Optimisation of enzyme-assisted extraction of camptothecin from Nothapodytes nimmoniana and its characterisation.

INTRODUCTION: Efficient extraction of camptothecin (CPT), an anticancer agent from the commercial source Nothapodytes nimmoniana (J. Graham) Mabb in India, is of paramount importance. CPT is present in the highest concentration in the stem portion, and the stem can be readily harvested without uprooting the plant. The fluorescence microscopy mapping of the bark matrix for CPT revealed its presence in a free form within both the outer (epidermal and cortical tissues) and inner (xylem and phloem tissues) sections. The bark matrix primarily consists of cellulose, hemicellulose, and lignin, rendering it woody, rigid, and resistant to efficient solvent penetration for CPT extraction. We proposed a hypothesis that subjecting it to disruption through treatment with hydrolytic enzymes like cellulase and xylanase could enhance solvent diffusion, thereby enabling a swift and effective extraction of CPT. OBJECTIVE: The present study was aimed at enzyme-assisted extraction, using cellulase and xylanase for hydrolytic disruption of the cells to readily access CPT from the stem of the plant N. nimmoniana (J. Graham) Mabb. METHODOLOGY: The hydrolytic cell disruption of ground powder from the stem bark was studied using cellulase and xylanase enzymes. The enzymatically pretreated stem bark powder was subsequently recovered by filtration, dried, and subjected to extraction with methanol to isolate CPT. This process was optimised through a Box-Behnken design, employing a one-factor-at-a-time approach to assess parameters such as enzyme concentration (2-10% w/w), pH (3-7), incubation time (6-24 h), and solid-to-solvent ratio (1:30-1:70 g/mL). CPT was characterised using proton nuclear magnetic resonance (1H-NMR) and Fourier transform infrared (FTIR) spectra, and a high-performance liquid chromatography (HPLC) method was developed for quantification. RESULTS: The cellulase and xylanase treatment resulted in the highest yields of 0.285% w/w and 0.343% w/w, with efficiencies of 67% and 81%, respectively, achieved in a significantly shorter time compared to the untreated material, which yielded 0.18% with an efficiency of only 42%. Extraction by utilising the predicted optimised process parameters, a nearly two-fold increase in the yield, was observed for xylanase, with incubation and solvent extraction times set at 16 and 2 h, respectively. Scanning electron microscopy (SEM) images of the spent material indicated perforations attributed to enzymatic action, suggesting that this could be a primary factor contributing to the enhanced extraction. CONCLUSION: Enzyme-mediated hydrolytic cell disruption could be a potential approach for efficient and rapid isolation of CPT from the bark of N. nimmoniana.

Increased neuronal expression of the early endosomal adaptor APPL1 leads to endosomal and synaptic dysfunction with cholinergic neurodegeneration.

