Increased metformin dosage suppresses pro-inflammatory cytokine levels in systemic circulation and might contribute to its beneficial effects.

Type 2 Diabetes Mellitus (T2DM) is a metabolic disorder, characterized by persistent elevation of blood glucose either due to insulin resistance or insulin insufficiency. Metformin is the recommended first choice of drug for the management of T2DM and is known to improve insulin sensitivity and prevents hyperglycemia by reducing chronic inflammation. T-helper type 1 (Th1) and type 17 (Th17) cells, are important pro-inflammatory CD4+ T cell subsets secreting TNF-α, and INF-γ (Th1), and interleukin 17 (Th17). These cytokines have been shown to play a crucial role in inflammation, insulin resistance, and the development of T2DM. Here, we explore the effect of different metformin dosages on pro-inflammatory cytokine (TNF-α, INF-γ, GM-CSF and IL-17) levels in systemic circulation among T2DM patients in Ghana, since inflammatory responses and cytokines play significant roles in the pathogenesis and progression of T2DM patients on metformin. Two hundred and nine (209) consenting T2DM patients receiving treatment at the Diabetic unit of the Komfo Anokye Teaching Hospital (KATH) in the Ashanti region of Ghana were recruited in a hospital-based cross-sectional study design. Blood samples were collected and serum obtained from each participant were analyzed for the concentrations of TNF-α, INF-γ, GM-CSF and IL-17 cytokine levels by solid-phase sandwich ELISA. We observed that participants on 3000 mg/day dose of metformin had significantly lower levels of TNF-α (p < .001) and IFN-γ (p = .014) compared to those on other dosages (1000 mg and 2000 mg/day). However, GM-CSF and IL-17 levels were not affected by increased metformin dosages. After adjusting for age, gender, dose and duration of metformin use, we observed that participants who took higher doses of metformin had significantly reduced levels of TNF-α (ß = -0.0297, 95% CI = (-0.005 to -0.002) p < .001. Metformin dosage independently predicted reduced TNF-α levels with 14.4% variations in the metformin dosage levels. Increased metformin dosage suppresses TNF-α levels in systemic circulation and hence might contribute to its beneficial effects.

Mammographic Breast Density Model Using Semi-Supervised Learning Reduces Inter-/Intra-Reader Variability.

Breast density is an important risk factor for breast cancer development; however, imager inconsistency in density reporting can lead to patient and clinician confusion. A deep learning (DL) model for mammographic density grading was examined in a retrospective multi-reader multi-case study consisting of 928 image pairs and assessed for impact on inter- and intra-reader variability and reading time. Seven readers assigned density categories to the images, then re-read the test set aided by the model after a 4-week washout. To measure intra-reader agreement, 100 image pairs were blindly double read in both sessions. Linear Cohen Kappa (κ) and Student's t-test were used to assess the model and reader performance. The model achieved a κ of 0.87 (95% CI: 0.84, 0.89) for four-class density assessment and a κ of 0.91 (95% CI: 0.88, 0.93) for binary non-dense/dense assessment. Superiority tests showed significant reduction in inter-reader variability (κ improved from 0.70 to 0.88, p ≤ 0.001) and intra-reader variability (κ improved from 0.83 to 0.95, p ≤ 0.01) for four-class density, and significant reduction in inter-reader variability (κ improved from 0.77 to 0.96, p ≤ 0.001) and intra-reader variability (κ improved from 0.89 to 0.97, p ≤ 0.01) for binary non-dense/dense assessment when aided by DL. The average reader mean reading time per image pair also decreased by 30%, 0.86 s (95% CI: 0.01, 1.71), with six of seven readers having reading time reductions.

Tissue inhibitor metalloproteinase 2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) best predicts the development of acute kidney injury.

BACKGROUND: Acute kidney injury (AKI) is routinely diagnosed by creatinine-based guidelines, which is sub-optimal marker after injury due to renal and non-renal factors. This has necessitated the need for more specific and sensitive biomarkers for early detection of AKI in at risk patients. This prospective cross-sectional study used the biomarkers of cell cycle arrest and Neutrophil Gelatinase Associated Lipocalin (NGAL) to assess AKI among hospitalized patients. METHODS: We conveniently enrolled 151 in-patients at the Trauma and Specialist Hospital, Winneba in Ghana. Socio-demographic and clinical information were collected using structured questionnaires. Blood samples were collected for the estimation of serum creatinine, and AKI diagnosed and staged using the KDIGO guideline. Fresh urine samples were collected and urinary NGAL, TIMP-2 (tissue inhibitor metalloproteinase 2) and IGFBP-7 (insulin-like growth factor binding protein 7) were estimated using ELISA kits. RESULTS: The cell cycle arrest biomarkers and NGAL were significantly (P < 0.001) higher among participants with AKI than those without AKI. [TIMP-2]∗[IGFBP-7] showed the best diagnostic performance (AUC = 0.94, CI = 0.90-0.98) followed by [IGFBP-7]∗NGAL] (AUC = 0.93, CI = 0.87-0.99), with NGAL having the least (AUC = 0.62, CI = 0.46-0.78). The cut-off for [TIMP-2]∗[IGFBP-7] showed the best predictive ability (95.8% sensitivity, 77.2% specificity, 44.2% PPV and 99% NPV). The cut-off for NGAL, on the other hand, showed the least predictive ability (62.5% sensitivity, 42.5% specificity, 17.0% PPV and 85.7% NPV). CONCLUSION: Tissue inhibitor metalloproteinase 2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) best predicts the development of AKI, and can be used in high risk patients for early diagnosis of AKI.

