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Hepatitis E virus (HEV) is the main cause of acute hepatitis worldwide. The viral infection caused by G1 HEV in pregnant women has become a major health concern in the past few years. The mechanism underlying the pathogenesis of viral infection in HEV G1 isolates is attributed to four different open-reading frames (ORFs) i.e., ORF1, ORF2, ORF3 and ORF4. The present analysis has considered ORF4 protein as the molecular target due to its intrinsic disorder propensity. Intrinsically disordered regions (IDRs) are regions in proteins that do not possess stable secondary and tertiary structure and are prevalent in eukaryotes. IDRs are found to be closely associated with numerous human diseases, for instance, Parkinson and Alzheimer disease. The extreme flexibility and random coiled conformations of IDR allow it to undergo protein-protein interaction (PPI). The 3-dimensional (3D) structures of the target protein were designed using homology modelling algorithms. The generated models were assessed through structure verification tool PROCHECK. In this paper, we provide an overview of ORF4 protein structure-function relationship and its involvement in several biological processes through PPIs. Our results suggest that ORF4 protein has the potential to act as drug molecule, thus can accelerate the process of drug designing strategies against HEV.

Abstract Background 6-Gingerol (6-G) is the primary active phytocomponent of ginger and has been shown to regulate multiple targets against cancer and its treatment. Androgen receptors (ARs) remain critical in the progression of prostate cancer (PCa). This study focuses on investigating 6-G as a promising anti-cancerous agent that inhibits AR activity significantly. Methods In this study, molecular docking simulation was done to investigate the binding affinity of 6-G and control drug Bicalutamide (BT) against oncogenic AR and tumor suppressor estrogen receptor β (ERβ). The crystal structure of AR and ERβ was retrieved from Protein Data Bank (PDB) and docked with 3D Pubchem structures of 6-G using iGEMDOCK and AutoDock. Further in vitro study was done to evaluate the antioxidant, anti-cancerous, apoptotic, and wound healing potential of 6-G. Results The result displays that 6-G shows good binding affinity with AR and ERβ. Condensation of the nucleus, change in mitochondrial membrane potential (MMP) and the ability to induce reactive oxygen species (ROS) were done in human PCa PC-3 cells. Results from the MTT assay demonstrated that 6-G and control drug BT showed significant (p

Abstract The most common denture material used for dentistry is poly-methyl-methacrylate (PMMA). Usually, the polymeric PMMA material has numerous biological, mechanical and cost-effective shortcomings. Hence, to resolve such types of drawbacks, attempts have been made to investigate fillers of the PMMA like alumina (Al2O3), silica (SiO2), zirconia (ZrO2) etc. For the enhancement of the PMMA properties a suitable additive is required for its orthopedic applications. Herein, the main motive of this study was to synthesize a magnesium oxide (MgO) reinforced polymer-based hybrid nano-composites by using heat cure method with superior optical, biological and mechanical characteristics. For the structural and vibrational studies of the composites, XRD and FT-IR were carried out. Herein, the percentage of crystallinity for all the fabricated composites were also calculated and found to be 14.79–30.31. Various physical and optical parameters such as density, band gap, Urbach energy, cutoff energy, cutoff wavelength, steepness parameter, electron–phonon interaction, refractive index, and optical dielectric constant were also studied and their values are found to be in the range of 1.21–1.394 g/cm3, 5.44–5.48 eV, 0.167–0.027 eV, 5.68 eV, 218 nm, 0.156–0.962, 4.273–0.693, 1.937–1.932, and 3.752–3.731 respectively. To evaluate the mechanical properties like compressive strength, flexural strength, and fracture toughness of the composites a Universal Testing Machine (UTM) was used and their values were 60.3 and 101 MPa, 78 and 40.3 MPa, 5.85 and 9.8 MPa-m1/2 respectively. Tribological tests of the composites were also carried out. In order to check the toxicity, MTT assay was also carried out for the PM0 and PM15 [(x)MgO + (100 − x) (C5O2H8)n] (x = 0 and 15) composites. This study provides a comprehensive insight into the structural, physical, optical, and biological features of the fabricated PMMA-MgO composites, highlighting the potential of the PM15 composite with its enhanced density, mechanical strength, and excellent biocompatibility for denture applications.

Among all diseases, cancer has the highest fatality rate in the world. Conventional therapies have come a long way, but they still are not a panacea for cancer. Nanotechnology-based compositions of several nanoparticles (NPs) hold promise as a cancer therapy. The co-delivery of amphiphilic molecules is feasible with lipid-polymer hybrid nanoparticles (LPHNPs). Increased encapsulation of pharmaceuticals with a regulated release profile is made possible by the physical benefits of polymers and the resembling features of lipids. For increased cytotoxicity against cancer, LPHNPs have been created for the co-transport of phytochemicals and other chemotherapeutic medicines. Specifically, LPHNPs are used as carrier systems to deliver drugs to tumour locations in a regulated fashion. This method makes use of the biocompatibility of lipids and the structural advantages of polymers to provide controlled drug administration. Co-loaded LPHNPs demonstrated improved cytotoxicity, and the incorporation of phytochemicals into various medications resulted in enhanced chemo-sensitization of carcinomas. The shortcomings of existing nano-delivery systems and the benefits of LPHNPs have been discussed in this review.

