted on inpatients from 5 Central European countries (Croatia, Czech Republic, Hungary, Slovakia and Slovenia) was 78% and ranged from 65% (Croatia) to 82% (Czech Republic) [11]. Our findings confirmed the high responsiveness to Polish hospital patients in respect to dignity is comparable to that of the OECD countries of the highest prevalence, and much higher of that of the Central European countries. In our study, the items of the questionnaire related to dignity presented the low diversity. Over 80% of patients experienced kindness, 162 L. Gromulska, P. Goryński, P. Supranowicz et al. empathy, care and gentleness, and over 90% of them had the sense of security in hospital, met with friendliness during admission to hospital and never encountered inappropriate comments from medical staff. Autonomy Autonomy got the sixth most important domain of responsiveness in the WHO study mentioned above and its mean was 11.7 points (in Poland – fifth rating and 12.0). The highest rating was reported in Austria, Netherlands, Switzerland, Sweden, China and Denmark, while the lowest in Egypt, Romania and Georgia [10]. The study of 16 OECD countries showed that the average prevalence of positive responses concerning autonomy was 72% (range 44-84%). The higher responsiveness (>80%) was reported in New Zealand, United States, Luxemburg, Sweden and United Kingdom, while the lower (<70) in Greece, Italy, Spain and Portugal [9]. In the study of the Central European countries, the average percentage of positive responses related to autonomy was 54% and ranged from 32% (Croatia) to 63% (Hungary) [11]. Our findings demonstrated that the responsiveness to the Polish hospital patients in respect to autonomy is slightly above the mean for the OECD countries and considerably higher than the mean for the Central European countries. Particular items of the questionnaire related to autonomy presented the high diversity. About 80% of patients accepted the active role of patients in hospital, they stated that they had influence on procedures related to staying in hospital and course of treatment, and they were given response to their requests and concerns from medical staff. Over 90% of patients had opportunity to communicate their concerns to medical staff and to discuss the course of treatment with the physician. On the other hand, the explanation of the reason for the refusal to address their requests was given to only 23 % of the patients interested. Confidentiality Confidentiality got the fourth rating in the WHO study and its mean was 12.4 (in Poland - sixth rating and a mean of 12.0). The highest rating was reported in Iceland, Germany, France, Belgium and Egypt, while the lowest in Lithuania, Indonesia and Romania [10]. The study of 16 OECD countries showed that the average prevalence of positive responses related to confidentiality was 82% (range 68-92%). The higher responsiveness (>90%) was reported in Ireland, Canada and United States, while the lower (70) in Italy and Portugal [9]. The average percentage of positive responses related to confidentiality was 70% and ranged from 54% (Croatia) to 78% (Czech Republic) [11]. Our findings demonstrated that the responsiveness to the Polish hospital patients in respect in terms of confidentiality is comparable with Nr 2 that for the OECD countries of the lowest prevalence and lowest than the mean for the Central European countries. The items of the questionnaire composing the domain of confidentiality presented the high diversity. Although 70-80 % of patients declared they experienced respect for privacy and confidentiality during collecting the health information and during medical examinations, and were not examined in presence of other people, nevertheless, only 23 % of those examined were asked of their consent. Communication Communication got the third rating in the WHO study and its mean was 14.0 (in Poland – fourth rating and a mean of 12.7). The highest rating was reported in Republic of Korea and Indonesia, while the lowest in Venezuela and Portugal [10]. The study of 16 OECD countries showed that the average prevalence of positive responses concerning communication was 82% (range 49-89%). The higher responsiveness (>85%) was reported in Sweden, New Zealand, France, United States and Finland and dramatically low (<50%) in Greece [9]. The average percentage of positive responses related to communication in the study of 5 Central was 72% and ranged from 56% (Croatia) to 80% (Slovakia) [11]. Our findings demonstrated that responsiveness to Polish hospital patients in respect to communication is slightly higher than the mean for OECD countries and considerably higher than that of the Central European countries. The items of communication presented