Hyperglycaemia cause vascular inflammation through advanced glycation end products/early growth response-1 axis in gestational diabetes mellitus.

Hyperglycaemia during pregnancy is the main reason for developing diabetes mediated vascular complications. Advanced glycation end products (AGEs) are formed due to non-enzymatic glycation of proteins, lipids and nucleic acids during hyperglycaemia. It has the potential to damage vasculature by modifying the substrate or by means of AGEs and receptor of AGE (RAGE) interaction. It has been linked with the pathogenesis of various vascular diseases including coronary heart disease, atherosclerosis, restenosis etc. This study was carried out to investigate the role of AGEs-EGR-1 pathway in gestational diabetes mellitus (GDM) vascular inflammation. Human umbilical vein endothelial cells (HuVECs) isolated from normal glucose tolerant mothers were subjected to various treatments including high glucose, silencing of early growth response (EGR)-1, blockade of protein kinase C (PKC) ß, blocking extracellular signal-regulated protein kinases 1 and 2 (ERK1/2), and treatment with AGEs and assayed for EGR-1, tissue factor (TF) and soluble intercellular adhesion molecule (sICAM)-1. Similarly, umbilical vein endothelial cells isolated from normal and GDM mothers were assayed for EGR-1, TF, and sICAM-1. There was a significant increase in EGR-1 and TF levels in HuVECs isolated form GDM mother's umbilical cord and normal HuVECs treated with high glucose condition. This was accompanied by elevated levels of sICAM-1 in high glucose treated cells. Our results revealed AGE-mediated activation of EGR-1 and its downstream genes via PKC ßII and ERK1/2 signaling pathway. The present study demonstrated a novel mechanism of AGEs/ PKC ßII/ ERK1/2/EGR-1 pathway in inducing vascular inflammation in GDM.

Role of VEGF165b/VEGFTOTAL ratio in gestational diabetes mellitus.

Proper vascular function is important for well-being of mother and growing fetus. VEGFTOTAL, and VEGF165b levels and its vascular endothelial complications in gestational diabetes mellitus (GDM) together with the association of inflammation and advanced glycation end products (AGEs) are less studied. VEGF165b/VEGFTOTAL (VEGF RATIO) in GDM pregnant women was investigated in this study. Plasma VEGFTOTAL was lower in GDM (17.68 ± 1.30 pg/mL) compared to non-GDM (25.69 ± 1.40 pg/mL). VEGF165b, ICAM-1, and AGEs were higher in GDM (9.9 ± 1.4 pg/mL, 201.04 ± 7.85 µg/mL, and 10.40 ± 0.98 µg/mL, respectively) and lower in non-GDM (6.47 ± 0.70 pg/mL, 174.1 ± 7.11 µg/mL, and 4.71 ± 0.39 µg/mL, respectively). Compared to non GDM (0.25 ± 0.02), VEGF RATIO was higher in GDM (0.45 ± 0.04) and correlated with -ICAM-1 (r = 0.375, p < .001) and AGEs (r = 0.199, p < .05). Tertile stratification of VEGF RATIO implied that frequency of GDM increases with increasing tertiles of VEGF RATIO (p for trend <.001). Association of VEGF RATIO with GDM was significant even after adjusting for AGEs (OR = 1.279, CI = 1.118-1.462, p < .0010) but it lost its significance when adjusted for ICAM-1 (OR = 1.006, CI = 0.995-1.017, p = .308). VEGF RATIO plays an important role in GDM in association with vascular inflammation.

Insulin Treatment Forces Arteriogenesis in Diabetes Mellitus by Upregulation of the Early Growth Response-1 (Egr-1) Pathway in Mice.

The process of arteriogenesis is severely compromised in patients with diabetes mellitus (DM). Earlier studies have reported the importance of Egr-1 in promoting collateral outward remodeling. However, the role of Egr-1 in the presence of DM in outward vessel remodeling was not studied. We hypothesized that Egr-1 expression may be compromised in DM which may lead to impaired collateral vessel growth. Here, we investigated the relevance of the transcription factor Egr-1 for the process of collateral artery growth in diabetic mice. Induction of arteriogenesis by femoral artery ligation resulted in an increased expression of Egr-1 on mRNA and protein level but was severely compromised in streptozotocin-induced diabetic mice. Diabetes mellitus mice showed a significantly reduced expression of Egr-1 endothelial downstream genes Intercellular Adhesion Molecule-1 (ICAM-1) and urokinase Plasminogen Activator (uPA), relevant for extravasation of leukocytes which promote arteriogenesis. Fluorescent-activated cell sorting analyses confirmed reduced leukocyte recruitment. Diabetes mellitus mice showed a reduced expression of the proliferation marker Ki-67 in growing collaterals whose luminal diameters were also reduced. The Splicing Factor-1 (SF-1), which is critical for smooth muscle cell proliferation and phenotype switch, was found to be elevated in collaterals of DM mice. Treatment of DM mice with insulin normalized the expression of Egr-1 and its downstream targets and restored leukocyte recruitment. SF-1 expression and the diameter of growing collaterals were normalized by insulin treatment as well. In summary, our results showed that Egr-1 signaling was impaired in DM mice; however, it can be rescued by insulin treatment.

