Suresh Kumar Jatawa

Occult hepatitis C virus elicits mitochondrial oxidative stress in lymphocytes and triggers PI3-kinase-mediated DNA damage response

Occult hepatitis C viral infection (OHCI) is a newly reported pathological entity associated with increased risk of developing hepatocellular carcinoma and lymphoproliferative disorders. Although hepatocytes are the primary sites of viral replication, hepatitis C virus is potentially lymphotropic, invading and propagating in cells of the immune system. Lymphocytes, the extrahepatic viral reservoirs, are differentially implicated in the occult and the active forms of the disease. This study aimed to elucidate the implications of mitochondrial oxidative stress on the immune pathophysiological mechanisms of OHCI. We herein report that OHCI induces mitochondrial oxidative stress, leading to DNA double-strand breaks and elicitation of a phosphoinositol 3-kinase-mediated cellular response in peripheral blood lymphocytes. Compared to controls, OHCI subjects showed higher accumulation of pATM, pATR, γH2AX, and p-p53, along with active recruitment of repair proteins (Mre11, Rad50, and Nbs1) and altered mitochondrial DNA content. Increased mitochondrial membrane depolarization and circulating nucleosome levels along with chromatid-type aberrations and decreased T-cell proliferative index observed in the OHCI group further indicated that this damage might lead to Bax-triggered mitochondria-mediated cellular apoptosis. Together our results provide the mechanistic underpinnings of mitochondrial dysfunction in OHCI, a previously unknown paradigm, for explaining the immune pathogenesis in a redox-dependent manner.

Induction of genomic instability in cultured human colon epithelial cells following exposure to isocyanates

The toxic response of cultured human colon epithelial‐FHC cells to methyl isocyanate was investigated with regard to genomic instability. Qualitative and quantitative assessments of the extent of phosphorylation of DNA damage signaling factors such as ATM, γH2AX and p53, was increased in treated cells compared to controls. At the same time, many treated cells were arrested at the G2/M phase of the cell cycle, and had an elevated apoptotic index and increased inflammatory cytokine levels. Cytogenetic analyses revealed varied chromosomal anomalies, with abnormal expression of pericentrin protein. Analysis through ISSR PCR demonstrated increased microsatellite instability. The results imply that isocyanates can cause genomic instability in colonocytes.

In silico docking of methyl isocyanate (MIC) and its hydrolytic product (1, 3-dimethylurea) shows significant interaction with DNA Methyltransferase 1 suggests cancer risk in Bhopal-Gas- Tragedy survivors.

DNA methyltransferase 1 (DNMT1) is a relatively large protein family responsible for maintenance of normal methylation, cell growth and survival in mammals. Toxic industrial chemical exposure associated methylation misregulation has been shown to have epigenetic influence. Such misregulation could effectively contribute to cancer development and progression. Methyl isocyanate (MIC) is a noxious industrial chemical used extensively in the production of carbamate pesticides. We here applied an in silico molecular docking approach to study the interaction of MIC with diverse domains of DNMT1, to predict cancer risk in the Bhopal population exposed to MIC during 1984. For the first time, we investigated the interaction of MIC and its hydrolytic product (1,3-dimethylurea) with DNMT1 interacting (such as DMAP1, RFTS, and CXXC) and catalytic (SAM, SAH, and Sinefungin) domains using computer simulations. The results of the present study showed a potential interaction of MIC and 1,3-dimethylurea with these domains. Obviously, strong binding of MIC with DNMT1 interrupting normal methylation will lead to epigenetic alterations in the exposed humans. We suggest therefore that the MIC- exposed individuals surviving after 1984 disaster have excess risk of cancer, which can be attributed to alterations in their epigenome. Our findings will help in better understanding the underlying epigenetic mechanisms in humans exposed to MIC.

