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 (25th - 75th
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.).