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In general parlance, the present study is an attempt to understand the root causes behind student migration from rural parts of West Bengal and its overall impacts on society as well as on the state. The number of students migrate from rural parts of Bengal is increasing tremendously. The aim and objective and ultimate preferences of studying outside the state, is that, students always prefer to get the option in which they are interested in and in which they want their specialization, which they might not be getting in their native colleges and Universities. The degree which they obtain from that universities or colleges from that states is considered to be worth when it comes to searching a good job in any place. The other benefit of obtaining the degree is most of the universities or colleges allow the students to go abroad for completing their final semester. A chance to get educated at any foreign land might be a distant dream for any middle class students. Moreover, apart from these factors there are some key factors like quality education, potential placement, a good standard of living, and especially a very poor job opportunities and lower pay scales in the domestic areas are among the most important reasons for thinking of migration. There is no doubt that the overall economic development of the rural parts of Bengal largely depends on the people, especially on the young generations to a great extent. If the generation leaves or migrate to other states for searching of a better future, It should be considered as brain drain of intellectual resource.Therefore the study initially indicated that constant and comprehensive monitoring of the trend, and if possible a regular evaluation process has to be implemented in the educational institutions to observe the parameter of the trend. The trend cannot be eradicated permanently , until and unless State Government, Panchayet, Bank assistance (Easiest way to get Student’s loan for higher studies),and above all a State-Central placement cell run and governed by affiliated Universities are not established immediately.

Adsorption and activation of CO2 is a key step in any chemical reaction, which aims to convert it to other useful chemicals. Therefore, it is important to understand the factors that drive the activation process and also search for materials that promote the process. We employ the density functional theory to explore the possibility of using small-sized bimetallic Cu-Zr clusters, Cu4-nZrn, with n = 1-3 for the above-mentioned key step. Our results suggest that after adsorption, a CO2 molecule preferably resides on Zr atoms or at the bridge and triangular faces formed by Zr atoms in bimetallic Cu-Zr clusters accompanied with its high degree of activation. Importantly, maximum activation occurs when CO2 is adsorbed on the CuZr3 cluster. Interestingly, we find that the adsorption energy of CO2 can be tuned by varying the extent of the Zr atom in Cu-Zr clusters. We rationalize the high adsorption of CO2 with the increase in the number of Zr atoms using the d-band center model and the concept of chemical hardness. The strong chemisorption and high activation of CO2 are ascribed to charge migration between Cu-Zr clusters and the CO2 molecule. We find an additional band in the infrared vibrational spectra of CO2 chemisorbed on all of the clusters, which is absent in the case of free CO2. We also observe that the energy barriers for the direct dissociation of the CO2 molecule to CO and O decrease significantly on bimetallic Cu-Zr clusters as compared to that on pure Cu4. In particular, the barrier heights are considerably small for Cu3Zr and CuZr3 clusters. This study demonstrates that Cu3Zr and CuZr3 clusters may serve as good candidates for activation and dissociation of the CO2 molecule.

Emerging mutations in the SARS-CoV-2 genome pose a challenge for vaccine development and antiviral therapy. The antiviral efficacy of Azadirachta indica bark extract (NBE) was assessed against SARS-CoV-2 and m-CoV-RSA59 infection. Effects of in vivo intranasal or oral NBE administration on viral load, inflammatory response, and histopathological changes were assessed in m-CoV-RSA59-infection. NBE administered inhibits SARS-CoV-2 and m-CoV-RSA59 infection and replication in vitro, reducing Envelope and Nucleocapsid gene expression. NBE ameliorates neuroinflammation and hepatitis in vivo by restricting viral replication and spread. Isolated fractions of NBE enriched in Nimbin isomers shows potent inhibition of m-CoV-RSA59 infection in vitro. In silico studies revealed that NBE could target Spike and RdRp of m-CoV and SARS-CoV-2 with high affinity. NBE has a triterpenoids origin that may allow them to competitively target panoply of viral proteins to inhibit mouse and different strains of human coronavirus infections, suggesting its potential as an antiviral against pan-β-Coronaviruses.

Interferon regulatory factor 8 (IRF8), a myeloid lineage transcription factor, emerges as an essential regulator for microglial activation. However, the precise role of IRF8 during Japanese encephalitis virus (JEV) infection in the brain remains elusive. Here, we report that JEV infection enhances IRF8 expression in the infected mouse brain. Comparative transcriptional profiling of whole-brain RNA analysis and validation by quantitative reverse transcription-PCR (qRT-PCR) reveals an impaired interferon gamma (IFN-γ) and related gene expression in Irf8 knockout (Irf8−/−)-infected mice. Further, Ifnγ knockout (Ifnγ−/−) mice exhibit a reduced level of Irf8. Both Ifnγ−/− and Irf8−/− mice exhibit significantly reduced levels of activated (CD11b+ CD45hi, CD11b+ CD45lo, Cd68, and CD86) and infiltrating immune cells (Ly6C+, CD4, and CD8) in the infected brain compared to those of wild-type (WT) mice. However, a higher level of granulocyte cell (Ly6G+) infiltration is evident in Irf8−/− mice as well as the increased concentration of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), monocyte chemoattractant protein 1 (MCP1) levels in the brain. Interestingly, neither the Irf8−/− nor the Ifnγ−/− conferred protection against lethal JEV challenge to mice and exhibit augmentation in JEV replication in the brain. The gain of function of Irf8 by overexpressing functional IRF8 in an IRF8-deficient cell line attenuates viral replication and enhances IFN-γ production. Overall, we summarize that in the murine model of JEV encephalitis, IRF8 modulation affects JEV replication. We also show that lack of Irf8 affects immune cell abundance in circulation and the infected brain, leading to a reduction in IFN-γ level and increased viral load in the brain. IMPORTANCE Microglial cells, the resident macrophages in the brain, play a vital role in Japanese encephalitis virus (JEV) pathogenesis. The deregulated activity of microglia can be lethal for the brain. Therefore, it is crucial to understand the regulators that drive microglia phenotype changes and induce inflammation in the brain. Interferon regulatory factor 8 (IRF8) is a myeloid lineage transcription factor involved in microglial activation. However, the impact of IRF8 modulation on JEV replication remains elusive. Moreover, the pathways regulated by IRF8 to initiate and amplify pathological neuroinflammation are not well understood. Here, we demonstrated the effect of IRF8 modulation on JEV replication, microglial activation, and immune cells infiltration in the brain.