Genetic mutations in cell cycle regulatory proteins, microsatellite loci and their association with gallbladder carcinogenesis in the population that exposed to methyl isocyanate

T novel lipases TALipA, TALipB and TALipC from opportunistic pathogen Trichosporon asahii MSR54 were isolated, sequenced and heterologously expressed in E.coli and Pichia pastoris X33. Both glycosylated and non-glycosylated proteins were characterized. These lipases are biotechnologically important because all the three lipases are enantioselective. Out of the three, TALipC has an asprich domain and shows enantioinversion in different solvents. It shows Mg activation and calcium inhibition. The role of these lipases was studied in growth and biofilm formation by knock-down strategy. The RNAi silencing and agrobacterium mediated transformation for developing knock-down strains of T. asahii was standardized in our laboratory for the first time. The lipase knock-down strains lost the property of yeast to fungal transition and predominantly retained the yeast stage. Detailed structural and functional comparisons of these lipases will be highlighted.M (miRNAs) are small noncoding RNA gene products about 18 to 24-nt-long. Mainly they negatively regulate protein expression of specific mRNA by either translational inhibition or mRNAs degradation. Mature miRNAs are the results of successive processing of primary transcripts (pri-miRNAs) which are facilitated by two RNase III enzymes, Drosha and Dicer with a characteristic hairpin secondary structure. Reporting of the aberrant expressions of miRNAs have been done in respect to different human diseases mainly in concern with cancer viz., lung cancer, brain cancer, chronic lymphocytic leukemia, neurodegenerative diseases etc. It is well established fact that miRNAs defines the expression patterns of the tissuespecificity. Neurodegenerative disease states the physiological nature of the cell, expression of miR-29a, miR-29b-1 and miR-9 are found significantly down regulated in Alzheimer’s disease. Biopsies of tumor from Parkinson’s disease patients exposed association of miR-30b, miR-30c and miR-26a. Phylogenetic analysis of miRNA of Alzheimer and Huntington diseases gives insight into evolutionary relationship and reveals regulation of Mir-22, Mir 29a and mir-128-1 in both diseases. Mir-22 shows down-regulation in Parkinson disease and Alzheimer disease while mir-128-1 and mir-29a shows the difference in their regulation pattern. In Alzheimer disease mir-128-1 is up regulated while in Huntington disease it is down regulated. Mir-29a is up regulated in Huntington disease while in Alzheimer disease, it is down regulated. These findings illustrates the importance of miRNA research in neurodegenerative diseases with reference to novel targets identification which can give a better lead in concern to protective or prophylective approaches.C cancer is one of the main causes of mortality worldwide and several anti-cancer drugs are known for ages but still new therapeutical research is ongoing to target the anti-cancer drug at tumour specific site with lesser side effect. We set out to investigate comparative role of biomolecules like luteolin, and lupeol in cancer therapy via its anti-angiogenic mechanism. We have compared the role of anti-angiogenic drug via chick chorioallantoic membrane assay, colon cancer cell line proliferation ability and cell migration assay. Upon comparing the anti-angiogenic ability luteolin followed by lupeol has been found to be the best in inhibiting angiogenesis, cell proliferation ability and cell migration ability. Colon tumour has been analyzed for FISH and immunohistochemistry. Expression of VEGF, EGFR, FGFR has been analyzed. VEGF and FGFR are overexpressed in colon tumour compared to EGFR. Hence effect of biomolecule luteolin is analyzed in colon cancer cell line via western blotting for these signaling pathway dissections. Therefore, it can be concluded that luteolin is the best screened anti-cancerous drug which targets several downstream targets of VEGF and FGF pathway.A is a branch of Ayurveda, the ancient 5,000 year old Indian system of Vedic healthcare. Loosely translated, “ayur” in Sanskrit is for health, “veda” means wisdom, and “vastra” is cloth or clothing. Ayurvastra clothing is made from organic cotton fabric that has been permeated with special herbs and oils that promote health and cure special diseases depending upon the blends of embedded herbs and oils. There is a whole spectrum of colors that can be obtained from a multitude of plants, insects and fungi; and these have been used across the centuries to dye textiles, color artefacts, pattern