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Background: Designing and development of molecules for cancer treatment useful and with no side effects are a big challenge for the researchers in the field of drug discovery. The use of phytochemicals for chemoprevention is gaining more advantages, and intake of flavonoids has proved to reduce the occurrence of various cancers. Objectives: The present study was focused on selecting eight flavonoids and study them by in silico methods to analyse the interactions, affinity and pharmacophoric features that participate in the interactions between the flavonoid and the active sites of different cancer targets. Methods: The cancer targets were downloaded from the protein data bank, and flavonoids from PubChem and were docked by Glide XP molecular docking method to find the molecular interactions. The binding energy was calculated by Prime MM-GBSA application and ADMET analysis by Qikprop of Schrodinger. The anticancer potential of flavonoids screening was based on an online tool, Pass predictor. Phase module was used to find the common pharmacophore features that participate in essential interactions between the flavonoid and the active site. Results: In this study, myricetin has proved to be the best flavonoid for the treatment of breast and lung cancer with docking score of -11.50 kcal/mol and -10.56 kcal/mol respectively, whereas, quercetin has proved to be the best for prostate and colorectal cancer with docking score of -14.18 kcal/mol and -12.94 kcal/mol, respectively. The responsible forces for the interaction of these flavonoids are hydrogen bond, hydrophobic interactions, polar and pi-pi stackings. The PASS tool predicted the anticancer potential for the flavonoids, in particular, myricetin had responded highly active for most cancer cells. The hypothesis AADRR_1 has the highest survival score, which indicates the best alignment of the active ligands and represents the best pharmacophore model for anticancer activity. Conclusion: This work has screened eight flavonoids against various cancer targets and shown the binding interactions between them, stating that myricetin is the suitable lead candidate for breast and lung cancer; whereas, quercetin is the best lead for prostate and colorectal cancer. And these data are about the results obtained from PASS predictor. Moreover, the pharmacophore model has generated for the flavonoids, which correlate activities with the spatial arrangement of various chemical features. Therefore, this investigation strongly suggests that these flavonoids can be used as leads as anticancer agents.

BACKGROUND COPD is characterized by chronic airflow limitation and a range of pathological changes in the lungs. In addition, COPD presents significant extrapulmonary effects and is associated with important comorbidities that may contribute to the disease severity. COPD affects pulmonary blood vessels, right ventricle, as well as left ventricle leading to the development of pulmonary hypertension, cor-pulmonale, right ventricular dysfunction, and left ventricular dysfunction. This study was undertaken to evaluate cardiac function with ECG and echocardiography in COPD patients which may further help to assess the prognosis and identifying the individuals likely to suffer increase morbidity and mortality. METHODS An observational, cross sectional study was conducted among 100 patients attending the Department of General Medicine of Burdwan Medical College and Hospital, diagnosed as having COPD wherein detailed history taking and clinical examination was done along with routine investigations such as complete blood count, renal functions, randomized blood sugar and further for a Chest X-ray, electrocardiography, and two-dimensional (2D)-echocardiography. Statistical analysis was done by SPSS version 19.0. RESULTS Majority of the COPD patients were male and belonged to GOLD stage 2. The most electrocardiographic finding was P-Pulmonale and Echocardiographic finding was PAH (Pulmonary Arterial Hypertension) with maximum patients having mild PAH. Most common clinical finding was pedal edema. Severity of the disease was found to have linear relation with severity of cardiac complications. CONCLUSION There is significant involvement of the cardiovascular system in COPD patients. The electrocardiographic and echocardiographic findings had linear relation with the severity of the disease. It is mandatory to investigate all the COPD patients with ECG and 2D ECHO as it aids in early detection and treatment of cor pulmonale.

Visceral leishmaniasis is characterized by mixed production of Th1/2 cytokines and the disease is established by an enhanced level of Th2 cytokine. CD4+ T cells are main cell type which produces Th1/2 cytokine in the host upon Leishmania infection. However, the regulatory mechanism for Th1/2 production is not well understood. In this study, we co-cultured mice CD4+ T cells with Leishmania donovani infected and uninfected macrophage for the identification of dysregulated miRNAs in CD4+ T cells by next-generation sequencing. Here, we identified 604 and 613 known miRNAs in CD4+ T cells in control and infected samples respectively and a total of only 503 miRNAs were common in both groups. The expression analysis revealed that 112 miRNAs were up and 96 were down-regulated in infected groups, compared to uninfected control. Nineteen up-regulated and 17 down-regulated miRNAs were statistically significant (p insilco approach, we identified the gene targets of significant miRNAs on the basis of CD4+ T cell biology. Eleven up-regulated miRNAs and 9 down-regulated miRNAs were associated with the cellular immune responses and Th1/2 dichotomy upon Leishmania donovani infection. The up-regulated miRNAs targeted transcription factors that promote differentiation of CD4+ T cells towards Th1 phenotype. While down-regulated miRNAs targeted the transcription factors that facilitate differentiation of CD4+ T cells towards Th2 populations. The GO and pathway enrichment analysis also showed that the identified miRNAs target the pathway and genes related to CD4+ T cell biology which plays important role in Leishmania donovani infection.

This paper discusses an ontology based clinical decision support system for the specialty of Geriatric Medicine. We created a domain level ontology based on Handbook of Geriatrics and then mapped it to an upper level Basic Formal Ontology. The decision support system has been developed in Prolog. For accessing this ontology, we created an interactive web and android application which acts as a front-end to the system. Both applications are able to display the ontology structure and to predict the disease based on the symptom entered. We have uploaded our ontology to Bio-Portal, which is one of the most comprehensive biomedical ontology repository. It can be accessed using the URL https://bioportal.bioontology.org/ontologies/G-O.

