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ENDOCRINE REGULATIONS

Volume 37 / No. 3 / 2003

• INSTRUCTIONS TO AUTHORS

• Original articles

• Groubet R, Pallet V, Delage B, Redonnet A, Higueret P, Cassand P (Bordeuax, France)
  Hyperlipidic diets induce early alterations of the vitamin A signalling pathway in rat colonic mucosa

• She-Fang YE, Saga I, Ichimura K, Nagai T, Shinoda M, Matsuzaki S (Mibu, Tochigi, Japan)
  Coumestrol as well as isoflavones in soybean extract prevent bone resorption in ovariectomized rats

• Lebl J, Schober E, Zidek T, Baldis S, Rami B, Pruhova S, Kolouskova S, Snajderova M, Frisch H (Prague, Czech Republic and Vienna, Austria)
  Growth data in large series of 587 children and adolescents with type 1 diabetes mellitus

• Ostrowska Z, Kos-Kudla B, Marek B, Kajdaniuk D (Zabrze, Poland)
  Influence of lighting conditions on daily rhythm of bone metabolism in rats and possible involvement of melatonin and other hormones in this process

• Langer P, Kocan A, Tajtakova M, Petrik J, Koska J, Huckova M, Hanzen E, Ksinantova L, Radikova Z, Imrich R, Trnovec T, Blazicek P, Sebokova E, Klimes I (Bratislava, Kosice and Lubochna, Slovakia)
  Thyroid function and cholesterol level: paradoxical findings in large groups of population with high cholesterol food intake

• Daniel Smutek D, Sara R, Sucharda P (Praha, Czech Republic)
  Relation between quantitative description of ultrasonographic image and clinical and laboratory findings in lymphocytic thyroiditis

• Minireview

• Radikova Z (Bratislava, Slovakia)
  Assessment of insulin sensitivity/resistance in epidemiological studies

Endocrine Regulations (since 1967 to 1990 Endocrinologia Experimentalis) is an international journal on experimental and clinical endocrinology edited quarterly in English by care of the Institute of Experimental Endocrinology, Slovak Academy of Sciences (Bratislava, Slovakia) and published by the Slovak Academic Press (Bratislava, Slovakia).

This journal aims to publish original manuscripts or minireviews on experimental and clinical endocrinology and diabetes.

The submission of a manuscript to Endocrine Regulations implies that it has not been previously published or is not being submitted for publication elsewhere and that the manuscript has been approved by all authors who are ready to take public responsibility for the content.

All materials relating to human investigation will be published upon the understanding that design of the work has been approved by the local Ethical Committee or that it conforms to ethical guidelines of the Declaration of Helsinki. The animal experiments should state the conformance to guidelines on animal care.

Manuscripts in triplicate with three sets of illustrations (of which one is an original) should be sent to:

Richard Kvetnansky, Ph.D., Dr.Sc., Chief Editor,

Institute of Experimental Endocrinology,

Vlárska 3, 833 06 Bratislava, Slovakia

All text must be printed on one side of the sheet only with appropriate margins and double spacing to give adequate space for editorial notes. The corresponding author should indicate his/her full mailing address including phone and fax numbers and the e-mail address.

Manuscripts on disc. The submissions of manuscripts prepared on 3.5 inch discs on IBM compatible computers is encouraged, the preferred word processors being Microsoft Word. However, also in this case the disc must be accompanied by three hard copies of the manuscript. The disk should be labelled by the name of the first author, type of word processor, its version and file name and must also accompany the final version of the manuscript.

Types of manuscripts. Standard original papers should contain following sections: * Title, * Abstract (divided into sections Objective, Methods, Results, Conclusions), * Key Words, * Introduction, * Materials and Methods (in clinical papers this section should read * Subjects and Methods), * Results, * Discussion, (* Acknowledgements), * References. There is no length limit for these papers.

Minireviews should give an overview of a defined field preferably of author,s own professional interest and experience. They should not exceed 25 typed pages including complete References and should usually contain * Abstract, * Key Words, * Individual sections and subsections, * References.

Title page should give * the title of the article (main key words should be preferably included into the title to give sufficient information to allow the reader to judge the relevance of a paper to his field), * full names of authors, * institute of origin, * short title (running head), * name and full address of corresponding author including phone and fax numbers and e-mail naddress as well.