Dysfunction of the endolysosomal system within neurons is a prominent feature of Alzheimer's disease (AD) pathology. Multiple AD-risk factors are known to cause hyper-activity of the early-endosome small GTPase rab5, resulting in neuronal endosomal pathway disruption. APPL1, an important rab5 effector protein, is an interface between endosomal and neuronal function through a rab5-activating interaction with the BACE1-generated C-terminal fragment (ßCTF or C99) of the amyloid precursor protein (APP), a pathogenic APP fragment generated within endolysosomal compartments. To better understand the role of APPL1 in the AD endosomal phenotype, we generated a transgenic mouse model over-expressing human APPL1 within neurons (Thy1-APPL1 mice). Consistent with the important endosomal regulatory role of APPL1, Thy1-APPL1 mice have enlarged neuronal early endosomes and increased synaptic endocytosis due to increased rab5 activation. We additionally demonstrate pathological consequences of APPL1 overexpression, including functional changes in hippocampal long-term potentiation (LTP) and long-term depression (LTD), as well as degeneration of the large projection cholinergic neurons of the basal forebrain and impairment of hippocampal-dependent memory. Our findings show that increased neuronal APPL1 levels lead to a cascade of pathological effects within neurons, including early endosomal alterations, synaptic dysfunction, and neurodegeneration. Multiple risk factors and molecular regulators, including APPL1 activity, are known to contribute to the endosomal dysregulation seen in the early stages of AD, and these findings further highlight the shared pathobiology and consequences to a neuron of early endosomal pathway disruption. Significance Statement: Dysfunction in the endolysosomal system within neurons is a key feature of Alzheimer's disease (AD). Multiple AD risk factors lead to hyperactivity of the early-endosome GTPase rab5, disrupting neuronal pathways including the cholinergic circuits involved early in memory decline. APPL1, a crucial rab5 effector, connects endosomal and neuronal functions through its interaction with a specific amyloid precursor protein (APP) fragment generated within endosomes. To understand APPL1's role, a transgenic mouse model over-expressing human APPL1 in neurons (Thy1-APPL1 mice) was developed. These mice show enlarged early endosomes and increased synaptic endocytosis due to rab5 activation, resulting in impaired hippocampal long-term potentiation and depression, the degeneration of basal forebrain cholinergic neurons, and memory deficits, highlighting a pathological cascade mediated through APPL1 at the early endosome.

Phytochemical profiling of Clerodendrum paniculatum leaf extracts: GC-MS, LC-MS analysis and comparative evaluation of antimicrobial, antioxidant & cytotoxic effects.

The present study evaluates the phytochemical content and biological activities of eight Clerodendrum paniculatum leaf extracts obtained using four solvents of varying polarity and two extraction methods. GC-HRMS analysis of the hexane extract predicted presence of phytol, 22-tritetracontanone, and 6,9,12-octadecatrienoic acid, phenyl methyl ester, (ZZZ)-as major compounds, and ethyl acetate extract was predicted to contain phytol. LC-HRMS analysis of methanolic extract exhibited the presence of 8',10'-dihydroxydihydroergotamine, Khayanthone, Galactonic acid, Calotropin, and 26,26,26,27,27,27-hexafluoro-1alpha,24-dihydroxy vitamin D3 as significant fractions. Ethyl acetate extract showed significant antimicrobial activity in-vitro. Methanolic extracts possess the highest radical scavenging activity (up to 87%) and antidiabetic activity (up to 49%) at 1 mg/mL concentration. Methanolic extracts from maceration and the Soxhlet method showed better cytotoxicity against breast and oral cancer cell lines, while the aqueous extract from maceration demonstrated better cytotoxicity against lung cancer cell lines. Methanolic extracts containing tannins, flavonoids, alkaloids, quinones, glycosides, terpenoids, diterpenoids, and phytosterols demonstrated significant bioactivity.

Diabetes disease detection and classification on Indian demographic and health survey data using machine learning methods.

BACKGROUND & AIM: Diabetes mellitus has become one of the out brakes causing major health issues in developing countries like India. The need for leveraging technology is felt in diabetes management. The main objective of this work is to deploy machine learning methods for the detection and classification of diabetes having clinical relevance. METHODS: Indian demographic and health survey-2016 dataset is considered and determined the risk factors for continuous and categorical data. Kernel entropy component analysis is used for the dimensionality reduction of the feature set. Predictive exploration-based machine learning methods like logistic regression, gaussian naive Bayes, linear discriminant analysis, support vector classifier, k-nearest neighbor, decision tree, extreme gradient boosting, kernel entropy component analysis, and random forest are deployed in the work. The deployed methodology has three phases: feature extraction, classification, and prediction. RESULTS: Random Forest gave the maximum classification accuracy of 99.84% and 96.75% for imbalanced and kernel entropy component analysis-induced balanced datasets (using synthetic minority oversampling technique) respectively. The maximum precision of 99.64% is obtained using a support vector classifier on the balanced dataset. The area under the curve is 99%, which is observed from kernel entropy component analysis induced random forest on the balanced dataset. All other models performed moderately when applied to kernel entropy component analysis trained dataset. CONCLUSIONS: Random Forest model performed better in comparison with other models. The overall performance of the machine learning models can be improved by training the diabetes dataset using kernel entropy component analysis.

