Differential activation and expression of insulin receptor substrate 1 (IRS1) in mononuclear cells of type 2 diabetes patients after insulin stimulation.

Insulin regulates the glucose homeostasis by inducing tyrosine phosphorylation of insulin receptor substrate (IRS) proteins. IRS1 is the best studied member of this family and insulin-induced Tyrosine phosphorylation of (YXXM) motifs provides docking site for SH2 domain-containing proteins. Recent studies have suggested that genetic and/or environmental factors may affect the expression and phosphorylation levels of IRS1, and these could be important for development of insulin resistance. To shed light to the molecular basis of type 2 diabetes we wanted to determine whether YXXM motifs are genetically modified in these patients. We have isolated mononuclear cells of eighteen type 2 diabetes patients and prepared genomic DNA and protein lysates from these cells. The genomic DNA was used to sequence IRS1 gene, and protein lysates were used to determine the expression and phosphotyrosine levels of IRS1 after insulin stimulation. Although, we did not detect any mutations at/or near the YXXM coding regions in patients' DNA, immunprecipitation analysis of IRS1 indicated decreased levels of expression and tyrosine phosphorylation of IRS1 in patient's samples compared to that of healthy controls. Our results suggest that mononuclear cells of patients can be used to test the levels of insulin responsiveness before therapy.

Serum level and placental expression of resistin in pregnancies complicated by preeclampsia: relationship with disease severity.

The authors aimed to compare the maternal serum level and placental expression of resistin in pregnancies complicated by preeclampsia and clarify their relationship with disease severity. This cross-sectional study included 50 healthy pregnant women, 50 women with mild preeclampsia, and 48 women with severe preeclampsia. Serum resistin levels were measured by enzyme immunoassay and placental resistin expression was determined by immunohistochemistry. Resistin levels were significantly higher in women with mild and severe preeclampsia than in the healthy controls (p = 0.012 andp < 0.001, respectively). Placental resistin expression was significantly higher in women with severe preeclampsia compared to women with mild preeclampsia (p = 0.003) and healthy controls (p < 0.001). Serum resistin levels were positively correlated with gestational age and umbilical and uterine artery Doppler indices, as well as systolic and diastolic blood pressure, but negatively correlated with birth weight (p < 0.05). On the other hand, placental resistin expression was positively correlated with systolic blood pressure and uterine artery indices, but negatively correlated with birth weight (p < 0.05). In conclusion, increased cir- culating levels and placental expression of resistin in pregnancies complicated by preeclampsia were correlated with disease severity.

RLIP76 Gene Variants are not Associated with Drug Response in Turkish Epilepsy Patients.

Approximately 30% of epileptic patients remain untreated, in spite of trials with maximum tolerable doses of more than one drug. The RalA binding protein 1 (RALBP1/RLIP76), a multifunctional, anti-apoptot-ic, multidrug transporter protein, has been proposed as being responsible for the drug resistance mechanism in epilepsy. We have investigated polymorphic differences in the coding regions and exonintron boundaries of the RLIP76 gene, between 146 refractory and 155 non refractory epileptic patients in Turkey, using denaturing high performance liquid chromatography (HPLC) and sequencing analysis techniques. We have detected the following sequence variants: c.160-4G>A, c.187C>G, c.1562-38G>A, c.1670+107G>A, c.1670+93G>A, c.1670+96G>A, c.1670+100C>T, c.1670+130C>T, c.1670+131G>C, c.1670+140 G>C, and found no statistically significant correlation between allele frequencies and drug response status. We conclude that sequence variants of this gene are not involved in drug resistance in epilepsy.

A comparison between finasteride, flutamide, and finasteride plus flutamide combination in the treatment of hirsutism.