AbstractIn this study, we have developed an environment friendly and novel approach for the synthesis of silver nanoparticles utilising plumeria pudica leaf extract. In this technique, the leaf extract was employed as both a reducing agent for the reduction of a silver nitrate (AgNO3) solution and a capping agent, leading to the synthesis of silver nanoparticles (Ag NPs). The outcomes of these analyses revealed an average particle size of ∼19 nm as determined by SEM, while XRD measurements indicated a crystalline domain size of ∼12 nm and a lattice parameter of approximately 4.087467 Å. Furthermore, the anti-cancer potential of the synthesised silver nanoparticles was evaluated, unveiling an IC50 value of around 28 µM. This suggests that the introduced silver nanoparticles may have triggered apoptosis, consequently inducing cell death. These findings underscore the potential of utilising environmentally benign silver nanoparticles in lung cancer chemotherapy.

Abstract Background Hepatitis E virus (HEV) is the cause of a liver disease hepatitis E. The translation product of HEV ORF2 has recently been demonstrated as a protein involved in multiple functions besides performing its major role of a viral capsid. As intrinsically disordered regions (IDRs) are linked to various essential roles in the virus’s life cycle, we analyzed the disorder pattern distribution of the retrieved ORF2 protein sequences by employing different online predictors. Our findings might provide some clues on the disorder-based functions of ORF2 protein that possibly help us in understanding its behavior other than as a HEV capsid protein. Results The modeled three dimensional (3D) structures of ORF2 showed the predominance of random coils or unstructured regions in addition to major secondary structure components (alpha helix and beta strand). After initial scrutinization, the predictors VLXT and VSL2 predicted ORF2 as a highly disordered protein while the predictors VL3 and DISOPRED3 predicted ORF2 as a moderately disordered protein, thus categorizing HEV-ORF2 into IDP (intrinsically disordered protein) or IDPR (intrinsically disordered protein region) respectively. Thus, our initial predicted disorderness in ORF2 protein 3D structures was in excellent agreement with their predicted disorder distribution patterns (evaluated through different predictors). The abundance of MoRFs (disorder-based protein binding sites) in ORF2 was observed that signified their interaction with binding partners which might further assist in viral infection. As IDPs/IDPRs are targets of regulation, we carried out the phosphorylation analysis to reveal the presence of post-translationally modified sites. Prevalence of several disordered-based phosphorylation sites further signified the involvement of ORF2 in diverse and significant biological processes. Furthermore, ORF2 structure-associated functions revealed its involvement in several crucial functions and biological processes like binding and catalytic activities. Conclusions The results predicted ORF2 as a protein with multiple functions besides its role as a capsid protein. Moreover, the occurrence of IDPR/IDP in ORF2 protein suggests that its disordered region might serve as novel drug targets via functioning as potential interacting domains. Our data collectively might provide significant implication in HEV vaccine search as disorderness in viral proteins is related to mechanisms involved in immune evasion.

Cervical cancer has historically been the deadliest malignancy in women. It continues to create several health issues, particularly in developing countries. Current management techniques include cisplatin-based chemoradiotherapy and surgical procedures. These treatments have some drawbacks, such as low absorption, side effects, systemic toxicity, the development of resistance to various therapeutics, and targeting that is too broad and insufficiently precise. To compensate for these shortcomings, researchers are still hunting for novel anticancer drugs. Plant-derived phytochemicals and their derivatives have promise for improving cancer treatment efficacy while reducing adverse effects. Phytomolecules are utilized to treat cancer, but they are difficult to work with since they have low bioavailability, excessive dosages, negative side effects and low therapeutic indices. They must be administered in high doses to be effective and nanotechnology can be utilized to overcome these obstructions. Piperlongumine, rutin, quercetin, lycopene, leutin, curcumin, green tea polyphenols, and other phytomolecules have been loaded into a carrier called nanophytomedicine to be beneficial in chemoprevention and chemotherapy. Nanocarriers have a high level of biocompatibility, biodegradability, and biological stability. Nano-based drug delivery systems are an excellent approach to improving therapeutic specificity, making it easier for the body to absorb it reducing the drug's breakdown and systemic toxicity. This review looks at phytonanomedicine and how it can be used to treat cervical cancer instead of traditional chemo-radiotherapy.

Purpose: To study the corelation between outer retinal layer thickness (ORL), outer photoreceptor segment thickness (PROS), and central macular thickness (CMT) with best-corrected visual acuity (BCVA) in patients having clinically significant macular edema (CSME) and compare these parameters with normal patients. Methods: This was a prospective, nonrandomized, observational, comparative study done during the period of January to May 2019. The study included 60 eyes of 36 patients. The patient population was segregated into two Groups: Group I (30 normal eyes of 15 normal patients) and Group II (30 eyes of 21 diabetic patients) with CSME. The comparison between ORL, PROS, and CMT was made between both the groups, and the correlation between ORL thickness, PROS thickness, and CMT with BCVA in Group II was studied. Results: The mean age in Group I was 52.6+10.66 years, and 53.42+8.15 years in Group II. The male/female ratio was 1.1:1 in Group I and 4:3 in Group II. The mean CMT was greater in Group II (330.13 ± 37.01) than in Group I (222.20 ± 12.30). The mean ORL thickness was greater in Group I (97.73 ± 6.92) than in Group II (80.63 ± 9.03). The PROS thickness was statistically significant in Group I (35.05 ± 3.4) than in Group II (28.57 ± 3.53). There was a strong correlation between BCVA and ORL thickness (r = −0.580, P < 0.001) and more strong correlation between BCVA and PROS thickness in Group II (r = −0.611, P < 0.000). There was a moderate correlation between BCVA and CMT (r = 0.410, P < 0.025), and all results were statistically significant. Conclusion: Both ORL and PROS thickness were greater in healthy normal eyes than in eyes with CSME. BCVA was strongly correlated with PROS and ORL thickness and moderately associated with CMT.

