Effect of heparin on high glucose induced proliferation and expression of matrix metalloproteinases in normal human mesangial cells

Background The pathogenesis of diabetic nephropathy (DN) is a complex pathophysiological process.Its precise mechanism is not fully known. In recent years it has been recognized that synthesis of various extracelluar matrix (ECM) components may increase, and that degradation of ECM may decrease in DN. It was reported heparin could inhibit mesangial cells proliferation in vitro. The main aim of this study is to explore whether heparin inhibits proliferation of mesangial cells grown in high glucose concentration and to measure the effect of heparin on matrix metalloproteinases (MMPs) expression in mesangial cells. Methods The medium contained either low glucose (5 mmol/L) or high glucose (25 mmol/L). The concentrations of heparin in the culture medium were 0, 25, 50,100, 200 or 400 μg/mL. A metabolic (WST-1) assay was used to measure mesangial cell proliferation and Western blot analysis was used to measure MMPs expression of mesangial cells. Results Normal human mesangial cell (NHMC) proliferation was higher in high glucose (HG) medium than in low glucose (LG) medium. They showed a 1.93 fold expansion after 72 h in high glucose in contrast to a 1.63 fold expansion in low glucose. In the presence of heparin, mesangial cells proliferation was inhibited, which was more obvious at high glucose concentrations than at low glucose concentrations. In high glucose, with heparin concentration of 50, 100, 200 and 400 μg/mL, the mesangial cells showed a 0. 61 fold, 0.52 fold, 0.52 fold and 0.41 fold reductions in cell number compared to cells grown without heparin. In low glucose, only concentrations of 200 μg/mL and 400 μg/mL showed reduction in cell number, namely 0.54 fold and 0.45 fold, when compared to cells grown without heparin. In Western blot analysis,MMP1, MMP2, MMP3 and MMP9 was expressed by mesangial cells expressed in both high and low glucose concentrations, which was more prominent in high glucose medium. Incubation of heparin further increased expression of MMP1, MMP2, MMP3 and MMP9. Conclusions This study suggests that glucose can accelerate mesangial cell proliferation while heparin can reduce proliferation, being more obvious at high glucose concentrations. Higher glucose concentrations led to increased MMP expression, which may take part in the regulation of mesangial matrix synthesis and degradation. Addition of heparin resulted in a corresponding increase in MMP expression, most notably at high glucose concentrations, indicating a potentially renoprotective role in DN.     

Protein Expression of Angiotensin-Converting Enzyme 2 (ACE2) is Upregulated in Brains with Alzheimer’s Disease

Alzheimer’s disease is a chronic neurodegenerative disorder and represents the main cause of dementia globally. Currently, the world is suffering from the coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a virus that uses angiotensin-converting enzyme 2 (ACE2) as a receptor to enter the host cells. In COVID-19, neurological manifestations have been reported to occur. The present study demonstrates that the protein expression level of ACE2 is upregulated in the brain of patients with Alzheimer’s disease. The increased ACE2 expression is not age-dependent, suggesting the direct relationship between Alzheimer’s disease and ACE2 expression. Oxidative stress has been implicated in the pathogenesis of Alzheimer’s disease, and brains with the disease examined in this study also exhibited higher carbonylated proteins, as well as an increased thiol oxidation state of peroxiredoxin 6 (Prx6). A moderate positive correlation was found between the increased ACE2 protein expression and oxidative stress in brains with Alzheimer’s disease. In summary, the present study reveals the relationships between Alzheimer’s disease and ACE2, the receptor for SARS-CoV-2. These results suggest the importance of carefully monitoring patients with both Alzheimer’s disease and COVID-19 in order to identify higher viral loads in the brain and long-term adverse neurological consequences.     

Generation of single-chain Fvs against detergent-solubilized recombinant antigens with a simple coating procedure

Antigen coating on polystyrene is prevented by detergent. We present here a simple procedure to coat detergent-solubilized antigen for subsequent panning selection of single-chain Fv (scFv), the target antigen of which was the hepatitis C virus (HCV) non-structural protein (NS) 4B, an integral membrane protein.     