Profiling immuno-metabolic mediators of vitamin B12 deficiency among metformin-treated type 2 diabetic patients in Ghana.

BACKGROUND: The association between prolong metformin usage and B12 deficiency has been documented. However, the prevalence estimates of metformin-induced vitamin B12 deficiency showed substantial disparity among studies due to varied study definitions of vitamin B12 deficiency. Metformin blocks the calcium dependent absorption of the vitamin B12-Intrinsic Factor complex at the terminal ileum. Lack of intrinsic factor due to the presence of auto-antibodies to parietal cells (IFA) could lead to vitamin B12 deficiency and subsequently cause peripheral neuropathy. We investigated the prevalence of vitamin B12 deficiency using more sensitive, combined markers of vitamin B12 status (4cB12) and the immuno-biochemical mediators of vitamin B12 deficiency. METHODS: In this observational study, 200 consecutive consenting metformin-treated T2DM patients, aged 35 and above, attending the diabetic clinic at KATH were recruited. Vitamin B12 deficiency was classified based on the Fedosov age-normalized wellness quotient. Anthropometric measurement was taken as well as blood samples for immunological and biochemical mediators. Peripheral neuropathy was assessed using the Michigan Neuropathy Screening Instrument (MNSI). Statistical analysis was performed using the R Language for Statistical Computing. RESULTS: Using the combined indicator (4cB12), the prevalence of metformin induced vitamin B12 deficiency was 40.5% whilst the prevalence of MNSI-Q and MNSI-PE diabetic neuropathy was 32.5% and 6.5% respectively. Participants with vitamin B12 deficiency had significantly higher levels of IFA, GPA, TNF-α, TC, LDL and albumin compared to those with normal vitamin B12 levels (p < 0.05). Correlation analysis revealed a statistically significant negative association between 4cB12 and the immunological markers [IFA (rs = -0.301, p<0.0001), GPA (rs = -0.244, p = 0.001), TNF-α (rs = -0.242, p = 0.001) and IL-6 (rs = -0.145, p = 0.041)]. Likewise, 4cB12 was negatively associated with TC (rs = -0.203, p = 0.004) and LDL (rs = -0.222, p = 0.002) but positively correlated with HDL (rs = 0.196, p = 0.005). CONCLUSION: Vitamin B12 deficiency and diabetic neuropathy are very high among metformin-treated T2DM patients and it is associated with increased GPA, IFA, TNF-α and cardiometabolic risk factors (higher LDL and TC and lower HDL). Upon verification of these findings in a prospective case-control study, it may be beneficial to include periodic measurement of Vitamin B12 using the more sensitive combined indicators (4cB 12) in the management of patients with T2DM treated with metformin in Ghana.

Intracytoplasmic Expression of IL-6 and IL-17A in Circulating CD4+ T Cells Are Strongly Associated with and Predict Disease Activity in Rheumatoid Arthritis: A Case-Control Study in Ghana.

BACKGROUND: T cell cytokines play important roles in the development and progression of rheumatoid arthritis (RA). Loss of Th1/Th2 and Th17/Treg balance has been reported in several inflammatory autoimmune diseases. However, their role in RA within hitherto rare Ghanaian context has not been explored. Here, we evaluated the intracytoplasmic CD4+ T cell cytokine patterns in rheumatoid arthritis patients in Ghana and determined their relationship with disease activity. METHODS: This case-control study included 48 newly diagnosed RA patients and 30 apparent healthy controls from two major hospitals in Ghana. Validated structured questionnaires were administered to obtain demographic data; blood samples were collected and processed for flow cytometric analysis. RESULTS: IFN-γ, TNF-α, IL-4, IL-6, IL-10, IL-17A, IL-6/IL-4, and IL-17/IL-10 expressions were significantly higher in RA cases compared to the healthy controls. The expression of IL-6 (0.00 (0.00-0.98) vs. 0.82 (0.34-1.10) vs. 1.56 (1.39-1.68), p < 0.0001), IL-17A (0.00 (0.00-0.02) vs. 0.19 (0.09-0.30) vs. 0.99 (0.64-1.25), p < 0.0001), and IL-17A/IL-10 (0.00 (0.00-0.39) vs. 0.15 (0.09-0.26) vs. 0.88 (0.41-1.47), p < 0.0001) increased significantly from the healthy controls through RA patients with low DAS scores to RA patients with moderate DAS scores. IL-6 (ß = 0.681, r 2 = 0.527, p < 0.0001), IL-17A (ß = 0.770, r 2 = 0.593, p < 0.0001), and IL-17A/IL-10 (ß = 0.677, r 2 = 0.452, p < 0.0001) expressions were significantly directly associated with DAS28 scores. IL-6 (cutoff = 1.32, sensitivity = 100.0%, specificity = 100.0%, accuracy = 100.0%, and AUC = 1.000) and IL-17A (cutoff = 0.58, sensitivity = 100.0%, specificity = 100.0%, accuracy = 100.0%, and AUC = 1.000) presented with the best discriminatory power in predicting moderate DAS scores from low DAS scores. CONCLUSION: Th1- and Th17-related cytokines predominate in the pathophysiology of RA, with IL-6 and IL-17 being principally and differentially expressed based on the severity of the disease. IL-6 and IL-17A could serve as useful prognostic and disease-monitoring markers in RA in the African context.