To investigate the clinical efficacy and safety of the finasteride (5 mg/day) plus flutamide (125 mg/day) combination therapy in unselected women with hirsutism, 44 women were involved in the study. The effects of such combination treatment have not been reported previously. The patients were assigned to 3 treatment groups: 14 patients (group 1) were treated with finasteride (5 mg per day), 16 patients (group 2) were treated with flutamide (125 mg per day), and 14 patients (group 3) were treated with finasteride (5 mg per day) plus flutamide (125 mg per day) for 12 months. Serum FSH, LH, estradiol, total testosterone, free testosterone, androstenedione, DHEAS, and SHBG were obtained. Hirsutism score was measured before and after treatment. Blood chemistry and side effects were evaluated during the study. The reductions in hirsutism score (% of the baseline) at 6 months were as follows: 24% for group 1, 35% for group 2, and 33% for group 3. Combination therapy resulted in (49%) similar improvement to flutamide alone (45%), but significantly (p<0.05) more efficacious improvement in hirsutism when compared to finasteride (32%) after 12 months of treatment. In conclusion, flutamide is more effective than finasteride and the combination of these two drugs is not better than flutamide alone, but better than finasteride in hirsute women.

Plasminogen activator inhibitor-1 (PAI-1) gene polymorphism (-675 4G/5G) associated with obesity and vascular risk in children.

UNLABELLED: Atherothrombotic complications in insulin resistance are partly attributed to impaired fibrinolysis caused by increased PAI-1 plasma levels, and 4G/5G promotor polymorphism of the PAI-1 gene may modulate PAI-1 transcription. OBJECTIVE: To investigate PAI-1-675 4G/5G allele gene polymorphism and its relationship with obesity in children. CHILDREN AND METHOD: The study participants were 133 apparently healthy non-obese children, 24 probable exogenously obese without family history (Group I), 66 probable familial obese (Group II), and 44 obese children who were referred to the pediatric endocrinology department with any complication of obesity (Group III). Group I and Group II obese children were gathered from a school-based epidemiological study. RESULTS: Incidence of obesity was 19% in a school with high socio-economic status, whereas it was 4% in a school with low socio-economic status. Frequencies of 4G/4G gene polymorphisms were 24.81%, 37.50%, 64.80% and 61.11% in the control group, and groups I, II, and III, respectively. In groups II and III, 4G/4G gene polymorphism, and in non-obese control children 5G/5G gene polymorphism, was common. In obese children in the presence of family history for obesity and metabolic syndrome (odds ratio [OR]: 4.48, 95% confidence interval [CI]: 1.26-15.82), carriage of the 4G allele either in heterozygous or homozygous state increased the risk of vascular disease (OR: 6.10, 95% CI 1.64-22.90). In patients with acanthosis nigricans, high HOMA-IR values, hypertriglyceridemia and elevated atherogenic index, 4G/4G genotype frequency was remarkably higher compared to patients with other features of metabolic syndrome. CONCLUSION: The increasing prevalence of childhood obesity in high socio-economic status is associated with health risks. In obese children with family history of obesity and cardiovascular disease or type 2 diabetes mellitus and in obese children who had any feature of metabolic syndrome, frequency of 4G/4G genotype was more than the 4G/5G and 5G/5G genotypes in the PAI-1 gene. These patients can be at increased risk for developing vascular disease. Acanthosis nigricans, high HOMA-IR value, hypertriglyceridemia and high atherogenic index can also reflect the high risk of vascular disease in metabolic syndrome.

Comparison of sedation method in pediatrics cardiac catheterization.

