Autoimmune insulitis leading to insulin dependent diabetes mellitus (IDDM, Type 1 Diabetes) is accompanied by autoantibodies as its invaluable markers. The aim of the study was to determine the frequency of autoantibodies against GAD65, IA2 and insulin in Czech diabetic children at the disease onset.Sera of 105 newly diagnosed children with IDDM drawn within 24 hours after the first insulin dose were investigated for anti-GAD65, anti-IA2 and insulin autoantibodies (IAA) using RIA methods. The cut-off normal levels were determined as the 99th percentile of 105 non-diabetic children. At given 99% specificity, the sensitivity was 71% for anti-GAD65, 73% for anti-IA2, and 46% for IAA. 29% diabetic children were positive for all three autoantibodies, 25% had anti-GAD65 and anti-IA2 (IAA negative), 5.7% anti-GAD65 and IAA (anti-IA2 negative), 7.6% anti-IA2 and IAA (anti-GAD65 negative). As the only positive autoantibody, anti-GAD65 was found in 12%, anti-IA2 in 11%, and IAA in 3.8% children. In 5.7% children, none of the investigated autoantibodies was positive. Diabetic children diagnosed before the age of 5 years had significantly higher prevalence of IAA than the older ones.We have determined normal levels in healthy children, and prevalence at childhood IDDM onset of autoantibodies against three main molecular-defined autoantigens.
Aim: Adiponectin is an adipocytokine secreted by adipose tissue which increases insuline sensitivity. It is a biomarker of metabolic syndrome in humans and its production is stimulated by insulin and insulin-like growth factor 1. Adiponectin serum levels are decreased in obese adults and in patients with type II diabetes in comparison to healthy controls. We measured adiponectin, insulin, and C-peptide serum levels and various anthropometrical parameters in obese children undergoing 5 weeks of weight-reduction therapy in spa. Methods: We measured serum adiponectin levels using human ELISA kit in 69 obese children (40 girls, 29 boys; mean age 13.1 ± 0.38 years; mean weight 74.2 ± 2.79 kg; mean weight-loss 6.8 kg) and in 32 age and gender matched healthy controls. We used the Matiegka´s anthropometrical method based on skinfold thickness to determine the amount of body fat. Results: Adiponectin levels before weight-reduction were 16.8 ± 0.85 ng/ml and after weight-reduction 17.0 ± 0.82 ng/ml (N.S., p = 0.6309). Levels in the control group were 22.4 ± 1.80 ng/ml and differed significantly from levels in obese children both before and after weight-reduction (p = 0.0017 and p = 0.0021 respectively). Adiponectin levels did not differ significantly between girls (17.3 ± 1.16 ng/ml) and boys (15.7 ± 1.07 ng/ml, p = 0.3385). We found a negative correlation between adiponectin and insulin levels (r = −0.3905; p = 0.0012), C-peptide levels (r = −0.3859; p = 0.0014) age (r = −0.4143; p = 0.0012), body height (r = −0.3684; p = 0.0044), body weight (r = −0.3888; p = 0.0026), body mass index (r = −0.3664; p = 0.0047) and amount of body fat (r = −0.272; p = 0.0388). Conclusion: Adiponectin levels correlate negatively with insulin, C-peptide, age, body height, body weight, BMI and amount of body fat, however they are not affected by middle-term changes of body weight in children. Supported by research grants IGA MZ NE/7443-3 and GAUK 59/2004C.
Early diagnosis of sepsis and its differentiation from non-infective SIRS is very important. The links between inflammation and coagulation play an important role in the SIRS/sepsis process. We investigated hematological and biochemical parameters (including thromboelastography (TEG)) in patients after surgical resection of esophagus. The aim of our project was to find out whether there are any changes in these parameters that could help in differentiation between SIRS and sepsis.