moderate diversity. About 90% of patients felt they could confide in their physician, received from him explanation regarding the course of treatment and information about further treatment after discharge from hospital, but physicians devoted sufficient time and attention to only 70% of them. Prompt attention Prompt attention was rated as first in the WHO study and its mean was 18.0 (in Poland - also the first and a mean of 19.5). The highest rating was reported in Indonesia and Italy, while the lowest in Lebanon and China [10]. The study of 16 OECD countries showed that the average prevalence of positive responses concerning prompt attention was 77% (range 61-85%). The higher responsiveness (>80%) was reported in Germany, Netherlands, Luxemburg, United Kingdom, Ireland and Finland, while the lower (<70%) in Greece [9]. The average percentage of positive responses related to prompt attention in the study of 5 Central European countries was 58%, and ranged from 43% (Croatia) to 74 % (Czech Republic) [11]. Our findings demonstrated that the responsiveness to the Polish hospital patients in respect to prompt attention is comparable to that of Nr 2 Responsiveness to hospital patient needs in Poland the OECD countries of the lowest responsiveness and higher than the mean for the Central European countries, however, lower than that of Czech Republic. The items of prompt attention demonstrated high diversity. Over 90% of patients experienced simplicity of arranging the formalities of admission to hospital and short waiting for treatment and additional tests in hospital (but only 50% received explanation for undergoing these procedures. 64% of the respondents declared they waited shortly for admission to hospital, and only 50% for admission to a ward. Social support Social support got the eighth rating in the WHO study and it mean was 6.3 (in Poland also eight rating and 4.7). The highest rating was reported in Canada and Kyrgyzstan, while the lowest in Republic of Korea , Hungary, Netherlands and Sweden [10]. The study of 16 OECD countries showed that the average prevalence of positive responses concerning social support was 88% (range 74-96%). The higher responsiveness (>90%) was reported in Netherlands, Canada, Sweden, United Kingdom and Luxemburg, while the lowest <80%) in Portugal, Greece and Italy [9]. The average percentage of positive responses related to social support in the study of 5 Central European countries was 80% and ranged from 61% (Croatia) to 93% (Hungary) [11]. Our findings demonstrated that responsiveness to Polish hospital patients in respect to social support is comparable to that of the OECD countries of the highest prevalence and higher than that of the Central European countries (except Hungary). The items of social support presented complete identity. The unlimited contact with family and friends by visits or phone declared 96% of patients. Basic amenities Basic amenities got seventh rating in the WHO study and it mean 10.6 (in Poland also seventh rating and 11.3). The highest ratings was reported in Turkey, Indonesia, Nigeria and Kyrgyzstan, while the lower in Canada and France [10]. The study of 16 OECD countries showed that the average prevalence of positive responses concerning basic amenities was 74% (range 59-88%). The higher responsiveness (>80%) was reported in Ireland, Germany and United Kingdom, while the lowest (<70%) in Italy, Greece and Portugal [9]. The average percentage of positive responses related to quality of basic amenities in the study of 5 Central European countries was 60% and ranged from 44% (Croatia) to 80% (Czech Republic) [11]. Our findings demonstrated that responsiveness to Polish hospital patients in respect to basic amenities is comparable to that of the OECD countries of the lowest prevalence and comparable with the mean for the Central European countries, but considerably lower of that in Czech 163 Republic. The items of the quality of basic amenities presented high diversity. The high percents of patients assessed positively the marking in hospital (97%) and cleanliness of linen (89%), followed by the appearance inside the hospital, room in which patient resides, lack of noise nuisance (70-80%), hospital meals, furniture (60-70%), availability of toilet paper and paper towels or hand dryers (50-60%), cleanliness of hospital room, toilet, showers and bathtubs , and soap available for patients(40-50%). Choice of provider Choice of provider got the fifth rating in The WHO study and its mean was 12.3 (in Poland - the second rating and a mean of 13.9). The highest rating was reported in United States, Estonia, Latvia, Cyprus and Czech Republic, while the lowest in Nigeria, India and Indonesia [10]. The study of 16 OECD