Network topology analysis of essential genes interactome of Helicobacter pylori to explore novel therapeutic targets

The Helicobacter pylori chronic colonization produces a wide range of gastric diseases in the gastric mucosa by abetting inflammation. Amidst coevolution and reorganization of its metabolism with humans, it has become difficult still imperative to understand and prevent its growth. This study focus to explore functional insights into identification of hub proteins/genes by aggregating the behavior of genes connected in a protein-protein interaction (PPI) network. We have constructed a PPI network of 123 essential genes along with 1213 interactions in H. pylori 26695. The degree and other centrality measures analysis assist in identifying the important hub nodes, which are top-ranked proteins. A total of nine proteins (recA, guaA, dnaK, rpsB, rplQ, rpmA, rpmC, rpmF, and rpsE) were obtained with high degree (k), betweenness centrality (BC) value. Gene ontology analysis reveals 8, 5 and 3 GO terms correspond to biological processes, cellular components and molecular function respectively. Gene complexes of hypothetical proteins (HPs) were related to aminoacyl-tRNA biosynthesis, biosynthesis of secondary metabolites, bacterial secretion system and protein export. The MCODE analysis revealed that protein from module M1, M3 and M6 include the proteins which have highest degree and BC values. It is noteworthy to mention that the bifunctional GMP synthase/glutamine amidotransferase protein (guaA), molecular chaperon (dnaK), recombinase A (recA) constitute as hub proteins. As a result, these genes are considered as network hub nodes that might be used as therapeutic targets. Our analysis affords a detailed understanding of the molecular process and pathways regulated by the essential genes in H. pylori 26695. © 2021 Elsevier Lt

Investigation on stabilization of ladle furnace slag with different additives

Ladle furnace slag disintegrates into fine powder during cooling due to phase transformations of di-calcium silicate. This creates an adverse impact on working conditions and the environment by dust generation. In this paper, a short overview on different studies to overcome the disintegration problem is provided. An attempt was also made to study the effects of several different additives and their mixtures on disintegration of slag. Phase equilibria calculations were carried out for some additives using FactSage® to understand the phase changes in the slag. Based on the phase equilibria calculations and literature data, initial laboratory experiments were conducted at 1650 °C with different additives such as boric acid, aluminium, and fly ash. Slag samples were analyzed with X-ray fluorescence and X-ray powder diffraction for chemical and phase analysis before and after treatment. The disintegration of slag can be prevented either by addition of 0.5 wt% or more of boric acid or 9 wt% of aluminium or 6 wt% of fly ash or 4–8 wt% fly ash along with 0.125–0.25 wt% of boric acid in slag. Based on the optimized conditions, industrial trials were conducted

LO STENT INTERNO-ESTERNO COME STRUMENTO DIAGNOSTICO PER L\u2019IDENTIFICAZIONE PRECOCE DELLA GLOMERULOSCLEROSI FOCALE SEGMENTALE NEI RICEVENTI IL TRAPIANTO DI RENE

Introduzione: La glomerulosclerosi focale segmentale (GSFS) \ue8 una delle pi\uf9 frequenti malattie glomerualri ricorrenti dopo trapianto renale (15-100%). Questo studio valuta l\u2019utilit\ue0 dello stent interno-esterno come mezzo diagnostico per monitorare la proteinuria dopo trapianto di rene nei pazienti con FSGS presso il nostro centro. Materiali e metodi: E\u2019 stata effettuata un\u2019analisi retrospettiva di 11 riceventi di trapianto di rene (7 da donatore viventi consanguinei, 4 da donatore cadavere) tra il 2000 e il 2008 con FSGS comprovata dalla biopsia che sono stati sottoposti al posizionamento di stent interno-esterno intraoperatorio. Tutti i pazienti ebbero una diuresi 65 500 ml. Un sondino da nutrizione pediatrica 6 Fr \ue8 stato usato come stent e posizionato nell\u2019uretere del rene trapiantato ed steriorizzato attraverso la vescica alla parete addominale. Fu effettuato l\u2019esame delle urine delle 12 ore giornalmente per lo studio del rapporto proteine/creatinina. Lo stent fu rimosso alla terza settimana. Risultati: L\u2019et\ue0 media alla diagnosi di GSFS fu di 42 anni (3-57). L\u2019intervallo medio dalla diagnosi istologica di GSFS all\u2019insufficienza renale terminale fu di 3,9 anni (1-8). Due pazienti ebbero GSFS ricorrente.Un paziente present\uf2 simultaneamente rigetto acuto di grado 1 b provato dalla biopsia. Gli altri pazienti ebbero proteinuria (> 3g) in prima giornata post-operatoria suggerendo la plasmaferesi. La biopsia dimostr\uf2 GSFS ricorrente. La proteinuria perdur\uf2 un anno e mezzo per poi sfociare in insufficienza renale terminale. Al ri-trapianto fu posizionato uno stent interno-esterno e non venne dimostrata alcuna recidiva. Al follow-up medio di 2 anni non c\u2019\ue8 ricorrenza di GSFS in 9 pazienti. La creatinina media ha valori di 1,44 mg/dl (0,9-2,8 mg/dl) e il rapporto medio proteine/creatinina \ue8 0,96 (0,1-2). Conclusioni: il posizionamento dello stent interno esterno nel rene trapiantato \ue8 utile come strumento diagnostico per valutare la ricorrenza precoce di GSFS dopo trapianto di rene a scopo preventivo nei pazienti con diuresi nel rene nativo. Gli outcomes a lungo termine sono migliorati con l\u2019identificazione e il trattament

