Regulation of protein kinase C delta degration

Reversible phosphorylation of proteins is a vital means of regulation in signal transduction processes. The PKC family of lipid dependent serine/threonine kinases is known to play key roles in a whole range of cellular functions. To comprehend the diverse roles of the different family members, it is important to define the regulation of each PKC isoform. While activation of PKC isoforms is critical for their function in targeting downstream substrates/effectors, of equal importance is the desensitisation of the active kinases in determining their consequential effects. The overall aim of this thesis was to explore the mechanism(s) causing the downregulation and degradation of PKCδ. The degradation of this novel isoform of PKC has been examined in NIH 3T3 cells, in response to chronic activation by the phorbol ester TPA. A significant part of this study focused on the dephosphorylation of PKCδ as a primary event predisposing the enzyme to degradation. This was addressed in vivo using cell permeable inhibitors of protein phosphatases. In addition, in vitro studies using purified forms of PP1, PP2A and PP2C were carried out to investigate the potential effects of the different protein phosphatases towards PKCδ as a substrate. Consistent with other data, PP2A was identified as the putative protein phosphatase involved in the dephosphorylation of PKCδ. Interestingly, studies revealed that either nucleotide binding or substrate binding to PKCδ protected the enzyme from the action of phosphatases. To further investigate the requirement of phosphorylation/dephosphorylation during the downregulation process in cells, phosphorylation site mutants of PKCδ were generated and studied in transient transfection experiments. While these provided useful information, it was concluded that ectopic expression of PKCδ did not entirely represent the behaviour of the endogenous protein. Nevertheless, information from mutant analyses together with data from in vivo studies of endogenous PKCδ, in contrast to the previous speculation, provides strong evidence for the role of phosphorylation in the targeting of PKCδ to degradation via the ubiquitin/proteasome pathway

Prognosis of Disease that may Occur with Growing Age using Confabulation Based Algorithm

The enduring diagnosis of patient’s medical records might be useful to determine the causes that are responsible for a particular disease. So that, one can take early preventive measures to curtail the risk of diseases that may occur with the growing age. Consequently, this can enhance the life expectancy probability. Here, a new algorithm CMARM is proposed for analysis of symptoms in order to find out the disease that may occur frequently and rarely with growing age. It uses map reduce paradigm inspired by cognitive learning. It is concerned with acquisition of problem-solving skills, intelligence and conscious thought and uses prevailing knowledge to generate new rules. It has been evaluated over synthetic data sets collected from the health data repository. Since, CMARM requires one-time file access therefore, it is consistently faster and also consuming less memory space than the FP tree based algorithm

ANTIBACTERIAL ACTIVITY IN DIFFERENT EXTRACTS OF LANTANA CAMARA AGAINST ENTEROPATHOGENS

Medicinal use of plants is the oldest form of healthcare known to mankind. India has a rich profusion of medicinal plants and 75% of its folk population is still using herbal preparations. Present study reports about antimicrobial potentiality of Lantana camara as its leaf extracts exhibit good antimicrobial, fungicidal, insecticidal and nematicidal properties and the plant might be a novel source of antimicrobial drug. Two enteropathogens as (A) - B. subtilis (positive, obligate aerobic) and (B) - E. coli (Gram-negative, facultative anaerobic) were used for the present study. Four solvent phases viz, methanol, ethanol, acetone and aqueous were used for extraction of antimicrobial agent. The screening of antimicrobial property was done by well diffusion method. Leaves of Lantana camara showed excellent antibacterial activity in all the solvent phases used against both E. coli as well as B. subtilis. Lantana is effective against both the bacteria.The aqueous extract showed minimum ineffective antimicrobial activity against E. coli and B. subtilis, where as acetone phase showed maximum activity against B. subtilis and ethanol against E. coli as shown in the terms of maximum zone of inhibition

Expressional dynamics of minisatellite 33.15 tagged spermatozoal transcriptome in Bubalus bubalis

<p>Abstract</p> <p>Background</p> <p>Transcriptionally quiescent spermatozoa have been established to be a repository of mRNA coding for several functionally essential cellular proteins. This entourage of mRNA is envisaged to be involved in post-fertilization and early embryogenesis. Minisatellites tagged with mRNA transcripts have been implicated with gene organization, regulation and function. However, the organization and expression of the minisatellite tagged transcript diversity, particularly in spermatozoa, remains unclear.</p> <p>Results</p> <p>In the present study, we identified and characterized 12 mRNA transcripts from the spermatozoa of water buffalo <it>Bubalus bubalis </it>employing minisatellite associated sequence amplification (MASA) and a consensus sequence of 33.15 repeat loci. Of these 33.15 tagged transcripts, only one was found to be homologous to Bovine steroid 21-hydroxylase (P-450-c21) gene. Other ten transcripts showed significant similarity with various mRNAs or chromosomal contigs across the species. The remaining one construed to be novel since this was unreported in the database (NCBI GenBank). All these uncharacterized and known transcripts showed highest expression in testis and spermatozoa compared to that in somatic tissues and ovary. Of these 12 mRNA transcripts, 4 showed differential expression in the forebrain and hindbrain of buffalo. Moreover, genes corresponding to all the 33.15 tagged spermatozoal transcripts were found to be conserved across 13 other species analyzed.</p> <p>Conclusion</p> <p>Our results show MASA as an important tool to capture mRNA transcript diversity tagged with minisatellites in the spermatozoa. Comprehensive characterization of these transcripts is envisaged to augment our understanding on the genes involved in testicular functions and sustenance of a viable paternal genome during pre- and post- fertilization events and early stages of development. Prospects of this approach in genome analysis in general and comparative genomics in particular are highlighted.</p