Purpose Amplifications of receptor tyrosine kinases (RTKS) are therapeutic targets in multiple tumor types (e.g. HER2 in breast cancer), and amplification of the chromosome 4 segment harboring the three RTKs KIT, PDGFRA, and KDR (4q12amp) may be similarly targetable. The presence of 4q12amp has been sporadically reported in small tumor specific series but a large‐scale analysis is lacking. We assess the pan‐cancer landscape of 4q12amp and provide early clinical support for the feasibility of targeting this amplicon. Experimental Design Tumor specimens from 132,872 patients with advanced cancer were assayed with hybrid capture based comprehensive genomic profiling which assays 186–315 genes for all classes of genomic alterations, including amplifications. Baseline demographic data were abstracted, and presence of 4q12amp was defined as 6 or more copies of KIT/KDR/PDGFRA. Concurrent alterations and treatment outcomes with matched therapies were explored in a subset of cases. Results Overall 0.65% of cases harbored 4q12amp at a median copy number of 10 (range 6–344). Among cancers with >100 cases in this series, glioblastomas, angiosarcomas, and osteosarcomas were enriched for 4q12amp at 4.7%, 4.8%, and 6.4%, respectively (all p < 0.001), giving an overall sarcoma (n = 6,885) incidence of 1.9%. Among 99 pulmonary adenocarcinoma cases harboring 4q12amp, 50 (50%) lacked any other known driver of NSLCC. Four index cases plus a previously reported case on treatment with empirical TKIs monotherapy had stable disease on average exceeding 20 months. Conclusion We define 4q12amp as a significant event across the pan‐cancer landscape, comparable to known pan‐cancer targets such as NTRK and microsatellite instability, with notable enrichment in several cancers such as osteosarcoma where standard treatment is limited. The responses to available TKIs observed in index cases strongly suggest 4q12amp is a druggable oncogenic target across cancers that warrants a focused drug development strategy. Implications for Practice Coamplification of the receptor tyrosine kinases (rtks) KIT/KDR/PDGFRA (4q12amp) is present broadly across cancers (0.65%), with enrichment in osteosarcoma and gliomas. Evidence for this amplicon having an oncogenic role is the mutual exclusivity of 4q12amp to other known drivers in 50% of pulmonary adenocarcinoma cases. Furthermore, preliminary clinical evidence for driver status comes from four index cases of patients empirically treated with commercially available tyrosine kinase inhibitors with activity against KIT/KDR/PDGFRA who had stable disease for 20 months on average. The sum of these lines of evidence suggests further clinical and preclinical investigation of 4q12amp is warranted as the possible basis for a pan‐cancer drug development strategy., Matching treatment strategy to tumor biology is central to precision medicine. The co‐amplification of three distinct RTK encoding genes (KIT, KDR, and PDGFRA) on chromosome 4q12 (4q12amp) may be an oncogenic driver and thus a target for therapy. This article assesses the pan‐cancer landscape of 4q12amp and provides clinical support for the feasibility of targeting this amplicon.

WHO has declared "Coronavirus disease 2019" (COVID-19), which is caused by "Severe Acute Respiratory Syndrome Coronavirus 2" (SARS-CoV-2), a worldwide pandemic in March 2020. With its advent in December 2019, it has affected over 86,095,614 people worldwide as of January 4, 2021. Medical workers and researchers are working towards developing a vaccine and improving diagnostic methods for early detection and disease progression monitoring methods. The objective of this study is to provide a robust "Convolutional Neural Network" (CNN) architecture for COVID-19 detection using "Chest X-Rays" (CXR) and Chest CT Scans in order to reduce the response time to diagnose infected patients. We developed deep learning image classification models using Dilated Convolutional Neural Networks as the backend for our model and utilized various fine-tuned pre-trained CNN models as the feature extractor for our model. For both Chest X-Ray and Chest CT, we created datasets by combining various publicly available databases. The Chest X-Ray dataset contains 196 COVID positive frontal CXR images and 196 normal images, and out of two Chest CT datasets, one contains 349 covid and 349 non-covid images and the other contains 1252 covid and 1230 non-covid images. We also utilized transfer learning because of less publicly available data. Image Enhancement Techniques were also used to improve image contrast. The best classification accuracy achieved on Chest X-Ray dataset is 100% and accuracies achieved on the two Chest CT datasets are 91.6% and 98% respectively.

Adult fish Heteropneustes fossilis were divided into 4 groups –(i) Group A: kept in artificial freshwater and daily injected intraperitoneally with vehicle; (ii) Group B: kept in artificial freshwater and were daily injected intraperitoneally with 0.1 mg/100 g body wt of oProlactin; (iii) Group C: maintained in calcium-deficient freshwater and daily injected intraperitoneally with vehicle; (iv) Group D: kept in calcium-deficient freshwater and daily injected intraperitoneally with 0.1 mg/100 g body wt of oProlactin. Blood samples were taken 2 h after the last injection on 1, 3, 5, 10 and 15 days of the treatment. Plasma calcium levels were analyzed. The corpuscles of Stannius (CS) were fixed for histological studies. Artificial freshwater: The plasma calcium levels of vehicle-injected specimens (group A) remained unaltered throughout the experiment. Following prolactin treatment (group B) the plasma calcium levels progressively increased from day 3 to day 5. The values became normocalcemic at day 10 and day 15. After day 5 following prolactin administration (group B), the nuclear volume of AF-positive cells increased and the cells were seen degranulated. After day 10, there was an increased dilatation of sinusoids and the nuclear volume of AF-positive cells showed further increase. On day 15, these changes were exaggerated. The AFnegative cells of the corpuscles of Stannius of prolactin-treated fish (group B) showed no change in their histological structure and nuclear volume. Calcium-deficient freshwater: The plasma calcium level decreased in vehicle-injected fish (group C) from day 1 to day 3 (as compared to level of the fish kept in artificial freshwater). Thereafter, the level increased from day 5 resulting in hypercalcemia at day 10 and day 15. In prolactin treated fish (group D) the plasma calcium level indicated progressive increase from day 5 to day 15. In the vehicle-injected fish (group C) the AF-positive cells of corpuscles of Stannius showed accumulati