Bat-associated microbes: Opportunities and perils, an overview.

The potential biotechnological uses of bat-associated bacteria are discussed briefly, indicating avenues for biotechnological applications of bat-associated microbes. The uniqueness of bats in terms of their lifestyle, genomes and molecular immunology may predispose bats to act as disease reservoirs. Molecular phylogenetic analysis has shown several instances of bats harbouring the ancestral lineages of bacterial (Bartonella), protozoal (Plasmodium, Trypanosoma cruzi) and viral (SARS-CoV2) pathogens infecting humans. Along with the transmission of viruses from bats, we also discuss the potential roles of bat-associated bacteria, fungi, and protozoan parasites in emerging diseases. Current evidence suggests that environmental changes and interactions between wildlife, livestock, and humans contribute to the spill-over of infectious agents from bats to other hosts. Domestic animals including livestock may act as intermediate amplifying hosts for bat-origin pathogens to transmit to humans. An increasing number of studies investigating bat pathogen diversity and infection dynamics have been published. However, whether or how these infectious agents are transmitted both within bat populations and to other hosts, including humans, often remains unknown. Metagenomic approaches are uncovering the dynamics and distribution of potential pathogens in bat microbiomes, which might improve the understanding of disease emergence and transmission. Here, we summarize the current knowledge on bat zoonoses of public health concern and flag the gaps in the knowledge to enable further research and allocation of resources for tackling future outbreaks.

Carboxymethyl Chitosan-Fluoride-doped Amorphous Calcium Phosphate: A Novel Remineralizing Gel.

Background: There is a need for innovative remineralizing gel formulations based on calcium and phosphates that can slowly release fluoride ions and enhance the formation of fluorapatite crystals that are more resistant to dissolution in an acidic environment. Aim: The aim of the work was to formulate a remineralizing agent that remineralizes enamel through the release of Ca2+, PO4³-, and F- ions for a prolonged period of time. Materials and methods: The gel was based on carboxymethyl chitosan (CMC) as a bioinspired gelling agent and on Fluoride-doped Amorphous Calcium Phosphate (F-ACP) as a remineralizing agent. This gel was tested in vitro on the enamel of extracted premolars after demineralization with methacrylic acid gel. Results: When compared to the control group and demineralized enamel group, the enamel slabs remineralized with CMC/F-ACP showed a higher calcium phosphate ratio in Energy-Dispersive X-ray (EDX) and better surface morphology under scanning electron microscope (SEM). Conclusion: Remineralization tests performed on demineralized human permanent teeth proved that CMC/F-ACP gel has excellent efficacy, inducing a complete remineralization of the outermost layers of enamel as well as a full restoration of lost mineral content. How to cite this article: Nimbeni BS, Nimbeni SB, Divakar DD, et al. Carboxymethyl Chitosan-Fluoride-doped Amorphous Calcium Phosphate: A Novel Remineralizing Gel. Int J Clin Pediatr Dent 2023;16(5):734-739.

Determinants of awareness on pictorial health warnings on tobacco products in an Eastern state of India.