Abstract Background Viral diseases are highly widespread infections caused by viruses. These viruses are passing from one human to other humans through a certain medium. The medium might be mosquito, animal, reservoir and food, etc. Here, the population of both human and mosquito vectors are important. Main body of the abstract The main objectives are here to introduce the historical perspective of mathematical modeling, enable the mathematical modeler to understand the basic mathematical theory behind this and present a systematic review on mathematical modeling for four vector-borne viral diseases using the deterministic approach. Furthermore, we also introduced other mathematical techniques to deal with vector-borne diseases. Mathematical models could help forecast the infectious population of humans and vectors during the outbreak. Short conclusion This study will be helpful for mathematical modelers in vector-borne diseases and ready-made material in the review for future advancement in the subject. This study will not only benefit vector-borne conditions but will enable ideas for other illnesses.

In the published publication [...]

The global health community is extremely concerned about the emergence and spread of antibiotic resistance as well as the evolution of new strains of disease-causing organisms. It takes the creation of novel pharmaceuticals or access to a supply of innovative therapeutics for a disease to be effectively treated. Commonly used medicinal herbs in our society could be a great source of medications to combat this issue. The antibacterial and anticancer capabilities of metallic nanoparticles made from plants are the main focus of this work. Two distinct nanoparticles were tested for their antibacterial and anticancer properties against four cancerous cell lines (prostate cancer, lung cancer A549, HeLa, and MCF-7) and five pathogenic microbes (Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli) bacteria, as well as one yeast (Candida albicans). Metallic nanoparticles from the plants Leucas cephalotes (AgNPs) and Ajuga macrosperma (FeONPs) were tested for their antibacterial capabilities using the agar well diffusion method and their ability to fight cancer using the MTS and MTT assays. According to the findings, AgNPs synthesized from Leucas cephalotes showed the strongest potential against Pseudomonas aeruginosa and Staphylococcus aureus. The zones of inhibition (ZOI) of Leucas cephalotes’ AgNPs were 19 mm and 18 mm respectively. It was also discovered that the FeONPs of Ajuga macrosperma had the most potential against the MCF-7 cancer cell line. Prostate cancer and lung cancer (A549) cell lines were the ones that responded most favorably to the AgNPs of Leucas cephalotes.

Abstract Background Hepatitis E virus (HEV) is a member of the family Hepeviridae and causes acute HEV infections resulting in thousands of deaths worldwide. The zoonotic nature of HEV in addition to its tendency from human to human transmission has led scientists across the globe to work on its different aspects. HEV also accounts for about 30% mortality rates in case of pregnant women. The genome of HEV is organized into three open reading frames (ORFs): ORF1 ORF2 and ORF3. A reading frame encoded protein ORF4 has recently been discovered which is exclusive to GT 1 isolates of HEV. The ORF4 is suggested to play crucial role in pregnancy-associated pathology and enhanced replication. Though studies have documented the ORF4’s importance, the genetic features of ORF4 protein genes in terms of compositional patterns have not been elucidated. As codon usage performs critical role in establishment of the host–pathogen relationship, therefore, the present study reports the codon usage analysis (based on nucleotide sequences of HEV ORF4 available in the public database) in three hosts along with the factors influencing the codon usage patterns of the protein genes of ORF4 of HEV. Results The nucleotide composition analysis indicated that ORF4 protein genes showed overrepresentation of C nucleotide and while A nucleotide was the least-represented, with random distribution of G and T(U) nucleotides. The relative synonymous codon usage (RSCU) analysis revealed biasness toward C/G-ended codons (over U/A) in all three natural HEV-hosts (human, rat and ferret). It was observed that all the ORF4 genes were richly endowed with GC content. Further, our results showed the occurrence of both coincidence and antagonistic codon usage patterns among HEV-hosts. The findings further emphasized that both mutational and selection forces influenced the codon usage patterns of ORF4 protein genes. Conclusions To the best of our knowledge, this is first bioinformatics study evaluating codon usage patterns in HEV ORF4 protein genes. The findings from this study are expected to increase our understanding toward significant factors involved in evolutionary changes of ORF4.

Abstract Background Hepatitis E virus (HEV) is a positive-sense RNA virus belonging to the family Hepeviridae. The genome of HEV is organized into three open-reading frames (ORFs): ORF1, ORF2, and ORF3. The ORF1 non-structural Y-domain region (YDR) has been demonstrated to play an important role in the HEV pathogenesis. The nucleotide composition, synonymous codon usage bias in conjunction with other factors influencing the viral YDR genes of HEV have not been studied. Codon usage represents a significant mechanism in establishing the host-pathogen relationship. The present study for the first time elucidates the detailed codon usage patterns of YDR among HEV and HEV-hosts (Human, Rabbit, Mongoose, Pig, Wild boar, Camel, Monkey). Results The overall nucleotide composition revealed the abundance of C and U nucleotides in YDR genomes. The relative synonymous codon usage (RSCU) analysis indicated biasness towards C and U over A and G ended codons in HEV across all hosts. Codon frequency comparative analyses among HEV-hosts showed both similarities and discrepancies in usage of preferred codons encoding amino acids, which revealed that HEV codon preference neither completely differed nor completely showed similarity with its hosts. Thus, our results clearly indicated that the synonymous codon usage of HEV is a mixture of the two types of codon usage: coincidence and antagonism. Mutation pressure from virus and natural selection from host seems to be accountable for shaping the codon usage patterns in YDR. The study emphasised that the influence of compositional constraints, codon usage biasness, mutational alongside the selective forces were reflected in the occurrence of YDR codon usage patterns. Conclusions Our study is the first in its kind to have reported the analysis of codon usage patterns on a total of seven different natural HEV hosts. Therefore, knowledge of preferred codons obtained from our study will not only augment our understanding towards molecular evolution but is also envisaged to provide insight into the efficient viral expression, viral adaptation, and host effects on the HEV YDR codon usage.