countries showed the average prevalence of positive responses concerning choice of provider was 87% (range 60-97%). The higher responsiveness (>90%) was noted in Belgium, France, New Zealand, Canada United States and United Kingdom, while the lowest in Finland (60%) [9]. The average percentage of positive responses related to choice of provider in the study of 5 Central European countries was 64% and ranged from 54% (Croatia) to 79% (Czech Republic) [11]. Our findings demonstrated that responsiveness to Polish hospital patients in respect to choice of provider is much lower than both: that of the OECD countries of the lowest prevalence and of the Central European countries (41% of patients declared that they had possibility to choose hospital), however, the underestimation due to only one item used for measuring should be taken into account. Total responsiveness The overall mean of positive responses of the total responsiveness for 16 OECD countries was 81% (range 62-88%). The higher responsiveness (>85%) was noted in United Kingdom, Ireland, Luxemburg, New Zealand, United States and Sweden, while the lower (<75%) in Greece, Portugal and Italy [9]. The overall mean for responsiveness for 5 Central European countries was 67% and ranged from 51% (Croatia) to 76% (Czech Republic) [11]. The study conducted by us showed that the overall mean of responsiveness of Polish hospital patient needs is similar to that of the OECD countries of the lowest health system responsiveness. Compared to the Central European countries, the responsiveness is lower than that of Czech Republic and only slightly higher of those of Slovenia, Slovakia and Hungary. Our findings are consistent with the common opinion on the healthcare in Poland confirmed by the population-based surveys [2, 7]. It should be noted, however, that 164 L. Gromulska, P. Goryński, P. Supranowicz et al. presented results are opposite to those of the patient satisfaction studies, which have permanently demonstrated the positive (even to 100% [3]) evaluation of medical services received in hospitals [4, 5]. The latter are undoubtedly very beneficial for hospitals when they apply for accreditation, but seem to be less sensitive to the actual patient interaction with health system. The use of the responsiveness measuring allows us to demonstrate authentic situation of the hospital patients in Poland. CONCLUSIONS Our findings showed that the responsiveness to Polish hospital patient needs is similar to the OECD countries of the lowest health system responsiveness. Compared to the Central European countries, the responsiveness is lower than that of Czech Republic and only slightly higher of those of Slovenia, Slovakia and Hungary. In particular: 1. the hospital patients legitimate needs and expectations were met sufficiently regarding the social support, dignity and communication; 2. the health system responsiveness was somewhat worse regarding the autonomy and prompt attention; 3. the patients’ rights were not respected enough regarding quality of basic amenities, confidentiality and choice of health providers. Acknowledgements The study was performed as the scientific project nr N N404 168540 ‘Analysis of factors influencing the opinions on treatment in Polish hospitals’ financed by National Science Centre, Poland. The authors would like to thank Bożena Moskalewicz, PhD and Elżbieta Buczak-Stec, MSc, for their invaluable contribution in the project. Conflict of interest The authors declare no conflict of interest Nr 2 REFERENCES 1. de Silva A.: A framework for measuring the responsiveness. Epi Discussion Paper No.32. Geneva, WHO, 2000 URL’ http://www3.who.int/whosis/discussion-papers/ discussion-papers.cfm# 2. Hipsz N.: Opinions of the functioning of the health care system. Warsaw, CBOS, 2012 (in Polish). 3. Kapica D., Orzeł Z., Draus J.: Evaluation of the level of patients’ satisfaction with hospital services. Zdr Publ 2001;111(1):26-30 (in Polish). 4. Marcinowicz L., Grębowski R.: Patient’s satisfaction in the light of the Polish empirical studies – an attempt to elucidate a secret of high satisfaction with care. Pol Merk Lek 2005;17(108):663-666 (in Polish). 5. Miller M., Supranowicz P., Gębska-Kuczerowska A., Car J.: Evaluation of medical service quality by medical patients. Przegl Epidemiol 2008;62(4):643-650 (in Polish). 6. Murray J.C.L., Frenk J.A.: A framework for assessing the performance of health systems. Bull World Health Org 2000;78(6):717-731. 7. Supranowicz P., Wysocki M.J., Car J., Dębska A., Gębska-Kuczerowska A.: Willingness of Warsaw inhabitants to cooperate with health services. II. Evaluation of health and retirement security. Przegl Epidemiol 2012;66(1):149-155. 