Background: Pictorial health warnings (PHW) are an effective strategy to deter or reduce tobacco use. This study was conducted to determine the level of awareness of PHWs on tobacco products and their correlates among adults attending the outpatient department (OPD) of a tertiary care facility in Eastern India. Materials and Methods: This cross-sectional study was conducted over a period of 3 months among 307 patients aged 18 years to 65 years. A pre-tested semi-structured questionnaire was used for the collection of data and inferential statistics were performed using JAMOVI version 2.3.21. Results: About 84% (95% CI, 78.9-87.2) of the participants were aware of PHW on tobacco products, 82.4% (95% CI, 77.8-86.3) for smoked forms and 51.8% (95% CI, 46.2-57.3) for smokeless forms. More than half of the participants felt that the current PHW were ineffective. Male [AOR, 3.13; 95% CI, 1.33-7.37], being educated [AOR, 3.37; 95% CI, 1.29-8.76], employment [AOR, 5.65; 95% CI, 1.21-26.30] and ever-tobacco use [AOR, 3.23; 95% CI, 1.43-7.2] were found to be independent correlates of PHW awareness for smoked tobacco products, whereas as being male [AOR, 2.01; 95% CI, 1.02-3.95] and being young (18-30 years) [AOR, 2.51; 95% CI, 1.08-5.84] were found to be the independent predictors of PHW awareness for smokeless tobacco products (SLT). Conclusion: Four out of every five individuals were aware of pictorial health warnings (PHW) on tobacco products; however, it was much less for SLT compared to smoked tobacco products. Male, being educated, employment and ever-tobacco use were independent correlates of awareness for the smoked form of tobacco products, whereas being young (18-30 years) was an independent predictor of awareness for SLT.

Acute and sub-chronic toxicity of Liberin, an anti-diabetic polyherbal formulation in rats.

BACKGROUND: The polyherbal formulation (PHF) liberin, is known to exert anti-hyperglycemic effects in type 2 diabetes mellitus. Hence, it is important to study the safety profile of PHF in the current study through acute and chronic toxicity evaluation. OBJECTIVES: This research aims to assess the acute and sub-chronic toxicity of PHF in rats. MATERIALS AND METHODS: PHF was administered once orally (1000 mg/kg body weight), and the rats (male and female) were monitored for toxicity signs for a 14-day period. For a 28-day chronic toxicity study, rats were daily administered with PHF dose of 500 mg/kg and 1000 mg/kg body weight. Rats were followed up for mortality, weight changes, and other morbidities. Further haematological, biochemical, and histopathological changes were assessed. RESULTS: No death related to treatment or toxicity signs were recorded in the acute single-dose administration group. The results showed that the PHF was tolerated well up to a dose of 1000 mg/kg body weight. Even at the high dose of 1000 mg/kg body weight, sub-chronic tests did not show any significant difference between the dosed and normal groups. No significant changes were seen in the histopathological analysis of the liver, spleen, and kidney as well as haematological and biochemical parameters in acute, sub-chronic and satellite groups following the administration of PHF. CONCLUSION: The results confirmed that there was no adverse effect of this PHF at the maximum dose of 1000 mg/kg body weight in Wistar rats. Further, no adverse delayed effects related to PHF were observed in the satellite group. Therefore, this PHF appears safe for therapeutic purposes in the Ayurvedic medicinal system.

Ingrained Insight into Nasal Polyposis; Role of Human Papillomavirus: An Indian Study.

Nasal polyposis (NP) are benign lesion of nasal cavity, affecting up to 1-4% of population. It is known to have up to 70% recurrence rate. The underlying pathophysiology is still undetermined. Recent literatures have thrown light on the viral aetiology for NP. To our knowledge this is the first Indian study done, that aims to evaluate the prevalence of Human papillomavirus (HPV) in NP. It was a prospective case control study done among 40 individuals with NP and 40 healthy controls in a tertiary care centre. All the patients were subjected to routine clinical evaluation, investigations prior to proposed surgeries. The mucosal samples after the surgery were subjected to HPV DNA analysis by RT-PCR. Among the cases, the male to female ratio was 1.3:1. The mean age of patients with NP was 39 ± 14.6. The mean serum IgE levels among the case were 154 IU/ml and was significantly higher as compared to controls. Other inflammatory markers such as absolute eosinophil count, erythrocyte sedimentation rate, and neutrophil to leukocyte ration were found to be not significant. There was no HPV DNA detected among both case as well as controls. There seems to be strong association of IgE and NP, suggesting an IgE mediated pathway for its Causation. There is no association of HPV in NP.

