Homeostatic Microbiome Disruption as a Cause of Insulin Secretion Disorders . Candida albicans, as a New Factor in Pathogenesis of Diabetes

Throat swabs and stool specimen were taken from 38 patients (36 women and two men, mean age 60.6 ± 10.4, min = 34, max = 72). Samples were collected at the Clinic for Endocrinology, Diabetes and Metabolic Diseases, Clinical Center of Serbia in Belgrade. All patients were informed about the purpose of this research and gave their oral consent for voluntary participation in this study, so it is conducted in accordance with the decision of the Ethics Committee of Medical Faculty in Belgrade. All parameters to determine the body mass index (BMI) of patients were obtained. Anthropometric measurements: body weight, body height, body mass index, waist circumference, hip circumference, waist to hip circumference ratio, were performed. Body mass index was calculated as body weight in kilograms divided by the square of the body height in meters. Glucose regulation was tested by oral glucose tolerance test (OGTT) which was performed on fasting patients by drinking 75gr of glucose dissolved in 300 ml of water. Then the values ​​of glycemia and insulin were measured after 0, 30 and 120 minutes. Insulin levels were determined by radioimmunoassay Microbiological analyses of samples (stool and throat swab) were performed by standard procedure at the Institute for Microbiology, Clinical Center in Serbia. Microbiological isolates were identified on the basis of microscopic, cultural and biochemical properties. Microscopic slides were prepared directly from patient samples, and also from colonies on culture plates, and then Gram stained. We used Columbia blood agar plates (7% sheep blood), MacConkey agar and XLD agar plates and Sabourand dextrose agar. Depending on the cultural and morphological characteristics of isolated microorganisms, we have prepared a small series of biochemical tests.

Results are presented as count (%), means ± standard deviation or median (25^th - 75^th Percentile) depending on data type and distribution. Groups are compared using parametric (t test) and nonparametric (Pearson Chi-square test, Fisher's exact test, Mann-Whitney U test) tests. All p values less than 0.05 were considered significant. All data were analyzed using SPSS 20.0 (IBM Corp. Released 2011. IBM SPSS Statistics for Windows, Version 20.0. Armonk, NY: IBM Corp.).