Development of Motor Stator with Rectangular‐Wire Lap Winding and an Automatic Process for Its Production

Aimed at providing small high‐power motors with excellent efficiency, a motor stator with rectangular‐wire lap windings and an automatic process for its production were developed. The structure of the windings, namely, two continuous “α‐shaped” coils, and the coil‐production method (involving simultaneous coil forming and coil insertion) enable mass production of this stator. The slot‐fill rate for a prototype stator was 80.5%, and the total height of the coil ends was 60 mm to 72 mm. On the basis of the ratio of the sectional areas of two types of the motors, its efficiency was estimated to be 3.7% higher than the efficiencies of motor stators with round‐wire concentric windings (at a rotation speed of 1000/min). In addition, the number of coil parts and the number of welding points were greatly reduced compared to those of a stator with segmented‐coil wave windings.     

Tracer monitored titrations: measurement of dissolved oxygen

The tracer monitored titration (TMT) technique is evaluated for measurement of dissolved oxygen. The TMT developed in this work uses a simple apparatus consisting of a low-precision pump for titrant delivery and an optical detector based on a white LED and two photodiodes with interference filters. It is shown that the classic Winkler method can be made free of routine volumetric and gravimetric measurements by application of TMT theory, which allows tracking the amounts of titrant and sample using a chemical tracer. The measurement precision of the prototype setup was 0.3% RSD.     

The High-Resolution X-Ray Microcalorimeter Spectrometer, SXS, on Astro-H

The science and an overview of the Soft X-ray Spectrometer onboard the STRO-H mission are presented. The SXS consists of X-ray focusing mirrors and a microcalorimeter array and is developed by international collaboration lead by JAXA and NASA with European participation. The detector is a 6×6 format microcalorimeter array operated at a cryogenic temperature of 50 mK and covers a 3′×3′ field of view of the X-ray telescope of 5.6 m focal length. We expect an energy resolution better than 7 eV (FWHM, requirement) with a goal of 4 eV. The effective area of the instrument will be 225 cm² at 7 keV; by a factor of about two larger than that of the X-ray microcalorimeter on board Suzaku. One of the main scientific objectives of the SXS is to investigate turbulent and/or macroscopic motions of hot gas in clusters of galaxies.     

Characterization of autoinducer 2 signal in Eikenella corrodens and its role in biofilm formation

Quorum sensing (QS) is a process by which bacteria communicate using secreted chemical signaling molecules called autoinducers (AIs). By this process, many bacterial species modulate the expression of a wide variety of physiological functions in response to changes in population density. In this study, the periodontal pathogen Eikenella corrodens was observed to secrete type 2 signaling molecules. An ortholog of luxS, the gene required for AI-2 synthesis in Vibrio harveyi, was isolated from the E. corrodens genome. A V. harveyi bioassay showed luxS functionality in E. corrodens and the ability of luxS to complement the luxS-negative phenotype of Escherichia coli DH5alpha. AI activity was detected in the supernatant, and the maximum expression of AI-2 was observed during the late exponential phase. To determine the potential role of luxS in the colonization processes, an E. corrodens luxS mutant was constructed and tested for its capacity to form an in vitro biofilm on a polystyrene surface. The biofilm forming efficiency of the luxS mutant was approximately 1.3-fold greater than that of the wild type. These data suggest that a LuxS-dependent signal plays a role in the biofilm formation by E. corrodens.     

Phase-separation behavior in a binary mixture fluid layer subjected to a vertical temperature gradient

The phase separation behavior in a layer of an isobutyric acid (IBA)-water binary mixture was observed from the bottom and side of the layer, where the layer was subjected to a vertical temperature gradient spanning the critical temperature. As a result, the following conclusions were obtained. (1) Patterns characterized by a polygonal cell including one droplet, similar to those observed by Assenheimer et al. (Physica A 208 (1994) 373), were formed in a relatively thick liquid layer, e.g., thick, in a limited range of the temperature gradient, although the patterns ultimately disappeared. (2) The growth and extinction rates of the phase-separated droplets increased with ΔT/d, and increased particularly rapidly at high ΔT/d. (3) The formation of the pattern of polygonal cells each with a droplet is considered to be due to thermocapillary motion along the droplet. (4) The concentration-driven buoyancy convection affects the growth rate of the phase-separated droplets. (5) The extinction rate of the phase-separated droplets at the late stage is affected by the thermocapillary motion along the droplet surface.