and color our skin/hair, and even color the food we eat. Various natural dyes and herbal products are found to be self-healing when they are applied on the fabrics. Ayurvastra cloth is used by Ayurveda Health Clinics in the treatment of a broad range of diseases such as diabetes, skin infections, eczema, psoriasis, hypertension and high blood pressure, asthma, arthritis, rheumatism and even some forms of cancer. Ayurvastra clothing is believed to help restore balance within the body’s system and strengthen the immune system. Indeed, the natural dyes are better products, simply because they do not contain chemicals harmful to health. Ayurvastra technology is beneficial to the mankind in curing diseases with the help of the medicinal characteristics associated with various herbs. It is environmental-friendly process and the usage of the cloth is based on the principle of touch. Today, the revival of this ancient technology is gaining importance all over the world. Other cultures and regions of the world are expressing their growing interest in more traditional and natural healthcare systems that are based upon restoring balance and health through natural methods rather than through Western medicines.The bacterial strains, Alcaligenes feacalis and Alcaligenes sp. DN25 which were isolated from the phyllosphere of four ornamental plant species, Ixora chinensis, Ervatamia divaricata, Hibiscus rosa-sinensis and Amaranthus cruentus in five highly polluted sites in Sri Lanka, showed the highest phenanthrene and naphthalene degradation ability. Transformation and plasmid curing results of them revealed, naphthalene and phenanthrene degradation ability of these bacterial strains were plasmid encoded character. The occurrence of naphthalene specific (nahR and nahU) genes and phenanthrene specific (phnAc and phnG) genes of these catabolic plasmids were analyzed by PCR using degenerate primers. According to the amplification results, plasmids of Alcaligenes faecalis and Alcaligenes sp. DN25 harbour nahR, nahU and phnG genes but, lack of phnAc gene. RFLP and sequence data of nahU and nahR amplicons revealed, both of these genes were homologous to these two bacterial strains. But, phnG gene of two phenanthrene and naphthalene degrading phyllosphere bacterial strains was coexistence as two distinct copies of alleles.B is caused by Fusarium fujikuroi (Nirenberg) is emerging as a serious disease of rice in India. Elongation and rotting of seedlings are distinguished symptoms of bakanae disease. Sixty three isolates of Fusarium fujikuroi were isolated from symptomatic diseased plants collected from Jammu and Kashmir, Punjab, Haryana, Uttar Pradesh, Uttarakhand and Bihar. Identities of the isolates were confirmed through translation elongation factor 1-α (tef-1α) gene sequence based identification. Out of 12 Universal Rice Primers (URP) used in the study, six primers effectively produced polymorphic bands in all the isolates. URP 17R and URP 6R primer produced 93% polymorphic bands. Mating type of the population was identified based on MAT-1 and MAT-2 region universal primers (GFmat1a, GFmat1b, GFmat2c and GFmat2d) for Gibberella fujikuroi. Among the 63 isolates 18 (28.57%) were identified as MAT-1 and 45 (71.42%) as MAT-2. Effective population number Ne(mt) for mating type was 89% of the count (total population). Since both the mating types were not present in equal frequencies in the F. fujikuroi population, asexual reproduction was likely to occur in the field isolates. However, the occurrence of both the mating types in the F. fujikuroi implied that the population members are capable of sexual reproduction. Therefore, it is important that programs to develop cultivars with the bakanae disease resistance considering other factors including the environmental conditions and variability of the pathogen in the area of intended cultivation.O the past few decades, microfluidic technology has laid the foundation for developing advanced and importantly more physiologically-realistic in vitro models for manipulating and analyzing cellular behaviors at both the tissue-specific and single-cell levels. Microfluidic devices have attracted much attention for cell-based studies because of their ability to use small quantities of cells and reagents, precisely control spatial and temporal microenvironments and facilitate highresolution visualization of cellular events in real-time. In this presentation, I will demonstrate the microfluidic-based analytical models, micro-fabrication methods, experimental validations and on-chip monitoring of cellular responses for a diverse set of biological studies namely nano toxicological analysis, synapse