Search

Micro RNAs (miRNAs) have been implicated in the regulation of maturation, proliferation, differentiation, and activation of immune cells. In this study, we demonstrated that miR-29a antagonizes IFN-γ production at early times post-LSDV infection in cattle. miR-29a was predicted to target upstream IFN-γ regulators, and its inhibition resulted in enhanced IFN-γ production in sensitized peripheral blood mononuclear cells (PBMCs). Further, stimulation of PBMCs with LSDV antigen exhibited lower levels of miR-29a, concomitant with a potent cell-mediated immune response (CMI), characterized by an increase in LSDV-specific CD8+ T cell counts and enhanced levels of IFN-γ, which eventually facilitated virus clearance. In addition, a few immunocompromised cattle (developed secondary LSDV infection at ~ 6 months) that failed to mount a potent cell-mediated immune response, were shown to maintain higher miR-29a levels. Furthermore, as compared to the sensitized crossbred cattle, PBMCs from sensitized Rathi (a native Indian breed) animals exhibited lower levels of miR-29a along with an increase in CD8+ T cell counts and enhanced levels of IFN-γ. Finally, we analysed that a ≥ 60% decrease in miR-29a expression levels in the PBMCs of sensitized cattle correlated with a potent CMI response. In conclusion, miR-29a expression is involved in antagonizing the IFN-γ response in LSDV-infected cattle and may serve as a novel biomarker for the acute phase of LSDV infection, as well as predicting the functionality of T cells in sensitized cattle. In addition, Rathi cattle mount a more potent CMI response against LSDV than crossbred cattle.

Johne's disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), poses significant challenges to the global livestock industry, particularly affecting bovine populations. To better understand the prevalence of paratuberculosis and its diagnostic nuances, a comprehensive meta-analysis was conducted. This analysis encompassed 21 studies involving 632,767 cows for milk enzyme-linked immunosorbent assay (ELISA) and 51 studies involving 256,409 cows for serum ELISA. The pooled prevalence estimate for paratuberculosis on a cow-basis was found to be 16% (95% CI: 14%; 18%) for milk ELISA and 8% (95% CI: 7%; 8%) for serum ELISA. Notably, higher confidence intervals (CI) were observed in milk ELISA, the Europe and Asia groups, suggesting variability in prevalence estimates within these regions. Conversely, lower CIs were noted in the USA and Canada groups, indicating greater consistency in prevalence estimates within these countries. However, serum ELISA exhibited high CI values across all regions, underscoring potential variability in diagnostic performance. These findings provide valuable insights for veterinarians, researchers, policymakers, and livestock producers in optimizing paratuberculosis detection and control strategies to mitigate its impact on bovine health and agricultural productivity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Since 2019, Lumpy skin disease (LSD) has suddenly spread in many Asian countries, including India. LSD primarily occurs in cattle. However, recent LSD outbreaks in India have also revealed significant morbidity and production losses in buffaloes. This has raised concerns about the role of buffaloes in the epidemiology and transmission of LSD and necessitates the inclusion of buffaloes in the mass vaccination program for the prevention and control of the disease in the country. However, there is no significant data on the immune response in buffaloes following vaccination with the LSD vaccine. In this study, we evaluated antibody- and cell-mediated immune responses following vaccination with a newly developed live-attenuated LSD vaccine (Lumpi-ProVac Ind ). The detectable amount of anti-LSDV antibodies was observed at 1-2 months following vaccination, with a peak antibody titer at 3 months. Upon stimulation of the peripheral blood mononuclear cells (PBMCs) with the UV-inactivated LSDV antigen, there was a significant increase in CD8 + T cell counts in vaccinated animals as compared to the unvaccinated animals. Besides, vaccinated animals also showed a significant increase in IFN-γ levels upon antigenic stimulation of their PBMCs with LSDV antigen. In conclusion, the buffaloes also mount a potent antibody- and cell-mediated immune response following vaccination with Lumpi-ProVac Ind ., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Allelopathic research is not getting the intended consideration because of the complexities involved in their isolation, identification, understanding their modes of action, interactions with other environmental factors, impacts on non-target organisms and exploration of their utility in diverse fields. Additionally, their variability and trace availability have presented hurdles in establishing future research utilities and their field applications. Exploring the historical context of allelopathic research is essential for obtaining a more profound understanding of the progression in this research domain and to identify the research gaps and potential future prospectives. Therefore, the current bibliometric review aims to examine the research advancements, trends, hotspots, research gaps and to identify future prospectives in allelopathic research. A Scopus database search was carried out to collect the bibliometric data using the combination of multiple search strings in advance search option. The outcomes of this study revealed a total of 5427 published articles, with an average of 19.12 citations per article. Despite the increasing trend in research and publications on allelopathy/allelochemicals over the last decade, the majority of allelopathic research remains focused on investigating novel allelochemicals and their potential for weed management. Other crucial considerations like their phytotoxicity and ecotoxicity, selectivity for crop growth, interactions with herbicides and their derivatives, biochemical signalling, identification of germplasm in allelopathic plants, inducing allelopathic trait into enhanced cultivars, their ultimate fate in the open environment are sparsely investigated. It is anticipated that this review will draw greater attention to some overlooked domains within allelopathic research., (© 2024 Scandinavian Plant Physiology Society.)