Neutrophils are versatile immune effector cells essential for mounting a first-line defense against invading pathogens. However, uncontrolled activation can lead to severe life-threatening complications. Neutrophils exist as a heterogeneous population, and their interaction with pathogens and other immune cells may shape the outcome of the host immune response. Diverse classes of viruses, including the recently identified novel SARS-CoV-2, have shown to alter the various aspects of neutrophil biology, offering possibilities for selective intervention. Here, we review heterogeneity within the neutrophil population, highlighting the functional consequences of circulating phenotypes and their critical involvement in exaggerating protective and pathological immune responses against the viruses. We discuss the recent findings of neutrophil extracellular traps (NETs) in COVID-19 pathology and cover other viruses, where neutrophil biology and NETs are crucial for developing disease severity. In the end, we have also pointed out the areas where neutrophil-mediated responses can be finely tuned to outline opportunities for therapeutic manipulation in controlling inflammation against viral infection.

Artificial microRNA (amiRNA)-mediated inhibition of viral replication has recently gained importance as a strategy for antiviral therapy. In this study, we evaluated the benefit of using the amiRNA vector against Japanese encephalitis virus (JEV). We designed three single amiRNA sequences against the consensus sequence of 3′ untranslated region (3′UTR) of JEV and tested their efficacy against cell culture-grown JEV Vellore strain (P20778) in neuronal cells. The binding ability of three amiRNAs on 3′UTR region was tested in vitro in HEK293T cells using a JEV 3′UTR tagged with luciferase reporter vector. Transient transfection of amiRNAs was nontoxic to cells as evident from the MTT assay and caused minimal induction in interferon-stimulated gene expression. Furthermore, our result suggested that transient expression of two amiRNAs (amiRNA #1 and amiRNA #2) significantly reduced intracellular viral RNA and nonstructural 1 (NS1) protein, as well as diminished infectious viral particle release up to 95% in the culture supernatant as evident from viral plaque reduction assay. Overall, our results indicated that RNA interference based on amiRNAs targeting viral conserved regions at 3′UTR was a useful approach for improvements of nucleic acid inhibitors against JEV.

Dengue virus (DENV) infection causes dengue fever in humans, which can lead to thrombocytopenia showing a marked reduction in platelet counts, and dengue hemorrhagic fever. The virus may cause thrombocytopenia either by destroying the platelets or by interfering with their generation via the process of megakaryopoiesis. MEG-01 is the human megakaryoblastic leukemia cell line that can be differentiated in vitro by phorbol-12-myristate-13-acetate (PMA) treatment to produce platelet-like-particles (PLPs). We have studied DENV infection of MEG-01 cells to understand its effect on megakaryopoiesis and the generation of PLPs. We observed that DENV could infect only naive MEG-01 cells, and differentiated cells were refractory to virus infection/replication. However, DENV-infected MEG-01 cells, when induced for differentiation with PMA, supported an enhanced viral replication. Following the virus infection, the MEG-01 cells showed a marked reduction in the surface expression of platelet markers (CD41, CD42a, and CD61), a decreased polyploidy, and significantly reduced PLP counts. DENV infection caused an enhanced Notch signaling in MEG-01 cells where the virus envelope protein was shown to interact with TAL-1, a host protein important for megakaryopoiesis. These observations provide new insight into the role of DENV in modulating the megakaryopoiesis and platelet production process.

Dengue virus (DENV) infection usually causes dengue fever (DF) with flu-like illness affecting infants, young children, and adults. The DF occasionally evolves into a potentially lethal complication called dengue severe (DS) leading to a rapid fall in platelet count along with plasma leakage, fluid accumulation, respiratory distress, and severe bleeding. The diverse clinical spectrum of dengue disease, as well as its significant similarity to other febrile viral illnesses, makes early identification more challenging in this high-risk group. microRNAs (miRNAs) are small (∼19 to 21 nucleotides [nt] in length), noncoding RNAs, extremely stable and easily detectable in the plasma; thus, they have potential as biomarkers for diagnosing and monitoring human diseases. This study provides a comprehensive analysis of miRNAs circulating in plasma of dengue virus-infected patients and identifies the miRNA signatures that have biomarker potential for dengue infection and disease progression., The circulating microRNA (miRNA) profile has been widely used for identifying potential biomarkers against viral infections. However, data on circulating microRNA expression patterns in dengue patients are scanty. Considering the impact of severity caused by dengue infection, circulating miRNA profiles in plasma of dengue patients may prove to be valuable for developing early prognostic markers for the disease severity. Here, we described an in-depth analytical study of small RNA sequencing data obtained from the plasma of 39 dengue patients. Integrating bioinformatics and in vitro studies, we identified differentially expressed miRNAs (DEMs) (log2 fold change ≥1.5, P

ACE2 receptor has been the focus of scientific community as receptor of COVID 19 and is been explored for therapeutic targeting. Recently, another receptor for COVID 19 was reported, the Glucose Regulated Protein 78 (GRP78). GRP78, a heat shock protein is an endoplasmic reticulum (ER) stress response protein. In cancer and certain autoimmune diseases there are reports of upregulation of GRP78. In breast cancer, prostate cancer, melanomas and malignant gliomas GRP78 overexpresses. Chemoresistance has been linked to upregulated GRP78. So, cancer patients might be at increased risk of COVID 19 infection owing to receptor abundance. We propose that tumour associated endothelial cells having upregulated GRP78 might also participate in dissemination of the virus. GRP78 also gets upregulated in rheumatoid arthritis and inflammatory bowel disease (IBD). Thus, both cancer patients as well as autoimmune diseases (Rheumatoid arthritis and IBD) should be considered as high-risk groups for COVID 19.