In this study, we describe changes of plasma levels of the hypothalamic neuropeptide orexin A in obese children during the reduction of body weight and its relationship to other biochemical and anthropometrical parameters. We measured orexin A fasting plasma levels by the RIA method in 58 obese children--33 girls and 25 boys; mean age 13.1+/-0.38 years (range 7-18.5) before and after 5 weeks of weight-reduction therapy. Leptin, IGF-1, and IGFBP-3 levels were measured in all the subjects and were compared to orexin A levels and anthropometrical data. Average weight in subjects before weight-reduction was 74.2+/-2.79 kg and after weight-loss 67.4+/-2.60 kg (p<0.0001). Orexin A levels before the therapy were 33.3+/-1.97 pg/ml and after the therapy 51.7+/-3.07 pg/ml (p<0.0001). Levels of orexin A were not significantly different between girls and boys (p=0.7842). We found negative correlation between orexin A and age (r = -0.5395; p<0.0001), body height (r = -0.4751; p=0.0002), body weight (r = -0.4030; p=0.0017) and BMI (r = -0.2607; p=0.0481). No correlation was found between orexin A and IGF-1, IGFBP-3 or leptin. Orexin A plasma levels increased during body weight loss, whereas the reverse was true for leptin levels. These findings support the hypothesis that orexin A may be involved in regulation of nutritional status in children.
Breastfeeding may protect children from developing metabolic syndrome and other diseases later in life. We investigated novel proteins in human breast milk that might play a role in this process.We used ELISA to measure adiponectin, adipocyte and epidermal fatty acid binding proteins (AFABP, EFABP), and leptin concentrations in human breast milk obtained from 59 mothers 48 h after initiation of lactation. Using a questionnaire and medical records, we collected information about the mothers and newborns.Mean (SE) adiponectin concentrations in breast milk were 13.7 (0.8), range 3.9-30.4 microg/L; AFABP concentrations 26.7 (4.4), range 1.2-137.0 microg/L; EFABP concentrations 18.1 (1.4), range 0.8-47.0 microg/L; and leptin concentrations 0.50 (0.05), range 0-1.37 microg/L. We found a significant correlation between AFABP and EFABP concentrations (r = 0.593, P <0.0001). Maternal EFABP concentrations were significantly higher in mothers who delivered boys than in those who delivered girls [21.7 (2.3) vs 15.4 (1.7) microg/L, P = 0.028] and correlated with newborn birth weight (r = 0.266, P = 0.045). Maternal leptin correlated with body weight before pregnancy (r = 0.272, P = 0.043) and at delivery (r = 0.370, P = 0.005), body mass index before pregnancy (r = 0.397, P = 0.003) and at delivery (r = 0.498, P <0.0001), body weight gain during pregnancy (r = 0.267, P = 0.047), and newborn gestational age (r = 0.266, P = 0.048). Leptin was significantly lower in mothers who delivered preterm vs term babies [0.30 (0.09) vs 0.60 (0.05) ug/L, P = 0.026].Concentrations of adiponectin, AFABP, and EFABP in human breast milk are related to nutritional variables of mothers and newborns and thus may play a role in the protective effects of breastfeeding.
Early diagnosis of sepsis and its differentiation from the noninfective SIRS is very important in order that treatment can be initiated in a timely and appropriate way. In this study we investigated standard haematological and biochemical parameters and thromboelastography (TEG) in patients who had undergone surgical resection of the oesophagus to find out if changes in any of these parameters could help in early differentiation between SIRS and sepsis development. We enrolled 43 patients (aged 41–74 years) of whom 38 were evaluable. Blood samples were obtained on the morning of surgery and then at 24-hour intervals for the next 6 days. Samples were analysed for procalcitonin (PCT), C-reactive protein (CRP), interleukin-6 (IL- 6), aspartate transaminase (AST), alanine transaminase (ALT) , lactate, white blood count (WBC), D-dimers, antithrombin (AT), international normalised ratio (INR), activated partial thromboplastin time (APTT) and parameters of TEG. Significant differences between patients who developed sepsis during this period (9 patients) and SIRS were found in ALT on Day 1, in AST on Days 1–4, in PCT on Days 2–6; in CRP on Days 3–6; in IL-6 on Days 2–5; in leucocytes on Days 2, 3 and 6; and in D-dimers on Days 2 and 4. Significance values ranged from p < 0.0001 to p < 0.05. Sequential measurements of ALT, AST, PCT and IL-6 during the early postoperative period can be used for early differentiation of sepsis and postoperative SIRS after oesophagectomy. Among the coagulation parameters measured, only D-dimer concentrations appeared to be helpful in this process. TEG does not seem to be a useful early predictor of sepsis development; however it can be used to differentiate sepsis and SIRS from Day 5 after surgery.