Mycobacterium avium subspecies paratuberculosis (MAP) infection leads to chronic, persistent granulomatous enteritis, causing prolonged diarrhoea and emaciation. The disease is managed using medications such as antibiotics, live vaccines, mycobacteriophage therapies and other treatments; however, a notable proportion of affected animals do not show improvement with this approach. We hypothesise that immunoinhibitory receptors TIM-3 (T cell immunoglobulin mucin protein-3) and PD-1 (Programmed death receptor 1) may be upregulated on Peripheral blood mononuclear cells (PBMCs) of MAP-seropositive bovines, potentially contributing to immune exhaustion. Samples (blood and faeces) were collected from 32 diarrhoeic bovines suspected of MAP infection; eight apparently healthy buffaloes from the dairy farm at Hisar, Haryana and from 14 cows (suffering from chronic diarrhoea, weakness and emaciation) housed in stray cattle shed. MAP infection was estimated using indigenous ELISA (i-ELISA), faecal IS 900 PCR, culture and acid-fast staining. TIM-3 and PD-1 gene expression on PBMCs were determined using qRT-PCR. TIM3 expression was relatively higher (~400-fold, 330-fold, 112-fold, 65-fold and 16-fold) in 5 chronically diarrhoeic PBMCs samples (MAP-seropositive), and higher PD-1 expression (around ~7-fold, 1.75-fold, 2.5-fold, 7.6-fold) was recorded in 4 diarrhoeic MAP-seropositive animals, compared to apparently healthy and other MAP-seronegative diarrhoeic animals. High co-expression of TIM-3 and PD-1 levels was also recorded in chronically diarrhoeic, emaciated stray cattle. Understanding immune responses in field conditions might aid in the therapeutic management of paratuberculosis.