With the advancement of isothermal nucleic acid amplification techniques, detection of the pathogenic DNA in clinical samples at point-of-need is no longer a dream. The newly developed recombinase polymerase amplification (RPA) assay incorporated in a suitcase laboratory has shown promising diagnostic efficacy over real-time PCR in detection of leishmania DNA from clinical samples. For broader application of this point-of-need system, we undertook a current multi-country diagnostic evaluation study towards establishing this technique in different endemic settings which would be beneficial for the ongoing elimination programs for leishmaniasis. For this study purpose, clinical samples from confirmed visceral leishmaniasis (VL) and post-kala-azar dermal leishmaniasis (PKDL) patients were subjected to both real-time PCR and RPA assay in Bangladesh, India, and Nepal. Further skin samples from confirmed cutaneous leishmaniasis (CL) patients were also included from Sri Lanka. A total of 450 clinical samples from VL patients, 429 from PKDL patients, 47 from CL patients, and 322 from endemic healthy/healthy controls were under investigation to determine the diagnostic efficacy of RPA assay in comparison to real-time PCR. A comparative sensitivity of both methods was found where real-time PCR and RPA assay showed 96.86% (95% CI: 94.45–98.42) and 88.85% (95% CI: 85.08–91.96) sensitivity respectively in the diagnosis of VL cases. This new isothermal method also exhibited promising diagnostic sensitivity (93.50%) for PKDL cases, when a skin sample was used. Due to variation in the sequence of target amplicons, RPA assay showed comparatively lower sensitivity (55.32%) than that of real-time PCR in Sri Lanka for the diagnosis of CL cases. Except for India, the assay presented absolute specificity in the rest of the sites. Excellent concordance between the two molecular methods towards detection of leishmania DNA in clinical samples substantiates the application of RPA assay incorporated in a suitcase laboratory for point-of-need diagnosis of VL and PKDL in low resource endemic settings. However, further improvisation of the method is necessary for diagnosis of CL.

Leishmaniasis comprises of a wide variety of diseases, caused by protozoan parasite belonging to the genus Leishmania. Leishmania parasites undergo different types of stress during their lifetime and have developed strategies to overcome this damage. Identifying the mechanistic approach used by the parasite in dealing with the stress is of immense importance for unfolding the survival strategy adopted by the parasite. Mevalonate kinase (MVK) is an important regulatory factor in the mevalonate pathway in both bacteria and eukaryotes. In this study, we explored the role of Leishmania donovani mevalonate kinase (LdMVK) in parasite survival under stress condition. Hydrogen peroxide (H2O2) and menadione, the two known oxidants were used to carry out the experiments. The MVK expression was found to be up regulated ∼2.1 fold and ∼2.3 fold under oxidative stress condition and under the effect of anti-Leishmania drug, AmBisome respectively. The cell viability declined under the effect of MVK inhibitor viz: vanadyl sulfate (VS). The level of intracellular ROS was also found to be increased under the effect of MVK inhibitor. To confirm the findings, LdMVK over expression (LdMVK OE) and LdMVK knockdown (LdMVK KD) parasites were generated. The level of ergosterol, an important component of plasma membrane in L. donovani, was observed and found to be reduced by nearly 60 % in LdMVK KD parasite and increased by nearly 30 % in LdMVK OE parasites as compared to wild type. However, the ergosterol content was found to be elevated under oxidative stress. Furthermore, LdMVK was also found to be associated with maintaining the plasma membrane integrity and also in preventing the peroxidation of cellular lipids when exposed to oxidative stress. The above data clearly suggests that MVK has a vital role in protecting the parasite from oxidative stress. These findings may also explore the contribution of LdMVK in drug unresponsiveness which may help in future rational drug designing for leishmaniasis.

Automated machine learning approaches to skin lesion diagnosis from images are approaching dermatologist-level performance. However, current machine learning approaches that suggest management decisions rely on predicting the underlying skin condition to infer a management decision without considering the variability of management decisions that may exist within a single condition. We present the first work to explore image-based prediction of clinical management decisions directly without explicitly predicting the diagnosis. In particular, we use clinical and dermoscopic images of skin lesions along with patient metadata from the Interactive Atlas of Dermoscopy dataset (1011 cases; 20 disease labels; 3 management decisions) and demonstrate that predicting management labels directly is more accurate than predicting the diagnosis and then inferring the management decision ($$13.73 \pm 3.93\%$$ 13.73 ± 3.93 % and $$6.59 \pm 2.86\%$$ 6.59 ± 2.86 % improvement in overall accuracy and AUROC respectively), statistically significant at $$p < 0.001$$ p < 0.001 . Directly predicting management decisions also considerably reduces the over-excision rate as compared to management decisions inferred from diagnosis predictions (24.56% fewer cases wrongly predicted to be excised). Furthermore, we show that training a model to also simultaneously predict the seven-point criteria and the diagnosis of skin lesions yields an even higher accuracy (improvements of $$4.68 \pm 1.89\%$$ 4.68 ± 1.89 % and $$2.24 \pm 2.04\%$$ 2.24 ± 2.04 % in overall accuracy and AUROC respectively) of management predictions. Finally, we demonstrate our model’s generalizability by evaluating on the publicly available MClass-D dataset and show that our model agrees with the clinical management recommendations of 157 dermatologists as much as they agree amongst each other.