Introduction Delhi is hyperendemic for dengue virus (DENV) where all the four DENV have previously been reported. A constant vigilance of circulating DENV serotypes is important in surveillance, since the introduction of a new variant to areas affected by preexisting serotypes constitutes a risk factor for dengue hemorrhagic fever and dengue shock syndrome. Objectives This retrospective study was performed with an objective to determine the circulating serotype and genotype of DENV in acute phase blood samples of patients who have reported to a tertiary liver care hospital in New Delhi during the last 2 years (2017–2018). Methods The data of clinician-initiated testing for dengue nonstructural protein 1 (NS1) antigen (Ag) was searched in the institutional hospital information system. The serum sample of dengue NS1 Ag-positive cases confirmed by enzyme-linked immunosorbent assay (ELISA; PANBIO, Gyeonggi-do, ROK) and a fever duration of less than 5 days were retrieved from the laboratory archive. The DENV serotyping on these sample was performed by reverse transcriptase polymerase chain reaction (RT-PCR). Sequencing and phylogenetic analysis was done for the capsid premembrane (CprM) region to determine the genotype. Results A total of 440 acute-phase samples were received. Twenty one (4.77%) were positive for dengue NS1 Ag with a mean age of 35.1 years and male-to-female ratio of 1.1:1. Eight cases (38.09%) were positive by dengue RT-PCR and all belonged to DENV-3 serotypes. Phylogenetic tree analysis revealed DENV-3 clustered to genotype III with 100% homology with 2008 Indian subcontinent strain. Conclusion This study revealed circulation of DENV-3, genotype III in Delhi from 2017 to 2018, similar to the 2008 viral type. Virological surveillance is an important exercise to be done for viral infections with public threat and outbreak potential.

Abstract Background Hepatitis E virus (HEV) of the family Hepeviridae is a major causative agent of acute hepatitis in developing countries. The Y-domain is derived from multi-domain non-structural polyprotein encoded by open reading frame 1 (ORF1). Previous studies have demonstrated the essentiality of Y-domain sequences in HEV life cycle; however, its function remains completely unexplored. The following study was thus conceptualized to examine the detailed computational investigation for the putative Y-domain to estimate its phylogenetic assessment, physiochemical properties, structural and functional characteristics using in silico analyses. Results The phylogenetic assessment of Y-domain with a vast range of hosts indicated that the protein was very well conserved throughout the course of evolution. The Y-domain was found to be unstable, hydrophilic and basic in nature with high thermostability value. Structural analysis of Y-domain revealed mixed α/β structural fold of the protein having higher percentage of alpha-helices. The three-dimensional (3D) protein model generated through homology modelling revealed the presence of clefts, tunnels and pore. Gene ontology analysis predicted Y-domain protein’s involvement in several binding and catalytic activities as well as significant biological processes. Mutations in the conserved amino acids of the Y-domain suggested that it may stabilize or de-stabilize the protein structure that might affect its structure–function relationship. Conclusions This theoretical study will facilitate towards deciphering the role of unexplored Y-domain, thereby providing better understanding towards the pathogenesis of HEV infection.

Abstract Background Hepatitis E is a liver disease caused by the pathogen hepatitis E virus (HEV). The largest polyprotein open reading frame 1 (ORF1) contains a nonstructural Y-domain region (YDR) whose activity in HEV adaptation remains uncharted. The specific role of disordered regions in several nonstructural proteins has been demonstrated to participate in the multiplication and multiple regulatory functions of the viruses. Thus, intrinsic disorder of YDR including its structural and functional annotation was comprehensively studied by exploiting computational methodologies to delineate its role in viral adaptation. Results Based on our findings, it was evident that YDR contains significantly higher levels of ordered regions with less prevalence of disordered residues. Sequence-based analysis of YDR revealed it as a “dual personality” (DP) protein due to the presence of both structured and unstructured (intrinsically disordered) regions. The evolution of YDR was shaped by pressures that lead towards predominance of both disordered and regularly folded amino acids (Ala, Arg, Gly, Ile, Leu, Phe, Pro, Ser, Tyr, Val). Additionally, the predominance of characteristic DP residues (Thr, Arg, Gly, and Pro) further showed the order as well as disorder characteristic possessed by YDR. The intrinsic disorder propensity analysis of YDR revealed it as a moderately disordered protein. All the YDR sequences consisted of molecular recognition features (MoRFs), i.e., intrinsic disorder-based protein–protein interaction (PPI) sites, in addition to several nucleotide-binding sites. Thus, the presence of molecular recognition (PPI, RNA binding, and DNA binding) signifies the YDR’s interaction with specific partners, host membranes leading to further viral infection. The presence of various disordered-based phosphorylation sites further signifies the role of YDR in various biological processes. Furthermore, functional annotation of YDR revealed it as a multifunctional-associated protein, due to its susceptibility in binding to a wide range of ligands and involvement in various catalytic activities. Conclusions As DP are targets for regulation, thus, YDR contributes to cellular signaling processes through PPIs. As YDR is incompletely understood, therefore, our data on disorder-based function could help in better understanding its associated functions. Collectively, our novel data from this comprehensive investigation is the first attempt to delineate YDR role in the regulation and pathogenesis of HEV.

