Redox regulation of type-I inositol trisphosphate receptors in intact mammalian cells.

A sensitization of inositol 1,4,5-trisphosphate receptor (IP3R)-mediated Ca2+ release is associated with oxidative stress in multiple cell types. These effects are thought to be mediated by alterations in the redox state of critical thiols in the IP3R, but this has not been directly demonstrated in intact cells. Here, we utilized a combination of gel-shift assays with MPEG-maleimides and LC-MS/MS to monitor the redox state of recombinant IP3R1 expressed in HEK293 cells. We found that under basal conditions, ∼5 of the 60 cysteines are oxidized in IP3R1. Cell treatment with 50 μm thimerosal altered gel shifts, indicating oxidation of ∼20 cysteines. By contrast, the shifts induced by 0.5 mm H2O2 or other oxidants were much smaller. Monitoring of biotin-maleimide attachment to IP3R1 by LC-MS/MS with 71% coverage of the receptor sequence revealed modification of two cytosolic (Cys-292 and Cys-1415) and two intraluminal cysteines (Cys-2496 and Cys-2533) under basal conditions. The thimerosal treatment modified an additional eleven cysteines, but only three (Cys-206, Cys-767, and Cys-1459) were consistently oxidized in multiple experiments. H2O2 also oxidized Cys-206 and additionally oxidized two residues not modified by thimerosal (Cys-214 and Cys-1397). Potentiation of IP3R channel function by oxidants was measured with cysteine variants transfected into a HEK293 IP3R triple-knockout cell line, indicating that the functionally relevant redox-sensitive cysteines are predominantly clustered within the N-terminal suppressor domain of IP3R. To our knowledge, this study is the first that has used proteomic methods to assess the redox state of individual thiols in IP3R in intact cells. © 2018 Joseph et al

Black carbon physical and optical properties across northern India during pre-monsoon and monsoon seasons

Black carbon (BC) is known to have major impacts on both climate and human health and is therefore of global importance, particularly in regions close to large populations that have strong sources. The size-resolved mixing state of BC-containing particles was characterised using a single-particle soot photometer (SP2). The study focusses on the Indo-Gangetic Plain (IGP) during the pre-monsoon and monsoon seasons. Data presented are from the UK Facility for Airborne Atmospheric Measurements BAe-146 research aircraft that performed flights during the pre-monsoon (11 and 12 June) and monsoon (30 June to 11 July) seasons of 2016. Over the IGP, BC mass concentrations were greater (1.95 µg m−3) compared to north-west India (1.50 µg m−3) and north-east India (0.70 µg m−3) during the pre-monsoon season. Across northern India, two distinct BC modes were recorded; a mode of small BC particles (core diameter <0.16 µm and coating thickness <50 nm) and a mode of moderately coated BC (core diameter <0.22 µm and coating thickness of 50–200 nm). The IGP and north-east India locations exhibited moderately coated black carbon particles with enhanced coating thicknesses, core sizes, mass absorption cross sections, and scattering enhancement values compared to much lower values present in the north-west. The coating thickness and mass absorption cross section increased with altitude (13 %) compared to those in the boundary layer. As the monsoon arrived across the region, mass concentration of BC decreased over the central IGP and north-east locations (38 % and 28 % respectively), whereas for the north-west location BC properties remained relatively consistent. Post-monsoon onset, the coating thickness, core size, mass absorption cross section, and scattering enhancement values were all greatest over the central IGP much like the pre-monsoon season but were considerably reduced over both north-east and north-west India. Increases in mass absorption cross section through the atmospheric column were still present during the monsoon for the north-west and central IGP locations, but less so over the north-east due to lack of long-range transport aerosol aloft. Across the Indo-Gangetic Plain and north-east India during the pre-monsoon and monsoon seasons, solid-fuel (wood burning) emissions form the greatest proportion of BC with moderately coated particles. However, as the monsoon develops in the north-east there was a switch to small uncoated BC particles indicative of traffic emissions, but the solid-fuel emissions remained in the IGP into the monsoon. For both seasons in the north-west, traffic emissions form the greatest proportion of BC particles. Our findings will prove important for greater understanding of the BC physical and optical properties, with important consequences for the atmospheric radiative forcing of BC-containing particles. The findings will also help constrain the regional aerosol models for a variety of applications such as space-based remote sensing, chemistry transport modelling, air quality, and BC source and emission inventories