Abstract should clearly indicate the purpose of the study (Objective), basic procedures (Methods), main findings (Results) and principal conclusions (Conclusions). New and original findings should be emphasized, clearly defined and defended. The abstract must be easily understood indepenently of the full text of the paper

Key Words. Up to 8 key words (in exceptional cases even more) should be carefully selected to give appropriate information to the users of international information networks.

Introduction should give a brief overview of background informations and clearly define the purpose of the study...

Materials and Methods (in clinical manuscripts Subjects and Methods) should give full informations sufficient to allow others to repeat the work. It is recommended to divide it into subsections. Established and routine methods (if not considerably modified) should be just cited by the appropriate references, the modifications being briefly but clearly described. Statistical methods should be clearly described.

Results should describe concisely and clearly the results in logical sequence. Any interpretations should be avoided and definitely shifted to the Discussion. Do not repeat Materials and Methods, and do not repeat the data presented in tables and figures.

Discussion. Do not simply repeat the data presented in Introduction and Results section. Define and emphasize the new and important aspects of the study and the conclusions that follow. Relate results to other relevant studies, interpret them and explain the differences, if any. Working hypotheses and theories may be briefly outlined.

Acknowledgements. This short section, if necessary, contains acknowledgements of personal and/or financial assistance.

References. Begin this section on a new page. References should be assembled in alphabetical order according to the first author. More than one paper from the same author(s) in the same year must be identified by the letters a, b, c etc. placed after the year of publication. All listed references must be cited in the text by the first author et al. and the year (in a case of two authors only cite both). Following possibilities are recommended: (1) Brown and White (1993) found that ...; (2) ... as observed by Black et al. (1992); (3) ... as previously reported by several authors (Black et al. 1992; Brown and White 1993; Green et al. 1995).

The names of authors in the text and in references should be typed in small letters and underlined (e.g. White and Brown). The volume should be typed in bold.

The style for the list of references is as follows:

Itoh M, Okugawa T, Shiratori N, Ohashi H: Treatment with triiodothyronine (T3) against multinodular goiter fails to prevent the onset of Graves disease. Endocrine Regul 29, 151-156, 1995

Mornex R, Orgiazzi JJ: Hyperthyroidism. In: The Thyroid Gland (Ed. M de Visscher), pp. 279-362, Raven Press, New York 1980

Podoba J: Endemic goiter in Slovakia. VEDA, Bratislava, 1962

The statement “in press” may be used only for a paper accepted for publication in the indicated journal. Unpublished data or Personal communication may be used in the text, but must not be listed in References.

Tables should be constructed as simply as possible, typed on separate sheets and numbered consecutively with Arabic numeral. There should be a short and descriptive heading and appropriate footnotes. Not more than 4 vertical rows should be used in a table planned to occupy one column and not more than 8-10 rows for that designed for two columns of a page.

Figures should be prepared in proportional way with lettering of appropriate size in order to permit such reduction in size to occupy either one or two columns on the page. Drawings (graphs, charts, diagrams etc.) should be submitted either as original or camera ready glossy photographs. Computer generated graphs must be printed by high quality laser printers on high quality camera ready paper. High quality photographs should be submitted on glossy paper.

Units of Measurement. Results should be expressed in SI units.

Abbreviations. Non-standard abbreviations should be properly defined in the text the first time they are used.

There are no page charges. Reprints order forms are sent to the corresponding author together with galley proofs. Color illustrations may be published for extra charges.

R. Groubet, V. Pallet, B. Delage , A. Redonnet, P. Higueret, P. Cassand

Objective. Dietary factors can be associated with colorectal cancer. Fatty acids modulate gene expression in various tissues, mediated by activation of the peroxisome proliferator activated receptor: PPAR. Vitamin A signalling is mediated by retinoic acid (RA) receptors (RAR) and retinoid X receptors (RXR). The steroid nuclear receptors PPAR, RAR, RXR, are DNA-binding proteins and they induce gene transcription upon activation by specific ligands and interacting with distinct promoter sequences in the target genes. The aim of this study was to investigate the impact of hyperlipidic diets on the expression of PPARg, RXRa and RARb mRNA in rat colon.

Methods. Rats were fed during 4 weeks with the following diets: a cafeteria diet where 60 % of the energy was supplied as lipids and a high fat diet (HFD) represented by 25 % of a safflower oil (w/w) rich in polyunsaturated fatty acids, mainly n-6. Nuclear receptors mRNA were quantified by real-time RT-PCR with TaqMan probe process or SYBRGreen I chemical.