formation, surfactant secretion in the lung alveoli and neural activity in the brain. Multi compartmented microfluidic platforms are fabricated for analyzing cytotoxic effects of quantum dots under physiologically relevant conditions. Notably, such devices show their potential applications in continuous real-time monitoring of neuronal processes involved in synapse formation. The role of flow-induced shear stress on the mechanisms regulating surfactant secretion in pulmonary alveolar type II epithelial cells is also investigated using microfluidic models. Furthermore, a microfluidic model of neural tissue that closely mimics the realistic and complex three-dimensional (3D) brain’s cyto-architecture is developed for measuring activity of the neurons in a 3D environment following site-specific chemical treatment of a brain-like neural network. Altogether, such integrated microfluidic platforms that combine bio-realistic growth conditions and optical access hold tremendous potential for high-throughput toxicity testing, tissue engineering and establishing mechanistic insights into respiratory physiology and neurodegenerative diseases.C born preterm are reported to be at increased risk of developing non communicable diseases in later life. Changes in placental DNA methylation patterns are implicated in fetal programming of adult diseases. Our earlier studies in animals have established that micronutrients (folic acid, vitamin B12) and LCPUFA are interlinked in the one carbon cycle influencing methylation reactions. Our studies in women delivering preterm show reduced levels of LCPUFA, altered levels of micronutrients and lowered placental global DNA methylation levels at delivery. We therefore postulate that alterations in the micronutrient metabolism may affect the regulation of the enzymes [methionine adenosyltransferase (MAT2a) and S-adenosyl homocysteine hydrolase (AHCY) in the placenta thereby affecting SAM synthesis and methylation potential in women delivering preterm. The present study examines the mRNA and protein levels of the enzymes [MAT2A and AHCY] along with SAM and SAH levels from preterm (n=73) and term (n=73) placenta. The mRNA levels of enzymes were analyzed by qRT-PCR, protein levels by ELISA and SAM-SAH levels by HPLC. The mRNA levels for MAT2a and AHCY are higher (p<0.05 for both) in the preterm group as compared to term group. SAM and SAH levels were similar in both groups, although SAM: SAH ratio was lower (p<0.05) in the preterm group as compared to term. This data indicates a lower methylation potential in the preterm placenta which may have implications for the epigenetic programming of the developing fetus.T final structure and connectivity of the nervous system depends upon the accurate guidance of axons and their cell bodies through complex environments during the process of development. Failure to achieve correct connectivity results in a dysfunctional nervous system which may be associated with disorders such as Autism and Down’s syndrome. An understanding of how neural connectivity is established helps to improve treatment of nervous system disorders. The leading edge of a cell or axon is a highly dynamic structure called the growth cone (GC). The process of GC guidance is intricately orchestrated and regulated by a plethora of extracellular and intracellular molecules along the dorsal-ventral (DV) axis and anterior-ior (AP) axis. UNC-6 was the first guidance clue discovered in C. elegans guiding GCs along the DV axis of the worm. Later the mammalian homolog Netrin was identified to perform similar function in vertebrates. UNC-6/Netrin serves as a bi-functional cue attracting the GCs along DV (Dorsal/Ventral) axis through UNC-40/DCC and repelling GCs through UNC5/UNC5 receptors. Another highly conserved guidance system working along the DV axis includes SLT-1/Slt acting though SAX-3/Robo receptors. The molecules acting in the AP guidance are few including Wnts are set of secreted glycoproteins, which steer GCs along the AP through frizzled receptor. The ligand receptor complex at the membrane transduces the signal to bring about reorganization of the actin cytoskeleton which steers the GCs either towards or away from the cues. My research work is focused on a cytoplasmic actin-binding scaffolding protein, UNC-53 required for GC migrations along the AP axis. Its mammalian homologs, the Neuron Navigators (NAVs) are also known to function in nervous system development. The unc-53 (uncoordinated) is required for GC migrations along the AP axis in C. elegans. This study will primarily focus on first finding novel molecular collaborators of UNC-53 and mechanism of its action.M