Resonant photoemission spectroscopy has been used to investigate the character of Fe 3d states in FeAl alloy. Fe 3d states have two different character, first is of itinerant nature located very close to the Fermi level, and second, is of less itinerant (relatively localized character), located beyond ∼2 eV below the Fermi level. These distinct states are clearly distinguishable in the resonant photoemission data. Comparison between the results obtained from experiments and first principle based electronic structure calculations show that the origin of the itinerant character of the Fe 3d states is due to the ordered B2 structure, whereas the relatively less itinerant (localized) Fe 3d states are from the disorders present in the sample. The exchange splitting of the Fe 3s core level peak confirms the presence of local moment in this system. It is found that the itinerant electrons arise due to the hybridization between Fe 3d and Al 3s and 3p states. Presence of hybridization is observed as a shift in the Al 2p core-level spectra as well as in the X-ray near edge absorption spectra towards lower binding energy. Our photoemission results are thus explained by the co-existence of ordered and disordered phases in the system.

In accordance with the well established gold-hydrogen analogy, a hydrogen atom mimics the properties of a gold atom in gold clusters. In a recent study it has been demonstrated that the properties of a hydrogen atom doped small gold cluster (Au7H) are not in conformity with the aforementioned analogy. In this paper we study the properties of the Au7H cluster exhaustively to re-examine the validity of the gold-hydrogen analogy in the context of adsorption of CO and O2 molecules on pristine gold and hydrogen atom doped gold clusters. For this purpose we first determine the most stable structure of the Au7H cluster by using an ab initio density functional theory based method with generalized gradient approximation (GGA) and Meta-GGA exchange-correlation functionals. We carry out geometry optimization by considering various planar and three-dimensional isomers of the Au7H cluster as initial geometries. We find that the lowest energy structure of Au7H is a planar one with C2 v symmetry, and it is very close to the structure of the Au8 cluster with D4 h symmetry. Furthermore, to examine the validity of the gold-hydrogen analogy we carry out a detailed investigation of the adsorption of CO and O2 molecules on the most stable as well as various other low energy isomers of the Au7H cluster. We find that the adsorption energies and the extent of activation of CO and O2 molecules on the most stable planar isomer of Au7H are almost the same as those on the parent Au8 cluster with D4 h symmetry proving the validity of the gold-hydrogen analogy. On the other hand, for the high energy three-dimensional isomers of the Au7H cluster obtained from the pristine Au8 cluster with T d symmetry, we find a significant enhancement in adsorption energy as well as the extent of activation of CO and O2 molecules as compared to those for the corresponding pristine cluster. Therefore, the high reactivity of the 3D isomer of the Au7H cluster may be attributed to its existence in a state which is higher in energy than its most stable planar isomer.

MicroRNAs (miRNAs) released from the activated microglia upon neurotropic virus infection may exacerbate the neuronal damage. Here, we identified let-7a and let-7b (let-7a/b) as one of the essential miRNAs over-expressed upon Japanese Encephalitis virus (JEV) infection and released in the culture supernatant of the JEV-infected microglial cells through extracellular vesicles. The let-7a/b was previously reported to modulate inflammation in microglial cells through Toll-like receptor 7 (TLR7) pathways; although their role in accelerating JEV pathogenesis remain unexplored. Therefore, we studied the role of let-7a/b in modulating microglia-mediated inflammation during JEV infection and investigated the effect of let-7a/b-containing exosomes on primary neurons. To this end, we examined let-7a/b and NOTCH signaling pathway in TLR7 knockdown (KD) mice. We observed that TLR7 KD or inhibition of let-7a/b suppressed the JEV-induced NOTCH activation possibly via NF-κB dependent manner and subsequently, attenuated JEV-induced TNFα production in microglial cells. Furthermore, exosomes secreted from let-7a/b over-expressed microglia when transferred to uninfected mice brain induced caspase activation. Exosomes secreted from virus-infected or let-7a/b over-expressed microglia when co-incubated with mouse neuronal (Neuro2a) cells or primary cortical neurons also facilitated caspase activation leading to neuronal death. Thus, our results provide evidence for the multifaceted role of let-7a/b miRNAs in JEV pathogenesis. Let-7a/b can interact with TLR7 and NOTCH signaling pathway and enhance TNFα release from microglia. On the other hand, the exosomes secreted by JEV-infected microglia can activate caspases in uninfected neuronal cells which possibly contribute to bystander neuronal death. Cover Image for this issue: doi: 10.1111/jnc.14506.

Raman spectroscopy/mapping is used to investigate the variation of Si phonon wavenumbers, i.e., lower wavenumber (LW ~ 495–510 cm−1) and higher wavenumber (HW ~ 515–519 cm−1) phonons, observed in Si–SiO2 multilayer nanocomposite (NCp) grown using pulsed laser deposition. Sensitivity of Raman spectroscopy as a local probe to surface/interface is effectively used to show that LW and HW phonons originate at surface (Si–SiO2 interface) and core of Si nanocrystals, respectively. The consistent picture of this understanding is developed using Raman spectroscopy monitored laser heating/annealing and cooling experiment at the site of the desired wavenumber, chosen with the help of Raman mapping. Raman spectra calculations for Si41 cluster with oxygen and hydrogen termination show strong mode at 512 cm−1 for oxygen terminated cluster corresponding to the vibration of surface Si atoms. This supports our attribution of LW phonons to be originating at the Si–SiO2 surface/interface. These results along with XPS show that nature of interface (oxygen bonding) in turn depends on the size of nanocrystals and LW phonons originate at the surface of smaller Si nanocrystals. The understanding developed can conclude the ongoing debate on large variation in Si phonon wavenumbers of Si–SiO2 NCps in the literature. Copyright © 2015 John Wiley & Sons, Ltd.