A corona virus disease 2019 (COVID-19) is a contagious disease which is caused by a novel corona virus. Human corona virus (HCoV) recognized as one of the most rapidly evolving viruses owing to its high genomic nucleotide substitution rates and recombination. Among the severe acute respiratory syndrome (SARS) and Middle-East respiratory syndrome (MERS), COVID-19 has spread more rapidly and increased the level of globalization and adaptation of the virus in every environmental condition due to their high rate of molecular diversity. The whole article highlights the general characteristics of corona virus, their molecular diversity, and molecular protein targeting against COVID-19 with their newer approaches. Through this review, an attempt has made to critically evaluate the recent advances and future aspects helpful to the treatment of COVID-19 based on the present understanding of SARS-CoV-2 infections, which may help offer new insights and potential therapeutic targets for the treatment of the COVID-19.

MicroRNAs are small ribonucleic acid that act as an important regulator of gene expression at the molecular level. However, there is no comparative data on the regulation of microRNAs (miRNAs) in visceral leishmaniasis (VL) and post-kala-azar dermal leishmaniasis (PKDL). In this current study, we compared the expression miRNA profile in host cells (GTHP), with VL strain (GVL) and PKDL strain-infected host cell (GPKDL). Normalized read count comparison between different conditions revealed that the miRNAs are indeed differentially expressed. In GPKDL with respect to GVL and GTHP, a total of 798 and 879 miRNAs were identified, out of which 349 and 518 are known miRNAs, respectively. Comparative analysis of changes in miRNA expression suggested that the involvement of differentially expressed miRNAs in various biological processes like PI3K pathway activation, cell cycle regulation, immunomodulation, apoptosis inhibition, different cytokine production, T-cell phenotypic transitions calcium regulation, and so on. A pathway enrichment study using in silico predicted gene targets of differentially expressed miRNAs showed evidence of potentially universal immune signaling pathway effects. Whereas cytokine–cytokine receptor interaction, phagocytosis, and transforming growth factor beta (TGF-β) signaling pathways were more highly enriched using targets of miRNAs upregulated in GPKDL. These findings could contribute to a better understanding of PKDL pathogenesis. Furthermore, the identified miRNAs could also be used as biomarkers in diagnosis, prognosis, and therapeutics of PKDL infection control.

Multiple myeloma is a malignant proliferation of the plasma cells mainly affecting bone marrow, but other organs may also be involved. Multiple myeloma is rarely associated with lung plasmacytoma. Patients with extramedullary plasmacytomas have coexistent multiple myeloma in only 5% of cases. The incidence of malignant pleural effusion in multiple myeloma is very low and is approximately 1%. In this case series, we report three patients with multiple myeloma, who presented with different thoracic manifestations such as left-sided myelomatous pleural effusion, posterior mediastinal mass with central airway obstruction, and pulmonary consolidation.

Primary brain tumors including gliomas continue to pose significant management challenges to clinicians. While the presentation, the pathology, and the clinical course of these lesions are variable, the initial investigations are usually similar. Patients who are suspected to have a brain tumor will be assessed with computed tomography (CT) and magnetic resonance imaging (MRI). The imaging findings are used by neurosurgeons to determine the feasibility of surgical resection and plan such an undertaking. Imaging studies are also an indispensable tool in tracking tumor progression or its response to treatment. As these imaging studies are non-invasive, relatively cheap and accessible to patients, there have been many efforts over the past two decades to increase the amount of clinically-relevant information that can be extracted from brain imaging. Most recently, artificial intelligence (AI) techniques have been employed to segment and characterize brain tumors, as well as to detect progression or treatment-response. However, the clinical utility of such endeavours remains limited due to challenges in data collection and annotation, model training, and the reliability of AI-generated information. We provide a review of recent advances in addressing the above challenges. First, to overcome the challenge of data paucity, different image imputation and synthesis techniques along with annotation collection efforts are summarized. Next, various training strategies are presented to meet multiple desiderata, such as model performance, generalization ability, data privacy protection, and learning with sparse annotations. Finally, standardized performance evaluation and model interpretability methods have been reviewed. We believe that these technical approaches will facilitate the development of a fully-functional AI tool in the clinical care of patients with gliomas., 31 pages, 6 figures. Accepted for publication in the Journal of Neural Engineering

Albeit, all forms of tuberculosis (TB) are endemic in India, spinal intramedullary TB and tubercular longitudinally extensive transverse myelitis (LETM) is deemed extremely rare. With recent advances in the field of neurology, autoimmune astrocytopathy (neuromyelitis optica spectrum disorders, NMOSD), myelin-oligodendrocyte glycoprotein associated encephalomyelitis (MOG-EM), metabolic myelopathy, connective tissue diseases and viral infections have gained considerable focus in the list of differentials of LETM whereas tubercular association is often forgotten. This report presents a rare case of acute transverse myelopathy which unveiled previously undiagnosed pulmonary tuberculosis in an adult rural Indian male. The patient responded well to anti-tubercular therapy and corticosteroids. Exact pathogenesis of LETM in TB remains elusive. Association of TB with MOG-EM has been one of the recent hot-cakes. However, an ill-defined immune-inflammatory response to the infectious agent is the likely cause of tubercular LETM. Hence, the primary care physicians who are the first medical contacts of acute LETM cases and in most cases due to diagnostic dilemma there is an unavoidable delay in accurate diagnosis and initiation of therapy. Primary care doctors should nurture a high index of suspicion to diagnose this potentially lifetime-debilitating yet absolutely treatable clinical condition i.e. tubercular LETM.