dependence of ZnO nanostructures on antibacterial activity: ZnO nanostructures with different morphologies (dumbbell, needles, petals, etc) were synthesized using chemical bath deposition (CBD) technique by varying the parameters like molar concentrations of Zn2+/OHand the pH value of the solution. Synthesized ZnO nanostructures were characterized structurally using XRD, SEM, TEM and optically using UV-spectrophotometer and Photoluminescence (PL) techniques. Morphological dependence of ZnO nanostructures on its antibacterial activity was studied using a gram negative bacterium Escherichia coli as a model microorganism. The antibacterial activity of ZnO nanoparticles was tested by spread plate method and the minimum inhibitory concentration (MIC) was determined. The antibacterial activity was found to depend strongly on the morphology of the nanostructures used. Unique nano sized petal shaped ZnO structures showed maximum antibacterial activity as compared to the other structures. The rupture/disruption of the bacterial cells after nanoparticle treatment was investigated by environmental SEM. The morphology-dependent antibacterial activity of ZnO observed in our case could be due to the higher surface-to-volume ratio of petal shape, specific surface area, surface modifications as well as defects associated with the nanostructures used.P aromatic hydrocarbons (PAHs) are toxic compounds. Due to leakage or spillage of crude oil, these toxic hydrocarbons may contaminate the sites. These compounds are low water soluble; as a result, they are not easily degraded by microorganisms. So, remediation of PAH’s through seeding only hydrocarbon degrading microorganisms take more time. Microbial produced surfactant can enhance the bioavailability of these hydrophobic compounds for bioremediation. Keeping this in mind, an investigation was carried out to develop an efficient consortium of biosurfactant producing bacterial strains that could be used for remediation of PAH from contaminated sites. For that, a total of twenty three bacterial strains were isolated from hydrocarbon contaminated soil. The ability of degradation of hydrocarbon of the isolates was tested individually and five of them were selected for further study on the basis of utilization of hydrocarbon. Among these five isolates, three were found biosurfactant producer and two non-producers. Ten different consortia were designed involving best hydrocarbon utilizing strains taking both biosurfactant producers and non-producers. The consortium consisting bacterial strain Bacillus pumilus KS2 and Bacillus cereus R2 (identified by 16s rDNA sequencing) had shown best results in degradation of crude oil. The consortium could degrade 82.15% of total petroleum hydrocarbon (TPH) from the crude oil in five weeks of incubation time. The consortium could degrade different PAH’s like naphthalene, fluorene, phenanthrene, anthracene, 3.beta.-Myristoyl Olean-12-en-1, 1H-indene, 2,3-dihydro-1,1,5,6 and their derivatives which indicates the prospective of the consortium to be used for remediation of petroleum hydrocarbon contaminated sites.R focusing on evaluating equine cytokines relied on the detection of mRNA rather than proteins due to limited availability of equine cytokines. Thus, the present work was undertaken to obtain biologically active recombinant equine IL-18 and IFN-γ. The coding sequences of equine IL-18 and IFN-γ gene were cloned and identity was confirmed by sequence analysis. Subsequently, recombinant equine IL-18 and IFN-γ were expressed in Escherichia coli and proteins were purified by Nickel-NTA chromatography. Biological activity of recombinant cytokines was assessed by lymphocyte proliferation assay, cytokine ELISA and real time RT-PCR. For lymphocyte proliferation assay, 4×105 equine peripheral blood mononuclear cells (PBMCs) were cultured per well in 96 well culture plate and stimulated with different concentration of recombinant cytokines and Concanavalin A (5 μg/ml) as positive control. At 72 hours post stimulation, cell proliferation assay was performed by XTT method. Recombinant equine IL-18 and IFN-γ induced significant proliferation at 500 ng/ml concentration. Secretion of IL10 and IFN-γ by activated PBMCs was assessed by ELISA at protein level and by real-time PCR at mRNA level. Recombinant IL-18 was able to induce the production of both IL-10 and IFN-γ (100 pg/ml) in equine PBMC while IFN-γ induced IL-10 secretion (100 pg/ml). On the other hand, IL-18 induced IFN-γ & IL-10 transcripts by 128 fold and IL-2 & IL-4 by 64 fold while IFN-γ induced transcription of IL-10 by 128 fold, IL-4 by 64 fold and IL-2 by 32 fold. Results indicate that