The major challenges faced by candidate electrode materials in lithium-ion batteries (LIBs) include their low electronic and ionic conductivities. 2D van der Waals materials with good electronic conductivity and weak interlayer interaction have been intensively studied in the electrochemical processes involving ion migrations. In particular, molybdenum ditelluride (MoTe2 ) has emerged as a new material for energy storage applications. Though 2H-MoTe2 with hexagonal semiconducting phase is expected to facilitate more efficient ion insertion/deinsertion than the monoclinic semi-metallic phase, its application as an anode in LIB has been elusive. Here, 2H-MoTe2 , prepared by a solid-state synthesis route, has been employed as an efficient anode with remarkable Li+ storage capacity. The as-prepared 2H-MoTe2 electrodes exhibit an initial specific capacity of 432 mAh g-1 and retain a high reversible specific capacity of 291 mAh g-1 after 260 cycles at 1.0 A g-1 . Further, a full-cell prototype is demonstrated by using 2H-MoTe2 anode with lithium cobalt oxide cathode, showing a high energy density of 454 Wh kg-1 (based on the MoTe2 mass) and capacity retention of 80% over 100 cycles. Synchrotron-based in situ X-ray absorption near-edge structures have revealed the unique lithium reaction pathway and storage mechanism, which is supported by density functional theory based calculations.

Imaging Infrared Spectrometer (IIRS) is an imaging hyperspectral instrument for mineralogy of the lunar surface (including the hydroxyl signature). IIRS operates in the 0.8–5 μm spectral range with about 250 contiguous bands. It has 80 m ground sampling distance and 20 km swath at nadir from 100 km orbit altitude. Optical design is based on fore-optics and Offner (convex multi-blazed grating)-type spectrometer. Focal plane array is HgCdTe (mercury–cadmium–telluride)- based actively cooled to 90 K, having 500 × 256 pixels format with 30 μm pixel size. Electronics comprises proximity, logic and control, power supply and cooler drive electronics. Mechanical system is realized to house various subsystems, namely optics, detector, electronics and thermal components meeting the structural, opto-mechanical thermal component and alignment requirements. Thermal system is designed such that the instrument is cooled and maintained at fixed temperature to reduce and control instrument background. Aluminum-based mirror, grating and housing are developed to maintain structural as well as opto-mechanical and thermal requirements. This article presents IIRS realization and spectroradoimetric performance.

Extracellular microRNAs (miRNAs) are essential for the cell to cell communication in the healthy and diseased brain. MicroRNAs released from the activated microglia upon neurotropic virus infection may exacerbate CNS damage. Here, we identified let-7a and let-7b (let-7a/b) as the overexpressed miRNAs in Japanese Encephalitis virus (JEV) infected microglia and assessed their role in JEV pathogenesis. We measured the let-7a/b expressions in JEV infected post-mortem human brains, mice brains and in mouse microglial N9 cells by the qRT-PCR and in situ hybridization assay. The interaction between let-7a/b and NOTCH signaling pathway further examined in Toll-like receptor 7 knockdown (TLR7 KD) mice to assess the functions. Exosomes released from JEV infected or let-7a/b mimic transfected N9, and HEK-293 cells were isolated and evaluated their function. We observed an upregulation of let-7a/b in the infected brains as well as in microglia. Knockdown of TLR7 or Inhibition of let-7a/b suppressed the JEV induced NOTCH activation possibly via NF-κB dependent manner and subsequently, attenuated JEV induced TNFα production in microglial cells. Further, exosomes secreted from JEV-infected microglial cells specifically contained let-7a/b. Exosomes overexpressed with let-7a/b were injected into BALB/c mice as well as co-incubated with mouse neuronal (Neuro2a) cells, or primary cortical neuron resulted in caspase activation leading to neuronal damage in the brain. Thus, our results provide evidence for the multifaceted role of let-7a/b miRNAs and unravel the exosomes mediated mechanism for JEV induced pathogenesis.

The conformational preferences of the industrially significant ethyl propionate (EP) molecule have been investigated from the Raman and FTIR spectra, aided by ab initio and Car-Parrinello molecular dynamics (CPMD) simulation studies. The vibrational signatures of various rotameric forms of the EP molecule have been assigned for the first time from the potential energy distributions (PEDs). The critical analyses of the vibrational signatures reveal the coexistences of the Trans-Trans (TT), Trans-Antigauche (TG(-)) [Trans-Gauche (TG(+))], Antigauche-Trans (G(-)T) [Gauche-Trans (G(+)T)], Antigauche-Antigauche (G(-)G(-)) [Gauche-Gauche (G(+)G(+))], and Gauche-Antigauche (G(+)G(-)) [Antigauche-Gauche (G(-)G(+))] forms of the EP molecule at room and at high temperatures. However, at low temperature (ca. 70 °C), the TT and TG(-) forms of the EP molecule is estimated to be preponderant. The Car-Parrinello molecular dynamics simulation studies of the EP molecule estimated at high, room, and low temperatures are also in harmony with our conjecture as suggested from the vibrational analyses. The ab intio molecular dynamics simulations are observed to be a useful tool for the conformational analyses of the molecule.

