Exploring the influence of adipokines on neuronal function in Alzheimer’s disease

Alzheimer’s disease (AD) is one of the major causes of dementia. Alzheimer’s disease is an irreversible, slow neurodegenerative disease leading to memory and language impairment and amyloid-β plaques and intraneuronal neurofibrillary tangles are the characteristic pathology of AD. Studies have shown that obesity is major risk factor in Alzheimer’s disease. While obesity related modulation has been suggested to influence AD pathogenesis, the molecular mechanisms involved are still unclear. Recent research has shown that adipocytes can actively secrete adipokines and these adipokines may possess the ability to influence AD pathology, but it remains poorly understood how adipokines impact neural functioning. The current study was focused on the potential neuroprotective effect of adipokines on the neurons during hydrogen peroxide (H2O2) induced oxidative stress and cell death. In this study, human neuroblastoma SHSY5Y cells were differentiated using retinoic acid to demonstrate neuronal-like characteristics which was confirmed by immunocytochemistry (ICC). The pre-adipocytes 3T3-L1 cells, were chemically differentiated to exhibit adipocyte-like characteristics (determined by Oil-Red-O staining). To understand the potential rescue effect of conditioned media from adipocyte-like cells on neuronal-like cells, and commercially available adipokines (chemerin, leptin and resistin) on neuronal-like cells and effect of chemerin on human skin fibroblast cells derived from AD and non-AD donors, MTT assay was performed in the presence and absence of H2O2 induced oxidative stress. Conclusively, the conditioned media and commercial adipokines showed protective effects on the neuronal-like cells (*p<0.05, **p<0.01, ***p<0.001) in comparison to the non-neuronal cells and chemerin showed no protective effect in AD patient derived cells under H2O2 induced oxidative stress in comparison to the non-AD donors derived fibroblast cells. Neuroprotection was not observed in the obesity mimicked induced oxidative stress condition. Our findings suggested that the adipokines might play vital role in neuronal-like cellular metabolic protection against oxidative stress and warrants further investigation of the role of adipokines dysregulation in Alzheimer’s disease

Functional Characterization of EngAMS, a P-Loop GTPase of Mycobacterium smegmatis

Bacterial P-loop GTPases belong to a family of proteins that selectively hydrolyze a small molecule guanosine tri-phosphate (GTP) to guanosine di-phosphate (GDP) and inorganic phosphate, and regulate several essential cellular activities such as cell division, chromosomal segregation and ribosomal assembly. A comparative genome sequence analysis of different mycobacterial species indicates the presence of multiple P-loop GTPases that exhibit highly conserved motifs. However, an exact function of most of these GTPases in mycobacteria remains elusive. In the present study we characterized the function of a P-loop GTPase in mycobacteria by employing an EngA homologue from Mycobacterium smegmatis, encoded by an open reading frame, designated as MSMEG_3738. Amino acid sequence alignment and phylogenetic analysis suggest that MSMEG_3738 (termed as EngAMS) is highly conserved in mycobacteria. Homology modeling of EngAMS reveals a cloverleaf structure comprising of α/β fold typical to EngA family of GTPases. Recombinant EngAMS purified from E. coli exhibits a GTP hydrolysis activity which is inhibited by the presence of GDP. Interestingly, the EngAMS protein is co-eluted with 16S and 23S ribosomal RNA during purification and exhibits association with 30S, 50S and 70S ribosomal subunits. Further studies demonstrate that GTP is essential for interaction of EngAMS with 50S subunit of ribosome and specifically C-terminal domains of EngAMS are required to facilitate this interaction. Moreover, EngAMS devoid of N-terminal region interacts well with 50S even in the absence of GTP, indicating a regulatory role of the N-terminal domain in EngAMS-50S interaction

SPARC 2018 Internationalisation and collaboration : Salford postgraduate annual research conference book of abstracts

Welcome to the Book of Abstracts for the 2018 SPARC conference. This year we not only celebrate the work of our PGRs but also the launch of our Doctoral School, which makes this year’s conference extra special. Once again we have received a tremendous contribution from our postgraduate research community; with over 100 presenters, the conference truly showcases a vibrant PGR community at Salford. These abstracts provide a taster of the research strengths of their works, and provide delegates with a reference point for networking and initiating critical debate. With such wide-ranging topics being showcased, we encourage you to take up this great opportunity to engage with researchers working in different subject areas from your own. To meet global challenges, high impact research inevitably requires interdisciplinary collaboration. This is recognised by all major research funders. Therefore engaging with the work of others and forging collaborations across subject areas is an essential skill for the next generation of researchers

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

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Not AvailableRESULTS OF A STUDY CONDUCTED AT DEHRADUN DURING 1986-91 ON RUNOFF AND SOIL LOSS ON 1.86 X 22.13 m RUNOFF PLOTS AT 8% SLOPE HAVE BEEN PRESENTED. METHODOLOGY TO ESTIMATE USLE- PARAMETERS IS DESCRIBED. THE AVERAGE RAINFALL EROSION INDEX ( R) FOR DEHRADUN IS 1048 AND SOIL ERODIBILITY FACTOR (K) IS 124 kg/ha /unit OF R. COWPEA COVER EFFECTIVELY REDUCED RUNOFF AND SOIL LOSS BY PROVIDING COVER NEAR THE LAND SURFACE AND AT EARLY STAGES OF CROP GROWTH. SIMILARLY , MANDUA AND JHINGORA , THE NATIVE HILL REGION CROPS, ARE EFFICIENT IN RESPECT OF SOIL AND WATER CONSERVATION. THE CROP MANAGEMENT FACTOR 'C' FOR COWPEA WAS 0.31 AND FOR MANDUA AND JHINGORA WAS 0.17 AND 0.18 , RESPECTIVELY. , WHICH IS CLOSE TO THE 'C' FACTOR FOR GRASS SPECIES. THE INFORMATION ON CANOPY OF DIFFERENT CROPS AND THEIR COMBINATION IS ALSO PRESENTED.Not Availabl

Detection of <i>Mycoplasma </i>species in cell culture by PCR and RFLP based method: Effect of BM-cyclin to cure infections

<b>Purpose:</b> A two-stage nested polymerase chain reaction (PCR) assay system was described that amplifies the 16S-23S rRNA spacer region sequences of <i> Mycoplasma</i> and <i> Acholeplasma</i> infections in cell cultures and virus stocks. <b> Methods:</b> Established cell lines and virus stocks were screened for the presence of <i> Mycoplasma</i> by using nested PCR using two sets of outer and inner primers, amplifies 16S-23S rRNA. PCR and restriction fragment length polymorphism (RFLP) assay was used to detect and identify most of the species-specific <i> Mycoplasmas</i> involved in cell cultures and virus stock contaminants. Infected cultures detected by PCR-RFLP were further treated with BM-cyclin (5 &#956;g/mL) and passaged for three times and tested for <i> Mycoplasma</i> infections by PCR-RFLP. <b> Results:</b> <i> Mycoplasma pirum</i> and <i> Mycoplasma orale</i> infections were detected by nested PCR. Species specificity was identified by using RFLP of <i> Vsp</i> I, <i> Cla</i> I and <i> Hin</i> dIII restriction enzymes. <i> Mycoplasma</i> infections were cured by treatment with BM-cyclin. This was further confirmed by non-amplification of PCR amplimers in BM-cyclin treated vs. non-treated cultures. <b> Conclusions:</b> Regular monitoring of cell cultures for <i> Mycoplasma</i> infections and identification of species-specific Mollicutes will identify the source of contaminations. This approach can be used for quality control of the biological reagents used in cell culture and virology laboratories