Background: Dengue fever has become a hampering menace in New Delhi India, since the disease has become hyperendemic, due to circulation of multiple serotypes of dengue virus (DENV). This hyperendemicity poses a greater risk of secondary infections in human health system. This is a major issue which leads to apprehension amongst the researchers and health organizations and thus requires regular epidemiological surveillance. Methods: We analyzed the prevalence and serotypic distribution of dengue fever cases reported from the Southern part of New Delhi during continued surveillance from 2011 to 2017. The blood samples for the investigation were obtained from the patients suspected with dengue fever attending the OPD at a local Health Centre. The data for 2011–2016 was already published from our laboratory. The samples collected during 2017 were serotyped and characterized in the present study. Results: A total of 565 samples (59%) were positive for DENV of 958 samples tested by RT-PCR during 7 years (2011–2017). Our study has shown that most infections were caused by DENV-2 during 2011−2015. The data has shown occurrence of all four serotypes of DENV during 2015 and predominance of DENV-3 in 2016 and 2017. Further, predominant combination of DENV-1 and DENV-2 was found in most of the co-infections. To the best of our knowledge this is the first study showing the epidemiological trend of dengue fever in reference to the circulating DENV serotypes and co-infections from a hyperendemic region of New Delhi during 2011–2017. Conclusions: This hyperendemic pattern of DENV and instantaneous shift in circulation of its serotypes is likely pose a greater risk of secondary infections. Inclusion of comprehensive community and hospital surveillance of dengue fever will assist in formulation and implementation of effective control measures.

The novel reactive methylmethacrylate (MMA) thermoplastic resin (commercially known as Elium® resin) is the first liquid thermoplastic resin which is curable at room temperature. This resin is a competitive solution against traditional epoxy-based composites. In this work, novel non-crimp (NC) glass fabric/MMA resin composites were manufactured using a vacuum infusion process. Cellulose microcrystals (CMCs) were dispersed in the resin to improve the fibre/matrix interface and the composite properties. CMCs were first dispersed in the resin using an ultrasonication process and then the CMC/MMA resin suspension, mixed with a peroxide initiator, was infused into the reinforcing fabric. The amounts of CMCs dispersed in the resin were 0.5%, 1% and 2% (of the weight of the resin). The influence of CMCs on the interlaminar shear strength (interface), flexural properties, abrasive wear and dynamic-mechanical behaviour was thoroughly investigated. From the mechanical characterisation, it was observed that the addition of 1% CMC to the NC glass/MMA resin composites improved the flexural strength, interlaminar shear strength and wear performance by 30.77%, 38.04% and 22.27%, respectively as compared to the neat glass/MMA resin composite. Above this amount of CMC (i.e., 1 wt%), the properties started to degrade as a result of CMC agglomeration.

Background: Dengue is a rapidly emerging arthropod borne viral infection affecting tropical and sub-tropical regions of the world. Dengue is an acute febrile illness but sometimes causes more fatal complications like dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS). Delhi, the capital of India has become hyper endemic for dengue virus because all the four serotypes are circulating here. Methods: The present study describes the identification of dengue virus from clinical samples collected from the suspected dengue patients from New Delhi, India during 2016. The CprM region of Dengue virus genome was analyzed for phylogenetic, selection pressure and Shannon entropy analyses. Results: The present study reports circulation of a single serotype (DENV-3) in New Delhi, during 2016. The phylogenetic analysis revealed that Indian subcontinent (genotype III) of DENV-3 was circulating in Delhi during this period. Neutral selection pressure in the analyzed region revealed relatively conserved nature of this part of the Dengue virus genome. Amino acid at 31 was positively selected and had high entropy value suggesting probability of variation at this position. Conclusions: The changing trend in circulation of dengue virus serotypes necessitates the continuous epidemiological surveillance for the dengue outbreaks in this region. Keywords: Dengue virus, Epidemiology, Phylogenetic analysis, Selection pressure, Entropy analysis

Quercetin is one of the most important plant flavanols, having several pharmacological and biological uses. Quercetin (Q) is an extremely hydrophobic phytochemical and has poor intracellular absorption, which makes its use limited. Present research demonstrates that quercetin-loaded PLGA nanoparticles (PLGA-QNPs) could overcome its low hydrophilicity and improve its anti-cancer potential. PLGA nanoparticles loaded with Q were prepared by the solvent evaporation technique and its anticancer activity was examined in vitro as well as in vivo. The cell viability was assessed through MTT assay and apoptosis was assayed through Hoechst-PI and EB/AO double staining followed by mitochondrial damage through Mito-tracker RMX-Ros. Gene expression was examined through RT-PCR. Cell cycle arrest in G2/M phase was analyzed through FACS. The results obtained revealed that PLGA-QNPs significantly reduced the viability of human cervical and breast cancer cell lines. PLGA-QNPs induced apoptosis in human cervical cancer cells in a dose dependent manner. The gene expression of PI3K/AKT was down-regulated and FoxO1 was upregulated in PLGA-QNP-treated cells, which showed a high expression level of active Caspase-3 and 7, which are responsible for apoptosis. In addition, PLGA-QNPs reduced the average number of tumors and prolonged the tumor latency period in DMBA-induced mammary adenocarcinoma SD rats. These findings suggest that PLGA-QNPs inhibit cervical and breast cancer progression via mitochondrial dependent Caspase-3 and 7 and mitochondrial independent FoxO1 activation with concomitant suppression of the PI3K/AKT pathway. For future studies, we suggest that potential druggability efficacy and clinical development of anticancer PLGA-QNPs need to be evaluated intensely for successful anticancer drug development.

