Ranitidine induces inhibition and structural changes in sucrase

Ranitidine is an antagonist of histamine-2 (H(2)) receptor. It is employed to treat peptic ulcer and other conditions in which gastric acidity must be reduced. Sucrase is a hydrolytic enzyme that catalyzes the breakdown of sucrose to its monomer content. A liquid of yeast sucrase was developed for treatment of congenital sucrase-isomaltase deficiency (CSID) in human. In this study, the effect of ranitidine on yeast sucrase activity was investigated. Our results showed that ranitidine binds to sucrase and inhibits the enzyme in a noncompetitive manner. The K(i) and IC(50) values were measured to be about 2.3 and 2.2 mM, respectively. Fluorescence measurement showed conformational changes after binding of ranitidine to the enzyme. The fluorescence spectra showed that ranitidine could bind to both free enzyme and enzyme-substrate complex, which was accompanied with reduction of emission intensity and red shift production.     

A comparative study on the B12N12, Al12N12, B12P12 and Al12P12 fullerene-like cages

The stability, geometry and electronic structure of the title nanoclusters were compared by using density functional theory (DFT) calculations. Their electrical property analysis showed that the relative magnitude of the HOMO-LUMO gaps (eV) that are average values from the calculated results with five different DFT functionals is as follows: B12N12(7:02)>Al12N12(4.09)>B12P12(3.80)>Al12P12(3.39). Computing the standard enthalpy and the Gibbs free energy of formation, it was found that the B(12)N(12) structure is thermodynamically stable at 298 K and 1 atmosphere of pressure, while the Al(12)N(12) structure may be stable at low temperatures. Due to positive values of change of enthalpy and entropy of formation for both the B(12)P(12) and Al(12)P(12) clusters, it seems that their formation from the consisting atoms is not spontaneous at any temperature.     

Mitochondrial fission triggered by hyperglycemia is mediated by ROCK1 activation in podocytes and endothelial cells

Several lines of evidence suggest that mitochondrial dysfunction plays a critical role in the pathogenesis of microvascular complications of diabetes, including diabetic nephropathy. However, the signaling pathways by which hyperglycemia leads to mitochondrial dysfunction are not fully understood. Here we examined the role of Rho-associated coiled coil-containing protein kinase 1 (ROCK1) on mitochondrial dynamics by generating two diabetic mouse models with targeted deletions of ROCK1 and an inducible podocyte-specific knockin mouse expressing a constitutively active (cA) mutant of ROCK1. Our findings suggest that ROCK1 mediates hyperglycemia-induced mitochondrial fission by promoting dynamin-related protein-1 (Drp1) recruitment to the mitochondria. Deletion of ROCK1 in diabetic mice prevented mitochondrial fission, whereas podocyte-specific cA-ROCK1 mice exhibited increased mitochondrial fission. Importantly, we found that ROCK1 triggers mitochondrial fission by phosphorylating Drp1 at serine 600 residue. These findings provide insights into the unexpected role of ROCK1 in a signaling cascade that regulates mitochondrial dynamics.     

Distribution of Metals (Cu,Zn, Pb,and Cd) in Sediments of the Anzali Lagoon,North Iran

Concentrations of four metals (Cu, Zn, Pb, and Cd) in the sediments of the Anzali Lagoon in the northern part of Iran were determined to evaluate the level of contamination and spatial distribution. The sediments were collected from 21 locations in the lagoon. At each lagoon site a core, 60 cm long, was taken. The ranges of the measured concentrations in the sediments are as follows: 17–140 mg kg^?1 for Cu, 20–113 mg kg^?1 for Zn, 1–37 mg kg^?1 for Pb and 0.1–3.5 mg kg^?1 for Cd in surficial (0-20 cm) and 16–87 mg kg^?1 for Cu, 28.5–118 mg kg^?1 for Zn, 3–20 mg kg^?1 for Pb and 0.1–3.5 mg kg^?1 for Cd in deep (40–60 cm) sediments. The results of the geoaccumulation index (I_geo) show that Cd causes moderate to heavy pollution in most of the study area. Environmental risk evaluation showed that the pollution in the Anzali Lagoon is moderate to considerable and the ranking of the contaminants followed the order: Cd > Cu > Pb > Zn. Some locations present severe pollution by metals depending on the sources, of which sewage outlets and phosphate fertilizers are the main sources of contaminants to the area.     