8. Valentine N.B., Lavalee R., Liu B., Bonsel G.J., Murray C.J.L.: Classical psychometric assessment of the responsiveness instrument in the WHO multi-country study on health and responsiveness. In: Murray C.J.L., Evans D.B, eds.: Health system performance assessment: debates, methods and empiricism. Geneva, WHO, 2003, 597-629. 9. Valentine N.B., Ortiz J.P., Tandon A., Kawabata K., Evans D.B., Murray C.J.L.: Patient experiences with health services: population survey from 16 OECD countries. In: Murray C.J.L., Evans D.B., eds.: Health system performance assessment: debates, methods and empiricism. Geneva, WHO, 2003, 643-652. 10. Valentine N.B., Salomon J.A.: Weights for responsiveness domains: analysis of country variation in 65 national sample surveys. In: Murray C.J.L., Evans D.B., eds.: Health system performance assessment: debates, methods and empiricism. Geneva, WHO, 2003, 631-642. 11. Vitrai J.: Inequalities in health system responsiveness: joint World Health Survey report based on data from selected Central European countries. Geneva, WHO, 2007. Received: 28.01.2014 Accepted: 28.04.2014 Rocz Panstw Zakl Hig 2014;65(2):165-168 INSTRUCTION FOR AUTHORS Quarterly Roczniki Państwowego Zakładu Higieny [Annals of the National Institute of Hygiene] is the peerreviewed scientific journal that publishes original articles, reviews, short communications, letters to the editor and book reviews. Since 2013, Volume 64, number 1 all the papers are published in English. Journal is devoted to the studies concerning scientific problems of food and water safety, nutrition, environmental hygiene, toxicology and risk assessment, public health and other related areas. Articles for publication in the Roczniki Państwowego Zakładu Higieny (Rocz Panstw Zakl Hig) should be sent to the following address: Roczniki Państwowego Zakładu Higieny Narodowy Instytut Zdrowia Publicznego - Państwowy Zakład Higieny 24 Chocimska street, 00-791 Warsaw, Poland or e-mailed: [email protected] RULES FOR THE ACCEPTANCE OF THE MANUSCRIPTS Editorial policy and basic information Only high scientific quality articles complying with the scope of the journal will be considered for the publication. Copyright. All the copyrights to the articles published in the Roczniki Państwowego Zakładu Higieny are reserved for the publisher: the National Institute of Public Health-National Institute of Hygiene. It means that after article is published the authors transfer the copyright to the publisher and can not publish this article or its parts elsewhere without the written permission from the publisher. Ethics. Clinical articles should comply with the generally accepted ethical standards and the Helsinki Declaration. For animal experiments reported in the articles the author(s) must obtain the acceptance by the relevant local Ethics Commission. Conflict of interest. Conflict of interest exists if authors or their institutions have financial or personal relationships with other people or organisations that could inappropriately influence their actions. Such relationships should be disclosed to the publisher. All authors should provide a signed statement of their conflict of interest as a part of the author’s statement form. Transparency. Openness of information on any party contributing to preparation of a publication (content related, financial, etc. input) is proof of ethical attitude of a researcher and of high editorial standards. “Ghostwriting” and “guest authorship” are indications of scientific dishonesty and all cases will be exposed and adequate institutions will be informed. “Ghostwriting” is a situation where a person contributes significantly to a publication and is not disclosed as one of the authors or named in the acknowledgments and “guest authorship” is a situation where an author’s contribution is insignificant or non existent and he is still listed as author/coauthor of a publication. The editor requires from the authors of the articles that they reveal the contribution of individual authors to the manuscript , i.e. who is the author of the concept and study design, data/material collection, study/analysis performance, statistical analysis, interpretation of the result, manuscript development etc. Authors of the manuscript should provide “Authors’ statement” form signed by the all authors. Also the information concerning the sources of financial support to the study presented in the submitted manuscript should be provided. 166 No 2 Peer review procedure for manuscripts 1. The manuscripts submitted to the Roczniki Państwowego Zakładu Higieny undergo preliminary evaluation in the editorial office to determine whether the topic is within the scope of the journal and to evaluate the adherence to the journal format, as well as to the rules of acceptance. In cases when the above prerequisites are not met the manuscript is not qualified, and the corresponding author is informed. 