Hepatitis B virus X protein C-terminal 127 amino acid truncation is often found expressed in hepatocellular carcinoma (HCC) tissue samples. The present in vitro study tried to determine the role of this truncation mutant in the hepatitis B–related liver diseases such as fibrosis, cirrhosis, HCC, and metastasis. HBx gene and its 127 amino acid truncation mutant were cloned in mammalian expression vectors and transfected in human hepatoma cell line. Changes in cell growth/proliferation, cell cycle phase distribution, expression of cell cycle regulatory genes, mitochondrial depolarization, and intracellular reactive oxygen species (ROS) level were analyzed. Green fluorescent protein (GFP)–tagged version of HBx and the truncation mutant were also created and the effects of truncation on HBx intracellular expression pattern and localization were studied. Effect of time lapse on protein expression pattern was also analyzed. The truncation mutant of HBx is more efficient in inducing cell proliferation, and causes more ROS production and less mitochondrial depolarization as compared with wild type (wt) HBx. In addition, gene expression is altered in favor of carcinogenesis in the presence of the truncation mutant. Furthermore, mitochondrial perinuclear aggregation is achieved earlier in the presence of the truncation mutant. Therefore, HBx C-terminal 127 amino acid truncation might be playing important roles in the development of hepatitis B–related liver diseases by inducing cell proliferation, altering gene expression, altering mitochondrial potential, inducing mitochondrial clustering and oxidative stress, and changing HBx expression pattern.

Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infection in children and infants. To date, there is no effective vaccine available against RSV. Heparan sulfate is a type of glycosaminoglycan that aids in the attachment of the RSV to the host cell membrane via the G protein. In the present study, the effect of amino acid substitution on the structure and stability of the ectodomain G protein was studied. Further, it was investigated whether mutation (K117A) in the CX3C motif of G protein alters the binding with heparan sulfate. The point mutation significantly affects the conformational stability of the G protein. The mutant protein showed a low binding affinity with heparan sulfate as compared to the wild-type G protein, as determined by fluorescence quenching, isothermal titration calorimetry (ITC), and molecular docking studies. The low binding affinity and decreased stability suggested that this mutation may play an important role in prevention of attachment of virion to the host cell receptors. Collectively, this investigation suggests that mutation in the CX3C motif of G protein may likely improve the efficacy and safety of the RSV vaccine.

This paper reports the use of cellulose microcrystals (CMCs) for improving fibre-matrix interface, mechanical, dynamic mechanical and thermal degradation behaviour of glass fibre reinforced epoxy composites. An ultrasonic treatment for 1 h was used to disperse CMCs (1–3 wt%) within an epoxy resin, which was subsequently infused through glass fabrics to develop hierarchical composites containing both macro and micro-scale reinforcements. It was observed that CMC dispersion in the epoxy resin was homogeneous at 1 wt% CMC and further increase in CMC concentrations led to linear increase in both agglomerate size and total agglomerated area. Addition of 1 wt% CMC to the composite matrix drastically changed the glass fibre-epoxy interface and led to a maximum improvement of 65% in interlaminar shear strength, 14% in tensile strength, 76% in flexural strength, 111% and 119% in fracture energy in tensile and flexural modes, 9.4% in impact strength, 13.5% in storage modulus, 21.9% in loss modulus and 13 °C in the glass transition temperature of composites. Therefore, the use of CMCs could be an industrially viable, economical and eco-friendly approach of developing hierarchical glass fibre composites with considerably improved performance. Keywords: Glass fibre composites, Cellulose microcrystals, Fibre-matrix interface, Mechanical properties, Fracture energy, Dynamic mechanical performance

Abstract Background Malaria, Dengue and Chikungunya are vector borne diseases with shared endemic profiles and symptoms. Coinfections with any of these diseases could have fatal outcomes if left undiagnosed. Understanding the prevalence and distribution of coinfections is necessary to improve diagnosis and designing therapeutic interventions. Methods We have carried out a systematic search of the published literature based on PRISMA guidelines to identify cases of Malaria, Dengue and Chikungunya coinfections. We systematically reviewed the literature to identify eligible studies and extracted data regarding cases of coinfection from cross sectional studies, case reports, retrospective studies, prospective observational studies and surveillance reports. Results Care full screening resulted in 104 publications that met the eligibility criteria and reported Malaria/Dengue, Dengue/Chikungunya, Malaria/Chikungunya and Malaria/Dengue/Chikungunya coinfections. These coinfections were spread over six geographical locations and 42 different countries and are reported more frequently in the last 15 years possibly due to expanding epidemiology of Dengue and Chikungunya. Few of these reports have also analysed distinguishing features of coinfections. Malaria/Dengue coinfections were the most common coinfection followed by Dengue/Chikungunya, Malaria/Chikungunya and Malaria/Dengue/Chikungunya coinfections. P. falciparum and P. vivax were the commonest species found in cases of malaria coinfections and Dengue serotype-4 commonest serotype in cases of dengue coinfections. Most studies were reported from India. Nigeria and India were the only two countries from where all possible combinations of coinfections were reported. Conclusion We have comprehensively reviewed the literature associated with cases of coinfections of three important vector borne diseases to present a clear picture of their prevalence and distribution across the globe. The frequency of coinfections presented in the study suggests proper diagnosis, surveillance and management of cases of coinfection to avoid poor prognosis of the underlying etiology.

The global burden of disease caused by a respiratory syncytial virus (RSV) is becoming more widely recognized in young children and adults. Heparan sulfate helps in attaching the virion through G protein with the host cell membrane. In this study, we examined the structural changes of ectodomain G protein (edG) in a wide pH range. The absorbance results revealed that protein maintains its tertiary structure at physiological and highly acidic and alkaline pH. However, visible aggregation of protein was observed in mild acidic pH. The intrinsic fluorescence study shows no significant change in the λmax except at pH 12.0. The ANS fluorescence of edG at pH 2.0 and 3.0 forms an acid-induced molten globule-like state. The denaturation transition curve monitored by fluorescence spectroscopy revealed that urea and GdmCl induced denaturation native (N) ↔ denatured (D) state follows a two-state process. The fluorescence quenching, molecular docking, and 50 ns simulation measurements suggested that heparan sulfate showed excellent binding affinity to edG. Our binding study provides a preliminary insight into the interaction of edG to the host cell membrane via heparan sulfate. This binding can be inhibited using experimental approaches at the molecular level leading to the prevention of effective host–pathogen interaction.

