Structural Basis for Dual-Inhibition Mechanism of a Non-Classical Kazal-Type Serine Protease Inhibitor from Horseshoe Crab in Complex with Subtilisin

Serine proteases play a crucial role in host-pathogen interactions. In the innate immune system of invertebrates, multi-domain protease inhibitors are important for the regulation of host-pathogen interactions and antimicrobial activities. Serine protease inhibitors, 9.3-kDa CrSPI isoforms 1 and 2, have been identified from the hepatopancreas of the horseshoe crab, Carcinoscorpius rotundicauda. The CrSPIs were biochemically active, especially CrSPI-1, which potently inhibited subtilisin (Ki = 1.43 nM). CrSPI has been grouped with the non-classical Kazal-type inhibitors due to its unusual cysteine distribution. Here we report the crystal structure of CrSPI-1 in complex with subtilisin at 2.6 Å resolution and the results of biophysical interaction studies. The CrSPI-1 molecule has two domains arranged in an extended conformation. These two domains act as heads that independently interact with two separate subtilisin molecules, resulting in the inhibition of subtilisin activity at a ratio of 1:2 (inhibitor to protease). Each subtilisin molecule interacts with the reactive site loop from each domain of CrSPI-1 through a standard canonical binding mode and forms a single ternary complex. In addition, we propose the substrate preferences of each domain of CrSPI-1. Domain 2 is specific towards the bacterial protease subtilisin, while domain 1 is likely to interact with the host protease, Furin. Elucidation of the structure of the CrSPI-1: subtilisin (1∶2) ternary complex increases our understanding of host-pathogen interactions in the innate immune system at the molecular level and provides new strategies for immunomodulation

An update on vitamin B12-related gene polymorphisms and B12 status.

Vitamin B12 is an essential micronutrient in humans needed for health maintenance. Deficiency of vitamin B12 has been linked to dietary, environmental and genetic factors. Evidence for the genetic basis of vitamin B12 status is poorly understood. However, advancements in genomic techniques have increased the knowledge-base of the genetics of vitamin B12 status. Based on the candidate gene and genome-wide association (GWA) studies, associations between genetic loci in several genes involved in vitamin B12 metabolism have been identified. The objective of this literature review was to identify and discuss reports of associations between single-nucleotide polymorphisms (SNPs) in vitamin B12 pathway genes and their influence on the circulating levels of vitamin B12. Relevant articles were obtained through a literature search on PubMed through to May 2017. An article was included if it examined an association of a SNP with serum or plasma vitamin B12 concentration. Beta coefficients and odds ratios were used to describe the strength of an association, and a  < 0.05 was considered as statistically significant. Two reviewers independently evaluated the eligibility for the inclusion criteria and extracted the data. From 23 studies which fulfilled the selection criteria, 16 studies identified SNPs that showed statistically significant associations with vitamin B12 concentrations. Fifty-nine vitamin B12-related gene polymorphisms associated with vitamin B12 status were identified in total, from the following populations: African American, Brazilian, Canadian, Chinese, Danish, English, European ancestry, Icelandic, Indian, Italian, Latino, Northern Irish, Portuguese and residents of the USA. Overall, the data analyzed suggests that ethnic-specific associations are involved in the genetic determination of vitamin B12 concentrations. However, despite recent success in genetic studies, the majority of identified genes that could explain variation in vitamin B12 concentrations were from Caucasian populations. Further research utilizing larger sample sizes of non-Caucasian populations is necessary in order to better understand these ethnic-specific associations

Galectin-3 alters the lateral mobility and clustering of beta 1-integrin receptors