Anti-hyperglycemic, anti-hyperlipidemic, and anti-inflammatory effect of the drug Guggulutiktaka ghrita on high-fat diet-induced obese rats.

BACKGROUND: Ayurveda is a holistic system of medicine and describes a vast array of herbs and herbal mixtures that are been demonstrated to possess efficacy in research investigations. Guggulutikthaka gritha (GTG) is one such drug evaluated for its role in skin and bone diseases. OBJECTIVE: In the current study, the hypoglycemic, hypolipidemic, and anti-inflammatory effect of the drug GTG was studied with the scope to treat dyslipidemia and thereby reduce the risk of cardiovascular disease. MATERIALS AND METHOD: The animals (Wistar rats) were fed a high-fat diet and dyslipidemia was induced. The control group was provided with a normal chow diet and had free access to water. The treatment with the drug GTG was given for 21 days after confirming dyslipidemia. The blood glucose was measured immediately using a glucometer. The serum was analyzed for lipid profile and Vascular Cell Adhesion Molecule - 1(VCAM 1) by ELISA method before and after treatment. The histopathology of the heart and liver was also performed. RESULTS: The abnormal change in lipid profile, blood glucose, and inflammatory marker along with the accumulation of intracellular fats in the arteries of the heart and liver confirmed dyslipidemia. A significant reduction in serum lipid profile (p < 0.05), blood glucose (p < 0.05), and VCAM 1 (p < 0.05) was noted after the treatment with significant histopathological changes in arteries of the heart and liver. CONCLUSION: The study provides scientific validation on the drug GTG being effective in hyperglycemia, hyperlipidemia, and inflammation in dyslipidemia.

In-silico screening and in-vitro assay show the antiviral effect of Indomethacin against SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the worldwide spread of coronavirus disease 19 (COVID-19), and till now, it has caused death to more than 6.2 million people. Although various vaccines and drug candidates are being tested globally with limited to moderate success, a comprehensive therapeutic cure is yet to be achieved. In this study, we applied computational drug repurposing methods complemented with the analyses of the already existing gene expression data to find better therapeutics in treatment and recovery. Primarily, we identified the most crucial proteins of SARS-CoV-2 and host human cells responsible for viral infection and host response. An in-silico screening of the existing drugs was performed against the crucial proteins for SARS-CoV-2 infection, and a few existing drugs were shortlisted. Further, we analyzed the gene expression data of SARS-CoV-2 in human lung epithelial cells and investigated the molecules that can reverse the cellular mRNA expression profiles in the diseased state. LINCS L1000 and Comparative Toxicogenomics Database (CTD) were utilized to obtain two sets of compounds that can be used to counter SARS-CoV-2 infection from the gene expression perspective. Indomethacin, a nonsteroidal anti-inflammatory drug (NSAID), and Vitamin-A were found in two sets of compounds, and in the in-silico screening of existing drugs to treat SARS-CoV-2. Our in-silico findings on Indomethacin were further successfully validated by in-vitro testing in Vero CCL-81 cells with an IC50 of 12 µM. Along with these findings, we briefly discuss the possible roles of Indomethacin and Vitamin-A to counter the SARS-CoV-2 infection in humans.

Development of a liquid chromatography/mass spectrometry methodology to separate, detect, characterize and quantify PEG-resveratrol prodrugs and the conjugation reaction precursors and intermediates.