AIMS: The prognosis of nonsmall cell lung cancer with cavitation (NSCLC-c) is not well-known. We compared the positron emission tomography/computed tomography (PET/CT) findings and survival data of patients with NSCLC-c patients with those without cavitation (NSCLC-nc). METHODS: Between 7/2004 and 6/2007, cavitary lung lesions were identified in 46/248 patients undergoing fluorodeoxyglucose (FDG) PET/CT for lung nodule characterization or lung cancer staging. Within the same period, 40 of 202 patients with NSCLC-nc were randomly selected for comparison. The primary was assessed by location, size, cell type, and standardized uptake value (SUV). Disease stage was determined according to American Joint Committee on Cancer guidelines for lung cancer. Kaplan–Meier method was used for survival analysis and Cox regression to assess the effect of clinical and imaging variables on survival. RESULTS: NSCLC-c was found in 87% of patients that had a cavitary lung lesion at PET/CT. Squamous cell carcinoma, primary size and primary-to-liver SUV ratio differed significantly between NSCLC-c and NSCLC-nc, whereas age, gender, primary location, primary SUV, type of treatment, and disease stage did not. Median survival and overall 5-year survival were 19 months and 24% for NSCLC-c, and 31 months and 31% for NSCLC-nc, P = 0.23. Disease stage was the only predictor of survival. CONCLUSION: Cavitary lung lesions in patients undergoing FDG PET/CT harbor a significant risk for cancer. NSCLC-c is associated with squamous cell carcinoma, larger size, and greater FDG metabolism compared with NSCLC-nc, although these variables may not be predictive of survival. Nonetheless, PET/CT contributes to accurate staging and has an indirect impact on prognosis.

Micro RNAs (miRNAs) have emerged as a critical regulator of several biological processes in both animals and plants. They have also been associated with regulation of immune responses in many human diseases during recent years. Visceral leishmaniasis (VL) is the most severe form of leishmaniasis, which is characterized by impairment of both innate and adaptive immune responses. In the present study, we observed that Leishmania establishes hypoxic environment in host macrophages that induces the expression of hypoxia inducible factor-1α (HIF-1α) and miRNA-210. Further, the expression of miRNA-210 was found to be dependent on activation of HIF-1α expression. The HIF-1α silencing by siRNA resulted in significantly (p < 0.001) decreased expression of miR-210 in parasites infected macrophages. We also observed that in siHIF-1α or antagomir-210 treated L. donovani infected macrophages, the parasitic load and percentage infectivity were significantly (p < 0.001) decreased. Furthermore, we found that inhibition of miR-210 leads to activation of NF-κB subunit p50, and it forms heterodimer with p65 and translocates into the nucleus from the cytoplasm. This significantly (p < 0.05) induced the transcription of pro-inflammatory cytokines genes such as TNF-α and IL-12 in miRNA-210 inhibited macrophages compared to uninhibited macrophages whereas the level of IL-10, an anti-inflammatory cytokine, was found to be significantly decreased (p < 0.001). These findings suggested that L. donovani infection induces hypoxic environment inside the macrophages that activates HIF-1α. Further, HIF-1α upregulates miR-210, which eventually establishes a suitable environment for the survival of parasite inside the host macrophages by downregulating NF-κB mediated pro-inflammatory immune responses.

Duodenal perforation is a common cause of acute abdomen which is complication of peptic ulcer disease. The pathogenesis is considered complex and involves imbalance between defensive mechanisms of intestinal mucosa and aggravating factors to which a person is exposed.

BACKGROUND: Laryngoscopy and tracheal intubation causes intense autonomic reflex responses consisting of increased circulating catacholamines, tachycardia, hypertension, myocardial oxygen demand, and dysarrythmias. To obtund haemodynamic response lignocaine, opiods, nitroprusside, nitroglycerine, vearpamil, nifedipine, esmolol, clonidine and recently, dexmedetomidine have been stu died. AIMS AND OBJECTIVES: We investigated whether dexmedetomidine a α2 agonist could attenuate sympathoadrenal response (Heart rate and MAP) to laryngoscopy and intubation. MATERIALS AND METHODS: Eighty patients, ASA grade I/II, undergoing routine general anesthesia were randomly premedicated by i . v . dexmedetomidine 0.6μg or saline. Heart rate (HR), mean arterial pressure (MAP), were measured before, after the premedication, after thiopental, after succinylcholine at laryngoscopy, immediately after intuba tion and then 1 min. 3 min. and 5 min after intubation. STATISTICAL ANALYSIS: Descriptive and inferential statistics using chi - square test, z - test and wilcoxon sign rank test was done. Software used in the analysis was SPSS 17.0 version and Graph Pad Prism 5.0. Data was reported as mean value ± SD & p - value

Malaria continues to be one of the major public health problem of India. Almost all deaths are caused by Plasmodium falciparum infected malaria. 1 Under normal physiological conditions, liver ensures homeostasis of lipid and lipoprotein metabolism. 2 Hepatocellular damage often associated with severe and acute P falciparum infections impairs these processes, leading to alteration in plasma lipid profile and lipoprotein patterns. An observation on the nature of dyslipidaemia in malaria patients with reference to correlation if any that exists between malaria and lipid profile in these patients was performed. Results revealed that the total cholesterol, HDL and LDL decreased while triglyceride and VLDL were found to increase in malaria patients. It can be concluded that alteration in lipid profile can be an index of malaria infection but specificity of malaria infection with lipid changes is questionable and need further studies. INTRODUCTION: Malaria is a protozoan disease transmitted by the bite of infected Anopheles mosquitoes. The most important of the parasitic diseases of humans, it is transmitted in 108 countries containing 3 billion people and causes nearly 1 million deaths each year. Almost all deaths are caused by falciparum malaria.3 India is a vast country with multiethnic society of 1.2 billion living in diverse geo-ecological paradigms and ecotype of malaria. Malaria continues to be one of the major public health problem of India with around 1.5-2 million confirmed cases/year, of which approximately 1000 reported malarial death per year. 4 As per World Health Organization report 2011-2012, South-east Asia region bear the second largest burden of malaria (13%), only being next to African region (81%). Among South-east Asia region, India shares two thirds of the burden (66%) followed by Myanmar (18%) and Indonesia (10%).5 Contrary to the African scenario, where much of the malaria mortality burden is borne by the infants and children, in India, it is the middle productive ages in both genders that suffer the most. 6 Under normal physiological conditions, liver ensures homeostasis of lipid and lipoprotein metabolism. 7 Hepatocellular damage often associated with severe and acute P falciparum infections impairs these processes, leading to alteration in plasma lipid and lipoprotein patterns.¶⁻· Parasites forage nutrients from their host as well as possess limited enzyme pathway for de novo synthesis of certain nutrients. So far, studies suggest that there may be some factors or enzymes, which allow the parasite to breakup and consume lipid/cholesterol from their host and utilize them for internalization of eukaryotic protozoa or for reproduction in case of helminthes.¸ Plasmodium is incapable of de novo synthesis of fatty acids and cholesterol; but it can fabricate its glycerides and phosphoglycerides with host-supplied nutrients, and can produce the glyceryl moiety during glycolysis.¹

