Intra-generational protein malnutrition impairs temporal astrogenesis in rat brain

The lack of information on astrogenesis following stressor effect, notwithstanding the imperative roles of astroglia in normal physiology and pathophysiology, incited us to assess temporal astrogenesis and astrocyte density in an intra-generational protein malnutrition (PMN) rat model. Standard immunohistochemical procedures for glial lineage markers and their intensity measurements, and qRT-PCR studies, were performed to reveal the spatio-temporal origin and density of astrocytes. Reduced A2B5+ glia restricted precursor population in ventricles and caused poor dissemination to cortex at embryonic days (E)11-14, and low BLBP+ secondary radial glia in the subventricular zone (SVZ) of E16 low protein (LP) brains reflect compromised progenitor pooling. Contrary to large-sized BLBP+ gliospheres in high protein (HP) brains at E16, small gliospheres and discrete BLBP+ cells in LP brains evidence loss of colonization and low proliferative potential. Delayed emergence of GFAP expression, precocious astrocyte maturation and significantly reduced astrocyte number suggest impaired temporal and compromised astrogenesis within LP-F1 brains. Our findings of protein deprivation induced impairments in temporal astrogenesis, compromised density and astrocytic dysfunction, strengthen the hypothesis of astrocytes as possible drivers of neurodevelopmental disorders. This study may increase our understanding of stressor-associated brain development, opening up windows for effective therapeutic interventions against debilitating neurodevelopmental disorders

A novel antimicrobial peptide derived from modified N-terminal domain of bovine lactoferrin: Design, synthesis, activity against multidrug-resistant bacteria and Candida

AbstractLactoferrin (LF) is believed to contribute to the host's defense against microbial infections. This work focuses on the antibacterial and antifungal activities of a designed peptide, L10 (WFRKQLKW) by modifying the first eight N-terminal residues of bovine LF by selective homologous substitution of amino acids on the basis of hydrophobicity, L10 has shown potent antibacterial and antifungal properties against clinically isolated extended spectrum beta lactamases (ESBL), producing gram-negative bacteria as well as Candida strains with minimal inhibitory concentrations (MIC) ranging from 1 to 8μg/mL and 6.5μg/mL, respectively. The peptide was found to be least hemolytic at a concentration of 800μg/mL. Interaction with lipopolysaccharide (LPS) and lipid A (LA) suggests that the peptide targets the membrane of gram-negative bacteria. The membrane interactive nature of the peptide, both antibacterial and antifungal, was further confirmed by visual observations employing electron microscopy. Further analyses, by means of propidium iodide based flow cytometry, also supported the membrane permeabilization of Candida cells. The peptide was also found to possess anti-inflammatory properties, by virtue of its ability to inhibit cyclooxygenase-2 (COX-2). L10 therefore emerges as a potential therapeutic remedial solution for infections caused by ESBL positive, gram-negative bacteria and multidrug-resistant (MDR) fungal strains, on account of its multifunctional activities. This study may open up new approach to develop and design novel antimicrobials

Molecular epidemiology and complete genome characterization of H1N1pdm virus from India.

BACKGROUND: Influenza A virus is one of world's major uncontrolled pathogen, causing seasonal epidemic as well as global pandemic. This was evidenced by recent emergence and continued prevalent 2009 swine origin pandemic H1N1 Influenza A virus, provoking first true pandemic in the past 40 years. In the course of its evolution, the virus acquired many mutations and multiple unidentified molecular determinants are likely responsible for the ability of the 2009 H1N1 virus to cause increased disease severity in humans. Availability of limited data on complete genome hampers the continuous monitoring of this type of events. Outbreaks with considerable morbidity and mortality have been reported from all parts of the country. METHODS/RESULTS: Considering a large number of clinical cases of infection complete genome based sequence characterization of Indian H1N1pdm virus and their phylogenetic analysis with respect to circulating global viruses was undertaken, to reveal the phylodynamic pattern of H1N1pdm virus in India from 2009-2011. The Clade VII was observed as a major circulating clade in phylogenetic analysis. Selection pressure analysis revealed 18 positively selected sites in major surface proteins of H1N1pdm virus. CONCLUSIONS: This study clearly revealed that clade VII has been identified as recent circulating clade in India as well globally. Few clade VII specific well identified markers undergone positive selection during virus evolution. Continuous monitoring of the H1N1pdm virus is warranted to track of the virus evolution and further transmission. This study will serve as a baseline data for future surveillance and also for development of suitable therapeutics

Phylogenetic tree of concatenated whole genome of representative global H1N1pdm viruses including four Indian viruses sequenced in this study generated by Bayesian method.

<p>Each strain is abbreviated with virus subtype, country of origin, strain name and year of isolation in parenthesis. Scale bar indicates number of nucleotide substitutions per site. The Indian isolates sequenced in this study are highlighted in different font in clade VII. Other Indian isolates are highlighted by solid diamond in respective clades. Each clade is defined by long branch and nodes supported by high Bayesian posterior probability (BPP) values (100%).</p

Confirmation of H1N1pdm virus.

<p>(<b>A</b>) Microscopic photograph of healthy and Influenza A (H1N1pdm) virus infected Madin Darby Canine Kidney Cells. (<b>B</b>) Immunofluorescence assay. (<b>C</b>) Haemagglutination assay. (<b>D</b>) WHO CDC Real-Time PCR amplification. Real time amplification curve of positive clinical samples showing amplification of all four probes.</p

Details of the genome sequences of the H1N1pdm virus isolates retrieved and investigated in the whole genome and complete HA gene based phylogenetic analysis in this study.

<p>Note: The isolates in bold font are sequenced in this study.</p

(A) Monthly trend of pandemic influenza A H1N1 and seasonal flu reported from September 2010 to December 2011.

<p>(B) Age and sex wise distribution of the influenza A H1N1 2010–2011 suspected ILI cases.</p

Description of major/important non-conservative and clade specific amino acid substitutions among the four Indian H1N1pdm virus (sequenced in this study) compared to prototype H1N1pdm strain (California/04/2009) and other Indian (A/Pune/NIV6447/2009) virus strain (sequenced previously).

<p>Note: Major non conservative changes involving basic to acidic amino acid are written in bold font and also underlined; The hydrophobic to hydrophilic amino acid substitutions and vice-versa are written in bold font. The substitutions involving charged residues to uncharged residues; cyclic to acyclic and vice versa are written in italics. The clade specific substitutions (NP:V100I; NA:V106I; NS1:I123V; HA:S220T, I338V) are written in normal font.</p>*<p>The residue position for the HA is the numbering considered inclusive of signal peptide.</p

Selection pressure analysis of HA protein (566 codons); NA protein (469 codons), M1 Protein (252 codons) and M2 Protein (97 codons) of H1N1pdm virus using SLAC, FEL,REL,MEME and FUBAR methods. (www.datamonkey.org).

<p>Note: The sites found under positive selection by atleast two methods are shown*. Site present in B-cell epitope region are highlighted in bold font.</p>*<p> <b>Significance value.</b></p><p>SLAC P value–0.5.</p><p>FEL P value- 0.25.</p><p>REL Bayes factor- 50.</p><p>MEME P value- 0.1.</p><p>FUBAR Posterior probability- 0.9.</p