A simple and reliable liquid chromatography/mass spectrometry (LC/MS) method to monitor pegylation of resveratrol is described. The developed LC/MS method can separate and quantify unmodified MeO-PEG-OH, carboxylic acid terminated PEG, resveratrol and PEG-resveratrol prodrugs. This methodology was able to monitor and determine the extent of conversion of MeO-PEG-OH into respective acidic functional derivatives such as MeO-PEG succinylester acid (MeO-PEGO-SuccOH), which was found to be complete. The developed method was also utilised to determine the extent of conjugation of resveratrol to carboxylic acid terminated PEG. The conversion of carboxylic acid terminated PEG into a PEG-resveratrol conjugate was found to be 100% and 73%, respectively, for MeO-PEG succinylamide resveratrol (MeO-PEGN-Succ-RSV) and MeO-PEG succinylester resveratrol (MeO-PEGO-Succ-RSV). The 100% conjugation of MeO-PEGN-Succ-RSV is consistent with the result obtained from a nuclear magnetic resonance (NMR) study. The average molecular weights determined by LC/MS for MeO-PEG-OH, MeO-PEGO-SuccOH and MeO-PEGO-Succ-RSV were found to be 2108, 2321 and 2423 Da, respectively. These data correlate well with the theoretical values. This methodology proved to be simple and effective in determining the extent of functionalisation of PEG and its conjugation to resveratrol. Overall our LC/MS method coupled with NMR permitted complete characterisation of the polymeric prodrug pegylated-resveratrol and the reaction precursors.

Isolation, identification and in vitro biological evaluation of phytochemicals from Memecylon randerianum: a medicinal plant endemic to Western Ghats of India.

The present investigation reports evaluation of phytochemical content, antimicrobial, antioxidant, antidiabetic and anticancer properties of the methanolic crude extracts prepared from Memecylon randerianum leaves. Qualitative evaluation showed the presence of carbohydrates, reducing sugars, alkaloids, phenols, flavonoids, cardiac glycosides, steroids, phytosterols, terpenoids, diterpenoids and coumarins. Quantitative estimation for phenols, flavonoids and alkaloids indicated total phenolic content 11.95 mg GAE/g, total flavonoid content 20.34 mg QE/g and total alkaloid content 316.68 mg AE/g dry weight of the crude extract respectively. The methanolic extract demonstrated effective antimicrobial activity against Escherichia coli, Staphylococcus aureus and Candida albicans. The preparation also demonstrated good antioxidant and antidiabetic activities. Anticancer activity was evaluated against breast (MCF), oral (KB) and lung (A-549) cancer cell lines and the respective IC50 values were found to be 159.81 ± 7.54 µg/mL, 240.21 ± 2.57 µg/mL and 124.17 ± 2.10 µg/mL which are comparable to IC50 values of Paclitaxel.

Role of Chitosan in Remineralization of Enamel and Dentin: A Systematic Review.

AIM AND OBJECTIVE: The purpose of this research was to analyze the role of chitosan in the remineralization of enamel and dentin. MATERIALS AND METHODS: An electronic search was done for articles published from January 2009 to January 2020. A manual search was done from bibliographies of selected articles for relevant articles that were unexplored. Only in vitro studies conducted on the application of chitosan for remineralization of enamel and dentin were included in the study. RESULTS: Of the 162 articles that were searched, only 15 in vitro studies were selected for the study. These studies met the inclusion criteria and were published from January 2009 to January 2020. CONCLUSION: The review provides insight into the mechanism of remineralization of enamel and dentin. The properties of chitosan make it an ideal biomaterial that can be employed in the formulation of a novel remineralizing gel. However, more in vivo studies, clinical trials, and research are essential to transform chitosan-based remineralizing gels from research to clinical use. CLINICAL SIGNIFICANCE: This review article opens a new window of opportunities for remineralizing enamel and dentin which have been long considered a challenging job. HOW TO CITE THIS ARTICLE: Nimbeni SB, Nimbeni BS, Divakar DD. Role of Chitosan in Remineralization of Enamel and Dentin: A Systematic Review. Int J Clin Pediatr Dent 2021;14(4):562-568.

Effectiveness of polyherbal formulations for the treatment of type 2 Diabetes mellitus - A systematic review and meta-analysis.