The survival of intracellular protozoan parasite, Leishmania donovani, the causative agent of Indian visceral leishmaniasis (VL), depends on the activation status of macrophages. l-Arginine, a semi-essential amino acid plays a crucial regulatory role for activation of macrophages. However, the role of l-arginine transport in VL still remains elusive. In this study, we demonstrated that intra-macrophage survival of L. donovani depends on the availability of extracellular l-arginine. Infection of THP-1-derived macrophage/human monocyte-derived macrophage (hMDM) with Leishmania, resulted in upregulation of l-arginine transport. While investigating the involvement of the transporters, we observed that Leishmania survival was greatly impaired when the transporters were blocked either using inhibitor or siRNA-mediated downregulation. CAT-2 was found to be the main isoform associated with l-arginine transport in L. donovani-infected macrophages. l-arginine availability and its transport regulated the host arginase in Leishmania infection. Arginase and inducible nitric oxide synthase (iNOS) expression were reciprocally regulated when assayed using specific inhibitors and siRNA-mediated downregulation. Interestingly, induction of iNOS expression and nitric oxide production were observed in case of inhibition of arginase in infected macrophages. Furthermore, inhibition of l-arginine transport as well as arginase resulted in decreased polyamine production, limiting parasite survival inside macrophages. l-arginine availability and transport regulated Th1/Th2 cytokine levels in case of Leishmania infection. Upregulation of l-arginine transport, induction of host arginase, and enhanced polyamine production were correlated with increased level of IL-10 and decreased level of IL-12 and TNF-α in L. donovani-infected macrophages. Our findings provide clear evidence for targeting the metabolism of l-arginine and l-arginine-metabolizing enzymes as an important therapeutic and prophylactic strategy to treat VL.

Background In vector-borne diseases such as leishmaniasis, the sand fly midgut is considered to be an important site for vector-parasite interaction. Digestive enzymes including serine peptidases such as trypsin and chymotrypsin, which are secreted in the midgut are one of the obstacles for Leishmania in establishing a successful infection. The presence of some natural inhibitors of serine peptidases (ISPs) has recently been reported in Leishmania. In the present study, we deciphered the role of these ISPs in the survival of Leishmania donovani in the hostile sand fly midgut environment. Methods In silico and co-immunoprecipitation studies were performed to observe the interaction of L. donovani ISPs with trypsin and chymotrypsin. Zymography and in vitro enzyme assays were carried out to observe the inhibitory effect of purified recombinant ISPs of L. donovani (rLdISPs) on trypsin, chymotrypsin and the sand fly midgut peptidases. The expression of ISPs in the amastigote to promastigote transition stages were studied by semi-quantitative RT-PCR and Western blot. The role of LdISP on the survival of ISP overexpressed (OE) and ISP knocked down (KD) Leishmania parasites inside the sand fly gut was investigated by in vitro and in vivo cell viability assays. Results We identified two ecotin-like genes in L. donovani, LdISP1 and LdISP2. In silico and co-immunoprecipitation results clearly suggest a strong interaction of LdISP molecules with trypsin and chymotrypsin. Zymography and in vitro enzyme assay confirmed the inhibitory effect of rLdISP on trypsin, chymotrypsin and the sand fly midgut peptidases. The expression of LdISP2 was found to be strongly associated with the amastigote to promastigote phase transition. The activities of the digestive enzymes were found to be significantly reduced in the infected sand flies when compared to uninfected. To our knowledge, our study is the first report showing the possible reduction of chymotrypsin activity in L. donovani infected sand flies compared to uninfected. Interestingly, during the early transition stage, substantial killing was observed in ISP2 knocked down (ISP2KD) parasites compared to wild type (WT), whereas ISP1 knocked down (ISP1KD) parasites remained viable. Therefore, our study clearly indicates that LdISP2 is a more effective inhibitor of serine peptidases than LdISP1. Conclusion Our results suggest that the lack of ISP2 is detrimental to the parasites during the early transition from amastigotes to promastigotes. Moreover, the results of the present study demonstrated for the first time that LdISP2 has an important role in the inhibition of peptidases and promoting L. donovani survival inside the Phlebotomus argentipes midgut.