Results. The cafeteria diet and the HFD induced a significant decrease in RARb mRNA: -36 % (p<0.02) and –64 % (P<0.001) respectively. Simultaneously, an increased expression of PPARg mRNA was observed for cafeteria diet +35% (P<0.05) and for HFD +45 % (P<0.05). The level of RXRa mRNA was significantly increased for cafeteria diet: +53% (P<0.0002), while no significant difference in RXRa mRNA was observed in colonic mucosa rats whose fed with the 25 % HFD.

Conclusions. These results showed that an hyperlipidic diet could induce early modifications in the pattern of expression of nuclear receptors in rat colon. Many mechanisms could be probably involved but one hypothesis is that a modification of the balance between the nuclear receptors, resulting from an increased expression of PPARg, could induce a decreased expression of RARb in rat colon.

Key words: Rat colon – Real-time RT-PCR – High polyunsaturated fatty acid diet – Nuclear receptors expression

ENDOCRINE REGULATIONS, Vol. 37, 137–144, 2003

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She-Fang YE, I. Saga, K. Ichimura, T. Nagai*, M. Shinoda**, S. Matsuzaki

Objective. Soybeans contain an abundance of phytoestrogens such as genistein and daidzein, and these compounds are thought to protect against bone loss under estrogen deficient conditions. It is possible, however, that phytoestrogens other than isoflavones may suppress bone resorption. The objective of the present study was to determine whether there are any phytoestrogens other than isoflavones in soybeans that can act as antiosteoporotic agents.

Methods. The isoflavones genistein and daidzein, and an extract of soybeans that contained very low levels of isoflavones were tested for their ability to reduce bone resorption in ovariectomized rats. The extract of soybeans was further analyzed for its main components by thin layer chromatography and gas chromatography-mass spectrometry.

Results. The soybean extract, as well as the two isoflavones, were effective to reduce urinary excretion of deoxypyridinoline and pyridinoline, typical markers of bone resorption in ovariectomized rats. However the extract, unlike the isoflavones, increased the uterine weight of ovariectomized rats significantly. Analysis by thin-layer and gas chromatography revealed that the main constituent of this extract exhibited a chromatographic profile corresponding to coumestrol. This substance was positively identified as coumestrol by gas chromatography-mass spectrometry.

Conclusion. These results suggest that the phytoestrogens including coumestrol and isoflavones in soybeans may exert effective prevention against bone resorption in estrogen deficient conditions.

Key word: soybean, urinary deoxypyridinoline, coumestrol, genistein, daidzein

ENDOCRINE REGULATIONS, Vol. 37, 145–152, 2003

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Jan Lebl¹, Edith Schober², Thomas Zidek³, Susana Baldis², Birgit Rami², Stepanka Pruhova¹,
Stanislava Kolouskova⁴, Marta Snajderova⁴, Herwig Frisch²

Objective. Discordant data were found in recent growth studies in children with type-1 diabetes mellitus. This study focuses on growth data and final height in the largest cohort of diabetic children studied so far.

Methods. 7598 growth data collected in a longitudinal / cross sectional way between 1971 and 1996 in 587 diabetic subjects (317 males, 270 females) were available for analysis of height and BMI, together with 3889 HbA1c measurements. Final height data were correlated with target height in 123 subjects. The individual growth and BMI linear regression curve of each patient was compared to growth standards and correlated with HbA_1c.

Results. Children of both sexes were taller at the first observation (males, SDS 0.15±1.10, mean±SD, P=0.02, females, SDS 0.74±1.46, P<0.001) and tended to lose height afterwards (males, P<0.001, females, n.s.). Males reached a final height of 176.5 cm (n=62, target height 176.8, n.s.) and females 167.0 cm (n=61, target height 165.6, n.s.). Children of both sexes had a higher than normal BMI at first observation (males, SDS 0.32±1.31, P<0.001, females, SDS 0.10±0.52, P=0.02). Females but not males gained weight over-proportionally afterwards. HbA_1c did not predict any of the variables.

Conclusions. Diabetic children are taller close to the diabetes onset, which may be due to the synchronization of onset of diabetic symptoms with the mid-childhood growth spurt or the pubertal growth spurt accompanied by elevated growth hormone and/or androgen levels and increased insulin resistance. The subsequent growth deceleration may represent a physiological lag-down growth. This concept is supported by normal adult heights following growth deceleration.