Organization and differential expression of the GACA/GATA tagged somatic and spermatozoal transcriptomes in Buffalo Bubalus bubalis

<p>Abstract</p> <p>Background</p> <p>Simple sequence repeats (SSRs) of GACA/GATA have been implicated with differentiation of sex-chromosomes and speciation. However, the organization of these repeats within genomes and transcriptomes, even in the best characterized organisms including human, remains unclear. The main objective of this study was to explore the buffalo transcriptome for its association with GACA/GATA repeats, and study the structural organization and differential expression of the GACA/GATA repeat tagged transcripts. Moreover, the distribution of GACA and GATA repeats in the prokaryotic and eukaryotic genomes was studied to highlight their significance in genome evolution.</p> <p>Results</p> <p>We explored several genomes and transcriptomes, and observed total absence of these repeats in the prokaryotes, with their gradual accumulation in higher eukaryotes. Further, employing novel microsatellite associated sequence amplification (MASA) approach using varying length oligos based on GACA and GATA repeats; we identified and characterized 44 types of known and novel mRNA transcripts tagged with these repeats from different somatic tissues, gonads and spermatozoa of water buffalo <it>Bubalus bubalis</it>. GACA was found to be associated with higher number of transcripts compared to that with GATA. Exclusive presence of several GACA-tagged transcripts in a tissue or spermatozoa, and absence of the GATA-tagged ones in lung/heart highlights their tissue-specific significance. Of all the GACA/GATA tagged transcripts, ~30% demonstrated inter-tissue and/or tissue-spermatozoal sequence polymorphisms. Significantly, ~60% of the GACA-tagged and all the GATA-tagged transcripts showed highest or unique expression in the testis and/or spermatozoa. Moreover, ~75% GACA-tagged and all the GATA-tagged transcripts were found to be conserved across the species.</p> <p>Conclusion</p> <p>Present study is a pioneer attempt exploring GACA/GATA tagged transcriptome in any mammalian species highlighting their tissue, stage and species-specific expression profiles. Comparative analysis suggests the gradual accumulation of these repeats in the higher eukaryotes, and establishes the GACA richness of the buffalo transcriptome. This is envisaged to establish the roles of integral simple sequence repeats and tagged transcripts in gene expression or regulation.</p

Synthesis and characterization of new thiazole involving isatin for studying their antimicrobial activity

485-4871-(Substituted-1-ylmethyl)indoline-2,3-dione 1a-c have been synthesized from different types of secondary amine with isatin and formaldehyde in alcohol. The compound 1 have then been converted to the respective compound 2a-g (Z)3-(4-subsitutedphenylimino)-1-(substituted-1-ylmethyl)indolin-2-one by treatment with different types of primary amines. Interaction of compound 2 with thioglycolic acid and chloro acetyl chloride results in cyclization to give compound spiro isatin derivatives compounds 3a-g. The anti-microbial activity screening of novel spiro isatin substituted compounds have also been carried out

Unveiling the Incipient Caries by Quantitative Light-induced Fluorescence

Dental caries is one of the most prevalent chronic diseases of&nbsp;humans worldwide. When different stages of the disease are&nbsp;taken into account, from the initial to the clinically manifesting&nbsp;lesion, very few individuals are truly unaffected. Quantitative&nbsp;light-induced fluorescence (QLF) is a prominent diagnostic&nbsp;technique in dentistry. The purpose of this article is to review&nbsp;the effectiveness of QLF system in early detection of carious&nbsp;lesions as well as its future clinical application in dentistry. The&nbsp;review article has been prepared doing a literature review from&nbsp;the world wide web and PubMed/medline

POLLUTION MANAGEMENT THROUGH INTERNET OF THINGS: A SUBSTANTIAL SOLUTION FOR SOCIETY

Purpose of Study: In this age where population and industrialization have extended exponentially, contamination is a noteworthy emergency looked by humanity and essential measures ought to be taken to control it quickly. High levels of pollutants in the atmosphere, which are responsible for causing pollution that causes human heart problems, breathing and other all health problems to our society. This paper focused to provide a substantial solution to pollution in our society as a result of which major social, personal health and environmental pollution problems can be monitored and by taking effective steps they can be reduced. Methodology: The proposed framework is an IOT unit for handling these issues, including powerful use of the different sensor, to capture the estimations of various sorts of toxins in the earth and show insights on a site which can be gotten to by all clients for nothing. The research data is collected from the different sensors installed at different locations in our surroundings. After collecting data from sensors, a comparative study of data is carried out with the standards provided by the Central Pollution Control Board (CPCB). Results: The proposed system helps to reduce the pollution in our environment and as a result of which pollution can be reduced by taking major steps towards it. Application/Implications: The framework we are proposing is a propelled answer for observing the ecological contamination at a specific spot and make data available for each person. Novelty/originality of the study: Our proposed system ensures that pollution which ultimately causes health, environmental and other social problems can be tracked from time to time and each and every person in the society can track and check pollution level in their surroundings. &nbsp