OBJECTIVE: Purpose of this study is to compare the effects of various anaesthetic combinations on hemodynamics, sedation level, recovery period and complications in the patients which undergo pediatric cardiac catheterization. PATIENTS AND METHODS: Four groups of anaesthetic combinations was created. The groups are classified as propofol-ketamine (group 1), propofol-dexmedetomidine (group 2), dexmedetomidine-ketamine (group 3), midazolam-ketamine (group 4) (for each group n=20). Baseline heart rate (HR), mean arterial blood pressure (MAP), respiratory rate (RR), peripheral oxygen saturation (SpO2) were recorded. This parameters values were recorded at 0., 5., 10., 15., 20., 25., 30. minutes; and the groups were compared according to these measurements data. RESULTS: For heart rate, Group 2 and 3 reduce the HR more than the drugs of Group 1 and 4 (p < 0.05). The SpO2 values of Group 1 were measured to have 5% further reduction compared to the Group 2 and 3; and Group 4 has the same SpO2 recordings compared to the Group 3 (p < 0.05). Comparing the recovery times; Group 4 was found to have the highest recovery time compared to the other drug groups. It is found that additional doses are needed for recovery in Group 4 (p < 0.008). Side effects were lowest for Group 3 and highest for Group 4. CONCLUSIONS: Considering the complication rates, it is concluded that Group 3 is spotted as the better sedation method among the other groups. In terms of additional propofol dose, Group 1 would be the better choice. Thus, the clinician should choose the suitable methods for the patient.

Common mutations at the homocysteine metabolism pathway and pediatric stroke.

Heterozygosity and/or homozygosity for mutations at the genes of the enzymes involved in homocysteine metabolism may confer an increased risk for thrombosis by causing hyperhomocysteinemia. Although the mutations related to homocysteine metabolism possibly increase the risk of stroke, the data are conflicting and there are very few reports linking these defects to acute stroke in children. We aimed to study the role of these mutations in Turkish children with ischemic stroke. Forty-six patients having cerebral infarct were clinically diagnosed, and the infarction verified with magnetic resonance imaging of the brain was included in the study. All patients were below the age of 18 (10 months to 18 years). Sixty-eight controls, consecutively selected among healthy unrelated subjects from the same geographic area of Turkey without personal and family history of thrombosis, stroke or Behest's disease, were included. Genotyping for the common mutations was carried out by the methods described previously. There was no difference between the pediatric stroke patients and controls for the distribution of methylene tetrahydrofolate reductase (MTHFR) 677 C-T, MTHFR 1298 A-C, methylene tetrahydrofolate dehydrogenase (MTHFD) 1958 G-A and methionine synthase reductase (MTRR) 66 A-G alleles. There was no risk for double gene alterations (MTHFR 677 C-T vs. 1298 A-C) after individuals with FV 1691 A mutation is excluded. Twelve of the 46 patients were found to carry FV 1691 A mutation (26.0%), one being homozygote. The cerebral infarct risk for FV 1691 A was found to be 6.4 (CI 95% 1.7-23.0). Eight of the 46 patients were found to carry PT 20210 A mutation (16.6%). Two of the FV 1691 A heterozygous patients carried PT 20210 A mutation at the same time (4.2%). As a conclusion, we can say that FV 1691 A and PT 20210 A mutations are important and must be included to the routine analysis of pediatric stroke patients.

Detection of virulence factors of Escherichia coli from children by multiplex polymerase chain reaction.

A total of 161 Escherichia coli (E. coli) strains isolated from children with urinary tract infection (UTI) were analysed for the genes encoding the virulence factors such as pyelonephritis (pap), s fimbriae (sfa), afimbrial adhesin I (afaI), haemolysin (hly), cytotoxic necrotising factor I (cnf I) and aerobactin (aer) by multiplex PCR. Ninety-four E. coli strains were found to carry at least one virulence factor. Therefore, 58.38% of total population was positive for one virulence gene at least. Percentage of genes within the total population for pap, sfa, afaI, hly, cnf I and aer was found as 22.98, 6.21, 9.94, 1.24, 9.94 and 39.75, respectively. Our analysis showed that sfa-pap (p < 0.001); pap-aer, afaI-aer and cnf I-pap (P < 0.05) and hly-sfa (p < 0.01) significantly co-occurred in their respective samples. In the light of these findings, we suggest an important role of pap causing UTI.