Combined Boyden-Flow Cytometry Assay Improves Quantification and Provides Phenotypification of Leukocyte Chemotaxis

Chemotaxis has been studied by classical methods that measure chemotactic and random motility responses in vitro, but these methods do not evaluate the total number and phenotype of migrating leukocytes simultaneously. Our objective was to develop and validate a novel assay, combined Boyden-flow cytometry chemotaxis assay (CBFCA), for simultaneous quantification and phenotypification of migrating leukocytes. CBFCA exhibited several important advantages in comparison to the classic Boyden chemotaxis assay (CBCA): 1) improved precision (intra-assay coefficients of variation (CVs): CBFCA-4.7 and 4.8% vs. CBCA-30.1 and 17.3%; inter-observer CVs: CBFCA-3.6% vs. CBCA 30.1%); 2) increased recovery of cells, which increased assay to provide increased sensitivity; 3) high specificity for determining the phenotype of migrating/attracted leukocytes; and 4) reduced performance time (CBFCA 120 min vs. CBCA 265 min). Other advantages of CBFCA are: 5) robustness, 6) linearity, 7) eliminated requirement for albumin and, importantly, 8) enabled recovery of migrating leukocytes for subsequent studies. This latter feature is of great benefit in the study of migrating leukocyte subsets. We conclude that the CBFCA is a novel and improved technique for experiments focused on understanding leukocyte trafficking during the inflammatory response

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049

C-reactive protein in type 2 diabetes and denture stomatitis

Objectives: Candida-associated denture stomatitis (CaDS), is a common disease in complete denture wearers with type 2 diabetes mellitus (T2DM). C-reactive protein (CRP), an acute phase reactant is a sensitive marker of inflammation associated with diabetes. The aim of this study was to determine the concentrations of CRP in patients with T2DM and CaDS, and investigate the relationship between CRP concentrations and glycated hemoglobin A1c (HbA1c) levels, which was used as an indicator of glycemic control. Materials and Methods: The study involved 110 T2DM patients (63 women and 47 men, mean age 63.2±10.5 years) with CaDS, and 20 patients (12 women and 8 men, mean age 65.8±12.9 years), with T2DM and healthy oral mucosa. A group of 20 non-diabetics (11 women and 9 men, mean age 59.2±9.9 years) with healthy oral mucosa served as a control. The yeasts were isolated by the culture method, and identified by microscopic examination and with the test kit, ID 32 C (bioMerieux SA, Marcy-l\u27Etoile, France). Serum concentrations of CRP were determined by rocket immunoelectrophoresis according to Laurell. Glycemic control was evaluated by measuring HbA1c levels using HPLC together with the Variant Hemoglobin A1c Program (Bio-Rad Laboratories, Hercules CA, USA). Results: The mean duration of diabetes was 10.6±5.1 years, and the mean HbA1c levels in T2DM subjects were 8.6 ±1.9%. Patients with T2DM had significantly elevated mean levels of CRP in comparison to patients with normal glucose metabolism (6.12±2.86 vs 2.57±0.96 mg/l, p\u3c0.05). The highest CRP levels were observed in diabetics with type II (Newton classification) CaDS (8.39±2.70 mg/l). A positive correlation was found between the concentrations of CRP and fasting plasma glucose levels (rS=0.517, p\u3c0.001) and HbA1c levels (rS=0.572, p\u3c0.001). Conclusions: The findings suggest that CRP concentrations can be used as a non-specific indicator of ongoing inflammation in patients with T2DM