Characterization of Smoc-1 uncovers two transcript variants showing differential tissue and age specific expression in Bubalus bubalis

<p>Abstract</p> <p>Background</p> <p>Secreted modular calcium binding protein-1 (<it>Smoc</it>-1) belongs to the BM-40 family which has been implicated with tissue remodeling, angiogenesis and bone mineralization. Besides its anticipated role in embryogenesis, <it>Smoc</it>-1 has been characterized only in a few mammalian species. We made use of the consensus sequence (5' CACCTCTCCACCTGCC 3') of 33.15 repeat loci to explore the buffalo transcriptome and uncovered the <it>Smoc</it>-1 transcript tagged with this repeat. The main objective of this study was to gain an insight into its structural and functional organization, and expressional status of <it>Smoc</it>-1 in water buffalo, <it>Bubalus bubalis</it>.</p> <p>Results</p> <p>We cloned and characterized the buffalo <it>Smoc</it>-1, including its copy number status, <it>in-vitro </it>protein expression, tissue & age specific transcription/translation, chromosomal mapping and localization to the basement membrane zone. Buffalo <it>Smoc</it>-1 was found to encode a secreted matricellular glycoprotein containing two EF-hand calcium binding motifs homologous to that of BM-40/SPARC family. In buffalo, this single copy gene consisted of 12 exons and was mapped onto the acrocentric chromosome 11. Though this gene was found to be evolutionarily conserved, the buffalo <it>Smoc</it>-1 showed conspicuous nucleotide/amino acid changes altering its secondary structure compared to that in other mammals. <it>In silico </it>analysis of the <it>Smoc</it>-1 proposed its glycoprotein nature with a calcium dependent conformation. Further, we unveiled two transcript variants of this gene, varying in their 3'UTR lengths but both coding for identical protein(s). <it>Smoc</it>-1 evinced highest expression of both the variants in liver and modest to negligible in other tissues. The relative expression of variant-02 was markedly higher compared to that of variant-01 in all the tissues examined. Moreover, expression of <it>Smoc</it>-1, though modest during the early ages, was conspicuously enhanced after 1 year and remained consistently higher during the entire life span of buffalo with gradual increment in expression of variant-02. Immunohistochemically, <it>Smoc</it>-1 was localized in the basement membrane zones and extracellular matrices of various tissues.</p> <p>Conclusion</p> <p>These data added to our understandings about the tissue, age and species specific functions of the <it>Smoc</it>-1. It also enabled us to demonstrate varying expression of the two transcript variants of <it>Smoc</it>-1 amongst different somatic tissues/gonads and ages, in spite of their identical coding frames. Pursuance of these variants for their roles in various disease phenotypes such as hepatocellular carcinoma and angiogenesis is envisaged to establish broader biological significance of this gene.</p

Liver function profile in thalassemic children receiving multiple blood transfusions

Background: Hepatic dysfunction is a frequent manifestation in thalassemic patients receiving multiple blood transfusions (BTs) as a part of treatment. Objective: The objective of the study was to study the liver function profile in thalassemic children and its correlation with the age of initiation of transfusion therapy. Materials and Methods: This cross-sectional study was done among 32 thalassemic patients in the age group of 1–18 years visiting a tertiary care hospital regularly for BTs at the Department of Pediatrics at the tertiary hospital of North India. Liver function tests (LFTs) were done in all thalassemic patients included total bilirubin, liver enzymes (serum glutamic oxaloacetic transaminase [SGOT], serum glutamic pyruvic transaminase [SGPT], and alkaline phosphatase [ALP]), total protein, serum albumin, serum ferritin, hepatitis B surface antigen (HBsAg), and anti-hepatitis C virus. The age of initiation of BT was also recorded. Derangement in LFTs and correlation between the age of initiation of transfusion therapy and derangement of liver function were studied. Results: Out of 32 patients, only 7 (21.87%) had normal LFT values. A total of 17 (53.12%) had increased SGOT, 15 (46.87%) had increased SGPT, and 25 (78.12%) had increased bilirubin levels. Total protein and serum albumin were below normal in 5 (15.65%) and 3 (9.3%) patients, respectively. ALP was increased in 24 (75%) patients. Majority of the patients (43.75%) had serum ferritin between 2000 and 2999 ng/ml. Only two patients had significantly deranged LFTs. No patient was positive for HBsAg. However, we did not find a significant correlation between age of initiation of transfusion therapy and derangement of liver enzymes in these patients. Conclusion: If thalassemic patients are given properly tested blood and regular chelation therapy, liver function remains normal. Immunization against hepatitis B and testing of blood bags is recommended. It is also recommended that LFT should be done regularly at 3 months interval to detect any abnormality