We have investigated the molecular diversity of Hepatitis B virus (HBV) among the HIV co-infected patients from eastern-India. HBsAg/HBV-DNA positive subjects (n=73) from 874 HIV-infected patients were analyzed by sequencing followed by genetic diversity quantification. HBV/genotype-D and HBV/sugenotype-D2 were predominant. HBV/D2 isolates from patients with low CD4 count manifested significantly lower non-synonymous substitutions (p

A previous study from West Bengal documented very high rate of occult HBV infection (OBI) among the HBsAg negative blood donors. This study was aimed to characterize the OBI strains circulating among the blood donors and to estimate the risk associated with the prevailing viral variants/mutants. Blood samples from 2195 voluntary blood donors were included in the study. HBsAg, HBeAg, anti-HBc, and anti-HBs statuses of the samples were done by ELISA based detection. PCR amplification and sequencing were done to determine HBV genotypes, basal core promoter (BCP), and precore (Pre-C) mutations. Among the study samples, 268 were anti-HBc positive/HBsAg negative, among which 65 (24.25%) were HBV DNA positive. Phylogenetic analysis revealed the presence of HBV/D (87.23%), HBV/A (8.51%), and HBV/C (4.26%) (P<0.0001).HBV/D3 (65.85%) was the significantly prevalent subgenotype over HBV/D2 (26.83%) and HBV/D1 (7.31%) (P=0.0003). Considerable prevalence of differential BCP (1752C, 1753C, 1762T/1764A, 1753C+1762T/1764A, 1773C, and 1814C) and reverse transcriptase (rt) gene (rtI91L, rtL93P, rtS106C, rtR110G, rtN118T, rtS119T, rtY126H, rtG127W/R, rtC136R, and rtY158H) mutations was identified. Association of specific HBV subgenotypes with OBI was interesting and needs further study. Clinically relevant mutations were prevalent among the OBI strains which are of serious concern.

Background Hepatitis B virus (HBV) X protein (HBx) reported to be associated with pathogenesis of hepatocellular carcinoma (HCC) and miR-122 expression is down regulated in HCC. Previous studies reported miR-122 targets cyclin G1 (CCNG1) expression and this in turn abolishes p53-mediated inhibition of HBV replication. Here we investigated the involvement of HBx protein in the modulation of miR-122 expression in hepatoblastoma cells. Methods Expression of miR-122 was measured in HepG2 cells transfected with HBx plasmid (HBx-HepG2), full length HBV genome (HBV-HepG2) and in constitutively HBV synthesizing HepG2.2.15 cells. CCNG1 mRNA (a direct target of miR-122) and protein expressions were also measured in both HBx-HepG2, HBV-HepG2 cells and in HepG2.2.15 cells. miR-122 expressions were analyzed in HBx-HepG2, HBV-HepG2 and in HepG2.2.15 cells after treatment with HBx mRNA specific siRNA. Expressions of p53 mRNA and protein which is negatively regulated by CCNG1 were analyzed in HBx transfected HepG2 cells; X silenced HBx-HepG2 cells and X silenced HepG2.2.15 cells. HBx induced cell proliferation in HepG2 cells was measured by cell proliferation assay. Flow cytometry was used to evaluate changes in cell cycle distribution. Expression of cell cycle markers were measured by real time PCR. Results Expression of miR-122 was down regulated in HBx-HepG2, HBV-HepG2 and also in HepG2.2.15 cell line compared to control HepG2 cells. CCNG1 expression was found to be up regulated in HBx-HepG2, HBV-HepG2 cells and in HepG2.2.15 cells. Following siRNA mediated silencing of HBx expression; increased miR-122 levels were documented in HBx-HepG2, HBV-HepG2 and in HepG2.2.15 cells. HBx silencing in HBx-HepG2 and HepG2.2.15 cells also resulted in increased p53 expression. FACS analysis and assessment of expressions of cell cycle markers revealed HBx induced a release from G1/S arrest in HepG2 cells. Further, cell proliferation assay showed HBx promoted proliferation of HepG2 cell. Conclusion Our study revealed that HBx induced down regulation of miR-122 expression that consequently increased CCNG1 expression. This subsequently caused cell proliferation and release from G1/S arrest in malignant hepatocytes. The study provides the potential to utilize the HBx-miR-122 interaction as a therapeutic target to limit the development of HBV related HCC.

Microglia cells in the brain play essential role during Japanese Encephalitis Virus (JEV) infection and may lead to change in microRNA (miRNA) and mRNA profile. These changes may together control disease outcome. Using Affymetrix microarray platform, we profiled cellular miRNA and mRNA expression at multiple time points during viral infection in human microglial (CHME3) cells. In silico analysis of microarray data revealed a phased pattern of miRNAs expression, associated with JEV replication and provided unique signatures of infection. Target prediction and pathway enrichment analysis identified anti correlation between differentially expressed miRNA and the gene expression at multiple time point which ultimately affected diverse signaling pathways including Notch signaling pathways in microglia. Activation of Notch pathway during JEV infection was demonstrated in vitro and in vivo. The expression of a subset of miRNAs that target multiple genes in Notch signaling pathways were suppressed and their overexpression could affect JEV induced immune response. Further analysis provided evidence for the possible presence of cellular competing endogenous RNA (ceRNA) associated with innate immune response. Collectively, our data provide a uniquely comprehensive view of the changes in the host miRNAs induced by JEV during cellular infection and identify Notch pathway in modulating microglia mediated inflammation.