Comparative efficacy of different methods in the generation of 12S particles from 146S particles of FMDV serotype A and their detection using serotype (146S) specific monoclonal antibodies

Foot-and-mouth disease [FMD] is a highly infectious and contagious viral disease of domestic and wild cloven hoofed animals. The disease is controlled by vaccination using inactivated vaccine as one of the important options. The integrity of 146S plays an important role in the efficacy of the vaccine. The currently applied methods like density gradient to check the integrity of the 146Sparticles are besotted with certain isadvantages. Therefore, we generated monoclonal antibodies (mAbs) specific to 146S of FMDV serotype A [Indianstrain]. To test these mAbs for their specificity to 146S, conversion of 146S into 12S is a must. Normally, three methods like strong [1N HCl] and weak acid [Na2HPO4] methods; and heat method are followed in the conversion of 146S into 12S. Upon comparison of these methods in the present study, consistent results were obtained using heat method at 560C and 600C each for half an hour and one hour. However, the former two methods were very inconsistent in yielding 12S from 146S particle due to slight variation in the pH. Hence, we optimized heat method for efficient conversion of 146S particle to 12S particle of FMDV serotype A [A/INDIA/40/00], an Indian vaccine strain. The results are ascertained applying serotype specific [146S] monoclonal antibody based double antibody sandwich ELISA. However, the method needs to be evaluated using more number of 146S samples

Evaluation of Different Methods for Conversion of Whole Virion Particle (146S) of FMDV into 12S Subunits and Application in Characterization of Monoclonal Antibodies

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven hoofed animals caused by FMD virus classified under genus Apthovirus and Picornaviridae family. Monoclonal antibodies (mAb) to FMDV provides best tool for the development of reliable diagnostics and there is a need for the development of mAbs that specifically recognizeeither 146S or 12S for the quality control analysis of FMDV vaccines. In the present study, we developed mAbs specific to FMDV serotype O and characterized their specificity in recognition of 146S or 12S particles. Three different methods were evaluated for the conversion of 146S into 12S subunits that includes heat method, strong acid and mild acid methods for testing the reactivity of mAbs with these antigens by double antibody sandwich ELISA. Mild heating of 146S antigens at 560C for 1hour and treatment of 146S antigen with weak acid (0.5M NaH2PO4, pH 4.5 - 5.5) resulted in complete conversion of 146S into 12S. Where as in strong acid method (1N HCl with pH below 4.5), 146S treated with 1N HCl resulted in loss of antigenicity of 12S subunits that reflected in absence of reactivity in ELISA. This may mislead to consider the mAb as 146S specific. This study revealed that mAb binding epitopes of all monoclonals are commonly shared among 146S and 12S antigens. Further it helps in understanding the specificity of mAbs to 146S and 12S particles which will be useful for application of mAbs in different diagnostic assays

Serum profiling of leptospirosis patients to investigate proteomic alterations

Leptospirosis is a zoonotic infectious disease of tropical, subtropical and temperate zones, which is caused by the pathogenic spirochetes of genus Leptospira. Although this zoonosis is generally not considered as fatal, the pathogen can eventually cause severe infection with septic shock, multi-organ failure and lethal pulmonary hemorrhages leading to mortality. In this study, we have performed a proteomic analysis of serum samples from leptospirosis patients (n=6), febrile controls (falciparum malaria) (n=8) and healthy subjects (n=18) to obtain an insight about disease pathogenesis and host immune responses in leptospiral infections. 2DE and 2D-DIGE analysis in combination with MALDI-TOF/TOF MS revealed differential expression of 22 serum proteins in leptospirosis patients compared to the healthy controls. Among the identified differentially expressed proteins, 8 candidates exhibited different trends compared to the febrile controls. Functional analysis suggested the involvement of differentially expressed proteins in vital physiological pathways, including acute phase response, complement and coagulation cascades and hemostasis. This is the first report of analysis of human serum proteome alterations in leptospirosis patients, which revealed several differentially expressed proteins, including alpha-1-antitrypsin, vitronectin, ceruloplasmin, G-protein signaling regulator, apolipoprotein A-IV, which have not been reported in context of leptospirosis previously. This study will enhance our understanding about leptospirosis pathogenesis and provide a glimpse of host immunological responses. Additionally, a few differentially expressed proteins identified in this study may further be investigated as diagnostic or prognostic serum biomarkers for leptospirosis. This article is part of a Special Issue entitled: Integrated omics. (C) 2012 Elsevier B.V. All rights reserved