In this work, multi-scale cementitious composites containing short carbon fibers (CFs) and carbon nanofibers (CNFs)/multi-walled carbon nanotubes (MWCNTs) were studied for their tensile stress sensing properties. CF-based composites were prepared by mixing 0.25, 0.5 and 0.75 wt.% CFs (of cement) with water using magnetic stirring and Pluronic F-127 surfactant and adding the mixture to the cement paste. In multi-scale composites, CNFs/MWCNTs (0.1 and 0.15 wt.% of cement) were dispersed in water using Pluronic F-127 and ultrasonication and CFs were then added before mixing with the cement paste. All composites showed a reversible change in the electrical resistivity with tensile loading; the electrical resistivity increased and decreased with the increase and decrease in the tensile load/stress, respectively. Although CF-based composites showed the highest stress sensitivity among all specimens at 0.25% CF content, the fractional change in resistivity (FCR) did not show a linear correlation with the tensile load/stress. On the contrary, multi-scale composites containing CNFs (0.15% CNFs with 0.75% CFs) and MWCNTs (0.1% MWCNTs with 0.5% CFs) showed good stress sensitivity, along with a linear correlation between FCR and tensile load/stress. Stress sensitivities of 6.36 and 11.82%/MPa were obtained for the best CNF and MWCNT-based multi-scale composite sensors, respectively.

The growing interest in wool fibres as an eco-friendly and sustainable material for diverse industrial applications requires an enhancement of their functional performance. To address this, wool fabrics were treated in the present research with low-pressure hexafluoroethane (C2F6) plasma to impart superhydrophobicity and improve their abrasion resistance. Unscoured and scoured wool fabrics were treated with C2F6 while varying plasma power (80 W and 150 W), gas flow rate (12 sccm and 50 sccm) and treatment time (6 min and 20 min), and the effect of plasma parameters on the abrasion resistance, water contact angle and dyeing behaviour of the wool fabrics was studied. Martindale abrasion testing showed that the surface abrasion of the wool fabrics increased with the number of abrasion cycles, and the samples treated with 150 W, 20 min, 12 sccm showed superior abrasion resistance. The scoured wool fabrics showed a contact angle of ~124°, which was stable for only 4 min 40 s, whereas the plasma-treated samples showed a stable contact angle of over 150°, exhibiting a stable superhydrophobic behaviour. The C2F6 plasma treatment also significantly reduced the exhaustion of an acid dye by wool fabrics. The EDX study confirmed the deposition of fluorine-containing elements on the wool fabrics significantly altering their properties.

Respiratory syncytial virus (RSV) is an important pathogen of global significance. The BA9 is one of the most predominant lineages of the BA genotype of group B RSV that has acquired a 60bp duplication in its G protein gene. We describe the local and global evolutionary dynamics of the second hyper variable region in the C- terminal of the G protein gene of the BA9 lineage. A total of 418 sequences (including 31 study and 387 GenBank strains) from 29 different countries were used for phylogenetic analysis. This analysis showed that the study strains clustered with BA (BA9 and BA8) and SAB4 genotype of group B RSV. We performed time-scaled evolutionary clock analyses using Bayesian Markov chain Monte Carlo methods. We also carried out glycosylation, selection pressure, mutational, entropy and Network analyses of the BA9 lineage. The time to the most recent common ancestor (tMRCA) of the BA genotype and BA9 lineage were estimated to be the years 1995 (95% HPD; 1987-1997) and 2000 (95% HPD; 1998-2001), respectively. The nucleotide substitution rate of the BA genotype [(4.58×10-3 (95% HPD; 3.89-5.29×10-3) substitution/site/year] was slightly faster than the BA9 lineage [4.03×10-3 (95% HPD; 4.65-5.2492×10-3)]. The BA9 lineage was categorized into 3 sub lineages (I, II and III) based on the Bayesian and Network analyses. The local transmission pattern suggested that BA9 is the predominant lineage of BA viruses that has been circulating in India since 2002 though showing fluctuations in its effective population size. The BA9 lineage established its global distribution with report from 23 different countries over the past 16 years. The present study augments our understanding of RSV infection, its epidemiological dynamics warranting steps towards its overall global surveillance.

Liquid chromatography tandem mass chromatography (LCâMS/MS) is an important hyphenated technique for quantitative analysis of drugs in biological fluids. Because of high sensitivity and selectivity, LCâMS/MS has been used for pharmacokinetic studies, metabolites identification in the plasma and urine. This manuscript gives comprehensive analytical review, focusing on chromatographic separation approaches (column packing materials, column length and mobile phase) as well as different acquisition modes (SIM, MRM) for quantitative analysis of glucocorticoids and stimulants. This review is not meant to be exhaustive but rather to provide a general overview for detection and confirmation of target drugs using LCâMS/MS and thus useful in the doping analysis, toxicological studies as well as in pharmaceutical analysis. Keywords: LCâMS/MS, Ionization techniques, Glucocorticoids, Stimulants, Hyphenated techniques, Biological fluid

