Evaluation of the frequency of ADIPOQ c.45 T>G and ADIPOQ c.276 G>T polymorphisms in adiponectin coding gene in girls with anorexia nervosa

Introduction: Anorexia nervosa (AN) is a serious chronic psychosomatic disorder, the essence of which are attempts by the sufferer to obtain a slim silhouette by deliberate weight loss (restrictive diet, strenuous physical exercise, provoking vomiting). The aetiology of this disorder is multifactorial. Genetic factors that influence the predisposition to AN have been sought. A broad meta-analysis points to a strong genetic correlation between AN and insulin resistance. Adiponectin (ADIPO) increases insulin sensitivity. In our pilot study we demonstrated that the TT genotype in locus ADIPOQ c.276 G>T of the ADIPO gene and a higher concentration of ADIPO in blood serum occurred significantly more frequently in 68 girls suffering from AN than in 38 healthy girls. The objective of this study was to evaluate the frequency of the occurrence of ADIPOQ c.45 T>G and ADIPOQ c.276 G>T in the ADIPO gene in a larger cohort of girls with AN and healthy girls, as well as an analysis of correlations between variants of the aforementioned polymorphisms and the levels of ADIPO in blood serum. Material and methods: The study covered 472 girls (age: 11–19 years): 308 with the restrictive form of AN (AN) and 164 healthy girls (C). The level of ADIPO in blood serum was determined by means of the ELISA method on a Bio-Vendor, LLC (Asheville, North Carolina, USA). The DNA isolation was carried out by means of Genomic Mini AX BLOOD (SPIN). The PCR reaction was carried out in a ThermoCycle T100 thermocycler. 80–150 ng of the studied DNA and relevant F and R starters were added to the reaction mixture. The reaction products were subjected to digestion by restriction enzymes and separated on agarose gels (RFLP). Results: The distribution of genotypes in the polymorphic site ADP c.45 of the ADIPO gene and ADP c.276 was similar in both groups. In both groups the T allele was most frequent in locus ADIPOQ c.45 and the G allele in locus ADIPOQ c.276. In all the study subjects collectively (AN and C) a statistically significant negative correlation between the levels of ADIPO in blood serum on one hand and body weight (r = –0.46; p < 0.0001) and BMI (r = –0.67; p < 0.0001) on the other was demonstrated. Exclusively in the AN group a significant correlation between the level of ADIPO in blood and the distribution of TG, TT, and GG alleles in loci ADIPOQ c.45 and ADIPOQ c.276 was demonstrated (p = 0.0052 and p < 0.0001, respectively). Conclusions: The genotype in loci ADIPOQ c.45 and ADIPOQ c.276 of the ADIPO gene seems to have no effect on the predisposition to AN. Girls suffering from AN with the TT genotype in loci ADIPOQ c.45 and ADIPOQ c. 276 may demonstrate higher insulin sensitivity because they have significantly higher levels of ADIPO than girls suffering from AN with other genotypes. This may be suggestive of their better adaptation to the state of malnutrition, and it has a potential effect on treatment results

Evaluation of the frequency of RETN c.62G>A and RETN c.-180C>G polymorphisms in the resistin coding gene in girls with anorexia nervosa

Introduction: Anorexia nervosa (AN) is a serious psychosomatic syndrome, classified as an eating disorder. AN patients strive to lose weight below the normal limits defined for a specific age and height, achieving their goal even at the expense of extreme emaciation. AN has a multifactorial aetiology. Genetic factors are believed to be significant in the predisposition to the development of AN. In girls suffering from AN significantly lower levels of resistin (RES) in blood serum are observed as compared to healthy girls. These differences may lead to a thesis that functional genetic polymorphisms in RES coding genes can be responsible for this phenomenon. In our pilot study we demonstrated significant differences in the distribution of genotypes in the locus RETN c.-180C>G of the RES gene in 67 girls with AN and 38 healthy girls. It seems reasonable to compare the frequency of polymorphisms of RETN c.62G>A and RETN c.-180C>G in the RES gene in girls with AN and in healthy subjects in a bigger cohort and to analyse correlations between individual variants of the polymorphisms referred to above and the RES levels in blood plasma. Material and methods: The study covered 308 girls with the restrictive form of AN (AN) and 164 healthy girls (C) (aged 11–19 years). The RES levels in blood serum were determined by means of the ELISA method on a Bio-Vendor machine from LLC (Asheville, North Carolina, USA). The DNA isolation was carried out by means of Genomic Mini AX BLOOD (SPIN). The PCR reaction was carried out on a ThermoCycle T100 thermocycler. 80–150 ng of the studied DNA and relevant F and R starters were added to the reaction mixture. The reaction products were subjected to digestion by restriction enzymes and separated on agarose gels (RFLP). Results: The average RES level in blood serum in the AN group was significantly lower (p < 0.0001) than in the C group. The distribution of genotypes in the locus RETN c.62 of the RES gene was similar in both groups. A significant difference was demonstrated in the distribution of genotypes in the polymorphic site RETN c.-180 of the RES gene between AN and C (p = 0.0145) and in the distribution of the C and G alleles in the locus RETN c.-180 (p < 0.0001). The C allele occurred significantly more frequently than the G allele in the C group as compared to the AN group. In all the study subjects jointly (AN and C) a significant positive correlation between the blood RES levels on one hand and the body mass (r = 0.42; p < 0.0001) and BMI (r = 0.61; p< 0.0001) on the other was observed. There was no correlation between the concentration of RES in blood serum and the distribution of genotypes in the loci of the resistin gene referred to above. Conclusions: The CG genotype in the locus RETN c.-180 C>G of the RES gene may constitute one of the factors predisposing to the development of AN in girls. The genotype in the loci RETN c.62 G>A and RETN c.-180 C>G of the resistin gene has no influence on the levels of this hormone in blood in AN patients