recombinant equine cytokines activate the immune cells and stimulate the secretion of natural cytokines.Introduction: Changes in the levels of various hormones during different phases of menstrual cycle are known to affect various functions of body apart from reproductive system. This study was planned to see effect of different phases of menstrual cycle on peak expiratory flow rate in normal healthy young females of age group between 18-24 years. Material & Method: Study was performed on 30 healthy normal regularly menstrual medical students of age group 18-24 years in different phases of menstrual cycle for single cycle. PEFR is recorded in different phases of menstrual cycle .menstrual phase(2nd to 4th day),proliferative phase(9th to 12th day)and luteal (19th to 21st day).Instrument used was WRIGHT’s peak expiratory flow meter. Three readings were taken in standing condition and maximum of three readings were considered in each phase. Results: Peak expiratory flow rate was significantly higher (p<0.05)during the luteal phase of menstrual cycle as compared to menstrual and proliferative phases. Conclusion: As PEFR was better during luteal phase so this suggests a possible role of increased levels of progesterone during the luteal phase on respiratory system.T common mechanism of resistance of Citrobacter freundii to β-lactam antibiotics is the presences of plasmid mediated blaAmpC. In this study we evaluate the emergence of ampC gene among clinical isolates of C. freundii isolated from urine of symptomatic UTI patients and also to assess mobile genetic elements. A total of 30 consecutive, clinical isolates of C. freundii were investigated for the presence of AmpC β-lactamase. Molecular characterization was studied by mPCR for gene of AmpC including the coproduction of other β-lactamase. Presence of integron was detected using integrase gene primers. Presence of integron to blaAmpC gene along with gene cassettes identified using specific primers. Genetic location on insertion sequence was also seen. A total of 16 isolates were phenotypically positive for AmpC. On performing their genotypic characterization, 5 isolates were found to be positive for blaCMY-2, 3 isolates having blaDHA-2 whereas one isolates coproducing blaCMY-2 and blaDHA-2. In which some AmpC positive isolates were coexisting blaNDM, blaIMP, blaVIM family of MBL and blaCTXM, blaSHV of ESBL. Among them 8 were harboring class 1 integron, linkage between integron and blaCMY-2 has been detected. Investigation of gene-cassettes revealed the presence of dihydrofolate reductase (dhfr) and aminoglycoside 6’-N-acetyltransferase (aac 6’) genes and presence of CMY-2 gene on ISEcp1 were also seen. The study showed that these genes have high ability to cross species barrier with ease and integrated other resistance genes. Eventually, AmpC gene highlights the potential risk in hospital environment. Thus the attainment of AmpC genes by C. freundii restricts therapeutic alternatives for combating infections.A is a naturally occurring biomolecule in Artemisia annua, known for its anti-malarial as well as anti-cancerous activity. In vitro enhancement in artemisinin content has been achieved employing biotic (Piriformospora indica and Agrobacterium rhizogenes strain A4) and abiotic (such as heavy metals and salicylic acid) elicitors with different explants in various MS media compositions were employed. A significant enhancement in artemisinin content of 60% was achieved in shoot cultures co-cultivated with P. indica. Hairy root cultures raised through leaf explants of A. annua when exposed to different concentration of Pb, Hg, Co and SA have shown a tremendous enhancement in artemisinin, the maximum elevation reaching to 1450% in lead nitrate (100 mg/L) supplemented medium over control. For abiotic stress, nodal explants if exposed to various heavy metal (Ag, Cu, Hg, Co and Zn) salts too revealed significant increase in artemisinin production, the optimum being 50% at 100 mg/L of Cu and Zn. The crude extract of A. annua has been fractionated, isolated and characterized through CC, TLC, FT-IR and NMR. Bioassays conducted with crude extract of leaves against larvae of malaria (Anopheles stephensi) and dengue (Aedesaegypti) vectors have shown a strong larvicidal activity and on human oral cancer cell line causing 98.5% mortality. This is our first report of elicitation of artemisinin in A. annua employing P. indica and hairy roots coupled with abiotic stresses.Forest cover in the hill regions is essential to maintain environmental, economic and ecological balances. North Bengal accounts for 3,086 sq km (26 %) of the 11,876 sq km area of classified forests in the