Reference intervals for six salivary cortisol measures based on the Croatian Late Adolescence Stress Study (CLASS)

IN ENGLISH: INTRODUCTION: The aim of this nested study is to provide the reference intervals for already published measurements of salivary cortisol from the Croatian Adolescence Stress Study (CLASS). MATERIAL AND METHODS: A total of 969 individuals (372 males and 597 females) were included in the reference sample (age range: 18-21 years). Salivary cortisol concentrations were determined by the enzyme immunoassay (LUCIO-Medical ELISA Salivary Cortisol Kit, Nal von Minden, Germany) in the Department of Medical Laboratory Diagnostics, University Hospital Split. Nonparametric statistics were used for calculating the reference intervals (RIs) and 90% confidence intervals (90% CIs). RESULTS: The lower limits of RIs determined by the direct method were higher in females (> 10%) than in males for the cortisol concentrations at awakening (SCC0), 30 to 45 after awakening (SCC30-45) and at bedtime (SCCbedtime). The upper limits of RIs for the SCCbedtime were higher (> 10%) in males than in females. Females also had higher upper limits of RIs for the cortisol awakening response (CAR) and the diurnal cortisol slope (DCS) and higher lower limits of RIs for the CAR and the area under the curve with respect to ground (AUCG). The lower limits of RIs for the DCS were higher in males than in females. CONCLUSIONS: Obtained reference values open the arena for introducing salivary bioscience in Croatian clinical laboratory practice and provide important data for better understanding of gender differences in adaptation to stress during late adolescence. --------------- IN CROATIAN: UVOD: Cilj ove studije bio je definirati referentne intervale za objavljene mjere salivarnog kortizola iz Hrvatskog istraživanja stresa u kasnoj adolescenciji (CLASS). MATERIJALI I METODE: Ukupno 969 ispitanika (372 djevojaka i 597 mladića) uključeno je u referentni uzorak (18-21 godine starosti). Koncentracije salivarnog kortizola izmjerene su imunokemijskom ELISA metodom (LUCIO-Medical ELISA Salivary Cortisol Kit, Nal von Minden, Germany) u Zavodu za medicinsko laboratorijsku dijagnostiku Kliničkog bolničkog centra Split. Referentni intervali (RI) i 90%-tni intervali pouzdanosti (90% CI) izračunati su neparametrijskim metodama. REZULTATI: Donja granica referentnog intervala određena izravnom metodom veća je kod djevojaka (>10%) nego mladića za koncentracije kortizola u sve tri vremenske točke (po buđenju, 30-45 minuta nakon buđenja i neposredno prije lijeganja). Gornja granica referentnog intervala za koncentracije salivarnog kortizola neposredno prije lijeganja veća je kod mladića (>10%) u odnosu na djevojke. Također, djevojke imaju veću gornju granicu referentnog intervala za odgovor kortizola na buđenje (engl. cortisol awakening response, CAR) i cirkadijano smanjenje kortizola (engl. diurnal cortisol slope, DCS) te veću donju granicu referentnog intervala za CAR i površinu ispod krivulje u odnosu na nultu razinu (engl. area under the curve with respect to ground, AUCG). Indeks cirkadijanog smanjenja kortizola, DCS je u donjoj granici referentnog intervala veći kod mladića u odnosu na djevojke. ZAKLJUČCI: Objavljeni referentni intervali mjera salivarnog kortizola važni su zbog uvođenje salivarnih analiza u rutinsku kliničku praksu u RH i pridonose detaljnijim spoznajama o spolnim razlikama u adaptacijskim mehanizmima na stresne poticaje tijekom kasne adolescencije

Analysis of leptin, adiponectin and adiponectin gene polymorphism and leptin receptor in obese children and adolescents