Restricted area heterojunctions, an array of lead sulfide colloidal quantum dots (PbS-CQDs) and crystalline silicon, are studied with a non-destructive remote contact light beam induced current (RC-LBIC) technique. As well as getting good quality active area images we observed an anomalous unipolar signal response for the PbS-CQD/n-Si devices and a conventionally expected bipolar signal profile for the PbS-CQD/p-Si devices. Interestingly, our simulation results consistently yielded a unipolar and bipolar nature in the signals related to the PbS-CQD/n-Si and PbS-CQD/p-Si heterostructures, respectively. In order to explain the physical mechanism involved in the unipolar signal response of the PbS-CQD/n-Si devices, we propose a model based on the band alignment in the heterojunctions, in addition to the distribution of photo-induced excess majority carriers across the junction. Given that the RC-LBIC technique is well suited to this context, the presence of these two distinct mechanisms (the bipolar and unipolar nature of the signals) needs to be considered in order to have a better interpretation of the data in the characterization of an array of homo/heterojunctions.

The presence of three different HBV genotypes (A, C and D) in Eastern India provided us a unique opportunity to study HBV pre-S mutants in these genotypes and subtypes among the same ethnic population. Furthermore, we also aimed to investigate the association of the HBV pre-S mutation with clinical outcome. Pre-S1-S2 and S gene was amplified and sequenced from 86 HBsAg positive study subjects with varying clinical manifestation. The genetic variability in the pre-S region (mutations) was studied with respect to different HBV genotypes, subtypes and different clinical categories. Six different types of HBV pre-S mutations were detected in 25 cases (29.07%), among which pre-S2 start codon mutation (28.0%) and pre-S2 deletion (24.0%) were most common. Pre-S mutation was highest in HBV/C (7/18; 38.89%) followed by HBV/A (9/27; 33.33%) and HBV/D (9/40; 22.50%). Pre-S1 deletion is common in HBV/D, whereas pre-S2 start codon mutation and pre-S2 deletions are frequent among HBV/A and HBV/C, respectively. Interestingly, in HBV/A and HBV/C the tendency of mutation/deletion increases from pre-S1 to pre-S2 region while in HBV/D the opposite tendency was observed. A significantly higher association of pre-S mutation (p=0.013) and pre-S2 deletion/ablation (p=0.016) was found among the HBeAg negative cases. Pre-S1 deletion and pre-S2 deletion were common among the ASC and CLD cases respectively, while pre-S2 start codon mutation was significantly associated with cirrhosis (p0.05). The study underscores the association of types of pre-S mutations with particular HBV genotype and clinical outcome in the study population.

The fourth component of human complement (C4) plays an important role in innate immune function. C4 activity has been observed to be significantly lower in patients with chronic hepatitis C virus (HCV) infections, although the mechanism remains unknown. In this study, we have examined the mechanisms of C4 regulation by HCV. Liver biopsy specimens from patients with chronic HCV infections displayed significantly lower C4 mRNA levels than liver tissue samples from patients with unrelated liver disease. Further, C4 mRNA levels of the two isoforms (C4A and C4B) were significantly reduced in hepatocytes transfected with RNA from HCV genotype 1a or 2a. Subsequently, a significant C4 regulatory role of HCV core or NS5A upon C4 promoter activity was observed. HCV core or NS5A transgenic mice displayed a reduction in C4 mRNA. Gamma interferon (IFN-γ)-induced C4 promoter activation was also impaired in the presence of HCV proteins. We further demonstrated that HCV core reduced the expression of upstream stimulating factor 1 (USF-1), a transcription factor important for basal C4 expression. On the other hand, the expression of interferon regulatory factor 1 (IRF-1), which is important for IFN-γ-induced C4 expression, was inhibited by hepatocytes expressing HCV NS5A. These results underscore the roles of HCV proteins in innate immune regulation in establishing a chronic infection.

Fibrinogen-beta (FBG-beta), an important acute-phase protein (APP), is generated by the liver as a target for inflammatory mediators. Here we identified FBG-beta as a hepatitis C virus (HCV) core interacting protein by screening a human liver complementary DNA (cDNA) library using mammalian two-hybrid analysis. An association between FBG-beta and HCV core protein was verified by confocal microscopy and coimmunoprecipitation from the transfected human hepatocyte (Huh-7) cell line. HCV core or genomic RNA transfected Huh-7 cells modestly increased FBG-beta protein expression when compared to the basal level in control hepatocytes. Transfection of HCV core or full-length (FL) gene into Huh-7 cells up-regulated basal FBG-beta promoter activity. Exogenous addition of IL-6 stimulates FBG-beta promoter activity in hepatocytes. However, ectopic expression of HCV core or FL in hepatocytes inhibited IL-6-stimulated FBG-beta promoter activation. Inhibition of endogenous FBG-beta expression following introduction of small interfering RNA (siRNA) into cells displayed a gain of function of promoter regulation by HCV core protein. Further studies suggested that HCV core gene expression in stable transfectants of Huh-7 cells resulted in a basal up-regulation of FBG-beta and other APPs. However, treatment with cytokines, interleukin-6 (IL-6), or tumor necrosis factor-alpha repressed FBG-beta and other acute-phase response (APR) genes.Our results reveal that the core/FBG-beta interaction may act as a regulatory feedback, allowing repression of IL-6-stimulated APR genes. Together, these data suggested a network of interactions between HCV core and the hepatic APR genes, and may contribute to impaired innate immunity for viral persistence.

The compartmentalization of viral variants in distinct host tissues is a frequent event in many viral infections. Although hepatitis B virus (HBV) classically is considered hepatotropic, it has strong lymphotropic properties as well. However, unlike other viruses, molecular evolutionary studies to characterize HBV variants in compartments other than hepatocytes or sera have not been performed. The present work attempted to characterize HBV sequences from the peripheral blood leukocytes (PBL) of a large set of subjects, using advanced molecular biology and computational methods. The results of this study revealed the exclusive compartmentalization of HBV subgenotype Ae/A2-specific sequences with a potent immune escape G145R mutation in the PBL of the majority of the subjects. Interestingly, entirely different HBV genotypes/subgenotypes (C, D, or Aa/A1) were found to predominate in the sera of the same study populations. These results suggest that subgenotype Ae/A2 is selectively archived in the PBL, and the high prevalence of G145R indicates high immune pressure and high evolutionary rates of HBV DNA in the PBL. The results are analogous to available literature on the compartmentalization of other viruses. The present work thus provides evidence in favor of the compartment-specific abundance, evolution, and emergence of the potent immune escape mutant. These findings have important implications in the field of HBV molecular epidemiology, transmission, transfusion medicine, organ transplantation, and vaccination strategies.