First surveillance of SARS-CoV-2 and organic tracers in community wastewater during post lockdown in Chennai, South India: Methods, occurrence and concurrence.

Surveillance of SARS-CoV-2 and organic tracers (OTs) were conducted in the community wastewater of Chennai city and the suburbs, South India, during partial and post lockdown phases (August-September 2020) as a response to the coronavirus disease 2019 (COVID-19) pandemic. Wastewater samples were collected from four sewage treatment plants (STPs), five sewage pumping stations (SPSs) and at different time intervals from a suburban hospital wastewater (HWW). Four different methods of wastewater concentrations viz., composite (COM), supernatant (SUP), sediment (SED), and syringe filtration (SYR) were subjected to quantitative real time-polymerase chain reaction (qRT-PCR). Unlike HWW, STP inlet, sludge and SPS samples were found with higher loading of SARS-CoV-2 by SED followed by SUP method. Given the higher levels of dissolved and suspended solids in STPs and SPSs over HWW, we suspect that this enveloped virus might exhibit the tendency of higher partitioning in solid phase. Cycle threshold (Ct) values were < 30 in 50% of the HWW samples indicating higher viral load from the COVID-19 infected patients. In the STP outlets, a strict decline of biochemical oxygen demand, >95% removal of caffeine, and absence of viral copies reflect the efficiency of the treatment plants in Chennai city. Among the detected OTs, a combination of maximum dynamic range and high concurrence percentage was observed for caffeine and N1 gene of SARS-CoV-2. Hence, we suggest that caffeine can be used as an indicator for the removal of SARS-CoV-2 by STPs. Our predicted estimated number of cases are in line with the available clinical data from the catchments. Densely distributed population of the Koyambedu catchment could be partly responsible for the high proportion of estimated infected individuals during the study period.

Association of HLA-A, -B, DRB, and DQB Alleles with Persistent HPV-16 Infection in Women from Tamil Nadu, India.

Women with persistent human papillomavirus (HPV) infections have a high risk of developing cervical cancer (CaCx). HPV-16 alone accounts for more than 60% of CaCx worldwide. Most of the HPV infections are transient and only a subset of women develop persistent HPV-16 infection. Many studies have shown associations of different human leukocyte antigen (HLA) alleles with HPV-mediated CaCx, but there are only a few studies globally that relate to persistent HPV-16 infection. Furthermore, such studies from India are sparse. Hence, we investigated the association of HLA-A, B, DRB, and DQB alleles with persistent HPV-16 infection and HPV-16-positive CaCx in south India (Tamil Nadu). HPV-16 persistent infection was observed in 7% of normal women. A total of 50 women with HPV-16-positive CaCx, 21 women with HPV-16 persistent infection, and 74 HPV-16-negative normal women were recruited for this study. Low-resolution typing of HLA-A, B, DRB, and DQB alleles was performed. HLA-B*44 and DRB1*07 showed a significant association with persistent HPV-16 infection (odds ratio, p-value = 26.3, 0.03 and 4.7, 0.01, respectively). HLA-B*27 and DRB1*12 were significantly associated with both HPV-16+ CaCx and persistent HPV-16 infection (23.8, 0.03; 52.9, 0.01; 9.8, 0.0009; and 13.8, 0.009; respectively). HLA-B*15 showed a negative association with HPV-16-positive CaCx (0.1, 0.01), whereas DRB1*04 exhibited protection to both HPV-16-positive CaCx and persistent HPV-16 infection (0.3, 0.0001 and 0.1, 0.0002, respectively). Thus, we show HLA allelic association with HPV-16 infection in Tamil Nadu. Larger studies on high-resolution HLA typing coupled with HPV-16 genome diversity will offer further insights into host/pathogen genome coevolution.