Clinical, biochemical and haematological changes in leptospirosis

Background: Leptospirosis is a globally important zoonotic disease caused by pathogenic leptospira. Leptospira species are spirochetes belonging to the order spirochetes and the family leptospiraceae. Present study is done to find out the incidence of thrombocytopenia in leptospirosis and to correlate it with other parameters like renal dysfunction, hepatic dysfunction and bleeding manifestation.Methods: Study includes 51 clinically suspected and diagnosed cases of leptospirosis, in Government hospital, South Gujarat, during a period from January 2017 to December 2017. Clinical signs and symptoms and complications, biochemical profile like bilirubin and creatinine, haematological profile like Hb, WBC count and platelet count were recorded. Thrombocytopenia was defined as a platelet count below 1,50,000/cmm.Results: The present study includes 51 cases of Leptospirosis. Age ranged from 16 years to 61 years (male-39 and Females-12) There were 38 (74.5%) cases with thrombocytopenia and 13 (25.4%) cases with normal platelet count. Out of 38 thrombocytopenic cases, 32 (84.2%) cases had renal dysfunction, 26 (68.4%) cases had hepatic dysfunction and 16 (42.1%) cases had pulmonary haemorrhage. Among 13 cases with normal platelet count, 8 (61.5%) cases had hepatic dysfunction and 7 (53.8%) cases had renal dysfunction and 3 (23%) cases had pulmonary haemorrhage.Conclusions: Thrombocytopenia is a frequent complication (present in more than half of the patient) in leptospirosis and associated with more frequent and more severe complications. Therefore, early recognition of thrombocytopenia is recommended to prevent complications and mortality in leptospirosis

In-vitro Toxicity Study of Cylindrospermopsin on Wistar Rats

Naturally derived cyanotoxin, Cylindrospermopsin (CYN) found in fresh water systems poses a threat to human health worldwide. Cylindrospermopsin (CYN) is most potent cyanotoxin which affects the functions of kidney and liver. The present study was carried out to determine the possible effects of Cylindrospermopsin on mammals using male Wistar rats as an animal model. All experimental protocols were approved by the ethics committee of Faculty of Medical Science, University of Sri Jayewardenepura (No. 17/18). Thirty-five Wistar rats were divided into five groups (n=7) and rats were orally treated with different concentrations of cyanotoxin; Cylindrospermopsin (2.5 μg/L, 2 μg/L, 1.5 μg/L) and control group was treated with distilled water for 90 days. Fifth group received environmental water sample contaminated with Cylindrospermopsin (2.3 μg/L) obtained from randomly selected well in Padaviya. Blood and urine samples from each individual were collected at 0, 7, 14, 28, 42, 60, 90 days intervals and the collected samples were subjected to serum creatinine and urine creatinine analysis using creatinine assay kits. Cylindrospermopsin in urine samples were quantified by the ELISA methods. Aspartate aminotransferase (AST), Aspartate alaninetransferase (ALT) and Full Blood Count (FBC) were analysed. The mean body weight of treated (200 to 310 g) and control groups (200-335 g) of the experiment gradually increased until fourteenth week. There was no significant difference of body weights between treated and control groups (p=0.08). The absolute and relative (% body weight) weights of liver and kidneys of the treated groups were less than control group. Cylindrospermopsin dose 2.5 μg/L, 2 μg/L , 1.5 μg/L, 2.3 μg/L and control showed increased serum creatinine levels from 0.62 to 0.87 mg/dL, 0.64 to 0.86 mg/dL, 0.64 to 0.86 mg/dL, 0.61 to 0.83 mg/dL and 0.6 to 0.79 mg/dL respectively after 90 days treatment. The control group did not show a significant cause. Rat treated with different concentrations of Cylindrospermopsin, 2.5 μg/L, 2 μg/L, 1.5 μg/L, 2.3 μg/L and control showed gradually decreased of urine creatinine level from 34 to 55 mg/dL, 40 to 54 mg/dL, 38 to 54 mg/dL, 32 to 53 mg/dL and 43 to 54 mg/dL respectively after 90 days treatment and statistically significant (p&lt;0.05) difference was found between treated and control groups. The highest Aspartate Aminotransferase (AST) and Aspartate Alaninetransferase (ALT) levels were obtained from Cylindrospermopsin dose 2.5 μg/L at 90 days exposure. Cylindrospermopsin concentration in urine gradually increased from 1.5 to 2.32 μg/L, 0.9 to 1.7 μg/L, 0.8 to 1.1 μg/L and 0.4 to 1.5 μg/L when animal exposed to 2.5 μg/L, 2 μg/L, 1.5 μg/L and 2.3 μg/L concentrations of Cylindrospermopsin where control did not show. Thus, the result of present study show consumption of Cylindrospermopsin contaminated water may lead to liver and kidney injuries.Keywords: Cylindrospermopsin (CYN), Wistar rats, ELISA, AST, AL

A comparison of outcomes in ultrasonography guided versus landmark guided corticosteroid injection for the treatment of adhesive capsulitis