Background: The aim of this study was to determine serum levels of leptin and adiponectin of obese children to identify the influence of leptin receptor gene polymorphisms on leptin resistance and leptin levels, as well as the association between the polymorphisms of adiponectin gene and adiponectin levels.Materials and methods: A case-control study comparing a study group of 74 obese children (age 13.34±2.60 years) to a normal weight-age matched (age 13.39±2.64 years) control group of 69 children. In both groups, body mass index (BMI) and waist/hip circumference, systolic and diastolic blood pressure were measured. Also, the leptin and adiponectin levels, as well as glucose and lipid metabolism parameters, and highly sensitive C-reactive protein (hs-CRP) were measured. Insulin sensitivity was evaluated using fasting insulinemia and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR). All subjects were tested for gene-tic polymorphisms in LEPRQ223R (rs1137101), ADIPOQ G276T (rs1501299) and ADIPOT45G (rs2241766).Results: The phenotypes of the obese children study group were significantly higher than in the control group in weight, BMI, waist/hip circumferences and systolic blood pressure (SBP) (P<0.001). We confirmed that in obese children the levels of leptin in the blood are increased and levels of adiponectin are decreased (P<0.001). The differences of the genotype distributions of leptin receptor (LEPRQ223R) and adiponectin (ADIPOG276T and ADIPOT45G) gene polymorphisms in the study group of obese chil-dren and a control group was not observed.Conclusion: In this study, we demonstrated increased leptin level and significantly decreased level of adiponectin in the obese children group compared with the control group. The results of the analysis of glucose metabolism and lipidogram between the two groups showed that insulin, HOMA-IR, and triglycerides, as well as hsCRP were increased and significantly different in the group of obese children compared to the control group, as expected. However, by including a significantly larger number of tested and control samples of both sexes and age-specific groups, with a larger number of tested SNPs, the genes investigated in this study would probably give better insight into a multicomplex disease such as obesity

Laboratory policies and practices for thyroid function tests in Croatia: survey on behalf of Working Group for Laboratory Endocrinology of the Croatian Society of Medical Biochemistry and Laboratory Medicine

IntroductionLaboratory plays important part in screening, diagnosis, and management of thyroid disorders. The aim of this study was to estimate current laboratory preanalytical, analytical and postanalytical practices and policies in Croatia. Materials and methodsWorking Group for Laboratory Endocrinology of the Croatian Society of Medical Biochemistry and Laboratory Medicine designed a questionnaire with 27 questions and statements regarding practices and protocols in measuring thyroid function tests. The survey was sent to 111 medical biochemistry laboratories participating in external quality assurance scheme for thyroid hormones organized by Croatian Centre for Quality Assessment in Laboratory Medicine. Data is presented as absolute numbers and proportions. ResultsFifty-three participants returned the questionnaire. Response rate varied depending on question, yielding a total survey response rate of 46-48%. All respondents perform thyroid stimulating hormone (TSH). From all other thyroid tests, most performed is free thyroxine (37/53) and least TSH-stimulating immunoglobulin (1/53). Laboratories are using nine different immunoassay methods. One tenth of laboratories is verifying manufacturer’s declared limit of quantification for TSH and one third is verifying implemented reference intervals for all performed tests. Most of laboratories (91%) adopt the manufacturer’s reference interval for adult population. Reference intervals for TSH are reported with different percentiles (90, 95 or 99 percentiles). ConclusionThis survey showed current practices and policies in Croatian laboratories regarding thyroid testing. The results identified some critical spots and will serve as a foundation in creating national guidelines in order to harmonize laboratory procedures in thyroid testing in Croatia

Testing the Institute of Medicine (IOM) recommendations on maternal reproductive health and associated neonatal characteristics in a transitional, Mediterranean population

Background:  High pre-pregnancy body mass index (BMI) and excessive gestational weight gain (GWG) are significant risk factors for maternal and neonatal health.  Aim:  To assess pre-pregnancy BMI and GWG during pregnancy and their association with different maternal and neonatal characteristics in the transitional Mediterranean population from the Eastern Adriatic islands.  Subjects and methods:  Two hundred and sixty-two mother–child dyads from the CRoatian Islands’ Birth Cohort Study (CRIBS) were included in the study. Chi-square test, ANOVA, and regression analysis were used to test the association between selected characteristics. Results: In total, 22% of women entered pregnancy as overweight/obese and 46.6% had excessive GWG. Pre-pregnancy overweight and obesity were significantly associated with elevated triglycerides uric acid levels, and decreased HDL cholesterol in pregnancy. Excessive GWG was associated with elevated fibrinogen and lipoprotein A levels. Women with high pre-pregnancy BMI and GWG values were more likely to give birth to babies that were large for gestational age (LGA), additionally confirmed in the multiple logistic regression model.  Conclusion:  High maternal pre-pregnancy BMI and excessive GWG were both significantly associated with deviated biochemical parameters and neonatal size. More careful monitoring of maternal nutritional status can lead to better pre- and perinatal maternal healthcare.</p