Nutritional Status and Selected Adipokines in Children with Irritable Bowel Syndrome

Background: The aim of this study was to assess the nutritional status and serum concentrations of adipokines in children with irritable bowel syndrome (IBS) and healthy controls. We also sought to evaluate their relation to metabolic parameters. Methods: We studied 33 IBS patients (11 girls, 22 boys) aged 5–17 years and 30 healthy age-matched controls (11 girls, 19 boys). The analysis included anthropometric measurements, body composition parameter measurements using bioimpedance, and biochemical tests and measurements of serum concentrations of leptin, adiponectin, chemerin, and omentin-1. Results: The results of the anthropometric measurements were comparable between the patients and the controls. The patients had higher triglycerides, HOMA-IRs, and chemerin concentrations than the healthy subjects. The HDL cholesterol and omentin-1 levels were lower than in the controls. Leptin and adiponectin did not differ significantly between the groups. An analysis of the receiver operator curves (ROCs) showed that serum concentrations of chemerin ≥ 232.8 ng/mL had 30% sensitivity and 87% specificity when they were used to differentiate between children with IBS and healthy subjects. In the case of serum omentin-1 concentrations ≤ 279.4 ng/mL, the sensitivity and specificity were 60% and 80%, respectively. Conclusions: The nutritional status of children with IBS did not differ from that of the healthy controls. We found significant differences in serum chemerin and omentin-1 concentrations between IBS patients and healthy children. These adipokines could be used as IBS biomarkers as they demonstrate good specificity and moderate sensitivity. The serum concentrations of chemerin and omentin-1 in IBS patients were related to nutritional status and insulin resistance

Wpływ wczesnych zakażeń na ekspresję molekuł CD11a, CD11b, CD11c, CD18, CD54 i CD62L na powierzchni neutrofili i monocytów obwodowej krwi żylnej u wcześniaków

INTRODUCTION: Cell adhesion molecules on neutrophils and monocytes play an important role in the immunity of newborns. MATERIAL AND METHODS: Eighty-two premature infants (47 with early-onset infection and 35 without infection, 37 male and 45 female, 40 delivered vaginally and 42 by Caesarean section) between the 3rd and 7th day of life were assessed for the expression rate of CD11a, CD11b, CD11c, CD18 CD54 and CD62L on granulocytes and monocytes using a Becton Dickinson FACS Canto II flow cytometer. Both groups were analysed for sex, body weight, gestational age, type of delivery and infection, and correlations between adhesion molecules were identified. RESULTS: Preterm infants with infection had a significantly higher mean CD62L level than preterm infants without infection. There was also a positive correlation between gestational age and CD11b and CD11c, as well as correlations between CD11b and CD11c, CD11b and CD54, and CD11c and CD54U. In preterm infants without infection, there was a significant correlation between body weight and CD11a level, and correlations between CD11a and CD18, as well as CD11a and CD54. The sex of the preterm infants and type of delivery in both groups had no significant effect on the mean levels of the analysed molecules. There was no significant relationship between the expression of the analysed molecules and the CRP concentration and platelet count in preterm infants with infection. CONCLUSIONS: 1. Early-onset infection in premature infants promotes increased expression of the CD62L molecule, which despite the lack of correlation with the CRP concentration and platelet count may prove useful in the diagnosis of infections. 2. CD11b expression in preterm infants with infections is affected by gestational age, while the expression of CD11a in preterm infants without infections is correlated with body weight. 3. Expression of the analysed molecules in premature neonates does not depend on sex or type of delivery.WSTĘP: Molekuły adhezji komórkowej na neutrofilach i monocytach odgrywają ważną rolę w odporności noworodków. MATERIAŁ I METODY: U 82 wcześniaków (47 z zakażeniem wczesnym i 35 bez zakażeń; 37 płci męskiej i 45 żeńskiej; 40 urodzonych siłami natury i 42 cięciem cesarskim) między 3 a 7 dobą życia dokonano oceny odsetka ekspresji CD11a, CD11b, CD11c, CD18, CD54 i CD62L na granulocytach i monocytach krwi cytometrem przepływowym Becton Dickinson FACS Canto II, z uwzględnieniem płci, masy ciała, wieku płodowego wcześniaków, typu porodu i zakażenia, oraz określono wzajemne korelacje między molekułami. WYNIKI: U wcześniaków z zakażeniem stwierdzono istotne statystycznie zwiększenie średniego odsetka CD62L niż u wcześniaków bez zakażeń, dodatnie korelacje między wiekiem płodowym a CD11b i CD11c oraz wzajemne korelacje między CD11b i CD11c, CD11b i CD54 oraz CD11c i CD54. U wcześniaków bez zakażeń wykazano znamienną zależność między masą ciała a CD11a oraz korelacje między CD11a i CD18 oraz CD11a i CD54. Płeć wcześniaków i typ porodu w obu grupach nie miały istotnego wpływu na średnie wartości badanych molekuł. Wykazano brak istotnych zależności między ekspresją badanych molekuł a stężeniem CRP i liczbą płytek krwi u chorych wcześniaków. WNIOSKI: 1. Wczesne zakażenia u wcześniaków sprzyjają zwiększeniu ekspresji molekuły CD62L, co mimo braku korelacji ze stężeniem CRP i liczbą płytek krwi może dowodzić przydatności oznaczania tej molekuły w diagnostyce zakażeń. 2. Na wartość ekspresji CD11b u wcześniaków z zakażeniem wpływa wiek płodowy, a na CD11a masa ciała u wcześniaków bez zakażeń. 3. Ekspresja badanych molekuł u wcześniaków nie zależy od ich płci i typu porodu