HBx genetic variability was explored in the Eastern Indian population with low HCC incidence. DNase I sensitive HBV DNA was detected in 53% samples, which differed significantly between clinical groups (P

We have studied the properties of various isomers of potassium clusters containing even number of atoms ranging from 2 to 20 at the ab initio level. The geometry optimization calculations of the isomers of each cluster are performed by using all-electron density functional theory with gradient corrected exchange-correlation functional. Using the optimized geometries of different isomers we investigate the evolution of binding energy, ionization potential, and static polarizability with the increasing size of the clusters. The polarizabilities are calculated by employing Möller-Plesset perturbation theory and time-dependent density functional theory. The polarizabilities of dimer and tetramer are also calculated by employing large basis set coupled cluster theory with single and double excitations and perturbative triple excitations. The time-dependent density functional theory calculations of polarizabilities are carried out with two different exchange-correlation potentials: (i) an asymptotically correct model potential and (ii) within the local density approximation. A systematic comparison with the other available theoretical and experimental data for various properties of small potassium clusters mentioned above has been performed. These comparisons reveal that both the binding energy and the ionization potential obtained with gradient-corrected potential match quite well with the already published data. Similarly, the polarizabilities obtained with Möller-Plesset perturbation theory and with model potential are quite close to each other and also close to experimental data.

In this paper we employ two distinct approaches - all-electron \textit{ab initio} method and the spherical jellium background model- within time dependent density functional theory to calculate the long range dipole-dipole dispersion coefficient (van der Waals coefficient) $C_{6}$ of sodium atom clusters containing even number of atoms ranging from 2 to 20 atoms. The dispersion coefficients are obtained via Casimir-Polder relation. All the calculations are carried out with local density approximation for exchange-correlation energy functional. These two sets of results are compared to assess the accuracy of jellium based results and to ascertain the effect of detail ionic structure of the clusters on the van der Waals coefficient., Comment: 15 pages including 2 figures. To be published in Journal of Computational Methods in Science and Engineering

We have theoretically studied the collective modes of a quasi two-dimensional (Q2D) Bose condensate in the large gas parameter regime by using a formalism which treats the interaction energy beyond the mean-field approximation. In the calculation we use the perturbative expansion for the interaction energy by incorporating the Lee, Huang and Yang (LHY) correction term. The results show that incorporation of this higher order term leads to detectable modifications in the mode frequencies., 10 pages, 2 figures

Objective: To study the prevalence of G1862T mutation in hepatitis B virus (HBV) isolates among Eastern Indian patients and its relationship with genotypes, HBeAg status and disease manifestation. Methods: HBV DNA was isolated from patients, amplified by nested PCR and sequenced directly. Results: Of the 102 patients, 32 were HBeAg positive and 70 HBeAg negative; 55, 24 and 23 isolates were infected with genotypes D, A and C, respectively. G1862T was detected in 18 samples, 15 (83%) of them belonged to genotype A (subgenotype HBV/A1), 3 (17%) to genotype D. This mutation was more frequent in HBeAg-negative than in HBeAg-positive patients (21 vs. 9%), whereas in HBV/A1 it was as common in HBeAg-positive as in HBeAg-negative patients and significantly associated with T1762/A1764 mutation. The mean viral load was lower in patients with G1862T mutation. Furthermore, this mutation was common in various clinical outcomes. Conclusion: In our community, G1862T mutation was predominantly found in HBV/A1 isolates irrespective of HBeAg status. Moreover this mutation could not be correlated to the clinical outcome. These findings indicate that the G1862T mutation is probably a part of the natural variability of HBV/A1.

Theoretical Study of Electronic and Response Properties of Large Metal Clusters

In this paper we employ the hydrodynamic formulation of time-dependent density functional theory to obtain the van der Waals coefficientsC 6 andC 8 of alkali metal clusters of various sizes including very large clusters. Such calculations become computationally very demanding in the orbital-based Kohn-Sham formalism, but are quite simple in the hydrodynamic approach. We show that for interactions between the clusters of the same sizes,C 6 andC 8 scale as the sixth and the eighth power of the cluster radius, respectively, and approach their classically predicted values for the large size clusters.

Occult hepatitis B is defined by the presence of hepatitis B virus (HBV) DNA in the serum in absence of hepatitis B surface antigen (HBsAg). Studies were conducted to screen for occult HBV infection among family members of HBV carriers, incidentally detected positive for HBV infection with a view to assess the pattern of virus transmission among them. Nested PCR assay, employing independent sets of primers to surface and core genes, was used for detection of HBV DNA in serum samples from 28 index cases with asymptomatic HBV infection, and in serum samples from 72 HBsAg negative/anti-HBc positive family members. HBV DNA was detected in 15 HBsAg negative family members of 10 HBsAg positive index patients and was studied in detail. Direct sequencing of S gene region of 25 isolates (10 index cases and 15 contacts) and phylogenetic analysis with data base sequences revealed that genotypes A, C, and D and subtype adw2, adr, and ayw3 were present among them. Evidence of transmission from outside family sources was found in addition to intrafamilial transmission among individuals with occult infection. Mutations in the major hydrophilic loop (MHL) of the S gene region were also detected, including the 'vaccine escape' mutation G145R in three cases. Although majority of the occult infection was associated with low viral load, 3/15 (20%) cases were with higher viral load and potential infectivity. These cases are especially notable in diagnostic, blood banking, and transplantation services.

