Prevalence and independent predictors of insulin resistance in children from Crete,Greece: The Children Study

Background It has been proposed that insulin resistance (IR) is associated with the development of Type 2 diabetes mellitus and cardiovascular disease. The aim of this study is to determine the prevalence of IR in Greek schoolchildren and to investigate factors associated with IR. Methods Between October 2005 and March 2006, 522 children were recruited from Crete. Physical activity and dietary habits, anthropometric and biochemical characteristics, as well as medical history of pupils’ parents were recorded. IR was estimated using the homeostasis model assessment (HOMA‐IR), fasting glucose‐to‐insulin ratio (FGIR) and quantitative insulin sensitivity check index (QUICKI). Multiple linear regression was used to determine independent predictors for IR. Results Fasting insulin levels and HOMA‐IR scores were higher in obese children and girls compared with their normal‐weight peers (P < 0.001). Moreover, the former had lower values in FGIR and QUICKI indices compared with the latter, indicating that obese children and girls are more insulin resistant compared with their counterparts (P < 0.001). The prevalence of IR was 9.2% (2.9% in normal‐weight, 10.5% in overweight and 31.0% in obese children), using as a threshold HOMA‐IR > 2.10 97.5th percentile of normal‐weight participants). Multiple linear regression revealed that central adiposity, female gender and intake of simple carbohydrates is associated positively with HOMA‐IR values, even after controlling for many other factors. Conclusion These findings demonstrate that girls and obese children, particularly those with central adiposity, are at high risk of developing IR. Therefore, these groups should be targets of Type 2 diabetes mellitus and cardiovascular disease preventive interventions.     

Determination of xylazine and its metabolites by GC-MS in equine urine for doping analysis

Xylazine and its main metabolites were detected in equine urine after a single-dose intravenous administration of 0.98 and 1.01 mg/kg body weight xylazine, respectively, in two horses, in order to be used for equine doping control routine analysis. The urine levels of the parent drug and its metabolites were determined using gas chromatography-mass spectrometry (GC-MS). Xylazine is metabolised rapidly, down to a concentration level of about 1.0 microg/ml after 1-3h administration. Seven metabolites were identified in urine. 4-Hydroxy-xylazine, the major metabolite, could be traced for 25 h and it is regarded as the long-term metabolite of xylazine in horse. 2,6-Dimethylaniline was, for the first time, reported as metabolite in equine.     

Isolation of Coxiella burnetii by a centrifugation shell-vial assay from ticks collected in Cyprus: detection by nested polymerase chain reaction (PCR) and by PCR-restriction fragment length polymorphism analyses

Ticks are the principal vectors and reservoirs of Coxiella burnetii. The identification of isolates is necessary for understanding the clinical diversity of Q fever in different geographic areas. This is the first report of isolation of C. burnetii from ticks by the shell-vial assay and by nested polymerase chain reaction (PCR) assay for the detection of this pathogen in ticks. Of 141 ticks collected in Cyprus (Rhipicephalus sanguineus and Hyalloma spp.), 10% were found to be infected with C. burnetii. Three ticks were positive by hemolymph test, and 11 triturated ticks were positive by nested PCR. Three isolates were obtained by the centrifugation shell-vial technique. Analysis by PCR, then restriction fragment length polymorphism showed that the 3 Cyprus isolates had identical restriction profiles to reference strains Nine Mile and Q212. The methods described are useful in studying the epidemiology and ecology of C. burnetii.     

Precise measurements of intrauterine vascular structures at hysteroscopy in menorrhagia and during Norplant use

Using currently available equipment for panoramic hysteroscopy, the size of images viewed cannot be accurately judged because of the magnifying and distorting effects of the objective lens. This study has demonstrated that magnification by the hysteroscope lens can alter the apparent size of images by up to 27%. An additional effect of lens distortion can alter the apparent size of images viewed by up to 28%, depending on the position of the image in the field of view. These effects are independent and may be additive. Thus, the apparent size of intrauterine structures at hysteroscopy may bear little resemblance to their actual size. Image-correction methods are described which reduce the effects of image magnification on the apparent size of an object viewed through the hysteroscope to 7%, and the effect of distortion to 3 %. This technique can greatly improve the accuracy of measurement at hysteroscopy, and has been utilized in this study for the precise measurement of superficial endometrial vascular diameter (mean+/-SEM) in 34 Norplant users (120+/-11.6 microm) and 20 women with spontaneous menorrhagia (74+/-7.2 microm). It has also confirmed the presence of scattered dilated vessels (up to 777 microm in diameter) on the endometrial surface in some Norplant users.      