Assessment of the Parameters Influencing Microbial Calcite Precipitation in Injection Experiments Using Taguchi Methodology

Soil improvement is one of the major concerns in civil engineering. Therefore, a variety of approaches have been employed for different soil types. The loose granular soils and sediments have always imposed challenges due to their low strength and bearing capacity as well as presenting difficulties in drilling and excavation. Biomediated soil improvement, i.e., utilizing some bacteria to precipitate calcite on soil particles, has recently been introduced as a novel link of biotechnology and civil engineering to improve the problematic soils. Biogrout as a branch of biomediated soil improvement is based upon microbial calcium carbonate precipitation (MICP). In the present study, the Taguchi method with the aim of optimizing the process was utilized to design the experiments (DOE). A standard L9 orthogonal array with four parameters comprising bacterial cell concentration, molar concentration ratio of nutrient solution, curing time, and flow rate, each assigned to three levels, was selected. In this regard, soil samples were stabilized in sandy soil columns. Two-phase injections were conducted by injecting the bacterium Sporosarcina pasteurii PTCC 1642 in the first phase and nutrient in the second phase. Specimens were subjected to an unconfined compressive strength (UCS) test. ANOVA pointed out how effectual each parameter was. The most effective parameter was curing time, which accounted for 45.97% of the overall variance of the experimental data followed by bacterial cell concentration (22.01%), nutrient strength (19.98%), and flow rate (12.04%). Predicted UCS values for the optimum condition were validated in a confirmation test. Indeed, the UCS of the soil increased from 85 kPa in the control sample to 930 kPa for the optimally treated specimen. It was concluded that rather than curing time, the other parameters are almost equally influential in the applied injection procedure.     

Fusion proteins comprising the catalytic domain of mutansucrase and a starch-binding domain can alter the morphology of amylose-free potato starch granules during biosynthesis

It has been shown previously that mutan can be co-synthesized with starch when a truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The mutan seemed to adhere to the isolated starch granules, but it was not incorporated in the starch granules. In this study, GtfICAT was fused to the N- or C-terminus of a starch-binding domain (SBD). These constructs were introduced into two genetically different potato backgrounds (cv. Kardal and amf), in order to bring GtfICAT in more intimate contact with growing starch granules, and to facilitate the incorporation of mutan polymers in starch. Fusion proteins of the appropriate size were evidenced in starch granules, particularly in the amf background. The starches from the various GtfICAT/SBD transformants seemed to contain less mutan than those from transformants with GtfICAT alone, suggesting that the appended SBD might inhibit the activity of GtfICAT in the engineered fusion proteins. Scanning electron microscopy showed that expression of SBD-GtfICAT resulted in alterations of granule morphology in both genetic backgrounds. Surprisingly, the amf starches containing SBD-GtfICAT had a spongeous appearance, i.e., the granule surface contained many small holes and grooves, suggesting that this fusion protein can interfere with the lateral interactions of amylopectin sidechains. No differences in physico-chemical properties of the transgenic starches were observed. Our results show that expression of granule-bound and “soluble” GtfICAT can affect starch biosynthesis differently.     

Oxytocin increases generosity in humans

Human beings routinely help strangers at costs to themselves. Sometimes the help offered is generous-offering more than the other expects. The proximate mechanisms supporting generosity are not well-understood, but several lines of research suggest a role for empathy. In this study, participants were infused with 40 IU oxytocin (OT) or placebo and engaged in a blinded, one-shot decision on how to split a sum of money with a stranger that could be rejected. Those on OT were 80% more generous than those given a placebo. OT had no effect on a unilateral monetary transfer task dissociating generosity from altruism. OT and altruism together predicted almost half the interpersonal variation in generosity. Notably, OT had twofold larger impact on generosity compared to altruism. This indicates that generosity is associated with both altruism as well as an emotional identification with another person.     

Peripheral Immunization Blocks Lethal Actions of Vesicular Stomatitis Virus within the Brain