2. Following the positive preliminary evaluation the paper is directed for peer review to at least two independent and recognized experts representing the scientific experience in the field covered by the manuscript and affiliated in the different institution than the institution(s) where the authors have been affiliated. The reviewers must ensure independence and lack of conflict of interests. 3. During the entire reviewing the double-blind review process is maintained. 4. The review in the written form is forwarded to the corresponding author with the accompanying statement suggesting status of the paper as: “accepted”, “accepted after revisions” or “not accepted”. The reviewers form is available on the journal’s website: http://www.pzh.gov.pl/roczniki_pzh/. 5. The list of reviewers is published once a year in the fourth number of each volume. Submission of manuscripts For the publication 1 paper copy of printed text and following attachments must be submitted: 1. Cover letter, in which the corresponding author applies for the publication of the paper in the Roczniki Państwowego Zakładu Higieny. The cover letter, signed by the corresponding author, should contain the name of the institution where he is affiliated, address, phone and fax numbers, e-mail. The corresponding author is requested to suggest two or three potential reviewers (including their e-mails) from different institutions than the author’s one. 2. Author(s)’ statements, signed by all authors, stating that the manuscript complies with the general rules set for the scientific articles and was not published and/or submitted for publication elsewhere and will not be send for publication to the other journal, and there is no infringement of property rights to any interested third parties. In the case of experiments on animals the approval of the relevant ethics commission is required. The clinical studies must be accompanied by the written statement, signed by the authors confirming that the research was conducted in accordance with the Helsinki Declaration. The contribution of the individual authors to preparation of a publication should be specified. All authors are required to sign the statement of their conflict of interest as a part of the author’s statement form. Author’s statement form is available on the website: http://www.pzh.gov.pl/roczniki_pzh/. 3. CD with the text. Text of the manuscript, tables and figures should be in the separate files. Arrangement of manuscripts Manuscripts submitted for publication should be typed by the editor Microsoft Word using Times New Roman 12 font and 1.5 space between lines on A4 paper size. Title page. It should contain the title of the paper in English and, the name(s) and surname(s) of the author(s), the complete name(s) of the institution(s) where the work was performed, and the exact postal address of the corresponding author, phone and/or fax numbers and the e-mail address inserted at the bottom of the title page. If appropriate, the number and the title of the project under which the study has been carried out should also be given on this page as a footnote. Abstract. It should contain 250-300 words and consist of the following sections: Background, Objective, Material and methods, Results, Discussion and Conclusions. The abstract of a review article should contain 250-300 words without division into sections. Keywords. Should be placed after the abstract. 3-6 words or short phrases according to the MeSH (Medical Subject Headings Index Medicus) catalogue available at http://www.nlm.nih.gov/mesh/meshhome.html Polish authors are asked to attach the abstract and keywords also in Polish language. No 2 167 Text. The text of the manuscript should be divided into the following sections: Introduction, Material and methods, Results, Discussion, Conclusions, Acknowledgements, References. Introduction. It should contain the scientific rationale and the aim of the study or in the case of a review the purpose of the article. Only references related to the paper should be cited. Material and methods. This section should provide detailed information on the subject of the study, methods, chemicals, apparatus and techniques used in sufficiently exhaustive way to enable readers to repeat the experiments or observations. For generally known methods references should be given together with name of the methods or statistical analysis used in the study. For new or substantially modified methods detailed descriptions are to be added. In the case of experimental studies on laboratory animals, the information should be provided on the approval by a local Ethics Commission, or