Determination of sialic acids in biological fluids using reversed-phase ion-pair high-performance liquid chromatography

A simple, rapid and sensitive reversed-phase ion-pair high-performance liquid chromatographic method for the determination of N-acetylneuraminic acid and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid in biological fluids is described. Determination of N-acetylneuraminic acid released by acidic hydrolysis, in serum, urine and saliva, and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid in urine, without hydrolysis, was accomplished by injecting the sample without derivatization, into the chromatograph. Measurements were carried out isocratically within 6 min using a C18 column and a mobile phase of aqueous solution of triisopropanolamine, as ion-pair reagent, 60 mM, pH 3.5 at room temperature with UV absorbance detection. The present method is reported for the first time for the determination of sialic acids in biological fluids. Recoveries in serum, urine and saliva ranged from 90 to 102% and the limits of detection were 60 nM and 20 nM for the two sialic acids, respectively. The method has been applied to normal and pathological sera from patients with breast, stomach, colon, ovarian and cervix cancers, to normal urine and urine from patient with sialuria and to normal saliva.     

云南省恶性疟原虫多药抗性基因Asn86Tyr多态性调查

目的检测云南恶性疟原虫多药抗性基因(Pfmdr1)Asn86Tyr的多态性,探索其与氯喹敏感性表现型的关系.方法用等位基因特异PCR和限制性酶切片段长度分析技术(AS-PCR/RFLP).结果在86%(19/22)的氯喹抗性分离株中检测到Pfmdr1基因的Asn86Tyr两种多态性,携带编码86Tyr多态变异的占50%,而含编码Asn86多态的也是50%.结论在云南氯喹抗性高度流行区,Pfmdr1基因Asn86Tyr的多态性是普遍存在的.     

Altered innate and adaptive immune responses in patients with hidradenitis suppurativa

BACKGROUND: The clinical improvement of hidradenitis suppurativa reported in a small number of patients with antitumour necrosis factor (anti-TNF)-alpha therapies supports the hypothesis for an altered immune response in these patients. OBJECTIVES: To evaluate the state of the innate and adaptive immune responses in patients with hidradenitis suppurativa. METHODS: Fifty-three patients and six healthy controls were studied. Blood was sampled and subpopulations of lymphocytes were analysed by flow cytometry; monocytes were isolated and their function was evaluated from the concentrations of TNF-alpha and interleukin (IL)-6 in supernatants of cell cultures after triggering with endotoxins (lipopolysaccharides). TNF-alpha and IL-6 were estimated by an enzyme immunoassay. RESULTS: CD3/CD8 lymphocytes were lower in patients with involvement of the perineum than in controls; patients with involvement of the breast had higher levels of natural killer (NK) cells than controls. A negative correlation was found between years lapsing since initial presentation of lesions of hidradenitis and the percentage of NK cells. Monocytes isolated from healthy volunteers were more active for the secretion of TNF-alpha and IL-6 than those of patients with hidradenitis suppurativa. CONCLUSIONS: A reduction in the percentage of NK cells over time and a lower monocyte response to triggering by bacterial components is observed in patients with hidradenitis suppurativa. Further research is needed to clarify if these changes are connected to an autoimmune mechanism in the pathogenesis of hidradenitis suppurativa.     

Evidence for the role of gastric mucosa at the secretion of soluble triggering receptor expressed on myeloid cells （strem-1） in peptic ulcer disease

AIM: To investigate the role of gastric mucosa at the secretion of sTREM-1 in peptic ulcer. METHODS: Seventy two patients were enrolled; 35 with duodenal, 21 with gastric ulcer and 16 with chronic gastritis. Patients were endoscoped and gastric juice was aspirated. Patients with duodenal and gastric ulcer underwent a second endoscopy post-treatment. Biopsies were incubated in the absence/presence of endotoxins or gastric juice. Supernatants were collected and sTREM-1 and TNFα were measured by enzyme immunoabsorbent assays. Scoring of gastritis was performed before and after treatment according to updated Sydney score. RESULTS: Patients with duodenal and gastric ulcer and those with chronic gastritis had similar scores of gastritis. sTREM-1 was higher in supernatants of tissue samples of H pylori-positive than of H pylori-negative patients with gastric ulcer. Median (± SE) sTREM-1 was found increased in supernatants of patients with gastric ulcer before treatment (203.21 ± 88.91 pg/1000 cells) compared to supernatants either from the same patients post-treatment (8.23 ± 5.79 pg/1000 cells) or from patients with chronic gastritis (6.21 ± 0.71 pg/1000 cells) (P < 0.001 and < 0.001, respectively). Similar differences for sTREM-1 were recorded among LPS-stimulated tissue samples of patients (P = 0.001). Similar differences were not found for TNFα. Positive correlations were found between sTREM-1 of supernatants from patients withboth duodenal and gastric ulcer before treatment and the degree of infiltration of neutrophils and monocytes. CONCLUSION: sTREM-1 secreted by the gastric mucosa is an independent mechanism connected to the pathogenesis of peptic ulcer. sTREM-1 was released at the presence of H pylori from the inflamed gastric mucosa in the field of gastric ulcer.