2.2. Study Design
obesity, T2D, bloating, fasting blood glucose, diarrhea, abdominal pain, constipation, Diabetes, diabetes
OBESITY, DIABETES, SECONDARY, DIABETES
This study was a single-center, double-blind, placebo-controlled, randomized clinical trial, with a 6-month duration, involving the administration of probiotic supplements to persons with T2D. Ethical approval was obtained from the Institution Review Board of Laiko General Hospital in Athens, Greece (approval number 306/26.04.2021). The trial protocol was registered on clinicaltrials.gov (identifier NCT06032988). Eligible participants were randomized using a computer generator (Participants were followed at the outpatient Diabetes Center every 3 months, aligning with standard practice for individuals with diabetes. At the baseline visit, and subsequently at the 3-month and 6-month intervals, anthropometric measurements were recorded, including height (in cm), weight (in kg), blood pressure (in mmHg), and waist circumference (in cm), and body mass index (BMI, kg/mAdditionally, blood biochemical parameters, including glycated hemoglobin (HbA1c), fasting blood glucose (FBG), fasting lipid panel (total cholesterol, triglycerides, HDL-cholesterol, and LDL-cholesterol), renal function indicators (blood urea and serum creatinine), and liver function tests (AST, ALT, γGT, and alkaline phosphatase), were extracted manually from participants’ medical records at the baseline, 3-month, and 6-month visits.Furthermore, participants were asked to give a stool sample at the beginning and the end (6 months) of the study for gut microbiome analysis. They were provided with instructions to collect samples at home in the morning. These samples were then transported to the clinic under refrigeration and stored in a freezer (−20 °C) until processing. Researchers responsible for the procedure utilized the Fecal Swab Collection and Preservation System/Stool DNA Isolation Kit provided to them. The collected samples were stored in the freezer until the entire collection process was completed at the follow-up visit of 6 months. This DNA isolation was analyzed through 16S rRNA Sequencing, and a microbial analysis through alpha and beta diversity was held for any quantitative or qualitative alterations, respectively [The primary outcome of this study was the change in HbA1c between the two groups (active probiotic treatment and control) at 6 months. Anthropometrics (especially obesity parameters), blood lipid levels, liver function tests, and gut microbiome changes were measured as secondary outcomes. A questionnaire was also administered at the last visit (6 months) to collect data on participants’ tolerance and satisfaction with the treatment, including the frequency of symptoms such as constipation, diarrhea, bowel function, bloating, gas production, and abdominal pain. 