in the case of clinical studies that they have been performed with the ethical standards according to the Helsinki Declaration. Results. These should be presented in a logical sequence in the text, the same applies to the tables and figures. The data from the tables and figures should not be repeated in the text, where only the most important observations from the studies are to be summarized. The place where the tables, figures or photographs should appear in the text should be marked. Discussion. Emphasise the new and important aspects of the results and a comprehensive interpretation of the results obtained against the background of results obtained by other authors. Quotations should be restricted to those with immediate relevance to the author’s findings. Conclusions. They should be stated precisely in points or descriptively and should be logically connected with the aims stated in the introduction. Statements and conclusions not derived from own observations should be avoided. If a hypothesis is proposed it must be stated clearly. Acknowledgements. These should be placed directly after the Conclusion section. One or more statements should specify: (1) persons who contributed substantially to the study but cannot be regarded as authorship, such as technical assistants, statisticians, data collectors etc. You should acknowledge their assistance for the sake of transparency. It must be clear that they are not responsible for the final version of the article. You must ensure you have the consent of all the persons named in the acknowledgements; (2) all sources of financial and material support, which should specify the nature of the support. The recommended form is: “This work was supported by: (name of the organization, project number xxxx)”; (3) relationships that may pose the conflict of interest. References. References in the Reference section should be presented in alphabetical order, according to author’s names. Each position in the list of references should start from the new line and contain: consecutive number, author’s (authors’) surname(s) and initials of name(s) (written in italic), full title of the paper, periodical’s title accepted abbreviation (according to the List Journal Indexed in Index Medicus), year, volume, the first and the last page number of the paper. When quoting them in the text only the number of the reference should be given in square brackets. If the article or book has a DOI number (“Digital Objects Identifier” number unique to the publication), it should be included in the references. The titles of the cited papers in other language than English should be translated into English and the information on the original language should be given in the brackets after the page numbers, for example: (in Polish). Article in a periodical: 1. Góralczyk K., Hernik A., Czaja K., Struciński P., Korcz W., Snopczyński T., Minorczyk M., Ludwicki J.K.: Organohalogen compounds – new and old hazards for people. Rocz Panstw Zakl Hig 2010;61(2):109-117 (in Polish). 2. Trucksess M.W., Scott P.M.: Mycotoxins in botanicals and dried fruits: a review. Food Addit Contam 2008;25:181-192. Books and chapter in a book: 3. Riley D.M., Fishbeck P.S.: History of methylene chloride in consumer products. In: Salem H., Olajos E.J. eds. Toxicology in Risk Assessment. London, Taylor & Francis, 2000. Legislative acts: 4. Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. Off J Eur Union L 364/5, 20.12.2006. 168 No 2 Internet source: 5. The Rapid Alert System for Food and Feed. Available from: http:/ec.europa.eu/food/food/rapidalert/index_ en.htm (18.10.2010) Tables. These should be printed on separate sheets and numbered using Arabic numerals. The title should be place directly above each table. Tables should always be cited in the text in consecutive numerical order. Each column in tables should have a brief heading, more extensive explanation, should be given below the table, if necessary. The number of tables should be limited to indispensable for the documentation of results. Figures and photographs. These should be numbered in Arabic numerals according to the sequence of their appearance in the text. Figures should be made by computer technique and the titles should be placed below the figures. Photographs must be of high quality, digital format is preferred. Tables, figures or photographs should be cited in the text in consecutive numerical order in the following way: (Table 1), (Figure 1), (Photo 1). Abbreviations, symbols, units. Generally known and used abbreviations may be left unexplained, others must be explained at the first use in the text. Metric SI units are recommended, however also other generally used units are accepted. General information The editor reserves the right for introducing the editorial corrections in the manuscript which will not influence the scientific contents of the article without prior informing the author. Publication of the papers in the quarterly Roczniki Państwowego Zakładu Higieny is free of charge. 