Currently, there is no approved vaccine for visceral leishmaniasis (VL) caused by L. donovani. The ability to manipulate Leishmania genome by eliminating or introducing genes necessary for parasites’ survival considered as the powerful strategy to generate the live attenuated vaccine. In the present study fructose-1,6-bisphosphatase (LdFBPase) gene deleted L. donovani (Δfbpase) was generated using homologous gene replacement strategy. Though LdFBPase gene deletion (Δfbpase) does not affect the growth of parasite in the promastigote form but axenic amastigotes display a marked reduction in their capacity to multiply in vitro inside macrophages and in vivo in Balb/c mice. Though Δfbpase L. donovani parasite persisted in BALB/c mice up to 12 weeks but was unable to cause infection, we tested its ability to protect against a virulent L. donovani challenge. Notably, intraperitoneal immunisation with live Δfbpase parasites displayed the reduction of parasites load in mice spleen and liver post challenge. Moreover, immunised BALB/c mice showed a reversal of T cell anergy and high levels of NO production that result in the killing of the parasite. A significant, correlation was found between parasite clearance and elevated IFNγ, IL12, and IFNγ/IL10 ratio compared to IL10 and TGFβ in immunised and challenged mice. Results suggested the generation of protective Th1 type immune response which induced significant parasite clearance at 12-week, as well as 16 weeks post, challenged immunised mice, signifying sustained immunity. Therefore, we propose that Δfbpase L. donovani parasites can be a live attenuated vaccine candidate for VL and a good model to understand the correlatives of protection in visceral leishmaniasis.

Amphotericin B (AmB), a polyene macrolide, is now a first-line treatment of visceral leishmaniasis cases refractory to antimonials in India. AmB relapse cases and the emergence of secondary resistance have now been reported. To understand the mechanism of AmB, differentially expressed genes in AmB resistance strains were identified by a DNA microarray and real-time reverse transcriptase PCR (RT-PCR) approach. Of the many genes functionally overexpressed in the presence of AmB, the ascorbate peroxidase gene from a resistant Leishmania donovani strain (LdAPx gene) was selected because the gene is present only in Leishmania , not in humans. Apoptosis-like cell death after exposure to AmB was investigated in a wild-type (WT) strain in which the LdAPx gene was overexpressed and in AmB-sensitive and -resistant strains. A higher percentage of apoptosis-like cell death after AmB treatment was noticed in the sensitive strain than in both the resistant isolate and the strain sensitive to LdAPx overexpression. This event is preceded by AmB-induced formation of reactive oxygen species and elevation of the cytosolic calcium level. Enhanced cytosolic calcium was found to be responsible for depolarization of the mitochondrial membrane potential and the release of cytochrome c (Cyt c ) into the cytosol. The redox behavior of Cyt c showed that it has a role in the regulation of apoptosis-like cell death by activating metacaspase- and caspase-like proteins and causing concomitant nuclear alterations, as determined by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) and DNA fragmentation in the resistant strain. The present study suggests that constitutive overexpression of LdAPx in the L. donovani AmB-resistant strain prevents cells from the deleterious effect of oxidative stress, i.e., mitochondrial dysfunction and cellular death induced by AmB.

Background: Laparotomy wound dehiscence is still a puzzle for most of the surgeons. Mortality associated with dehiscence has been estimated at 10-30%. Patients undergoing emergency laparotomy suffer from one of these comorbid conditions which are detrimental to healing. In this scenario interrupted suturing has been found to give good strength and have less incidence of wound dehiscence. The objective of the study was to compare the incidence of abdominal wound dehiscence in emergency midline laparotomy.Methods: This study was conducted on 300 consecutive patients undergoing emergency midline laparotomy in the Department of Surgery, Government Medical College and Hospital. Methods group-A: closed by suturing the rectus sheath using polydioxanone suture 1-0 (PDS) in continuous layer suturing method. group-B: closed by suturing the rectus sheath using polydioxanone suture 1-0 in interrupted layer suturing method.Results: The mean age in group A was 40.47 years and 37.47 in group B. In Group A 20.1% patients had burst abdomen and 5.4% in group B.Conclusions: Interrupted closure of abdominal wall fascia is better in emergency laparotomy as compared to continuous closure.

Objectives Indian Post kala-azar dermal leishmaniasis (PKDL) is the cutaneous aftermath of visceral leishmaniasis (VL) caused by L . donovani . Vitamin D-regulated cationic antimicrobial peptide cathelicidin (hCAP-18/LL-37) has microbicidal and immunomodulatory role against cutaneous infections, but its role in PKDL remains elusive. Methods Skin snips and blood-derived monocytes of PKDL patients ( n = 46), before (BT) and after (AT) chemotherapy, were used for this study. Serum vitamin D3 level was evaluated by ELISA. Cathelicidin and vitamin D receptor (VDR) levels were analyzed by real-time PCR and flowcytometry in PKDL patients. The mechanistic effect of cathelicidin on macrophage differentiation and anti-leishmanial activity was assessed through RNA interference techniques followed by subsequent microscopic evaluation of in vitro parasite killing and Th1/Th2 counter-regulation by ELISA/RT-PCR. Results Low vitamin D3 levels were accompanied with decreased expression of cathelicidin and VDR in PKDL-BT patients. Results suggested positive induction of VDR-dependent cathelicidin in PKDL macrophages by Amphotericin B treatment, which could be due to indirect effect of drug-induced IL12 upregulation. 1,25-Vitamin D3 stimulation induced cathelicidin in PKDL-BT patients through involvement of TLR2/IL-1β, but not TLR4. Cathelicidin also augmented the anti-leishmanial effect and macrophage activating potential of Amphotericin B, attributable to regulation of VDR-dependent enhancement of CD40, p-STAT-I and MHC-II expression leading to regulation of IL10/IL12 balance in PKDL-BT patient macrophages. Conclusions This study indicates that cathelicidin augments anti-leishmanial macrophage activating property of Amphotericin B in a TLR2/VDR dependent mechanism, and advocate the development of novel adjunct treatment modality of cathelicidin with conventional Amphotericin B in PKDL patients.