Dengue fever is the most important arboviral disease in the tropical and sub-tropical countries of the world. Delhi, the metropolitan capital state of India, has reported many dengue outbreaks, with the last outbreak occurring in 2013. We have recently reported predominance of dengue virus serotype 2 during 2011-2014 in Delhi. In the present study, we report molecular characterization and evolutionary analysis of dengue serotype 2 viruses which were detected in 2011-2014 in Delhi. Envelope genes of 42 DENV-2 strains were sequenced in the study. All DENV-2 strains grouped within the Cosmopolitan genotype and further clustered into three lineages; Lineage I, II and III. Lineage III replaced lineage I during dengue fever outbreak of 2013. Further, a novel mutation Thr404Ile was detected in the stem region of the envelope protein of a single DENV-2 strain in 2014. Nucleotide substitution rate and time to the most recent common ancestor were determined by molecular clock analysis using Bayesian methods. A change in effective population size of Indian DENV-2 viruses was investigated through Bayesian skyline plot. The study will be a vital road map for investigation of epidemiology and evolutionary pattern of dengue viruses in India.

Respiratory syncytial virus (RSV) is an important viral pathogen of acute respiratory tract infection (ARI). Limited data are available on molecular epidemiology of RSV from Saudi Arabia. A total of 130 nasopharyngeal aspirates were collected from children less than 5 years of age with ARI symptoms attending the Emergency Department at King Khalid University Hospital and King Fahad Medical City, Riyadh, Saudi Arabia between October and December, 2014. RSV was identified in the 26% of the hospitalized children by reverse transcriptase PCR. Group A RSV (77%) predominated during the study as compared to group B RSV (23%). The phylogenetic analysis of 28 study strains clustered group A RSV in NA1 and ON1 genotypes and group B viruses in BA (BA9) genotype. Interestingly, 26% of the positive samples clustered in genotypes with duplication in the G protein gene (ON1 for group A and BA for group B). Both the genotypes showed enhanced O-linked glycosylation in the duplicated region, with 10 and 2 additional sites in ON1 and BA respectively. Selection pressure analysis revealed purifying selection in both the ON1 and BA genotypes. One codon each in the ON1 (position 274) and BA genotypes (position 219) were positively selected and had high entropy values indicating variations at these amino acid positions. This is the first report describing the presence of ON1 genotype and the first report on co-circulation of two different genotypes of RSV with duplication in the G protein gene from Saudi Arabia. The clinical implications of the simultaneous occurrence of genotypes with duplication in G protein gene in a given population especially in the concurrent infections should be investigated in future. Further, the ongoing surveillance of RSV in this region will reveal the evolutionary trajectory of these two genotypes with duplication in G protein gene from largest country in the Middle East.

Dengue fever is the most prevalent arboviral disease in the tropical and sub-tropical regions of the world. The present report describes molecular detection and serotyping of dengue viruses in acute phase blood samples collected from New Delhi, India. Phylogenetic and molecular clock analysis of dengue virus serotype 1 and 3 strains were also investigated. Dengue virus infection was detected in 68.87% out of 604 samples tested by RT-PCR between 2011 & 2014. Dengue serotype 1 was detected in 25.48% samples, dengue serotype 2 in 79.56% samples and dengue serotype 3 in 11.29% samples. Dengue serotype 4 was not detected. Co-infection by more than one dengue serotype was detected in 18.26% samples. Envelope gene of 29 DENV-1 and 14 DENV-3 strains were sequenced in the study. All the DENV-1 strains grouped with the American African genotype. All DENV-3 strains were found to belong to Genotype III. Nucleotide substitution rates of dengue 1 and 3 viruses were determined in the study. Time to the most recent common ancestor (TMRCA) of dengue 1 viruses was determined to be 132 years. TMRCA of DENV-3 viruses was estimated to be 149 years. Bayesian skyline plots were constructed for Indian DENV-1 and 3 strains which showed a decrease in population size since 2005 in case of DENV- 1 strains while no change was observed in recent years in case of DENV-3 strains. The study also revealed a change in the dominating serotype in Delhi, India in recent years. The study will be helpful in formulating control strategies for the outbreaks. In addition, it will also assist in tracking the movement and evolution of this emerging virus.

BackgroundAcute respiratory infection (ARI) is a major killer of children in developing countries. Although the frequency of ARI is similar in both developed and developing countries, mortality due to ARI is 10-50 times higher in developing countries. Viruses are common causes of ARI among such children, yet the disease burden of these infections in rural communities is unknown.Methodology/principal findingsA prospective longitudinal study was carried out in children enrolled from two rural Indian villages at birth and followed weekly for the development of ARI, classified as upper respiratory infection, acute lower respiratory infection (ALRI), or severe ALRI. Respiratory syncytial virus (RSV), influenza, parainfluenza viruses and adenoviruses in nasopharyngeal aspirates were detected by direct fluorescent antibody testing (DFA) and, in addition, centrifugation enhanced culture for RSV was done. 281 infants enrolled in 39 months and followed until 42 months. During 440 child years of follow-up there were 1307 ARIs, including 236 ALRIs and 19 severe ALRIs. Virus specific incidence rates per 1000 child years for RSV were total ARI 234, ALRI 39, and severe ALRI 9; for influenza A total ARI 141, ALRI 39; for INF B total ARI 37; for PIV1 total ARI 23, for PIV2 total ARI 28, ALRI 5; for parainfluenza virus 3 total ARI 229, ALRI 48, and severe ALRI 5 and for adenovirus total ARI 18, ALRI 5. Repeat infections with RSV were seen in 18 children.Conclusions/significanceRSV, influenza A and parainfluenza virus 3 were important causes of ARI among children in rural communities in India. These data will be useful for vaccine design, development and implementation purposes.