15 reprints free of charge are provided to the corresponding author. SUBSCRIPTION The subscription information is available at: Library of the National Institute of Public Health - National Institute of Hygiene 24 Chocimska street, 00-791 Warsaw, Poland Phone: +48 22 54 21 262 or +48 22 54 21 264 e-mail: [email protected] Subscription Roczniki Państwowego Zakładu Higieny [Annals of the National Institute of Hygiene] may be ordered through Library of the National Institute of Public Health - National Institute of Hygiene mailing address: [email protected] Payment The payment should be made after invoice receipt to the following account: Bank PKO BP S.A. 98 1020 1042 0000 8302 0200 8027 National Institute of Public Health - National Institute of Hygiene 24 Chocimska street, 00-791 Warsaw, Poland Annual subscription rate in Poland in year 2014 (Vol. 65, 4 issues): 120,00 PLN Indeks 37468 Printing house: Libra-Print, Al. Legionów 114b, Łomża, Poland, http://www.libra-print.pl ROCZNIKI PAŃSTWOWEGO ZAKŁADU HIGIENY [ANNALS OF THE NATIONAL INSTITUTE OF HYGIENE] Volume 65 2014 Number 2 CONTENTS REVIEW ARTICLES Flavonoids – food sources and health benefits. A. Kozłowska, D. Szostak-Węgierek .................................................................................................................................. 79 Diacetyl exposure as a pneumotoxic factor: a review. B. Starek-Świechowicz, A. Starek ...................................................................................................................................... 87 ORIGINAL ARTICLES Development and validation of a method for determination of selected polybrominated diphenyl ether congeners in household dust. W. Korcz, P. Struciński, K. Góralczyk, A. Hernik, M. Łyczewska, K. Czaja, M. Matuszak, M. Minorczyk, J. K. Ludwicki ................................................................................................................................................................... 93 Variations of niacin content in saltwater fish and their relation with dietary RDA in Polish subjects grouped by age. M. Majewski, A. Lebiedzińska ......................................................................................................................................... 101 Evaluating adult dietary intakes of nitrate and nitrite in Polish households during 2006-2012. A. Anyżewska, A. Wawrzyniak ......................................................................................................................................... 107 School pupils and university students surveyed for drinking beverages containing caffeine. M. Górnicka, J. Pierzynowska, E. Kaniewska, K. Kossakowska, A. Woźniak ................................................................ 113 The use of vitamin supplements among adults in Warsaw: is there any nutritional benefit? A. Waśkiewicz, E. Sygnowska, G. Broda , Z. Chwojnowska ........................................................................................... 119 Energy and nutritional value of the meals in kindergartens in Niš (Serbia). K. Lazarevic, D. Stojanovic, D. Bogdanović .................................................................................................................. 127 Comparing diabetic with non-diabetic overweight subjects through assessing dietary intakes and key parameters of blood biochemistry and haematology. K. Gajda, A. Sulich, J. Hamułka, A. Białkowska ............................................................................................................ 133 Nutritional values of diets consumed by women suffering unipolar depression. E. Stefańska, A. Wendołowicz, U. Kowzan, B. Konarzewska, A. Szulc, L. Ostrowska .................................................... 139 Awareness of factors affecting osteoporosis obtained from a survey on retired Polish subjects. N. Ciesielczuk, P. Glibowski, J. Szczepanik .................................................................................................................... 147 Responsiveness to the hospital patient needs in Poland. L. Gromulska, P. Goryński, P. Supranowicz, M.J. Wysocki ............................................................................................ 155 Instruction for authors ................................................................................................................................................. 165 Abstracts and full texts: http:// www. pzh.gov.pl/roczniki_pzh/