state, and for nearly 5,000 sq km (40 %) of all land under tree cover. Khumani is a Village (26.84 o N, 88.60 o E) in Gorubathan Block in Darjeeling District of West Bengal State, India. The survey work was done in December, 2014 by visiting the Khumani Forest Village (established in 1949) of upper Kumai and the primary data were gathered through field survey and direct contact with common people and authorized centers of the region. Surveys on the demography, agriculture, livestock management, water management, education, culture, health, waste management, disaster management, transport, biodiversity, human animal conflict were done in this area. Topographic map of the area was prepared by using the database of National Atlas and Thematic Mapping Organization (NATMO), Kolkata office. In every phase of the survey work, pictorial documentation was done. In spite of being positioned in a diverse and sensitive ecological zone, the village is not adequately managed. There is an urgent need for implementing sustainable management systems in the areas for the betterment of the socio-environmental structures. Some of the possible management strategies have been suggested for maintaining the social, environmental, economic and ecological balance of the region. Proper funds should be raised to conserve and manage these ecologically important zones of north Bengal, which should be one of the main focuses of future researches.P root-knot nematodes (Meloidogyne spp.) are arguably the most damaging genus of biotrophic pests of vascular plants and thus have a major impact on global agricultural production. Due to the changing climate and agricultural practices RKNs are becoming a menace in newer crops and geographical localities. Currently available management practices have failed to contain the problem; hence, there is a critical need to develop environmentally-friendly and smart approaches tailor-made to reduce the nematode disease burden in Indian agriculture. Utility of host-delivered RNAi has been demonstrated in several plants (Arabidopsis, tobacco and soybean) that exhibited resistance against root-knot and cyst nematodes. In the present study, a M. incognita-specific protease gene, cathepsin L cysteine proteinase (Mi-cpl-1) was targeted to generate tomato transgenic lines to evaluate the genetically modified nematode resistance. In vitro knockdown of Mi-cpl-1 gene led to the reduced attraction and penetration of M. incognita in tomato suggesting the involvement of Mi-cpl-1 in nematode parasitism. Transgenic expression of the dsRNA of Mi-cpl-1 gene resulted in 60-80% reduction in infection and multiplication of M. incognita in tomato. Evidence for in vitro and in planta silencing of Mi-cpl-1 was confirmed by expression analysis using quantitative RT-PCR. Our study demonstrates that Mi-cpl-1 plays crucial role during plant-nematode interaction and plantmediated down regulation of this gene elicits detrimental effect on M. incognita development, reinforcing the potential of RNAi technology for management of phytonematodes in crop plants. The findings of the present study lead to the better understanding of the mechanism of nematode parasitism which ultimately helps in designing smarter nematode management options.C and behavioral disorders are thought to be a result of neuronal dysfunction but the underlying molecular defects remain largely unknown. The cAMP/Protein kinase A (PKA) pathway is known to be a key factor regulating neuronal behaviour. Genetic associations with neurobehavioral dysfunction include a chromosome region containing the gene coding for coronin 1. Our studies show that coronin 1, which was identified as a lymphocyte specific protein important for mycobacterial survival in macrophages as well as naive T cell survival is also expressed in excitatory neurons of the brain. Interestingly, deficiency of coronin 1 results in severe behavioral abnormalities such as reduced socialization and anxiety, increased aggression and disabilities in learning. Electrophysiological studies show that coronin 1 regulates cAMP/PKA dependent synaptic plasticity at excitatory synapses. Specifically, protein kinase A-dependent cortico-lateral amygdalary long‐term potentiation was completely absent in coronin 1‐deficient mice. The coronin 1-mediated cAMP/PKA response was dependent on the stimulus dependent interaction of coronin 1 with the stimulatory G alpha subunit (Gαs). Deletion of coronin 1 or expression of coronin 1 mutants incapable of interacting with Gαs, fails to stimulate cAMP signaling. In vivo infusion of a membrane–permeable cAMP analogue, 8-Br-cAMP into coronin 1-deficient mice brain effectively rescues