Keywords: Type 1 diabetes mellitus – Children – Growth – Final height – BMI – HbA_1c

ENDOCRINE REGULATIONS, Vol. 37, 153–161, 2003

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Z. Ostrowska, B. Kos-Kudla¹, B. Marek¹, D. Kajdaniuk¹

Department of Clinical Biochemistry, ¹Department of Pathophysiology and Endocrinology, The Medical University of Silesia, Zabrze, Poland
E-mail: ozdrasiek@poczta.onet.pl

Objective. To evaluate the influence of lighting conditions on daily fluctuations of biochemical markers of bone and mineral metabolism such as the activity of serum alkaline phosphatase (ALP), the concentration of carboxyterminal propeptide of type I procollagen (PICP), cross-linked carboxyterminal telopeptide of type I collagen (ICTP) and inorganic phosphorus (iP) as well as urinary excretion of hydroxyproline (HYP) and calcium (Ca). In addition, possible involvement of daily secretion of melatonin (MEL), GH/IGF-I axis activity and parathyroid, thyroid, adrenocortical and gonadal function in this process.

Methods. Three series of 48 adult male Wistar rats weighing 180±10 g were conditioned to different lighting conditions for 4 weeks: 1. control group (LD 12:12 h, light on from 7.00 to 19.00); 2. short-day group (LD 0.5:23.5 h, light on from 7.00 to 7.30) ; 3. long-day group (LD 23.5:0.5 h, light on from 19.30 to 19.00). The blood was obtained by decapitation. Material for studies was collected every 3 hours during a day. The concentration of hormones, PICP and ICTP was determined with the use of RIA method, whereas ALP, iP, HYP and Ca levels were estimated spectrophotometrically.

Results. A significant influence of different lighting conditions on the daily profile of investigated markers of bone metabolism in rats was confirmed. Thus a short day was found to have an inducing effect upon the level of these markers, while a long day showed a suppressing effect. Furthermore, substantial changes in the values of amplitude and phase of their daily rhythms were found. Also some distinct anomalies in daily oscillations of ALP, PICP, ICTP, HYP and Ca were found which were dependent on the time of the day and showed a negative correlation with the changes in endogenous MEL concentrations. Moreover, ICTP, HYP and Ca concentrations correlated positively with daily fluctuations of IGF-I and triiodothyronine.

Conclusions. Lighting conditions can influence daily fluctuations of studied bone metabolism markers in rats, and in the mechanism of this dependence the changes in endogenous MEL concentrations seem to play an important role. Secondary changes in daily IGF-I and triiodothyronine oscillations, caused by short- and long-day conditions, result in altered daily bone resorption rhythmicity.

Key Words: Lighting conditions – Bone metabolism – Hormones – Daily rhythm – Male rats

ENDOCRINE REGULATIONS, Vol. 37, 163–174, 2003

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¹P. Langer, ²A. Kocan, ³M. Tajtakova, ²J. Petrik, ¹J. Koska, ¹M. Hucková, ⁴E. Hanzen, ¹L. Ksinantova, ¹Z. Radikova, ¹R. Imrich, ²T. Trnovec, ⁵P. Blazicek, ¹E. Sebokova, ¹I. Klimes

Objective. To compare the levels of serum cholesterol with thyroid function as estimated by the level of thyrotropin and free thyroxine with possible participation of thyroperoxidase antibodies in large number of adults examined within large field surveys focused on the evaluation of thyroid status of Slovak rural population.

Subjects and Methods. Serum level of cholesterol and thyrotropin (TSH) was estimated in a total of 2786 adults. In addition, in 2038 of them also the level of free thyroxine (FT4), total triiodothyronine (TT3), cholesterol, triglycerides and phospholipids was measured. The levels of TSH, anti-TPO and FT4 were estimated by supersensitive electrochemiluminiscent immunoassay using the automatic system Elecsys (Roche, Switzerland).