Glycoprotein receptors are influenced by myriad intermolecular interactions at the cell surface. Specific glycan structures may interact with endogenous lectins that enforce or disrupt receptor-receptor interactions. Glycoproteins bound by multivalent lectins may form extended oligomers or lattices, altering the lateral mobility of the receptor and influencing its function through endocytosis or changes in activation. In this study, we have examined the interaction of Galectin-3 (Gal-3), a human lectin, with adhesion receptors. We measured the effect of recombinant Gal-3 added exogenously on the lateral mobility of the alpha 5 beta 1 integrin on HeLa cells. Using single-particle tracking (SPT) we detected increased lateral mobility of the integrin in the presence of Gal-3, while its truncated C-terminal domain (Gal-3C) showed only minor reductions in lateral mobility. Treatment of cells with Gal-3 increased beta 1-integrin mediated migration with no apparent changes in viability. In contrast, Gal-3C decreased both cell migration and viability. Fluorescence microscopy allowed us to confirm that exogenous Gal-3 resulted in reorganization of the integrin into larger clusters. We used a proteomics analysis to confirm that cells expressed endogenous Gal-3, and found that addition of competitive oligosaccharide ligands for the lectin altered the lateral mobility of the integrin. Together, our results are consistent with a Gal-3-integrin lattice model of binding and confirm that the lateral mobility of integrins is natively regulated, in part, by galectins

Effect of plasma immersion ion implantation of nitrogen on the wear and corrosion behavior of 316LVM stainless steel

Low energy plasma immersion ion implantation (PIII) of nitrogen ions on vacuum arc melted 316L (316LVM) austenitic stainless steel has been carried out at three different temperatures namely, 250 °C, 380 °C and 500 °C. X-ray diffraction analysis indicated that PIII results in the formation mixed iron-nitrides along with expanded austenite phase at all temperatures. Microhardness measurements revealed a significant increase in hardness after PIII treatment. Corrosion resistance in 3.5% NaCl increases when implanted for 3 h. The passive current density seems to increase with treatment temperature. Wear measurements carried out using a pin-on-disc machine show an increase in wear resistance with rise in treatment temperature.© Elsevie

Effect of alloyed molybdenum on corrosion behavior of plasma immersion nitrogen ion implanted austenitic stainless steel

Plasma immersion ion implantation (PIII) of nitrogen has been performed on two austenitic stainless steels (with and without Mo addition) at three different temperatures namely; 250, 380 and 500 degrees C for 3 h. Grazing angle X-ray diffraction (GXRD) was carried out on the surface of the steels (both PIII treated and untreated). GXRD results suggest that PIII is more effective in Mo containing stainless steel (SS). The electrochemical corrosion studies examined through both by DC polarization and EIS technique in 3.5 wt.% NaCl reveals that, 3 h N-implantation at 250 and 380 degrees C improves the corrosion and pitting resistance of both the austenitic stainless steels under investigation. The effect N implantation on pitting resistance is seen more in the presence of Mo, than when it is not present in the SS. It is further emphasized that the pitting resistance of the alloys significantly deteriorates, when they are implanted at 500 degrees C. (C) 2013 Elsevier Ltd. All rights reserved

The Water and Sanitation Scenario in Indian Metropolitan Cities: Resources and Management in Delhi, Calcutta, Chennai, Mumbai

This paper reviews the urban water and sanitation scenario in metropolitan cities. Section 1 focuses on the institutional and organizational structure of the service providers by looking at the level of technical and managerial decentralization reached in the four cities (Calcutta, Chennai, Delhi, Mumbai) Section 2 focuses on the question of property rights and the debate on usage conflicts in order to fill the gap for the future demand. Section 3 revisits the question of reforms that were launched in the 1990s for all infrastructure sectors. Section 4 and 5 concentrate on two directions the sector could look at for changes: the development of conservation based strategy and the need for a more participative approach by involving the civil society. This would mean a paradigm shift for the sector. Indeed, demand side solutions are rarely considered and the problem of water supply is mostly addressed by the supply angle.Water supply, Sanitation, Chennai, Mumbai, Delhi, Calcutta, Resources management; Urban India; Potable water; Water demand; Metropolitan cities; Water harvesting; Sewage water treatment

Policies are never implemented, but negotiated : analyzing integration of policies in managing water resources in the Indian Himalayas using a Bayesian network

Author name used in this publication: Saravanan. V.SubramanianAuthor name used in this publication: David, Ip2009-2010 > Academic research: refereed > Publication in refereed journalAccepted ManuscriptPublishe