The transformation of Leishmania donovani from a promastigote to an amastigote during mammalian host infection displays the immense adaptability of the parasite to survival under stress. Induction of translation initiation factor 2-alpha (eIF2α) phosphorylation by stress-specific eIF2α kinases is the basic stress-perceiving signal in eukaryotes to counter stress. Here, we demonstrate that elevated temperature and acidic pH induce the phosphorylation of Leishmania donovani eIF2α (LdeIF2α). In vitro inhibition experiments suggest that interference of LdeIF2α phosphorylation under conditions of elevated temperature and acidic pH debilitates parasite differentiation and reduces parasite viability (P < 0.05). Furthermore, inhibition of LdeIF2α phosphorylation significantly reduced the infection rate (P < 0.05), emphasizing its deciding role in successful invasion and infection establishment. Notably, our findings suggested the phosphorylation of LdeIF2α under H2O2-induced oxidative stress. Inhibition of H2O2-induced LdeIF2α phosphorylation hampered antioxidant balance by impaired redox homeostasis gene expression, resulting in increased reactive oxygen species accumulation (P < 0.05) and finally leading to decreased parasite viability (P < 0.05). Interestingly, exposure to sodium antimony glucamate and amphotericin B induces LdeIF2α phosphorylation, indicating its possible contribution to protection against antileishmanial drugs in common use. Overall, the results strongly suggest that stress-induced LdeIF2α phosphorylation is a necessary event for the parasite life cycle under stressed conditions for survival.

Various physiological stimuli trigger the conversion of noninfective Leishmania donovani promastigotes to the infective form. Here, we present the first evidence of the effect of glucose starvation, on virulence and survival of these parasites. Glucose starvation resulted in a decrease in metabolically active parasites and their proliferation. However, this was reversed by supplementation of gluconeogenic amino acids. Glucose starvation induced metacyclogenesis and enhanced virulence through protein kinase A regulatory subunit (LdPKAR1) mediated autophagy. Glucose starvation driven oxidative stress upregulated the antioxidant machinery, culminating in increased infectivity and greater parasitic load in primary macrophages. Interestingly, phosphoenolpyruvate carboxykinase (LdPEPCK), a gluconeogenic enzyme, exhibited the highest activity under glucose starvation to regulate growth of L. donovani by alternatively utilising amino acids. Deletion of LdPEPCK (Δpepck) decreased virulent traits and parasitic load in primary macrophages but increased autophagosome formation in the mutant parasites. Furthermore, Δpepck parasites failed to activate the Pentose Phosphate Pathway shunt, abrogating NADPH/NADP+ homoeostasis, conferring increased susceptibility towards oxidants following glucose starvation. In conclusion, this study showed that L. donovani undertakes metabolic rearrangements via gluconeogenesis under glucose starvation for acquiring virulence and its survival in the hostile environment.

Percutaneous balloon mitral valvuloplasty (BMV)using an Accura balloon is an effective procedure for the management of patients with rheumatic mitral stenosis.The inability to cross the mitral valve is one of the prime reasons for procedural failure. Here, we describe the case of a 24 year-old male with severe mitral stenosis with agiant, aneurysmal left atrium (left atrium12.3 × 10.4 cm) where the traditionally described technique of mitral valve entry failed. BMV was successfully performed by a U-shape modification of hairpin loop entry (the modified hairpin entry method), expanding the mitral valve area from 0.8 cm 2 to 1.9 cm 2 .

Manas R Dikhit,1,* Bidyut Purkait,1,* Ruby Singh,1 Bikash Ranjan Sahoo,2 Ashish Kumar,1 Rajiv K Kar,3 MdYousuf Ansari,1,4 SavitaSaini,1,5 KumarAbhishek,1 Ganesh CSahoo,1 Sushmita Das,6 Pradeep Das11Department of Molecular Parasitology and Biomedical Informatics, Rajendra Memorial Research Institute of Medical Sciences, Indian Council of Medical Research, Agamkuan, Patna, Bihar, India; 2Laboratory of Molecular Biophysics, Institute for Protein Research, Osaka University, Japan; 3Biomolecular Nuclear Magnetic Resonance and Drug Design Laboratory, Department of Biophysics, Bose Institute, Kolkata, West Bengal, 4Department of Pharmacoinformatics, 5Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Hajipur, 6Department of Microbiology, All India Institute of Medical Sciences, Patna, Bihar, India*These authors contributed equally to this workAbstract: New treatments for visceral leishmaniasis, caused by Leishmania donovani, are needed to overcome sustained toxicity, cost, and drug resistance. The aim of this study was to evaluate the therapeutic effects of 2-nitro-N-(pyridin-2-ylmethyl)benzenesulfonamide (2NB) against promastigote and amastigote forms of L. donovani and examine its effect in combination with amphotericin B (AmB) against AmB-resistant clinical isolates. Effects were assessed against extracellular promastigotes in vitro and intracellular amastigotes in L. donovani-infected macrophages. Levels of inducible nitric oxide and Th1 and Th2 cytokines were measured in infected 2NB-treated macrophages, and levels of reactive oxygen species and NO were measured in 2NB-treated macrophages. 2NB was active against promastigotes and intracellular amastigotes with 50% inhibitory concentration values of 38.5±1.5µg/mL and 86.4±2.4µg/mL, respectively. 2NB was not toxic to macrophages. Parasite titer was reduced by >85% in infected versus uninfected macrophages at a 2NB concentration of 120µg/mL. The parasiticidal activity was associated with increased levels of Th1 cytokines, NO, and reactive oxygen species. Finally, 2NB increased the efficacy of AmB against AmB-resistant L. donovani. These results demonstrate 2NB to be an antileishmanial agent, opening up a new avenue for the development of alternative chemotherapies against visceral leishmaniasis.Keywords: visceral leishmaniasis, AmB resistance, benzenesulfonamide, ROS, NO, Th1/Th2 cytokines