the synaptic and behavioral defects. Therefore, we conclude that coronin 1 regulates synaptic plasticity and neuronal behavior through its interaction with Gαs upon cell surface stimulation.I accumulation of Aβ42 is an early event in the pathogenesis cascade of Alzheimer’s disease (AD) causing in neuronal dysfunction, synaptic and neuronal loss together with dementia. A bifunctional protein C-terminus Hsp70 interactive protein (CHIP) is considered as a connecting link between molecular chaperones and Ubiquitin Proteasome System (UPS) while another E3 ligase Parkin targets several proteins for UPS degradation. Further Parkin’s mutations are the major cause of autosomal recessive Parkinson’s disease (ARPD) where Parkin catalyzes the post-translational modification of proteins with polyubiquitin targeting them to the 26S proteasome. In addition, Parkin together with CHIP reduces intracellular Aβ1-42 peptide levels, an important peptide that shares a cross talk between Alzheimer’s and Parkinson’s diseases. These ligases counteract its effects on cell death and reverse its effect to inhibit the proteasome. Herein, we reported the implication of CHIP and Parkin in the metabolism of β-amyloid precursor proteins (βAPP) and its derivative β-amyloid. We also proved a strong interaction between β-APP, CHIP and HSPs. Interestingly CHIP also promotes the association of ubiquitin with β-APP in proteasomal dependent manner. CHIP together with another Parkin enhances the Aβ degradation and eliciting neuro protective properties while Parkin alone consider as an inducers of amyloid clearance, a cryoprotectant and in the suppression of reactive inflammation. Furthermore, we have shown the effect of bioflavonoid in the attenuation of hypoxia and neurotoxin induced neurodegeneration in the rat model.Cervical cancer is the second most common malignancy in women worldwide and thus one of the leading causes of mortality in women. Lovastatin, a non polar, anticholesterol drug has previously been reported to exert antitumour activity in vitro. In the present study, lovastatin from Aspergillus terreus (KM017963) was purified by adsorption chromatography and evaluated for its anticancer and anti-oxidant properties with a human cervical cancer cell line (HeLa). Growth inhibitory and proapoptotic effects of purified lovastatin on HeLa cells were investigated by determining its influence on cell numbers, mitochondrial membrane potential (MMP), DNA fragmentation and antioxidant properties in terms of hydroxy radical scavenging effects as well as levels of total reduced glutathione. Cell cycle analysis by ow cytometry (propidium iodide staining) confirmed induction of apoptotic cell death and revealed cell cycle arrest in the G0/G1 phase. The results of the study give leads for the anticancer effects of lovastatin and its potential usefulness in the chemotherapy of cervical cancer.

Evaluation of Microsatellite Instability and Apoptosis in Gall Bladder Malignancy from Patients of a Cohort Exposed to Methylisocyanate

The molecular alterations are considered to play an important role in the both carcinogenesis and biological behavior of a variety of human malignancies. However, cancer of gallbladder is an obscured phenomenon and highly malignant with a poor survival due to underprivileged diagnosis. Tissues of 92 cases of gallbladder cancer patients (31 men and 61 women, age range 16-85 yrs, mean age 45.83 ± 1.50 yrs) were examined for microsatellite instability (MSI) of six microsatellite markers (D16S539, D13S317, D7S820, F13A01, FES/FPS, vWA) and apoptosis of malignant epithelial cells through M30CytoDEATH assay. Analysis of microsatellite markers revealed 08.7% (08/92) in gallbladder cancer, in which 10.0% (07/70) instability found in adenocarcinoma. The sensitivity of this test in adenocarcinoma, adenosquamous carcinaoma and adenoma with dysplasia was found to be 10.0%, 00.0% and 08.3% respectively suggesting its role in the multistage disease invasiveness. The immunohistochemical examination confirmed the presence of CK18 in moderately, well and poorly differentiated adenocarcinoma with the frequency of viz., 18.8%, 15.4% and 11.1% respectively showing positive sign of apoptosis. The mixed chimerism of STR loci and positive staining of caspase cleaved CK18 in epithelial cells of the gallbladder cancer tissues showed their independent and noteworthy character in the gallbladder carcinogenesis. Further investigations are in progress to undertake similar studies on archived tumor tissues of varied origins and forms. These might also provide modalities to translate forceful and reproducible strategies for defined clinical utility.