Aim: To compare aspects of wound healing after cleft lip surgery performed within one week of age and wound healing after surgery performed within 2 - 4 months of age, especially concentrations of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in tissue removed during surgery. Methods: 34 tissue samples (26 boys and 8 girls) were removed during surgery within one week of age (n=19) or within 2 - 4 months of age (n=15). Tissue samples were separated into epidermis, dermis and mucous membrane. Proteins were extracted in cacodylic buffer for 24 h at a temperature 2 - 8 ºC. Total protein concentrations were examined using a modification of the Lowry method. Samples were examined using ELISA kit Amersham Biotrak Activity Assay (GE Healthcare UK) for detection of MMP-9 and TIMP-1 concentrations. Results: MMP-9: early surgery - epidermis 2.168±3.303 μg/g of protein (mean±SD), dermis 1.251±1.848 µg/g, 2 - 4 months surgery - epidermis 0.347±0.212 μg/g, dermis 0.555±0.276 µg/g. TIMP-1: early surgery - epidermis 1.762±2.162 μg/g, dermis 1.628±0.822 µg/g, mucous membrane 2.066±1.717 µg/g, 2 - 4 months surgery - epidermis 1.881±2.810 μg/g, dermis 3.117±1.540 µg/g, mucous membrane 4.833±6.550 µg/g. Conclusions: There were no significant differences in concentrations of protein MMP-9 in epidermis and dermis and TIMP-1 in epidermis and mucous membrane according to time of surgery. Significantly decreased levels of TIMP-1 in dermis were found in samples obtained from early surgery compared to levels in samples obtained from 2 - 4 months surgery.
Individuals at risk for insulin dependent diabetes mellitus (IDDM) can be identified using a combination of genetic, immunological and metabolic markers. Our study was aimed at prediction of IDDM in a cohort of children having a first-degree relative with IDDM.In the period of three years, we investigated 208 non-diabetic children and adolescents, aged 10.0 +/- 5.3 (mean +/- SD), mostly siblings of diabetic children. The genetic risk was determined by the HLA-DQB1, -DQA1 genotyping and subtyping of the DRB1*04 alleles carried on the DQB1*0302 haplotypes. Insulitis was detected using a combination of autoantibody tests against three molecular-defined antigens (insulin, GAD65, IA-2). Prevalence of insulitis (defined as confirmed positivity of at least one autoantibody) was 9/208 (4.3%). In children carrying the IDDM highest-risk genotype (HLA-DQB1*0201-DQA1*05/DQB1*0302-DQA1*03), insulitis was almost 10 times more frequent (5/24, 21%) than in children with other genotypes (4/184, 2.2%, P = 0.003). In all subjects with insulitis, the first phase insulin response (FPIR) was determined by the intravenous glucose tolerance test. Three of the nine children had decreased FPIR, of whom two were later diagnosed with IDDM. None of the remaining children developed IDDM.We present the first IDDM prediction study in the Czech population, emphasising the utility of genetic risk investigation in the prediction scheme.
The aim of the study was to evaluate serum a-glutathione S-transferase (s-GSTA) levels in patients with cystic fibrosis (CF) and to compare s-GSTA with other liver function tests and with a hepatic ultrasound scan (US). The cytosolic enzyme, alpha-glutathione S-transferase is predominantly found in the liver and is distributed uniformly in the liver tissue. In our study s-GSTA levels were measured in 37 CF patients aged 1 to 28 years (mean age 10.4 years, 24 males). The control group consisted of 27 patients aged 2 to 17 years (mean age 8.5 years, 18 males). The presence of hepatobiliary abnormalities was assessed by clinical examination, ultrasound scan, s-GSTA, and conventional liver enzymes: alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST) and gama-glutamyl transferase (GMT). The calculated 5-95 % range of s-GSTA for the control group was 0.098-2.54 microg/l, for the CF group 0.43-9.76 microg/l. Mean s-GSTA level in the control group was 1.55 microg/l (S.D.=1.57), and 2.05 micro/l (S.D.=2.60) in the CF group. In the group of CF patients, the serum levels were significantly higher than in the control group (P<0.01). No significant correlation existed in the CF group between s-GSTA and conventional liver tests (ALT, AST, ALP and GMT). Four patients in the CF group had hepatobiliary abnormalities detectable by conventional liver tests, s-GSTA and US. Four patients had abnormal s-GSTA, while conventional liver tests and US were normal. One other patient had abnormal hepatic US, but normal standard liver tests and s-GSTA. The study has suggested that a raised s-GSTA level might be a marker of possible pathological changes of the hepatobiliar system in CF patients. Serum GSTA seems to be a more sensitive marker than transaminases for the monitoring of hepatocellular integrity and as an early predictor of hepatic damage.