Objective: The aim of the present study was to characterize the predominant hepatitis B virus (HBV) strains and their molecular variants present in the HBV isolates of the different genotypes found among the chronic carriers of the virus in our community. Methods: Precore/core and core promoter regions of HBV DNA were amplified by polymerase chain reaction and then subjected to direct sequencing. Of the 64 hepatitis B surface antigen (HBsAg)-positive chronic HBV carriers investigated, 44 were HBeAg negative and 20 were HBeAg positive. Results: In addition to genotype D, which was the predominant genotype, 12 genotype C (18.7%) and 6 genotype A (9.4%) were also detected. Presence of T at nt 1858 has often been related to the development of precore stop mutation at nt 1896, while that of C has been related to the development of 1762–1764 double mutation. In our study group, 39 of the 44 HBeAg-negative samples have T1858. The precore stop codon mutation was found in only 8 (18%) of the HBeAg-negative samples. More than half of the HBeAg-negative samples had wild-type sequence in the precore region. The core promoter region could be sequenced from 40 samples, and 1762–1764 double mutation was detected in 13 (32.5%) of them. No significant changes could be detected in the core amino acid sequence of these isolates. Conclusion: The pattern of core promoter and precore mutation of HBV isolates in the present study is atypical and not in accordance with reports from other parts of the world, where genotype D and genotype C with T at codon 1858 are common.

A procedure for calculating the A term and the A/D ratio of magnetic circular dichroism (MCD) within time-dependent density functional theory (TD-DFT) is described. Utilizing an implementation of the MCD theory within the Amsterdam Density Functional program, the A term contributions to the MCD spectra of MnO(4) (-), CrO(4) (2-), VO(4) (3-), MoO(4) (2-), VO(4) (3-), MoS(4) (2-), Se(4) (2+), Te(4) (2+), Fe(CN)(6) (4-), Ni(CN)(4) (2-), trichlorobenzene, hexachlorobenzene, tribromobenzene, and hexabromobenzene are calculated. For the most part, agreement between theory and experiment for A/D ratios and the relative magnitude of A terms is found to be good, leading to simulated spectra that are similar in appearance to those derived from measurements. The A terms are found to be too small whenever comparison with experiment was possible, probably due to the neglect of environment effects on the incident radiation and the relative low accuracy of dipole strengths calculated within TD-DFT.

Background Hepatitis B Virus (HBV) X protein (HBx) is known to be involved in the initiation and progression of hepatocellular carcinoma (HCC) through modulation of host gene response. Alterations in miRNA expressions are frequently noted in HCC. This study is aimed to examine the role of HBx protein in the modulation of oncogenic miRNA-21, miRNA-222 and tumor suppressor miRNA-145 in malignant hepatocytes. Methods Expressions of miRNA-21, miRNA-222 and miRNA-145 were measured in HepG2 cells transfected with HBx-plasmid (genotype D) and with full length HBV genome (genotype D) and also in stably HBV producing HepG2.2.15 cells using real time PCR. Their target mRNAs and proteins - PTEN, p27 and MAP3K - were analyzed by real time PCR and western blot respectively. miRNA expressions were measured after HBx/D mRNA specific siRNA treatment. The expressions of these miRNAs were analyzed in liver cirrhosis and HCC patients also. Results The study revealed a down-regulation of miRNA-21 and miRNA-222 expressions in HBx transfected HepG2 cells, pUC-HBV 1.3 plasmid transfected HepG2 cells as well as in HepG2.2.15 cells. Down regulation of miRNA-21 and miRNA-222 expression was observed in patient serum samples. Down regulation of miRNA-145 expression was observed in HepG2 cells transiently transfected with HBx and pUC-HBV1.3 plasmid as well as in patient samples but the expression of miRNA-145 was increased in HepG2.2.15 cells. Target mRNA and protein expressions were modulated in HepG2 cells and in HepG2.2.15 cell line consistent with the modulation of miRNA expressions. Conclusion Thus, HBx protein differentially modulated the expression of miRNAs. The study throws light into possible way by which HBx protein acts through microRNA and thereby regulates host functioning. It might suggest new therapeutic strategies against hepatic cancer.

Background Microglial cells, which are resident macrophages of the central nervous system, play important roles in immune responses and pathogenesis. Japanese encephalitis virus (JEV) is a neurotropic virus that infects microglial cells in brain. Several microRNAs including miR-155 and miR-146a play an important role in defining the microglia inflammatory profile. In this study, we have investigated the effect of miR-155 and miR-146a modulation on JEV infection as well as innate immune responses in human microglial cells. Methods In vitro studies were performed in JEV-infected human microglial CHME3 cells. miR-155 or miR-146a were overexpressed and total RNA and protein were extracted following JEV-infection. Expression of genes involved in innate immune responses was studied by PCR array, quantitative real-time PCR (qPCR), western blot and Fluorescence activated cell sorter (FACS). JEV replication was monitored by studying the viral RNA by qPCR, protein by western blot, and titres by plaque assay. Results Overexpression of miR-155 in CHME3 cells resulted in significantly reduced JEV replication whereas miR-146a overexpression had an insignificant effect. Additionally, interferon regulatory factor 8 (IRF8) and complement factor H (CFH) were induced during JEV infection; however, this induction was attenuated in miR-155 overexpressing cells following JEV infection. Further, JEV-induced NF-κB regulated downstream gene expression was attenuated. Interestingly, an increased level of CD45, a negative regulator of microglia activation and a reduced phosphorylated-Signal Transducers and Activators of Transcription (p-STAT1) expression was observed in miR-155 overexpressing cells upon JEV infection. Conclusion Induction of miR-155 in human microglial cells may negatively modulate JEV-induced innate immune gene expression and may have a beneficial role in limiting JEV replication in human microglial cells.