BACKGROUND: The mortality and morbidity rate of diabetes patients is increasing worldwide which requires an ideal treatment to prevent the disease worsening. Traditional medicine is gaining more attention in diabetes due to its efficacy and safety. We, therefore performed a systematic review study of clinical trials to assess the comparative effect of polyherbal formulations in type 2 Diabetes mellitus. OBJECTIVES: To find the effectiveness of polyherbal formulations in blood sugar and lipid level for type 2 Diabetes mellitus. MATERIAL AND METHODS: PubMed, Scopus and CINAHL databases for clinical trials investigating the effect of polyherbal formulations in Type 2 Diabetes mellitus patients were searched. Meta-analysis of eligible trials was conducted employing Revman 5.2 software. RESULTS: Fourteen randomized controlled trials were found eligible for meta-analysis. Meta-analysis of findings showed a significant effect of polyherbal formulations on blood sugar level compared to control group. The estimated standard mean changes at 95% confidence interval, following polyherbal formulations treatment were -0.59, (-0.91 to - 0.27) mg/dL; for fasting blood sugar(p < 0.001), -0.69, (-1.18 to -0.21) mg/dL; for postprandial blood sugar (p = 0.005) and -0.46, (-0.88 to -0.04) gm%; for glycated haemoglobin (p = 0.03). The reduction in postprandial sugar and glycated haemoglobin was statistically significant with polyherbal formulations compared to metformin treatment but not for fasting sugar. Similarly in lipid profile the reduction for total cholesterol and triglycerides was statistically significant with polyherbal formulations compared to control group but was not significant for HDL and LDL whereas in other group of polyherbal formulations and metformin only HDL was favouring polyherbal formulations. CONCLUSION: Polyherbal formulations occurred to be effective in lowering blood sugar level in Type 2 diabetes but their further efficacy in managing diabetes needs to be validated. Therefore, a qualitative, long term, randomized placebo-controlled trials of adequate sample size are necessary to determine the efficacy of polyherbal formulation in managing diabetes.

Molecular phylogeny of Capripoxviruses based on major immunodominant protein (P32) reveals circulation of host specific sheeppox and goatpox viruses in small ruminants of India.

Sheeppox and goatpox are highly contagious viral diseases of small ruminants causing severe economic losses to the livestock farmers. The disease is enzootic in Asia including India, Middle East and African countries. In the present study, a total of 28 isolates from twenty five sheeppox and goatpox disease outbreaks were phylogenetically analyzed based on P32 gene/protein along with homology modeling and docking using heparan sulfate and UDP-glucose. Three distinct lineage-specific clusters as per their host origin were recorded. Multiple sequence analysis of P32 gene revealed that genetically similar sheeppox virus (SPPV) and goatpox virus (GTPV) strains are circulating in India. Phylogenetically, Lumpy skin disease (LSDV) and SPPV had a closer genetic relationship than GTPV. Comparative sequence alignment indicated conservation of various motifs such as glycosaminoglycan (GAG), chemokine like motif (CX3C) and Asp-Glu-any other residue-Asp (D/ExD), as well as viral specific signature residues in SPPV and GTPV isolates. Structurally, P32 protein of SPPV and GTPV with mixed α helices and ß sheets resembled with crystal structure of homologue vaccinia virus H3L protein. Docking studies in P32 protein of SPPV and GTPV revealed conserved binding pattern with heparan sulfate which is involved in the virus attachment and varied glycosyltransferase fold with UDP-glucose. These findings may help in development of suitable vaccines/diagnostics and therapeutics against capripoxviruses.

Voice Analysis in Post Tonsillectomy Patients.

The main aim of this study was to analyse the change in voice in terms of acoustic parameters and its perceptual impact in patients who have undergone tonsillectomy. A prospective study was conducted in our institution-JSS Hospital and JSS institute of speech and hearing, Mysore for a duration of 1 year (December 2015-December 2016). 50 post tonsillectomy cases were selected randomly and subjected to acoustic analysis. It was inferred that situation of vocal analysis and assessment for the vowels 'a', 'i' and 'u' under the categories hoarse, harsh and breathy remain more or less the same during preoperative stages, first preoperative follow up and the second post operative follow up. It was concluded that tonsillectomy did not appear to change the acoustic features of vowels remarkably. It was assumed that the subject may adjust the shape of the vocal tract to produce consistent speech sound after surgery using auditory feedback.