Results. A total of 2786 adults was stratified into 7 groups according to the range of TSH level as related to generally recognized level of thyroid function, e.g. 1. TSH <0.10 mU/L (overt hyperthyroidism, N=41), 2. TSH 0.11-0.30 mU/L (overt or subclinical hyperthyroidism, N=149), 3. TSH 0.31-2.50 mU/L (normal level, N=1750), 4. TSH 2.51-4.50 (”high normal” level, N=607), 5 TSH 4.51-6.50 (mild or incipient subclinical hypothyroidism, N=137), 6. TSH 6.51-10.00 mU/L (mild hypothyroidism, N=50), 7. TSH 10.01-99.00 mU/L (severe hypothyroidism, N=53). The average levels of cholesterol in all groups were very similar ranging from 5.53 to 6.17 mmol/L and no interrelations with TSH level were found. In addition, no considerable differences between these groups were found when considering the levels of medians, upper quartiles and 90th percentiles of individual groups. When male and female subjects were divided into age groups according to the decades, an age dependent increase of cholesterol level was found in both sexes.

The fraction of 2038 subjects was divided into the same TSH related groups as defined above. Similarly as above, no considerable differences in cholesterol, triglycerides and phospholipids level were observed. However, the levels of FT4 and TT3 were significantly decreasing with the increase of TSH level which confirmed the continuing decrease of thyroid function. The frequency of positive anti-TPO in subjects with TSH >6.5 mU/l (71/86 = 82.5 %) was significantly higher than that in subjects with TSH <6.5 mU/l (468/1952 = 23.9 %).

Conclusions. No difference in the level of cholesterol and triglycerides was found in large groups of rural adults from Slovakia with various thyroid function as estimated by the level of TSH, FT4, TT3 and anti-TPO. It is assumed that this interrelation resulted from very high cholesterol intake due to inappropriate general nutritional status of rural population resulting from the consumption of unhealthy foods.

Key words: Thyroid function – TSH – Thyroid hormones – Autoantibodies – Serum lipids

ENDOCRINE REGULATIONS, Vol. 37, 175–180, 2003

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Daniel Smutek, Radim Sara¹, Petr Sucharda

Objective. Relations between measurable properties of B-mode ultrasound images of thyroid gland and clinical and laboratory findings in patients with chronic inflammation of thyroid gland were studied.

Methods. Data from 65 patients with lymphocytic thyroiditis (LT) and 38 control subjects were analysed. Raw values of individual B-mode image pixels and standard co-occurrence second-order texture features were selected as quantitative image features. Thyroid antibodies, thyrotropin level, thyroxine replacement therapy, and body mass index were used as clinical variables.

Results. In the LT group, significant differences (t-tests, p<0.05) in image features were found for body mass indices (BMI) under and over 25 kg.m-2, for thyroxine replacement therapy, and for the presence and absence of thyroid antibodies. Forward stepwise multiple regression was performed for the clinical or laboratory values as dependent variables and image features as independent variables. The following correlations were found: 1. between BMI and four image features in the normal group; 2. between the dose of thyroxine replacement therapy and two of image features in the LT group; and 3. for the level of thyroid antibodies in the LT group: five image features have correlated with the level of anti-thyroglobulin and three image features with level of anti-thyroperoxidase.

Conclusion. These findings suggest the possibility of using quantitative indicators of ultrasound image of thyroid gland as predictors of the presence or absence of thyroid antibodies in patient’s blood or as an auxiliary tool for dose recommendation of thyroxine replacement therapy.

Key words: Thyroid gland – Lymphocytic thyroiditis – Ultrasound – Sonography – Image features – Texture analysis

ENDOCRINE REGULATIONS, Vol. 37, 181–187, 2003

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Zofia Radikova

Hyperinsulinemic euglycemic clamp is known to be the “gold standard” in measurement insulin sensitivity. However, its time and financially consuming realization led to a simplified approach in quantification of insulin sensitivity. Various indices of insulin sensitivity/resistance using the data from an oral glucose tolerance test were proposed in last 20 years. The aim of this review is to evaluate critically the use of some of the proposed indices in insulin sensitivity estimation.

There are two groups of insulin sensitivity indices: 1. indices calculated by using fasting plasma concentrations of insulin, glucose and triglycerides, 2. indices calculated by using plasma concentrations of insulin and glucose obtained during 120 min of a standard (75g glucose) oral glucose tolerance test. Some authors used demographic parameters (BMI, age, weight) in their formulas to achieve the best correlation with euglycemic clamp data.

These indices are conveniently used in epidemiological and clinical studies to predict diabetes development in a non-diabetic population. Their use in clinical practice is limited because of the absence of reference values for normal and impaired insulin sensitivity.

Key words: Insulin sensitivity index – Oral glucose tolerance test – Insulin – Glucose

ENDOCRINE REGULATIONS, Vol. 37, 189–194, 2003

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