Background: Adhesive capsulitis is a debilitating disease in an otherwise healthy individual. Intra-articular corticosteroid injections offer a cost-effective, non-operative treatment option. However, it is currently unclear whether an ultrasound-guided injection relieves the symptoms of shoulder pain more effectively than if the injection was delivered landmark-guided. Methods: Eighty patients with adhesive capsulitis were randomized to two intervention groups - landmark guided and ultrasound (USG) guided. The functional status of the patients was documented prior to the intervention. Following allocation, the intra-articular steroid was administered either under USG guidance or following identification of the site of injection using landmarks. Follow-up was done on day 5, 3 weeks, 6 weeks, and 12 weeks post procedure to document the functional status. Results: The difference in visual analogue score (VAS) between the two arms was found to be statistically significant in favour of the ultrasound guided technique only on day 5 and day 21. On the other hand, the difference in disability of arm, shoulder, and hand (DASH) score between the 2 arms was found to be statistically significant in favour of the ultrasound guided technique on day 5, 21, 42 and 84. Finally, in our study, both shoulder flexion and abduction on day 84 achieved a statistically significant improvement, favouring the ultrasound guided arm. Conclusions: Ultrasound guided corticosteroid injections may offer modestly better short-term functional outcome and symptom relief when compared with landmark guided corticosteroids

Increased use of hypnotics in individuals with celiac disease: A nationwide case-control study

BACKGROUND: Although poor sleep is common in numerous gastrointestinal diseases, data are scarce on the risk of poor sleep in celiac disease. The objective of this study was to estimate the risk of repeated use of hypnotics among individuals with celiac disease as a proxy measure for poor sleep. METHODS: This is a nationwide case–control study including 2933 individuals with celiac disease and 14,571 matched controls from the general Swedish population. Poor sleep was defined as ≥2 prescriptions of hypnotics using prospective data from the National Prescribed Drug Register (data capture: July 2005-January 2008). We estimated odds ratios and hazard ratios for poor sleep before and after celiac disease diagnosis respectively. RESULTS: In this study, poor sleep was seen in 129/2933 individuals (4.4%) with celiac disease, as compared with 487/14,571 controls (3.3%) (odds ratio = 1.33; 95% CI = 1.08-1.62). Data restricted to sleep complaints starting ≥1 year before celiac disease diagnosis revealed largely unchanged risk estimates (odds ratio = 1.23; 95% CI = 0.88-1.71) as compared with the overall risk (odds ratio 1.33). The risk of poor sleep in celiac disease was essentially not influenced by adjustment for concomitant psychiatric comorbidity (n = 1744, adjusted odds ratio =1.26; 95% CI = 1.02-1.54) or restless legs syndrome (n = 108, adjusted odds ratio = 1.33; 95% CI = 1.08-1.63). Poor sleep was also more common after celiac disease diagnosis as compared with matched controls (hazard ratio = 1.36; 95% CI = 1.30-1.41). CONCLUSIONS: In conclusion, individuals with celiac disease suffer an increased risk of poor sleep, both before and after diagnosis. Although we cannot rule out that surveillance bias has contributed to our findings, our results are consistent with previous data suggesting that sleep complaints may be a manifestation of celiac disease. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12876-015-0236-z) contains supplementary material, which is available to authorized users

IP3 receptor isoforms differently regulate ER-mitochondrial contacts and local calcium transfer

Contact sites of endoplasmic reticulum (ER) and mitochondria locally convey calcium signals between the IP3 receptors (IP3R) and the mitochondrial calcium uniporter, and are central to cell survival. It remains unclear whether IP3Rs also have a structural role in contact formation and whether the different IP3R isoforms have redundant functions. Using an IP3R-deficient cell model rescued with each of the three IP3R isoforms and an array of super-resolution and ultrastructural approaches we demonstrate that IP3Rs are required for maintaining ER-mitochondrial contacts. This role is independent of calcium fluxes. We also show that, while each isoform can support contacts, type 2 IP3R is the most effective in delivering calcium to the mitochondria. Thus, these studies reveal a non-canonical, structural role for the IP3Rs and direct attention towards the type 2 IP3R that was previously neglected in the context of ER-mitochondrial calcium signaling

On the diurnal ranges of Sea Surface Temperature (SST) in the North Indian Ocean

This paper describes the variability in the diurnal range of SST in the north Indian Ocean using in situ measurements and tests the suitability of simple regression models in estimating the diurnal range. SST measurements obtained from 1556 drifting and 25 moored buoys were used to determine the diurnal range of SSTs. The magnitude of diurnal range of SST was highest in spring and lowest in summer monsoon. Except in spring, nearly 75-80% of the observations reported diurnal range below 0.5°C. The distributions of the magnitudes of diurnal warming across the three basins of north Indian Ocean (Arabian Sea, Bay of Bengal and Equatorial Indian Ocean) were similar except for the differences between the Arabian Sea and the other two basins during November-February (winter monsoon) and May. The magnitude of diurnal warming that depended on the location of temperature sensor below the water level varied with seasons. In spring, the magnitude of diurnal warming diminished drastically with the increase in the depth of temperature sensor. The diurnal range estimated using the drifting buoy data was higher than the diurnal range estimated using moored buoys fitted with temperature sensors at greater depths. A simple regression model based on the peak solar radiation and average wind speed was good enough to estimate the diurnal range of SST at ~1.0 m in the north Indian Ocean during most of the seasons except under low wind-high solar radiation conditions that occur mostly during spring. The additional information on the rate of precipitation is found to be redundant for the estimation of the magnitude of diurnal warming at those depths