Effect of piperine and quercetin alone or in combination with marbofloxacin on CYP3A37 and MDR1 mRNA expression levels in broiler chickens.

After oral route of administration, drug absorption is unpredictable and is governed by various factors such as multi drug resistance-1 (MDR1) an efflux transporter and drug metabolizing enzymes (like CYP3A4, CYP3A37, CYP2D6) at intestine and liver. Naturally available phyto chemicals like piperine and quercetin as well as some floroquinolones are known to inhibit MDR1 and CYP3A37 activity and increases bioavailability of co-administered drugs. This study was carried out to investigate the effect of piperine and quercetin alone or in combination with marbofloxacin on CYP3A37 and MDR1 mRNA expression levels in liver and intestine of broiler chicken. After oral administration of piperine and quercetin for 3 consecutive days followed by with or without oral administration of marbofloxacin for 5 days, CYP3A37 and MDR1 mRNA expression levels were determined using quantitative real-time PCR. Total of 36 broiler chickens in seven individual groups were treated with different regimen and the mRNA expression levels at duodenum and liver were analyzed with apt statistical tools. After piperine and quercetin combined treatment with marbofloxacin, CYP3A37 mRNA expression levels were significantly down regulated by 20.57 (p = .034) and 25.95 (p = .003) folds; and MDR1 mRNA expression levels were also significantly down regulated by 11.33 (p = .012) and 33.59 (p = .006) folds in liver and duodenum, respectively. Down regulation of CYP3A37 and MDR1 mRNA in liver and duodenum indicate the combined pretreatment of piperine and quercetin may be useful for improving the pharmacokinetics of orally administered drugs which are substrates for CYP3A37 and MDR1.

Elevation of Oxidative Stress and Decline in Endogenous Antioxidant Defense in Elderly Individuals with Hypertension.

INTRODUCTION: Hypertension is becoming an important medical and public health problem all over the world and is the most common disorder of ageing. There is a growing evidence of involvement of vascular oxidative stress in the development of hypertension from animal studies. However, studies on humans with hypertension, particularly in elderly are least and data remained controversial. Moreover, studies in elderly people with hypertension are scarce. AIM: To investigate the possible role of oxidative stress and antioxidant defense in the pathogenesis of hypertension in elderly. MATERIALS AND METHODS: A cross-sectional study was conducted on elderly males (n=60) with newly diagnosed hypertension and with normal blood pressure. Oxidative stress and antioxidant status were evaluated by assessing the following parameters: plasma Malondialdehyde (MDA), and antioxidants: Superoxide Dismutase (SOD) activity, reduced Glutathione (GSH), and vitamin C levels; and total Nitric Oxide concentration in plasma (NOx). Difference between groups was determined by using unpaired t-test/Mann-Whitney U test. Bivariate correlation and multiple regression analysis were used to determine the relationship between variables. RESULTS: A significant rise in plasma MDA (p-value=0.013) and lower levels of endogenous antioxidants: SOD (p-value≤0.001) and GSH (p-value≤0.001) were observed in elderly individuals with hypertension when compared to healthy controls. Though not significant, there was a mean decrease in plasma NOx in hypertensive subjects than normotensive ones. While vitamin C showed no significant difference between two groups. Decrease in GSH (ß=-0.398; p-value=0.001) and SOD (ß=-0.423; p-value≤0.001) were the significant determinants of hypertension in elderly individuals. CONCLUSION: Above findings indicate that elevation in oxidative stress and decrease in endogenous antioxidant level may be involved in the pathogenesis of hypertension. However, it remains unclear whether oxidative stress causes or augments hypertension.