Vesicular stomatitis virus (VSV) is the prototype virus for 75 or more negative-strand RNA viruses in the rhabdovirus family. Some of these viruses, including VSV, can cause neurological impairment or death upon brain infection. VSV has shown promise in the prevention and treatment of disease as a vaccine vector and an oncolytic virus, but infection of the brain remains a concern. Three VSV variants, the wild-type-related VSV-G/GFP and two attenuated viruses, VSV-CT1 and VSV-CT9-M51, were compared for neuroinvasiveness and neuromorbidity. In nonimmunized mice, direct VSV-G/GFP injection into the brain invariably resulted in lethal encephalitis; in contrast, partial survival was seen after direct injection of the attenuated VSV strains. In addition, both attenuated VSV strains showed significantly reduced neuroinvasiveness after intranasal inoculation of young postnatal day 16 mice. Of the three tested variants, VSV-CT9-M51 generated the lowest degree of neuropathology. Despite its attenuated state, peripheral inoculations of VSV-CT9-M51 targeted and killed human glioblastoma implanted into the mouse brain. Importantly, we show here that intranasal or intramuscular immunization prevents the lethal effects of subsequent VSV-G/GFP, VSV-CT1, and VSV-CT9-M51 injections into the brain. These results indicate that attenuated recombinant viruses show reduced neurovirulence and that peripheral immunization blocks the lethal actions of all VSVs tested.The brain occupies a special niche in viral immunity, and due to a number of mechanisms, viruses in the periphery generally do not enter the brain. However, the same mechanisms that give the brain a special protected status can also impede an immune response against intracerebral infection by viruses. Although many negative-strand RNA viruses can be tolerated peripherally, central nervous system (CNS) infection with vesicular stomatitis virus (VSV), rabies virus, measles virus, influenza virus, and others (14, 24, 28, 30, 34) can be fatal for rodents and for humans. Peripheral immunization does protect the brain from virus infections, but in most studies, it does so by eliminating viruses before they penetrate the blood-brain barrier and enter the brain (4, 25, 29, 38). In contrast, the set of experiments described here address the question of whether peripheral immunization can block the lethal consequences of direct VSV infections within the brain. When injected into the brain, VSV can cause permanent neurological dysfunction in rodents or primates (19, 28) or lethal encephalitis (11, 15). VSV can also enter the brain from the periphery along a cranial nerve, for instance, the olfactory nerve after intranasal administration, and can subsequently spread from the olfactory system to other regions of the brain (24, 36).Recombinant VSVs have shown promise in two respects: VSV can serve as a robust vaccine vector (26, 27, 16) and as a potent oncolytic virus against a variety of peripheral (1, 3, 10, 33) or CNS (9, 18, 22, 39, 40) tumors. A number of studies have shown the protective effects of peripheral immunization with VSV on peripheral viral infections (12, 13). In contrast, the effect of peripheral immunization on viral infections within the brain has received considerably less attention.Both as a vaccine vector and as an oncolytic virus, VSV infection of normal brain cells remains a concern. The set of experiments presented here addressed the primary question of whether peripheral immunization can protect the brain from subsequent direct exposure to VSV. A secondary question was whether recombinant VSVs with an attenuated phenotype in culture would also show reduced neurovirulence in the brain.VSV is an enveloped negative-strand RNA virus, and its 11.2-kb genome encodes five viral proteins (N, P, M, G, and L). VSV is a nonhuman pathogen that can cause a typically self-limiting disease in livestock with flu-like symptoms (20). Limiting factors of VSV for clinical use are its neurotropic properties and the still little understood potential of the brain to fight off a potential infection (5, 6, 15). The brain is largely protected from virus entry through the blood-brain barrier. Mice do not show signs of CNS infection after peripheral VSV application. In contrast, VSV with direct access to the CNS, either experimentally through direct injection or through the intranasal path, can spread through the brain, resulting in encephalitis with high mortality in mice. VSV spread through the brain after intranasal application is age dependent, with mature mice showing little or no spread beyond the olfactory nerve compared to young mice, which succumb to widespread viral infection throughout the brain (19, 36). Peripheral VSV infection triggers fast and effective upregulation of interferon-inducible genes, followed by induction of both the cellular and humoral branches of the systemic immune system.The extent of VSV pathogenesis in the brain is determined by the replicative efficacy of the virus and the efficiency of the host immune response in curbing the infection. Modification of either of these components can alter the course and extent of CNS damage. In the current work, we used a dual viral mutation that enhances the host innate cellular immune response (VSV-M51) and truncates the VSV-G cytoplasmic tail from 29 to 9 amino acids (VSV-CT9). Another VSV with a VSV-G truncation to 1 cytoplasmic amino acid (VSV-CT1), resulting in viral attenuation in vitro and in vivo, was also used (23, 31).Little is known about the extent to which the adaptive immune response can influence VSV within the brain. Here, we show that peripheral VSV immunization prior to intracerebral inoculation prevented lethal encephalitis in adult mice of the strongly attenuated VSV variants, VSV-CT9-M51 and VSV-CT1, as well as a wild-type VSV bearing a green fluorescent protein (GFP) reporter.     

Intracellular IL-15 controls mast cell survival

The regulation of mast cell activities and survival is a central issue in inflammatory immune responses. Here, we have investigated the role of mouse interleukin-15, a pro-inflammatory and pleiotropic cytokine, in the control of mast cell survival and homeostasis. We report that aged IL-15−/− mice show a reduced number of peritoneal mast cells compared to WT mice. Furthermore, IL-15 deficiency in bone marrow derived mouse mast cells (BMMCs) results in increased susceptibility to apoptosis mediated by growth factor deprivation and A-SMase-treatment. IL-15−/− BMMCs show a constitutive stronger mRNA and protein expression as well as enzymatic activity of the members of the mitochondrial apoptotic pathways including acidic lysosomal aspartate protease cathepsin D (CTSD), endogenous acid sphingomyelinase (A-SMase), caspase-3 and -7 compared to wild type (WT) BMMCs. Furthermore, IL-15−/− BMMCs constitutively generate more A-SMase-derived ceramide than WT controls and display a decreased expression of pro-survival sphingosin-1-phosphate (SPP) both in cytosol and membrane cell fractions. Furthermore, pre-treatment of mast cells with imipramine or pepstatin A, inhibitors of the intracellular acid sphingomyelinase and cathepsin D pathways respectively, increases survival in IL-15−/− BMMCs. These findings suggest that intracellular IL-15 is a key regulator of pathways controlling primary mouse mast cell homeostasis.