Purpose: As part of the European risk management plan of a 91-day extended levonorgestrel-containing combined oral contraceptive (COC LNG ), a study was performed to assess its safety. This analysis was conducted to examine delayed pregnancy detection and return to fertility with extended combined oral contraceptives (COC)., Methods: We conducted a retrospective cohort study in new users of 91-day COC LNG or 28-day COC LNG within a US-based healthcare claims database from 2006 to 2017. Delayed pregnancy detection during current COC LNG exposure was defined as the time between estimated pregnancy start and first prenatal care encounter. Additionally, the time between estimated pregnancy start and COC LNG discontinuation was measured. To measure return to fertility, pregnancy rates were estimated among females who discontinued treatment. 91-day COC LNG users were propensity score-matched to 28-day COC LNG users. Hazard ratio for pregnancy was calculated using Cox proportional hazards models., Results: The 91-day and 28-day COC LNG users had 25 593 and 76 586 treatment episodes, respectively. The median time to pregnancy detection was 64.5 and 61.0 days (p = 0.24) in users of 91-day COC LNG and 28-day COC LNG . The median exposure time to treatment after estimated pregnancy start was 54.0 and 38.0 days (p < 0.01). In the fertility analysis, pregnancy rates were 54.82 (95% CI, 50.05-59.93) and 69.30 (95% CI, 64.98-73.82) per 1000 person-years in extended COC LNG discontinuers and 28-day COC LNG discontinuers. The adjusted hazard ratio of pregnancy was 0.77 (95% CI, 0.69-0.85)., Conclusions: Small differences were observed for pregnancy rates and delayed pregnancy detection between 91-day extended COC LNG and 28-day COC LNG , which may be related to the longer days' supply of extended COC LNG . Differences in the fertility analysis may be related to unmeasured residual confounding., (© 2022 John Wiley & Sons Ltd.)

Relationship between various combinations of KIR ligands and HLA alleles have been studied in several diseases. The aim of this retrospective study was to estimate the frequency of HLA alleles and KIR ligands among acute myeloid leukemia patients and healthy controls in order to examine the possible association of HLA alleles and KIR ligands with AML. A total of 439 acute myeloid leukemia patients and 1317 unrelated, healthy ethnic Indian controls were included in the study. HLA typing was performed using PCR-SSP. KIR ligands were assigned by using the KIR ligand Calculator. The frequency of HLA alleles and KIR ligands in patients was then compared with the controls. As compared to controls, frequencies of HLA-A*03 and HLA-B*35 were increased in AML patients, whereas, that of HLA-C*03 was decreased. Frequencies of HLA-A*03 and HLA-C*15 were increased in male patients, however, no significant difference was observed in female patients as compared to controls. In the pediatric group, the frequencies of HLA-A*01 was decreased and that of HLA-A*03 and HLA-B*18 were increased, whereas, frequencies of HLA-B*13 was decreased and that of HLA-B*27 was increased in the adult patients. In the haplotype analysis, the frequency of HLA-A*24/B*35/DRB1*15 was increased in overall patients. In adult group, the frequency of HLA-A*01/B*44/DRB1*07 was increased in patients than in controls. No significant association was observed between KIR ligands and susceptibility/ protection to AML. Our results indicate that certain HLA alleles and haplotypes have presumptive positive or negative role in conferring protection/susceptibility to AML., Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-022-01550-0., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Author(s), under exclusive licence to Indian Society of Hematology and Blood Transfusion 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Madagascar's unique biota is heavily affected by human activity and is under intense threat. Here, we review the current state of knowledge on the conservation status of Madagascar's terrestrial and freshwater biodiversity by presenting data and analyses on documented and predicted species-level conservation statuses, the most prevalent and relevant threats, ex situ collections and programs, and the coverage and comprehensiveness of protected areas. The existing terrestrial protected area network in Madagascar covers 10.4% of its land area and includes at least part of the range of the majority of described native species of vertebrates with known distributions (97.1% of freshwater fishes, amphibians, reptiles, birds, and mammals combined) and plants (67.7%). The overall figures are higher for threatened species (97.7% of threatened vertebrates and 79.6% of threatened plants occurring within at least one protected area). International Union for Conservation of Nature (IUCN) Red List assessments and Bayesian neural network analyses for plants identify overexploitation of biological resources and unsustainable agriculture as the most prominent threats to biodiversity. We highlight five opportunities for action at multiple levels to ensure that conservation and ecological restoration objectives, programs, and activities take account of complex underlying and interacting factors and produce tangible benefits for the biodiversity and people of Madagascar.

Madagascar's biota is hyperdiverse and includes exceptional levels of endemicity. We review the current state of knowledge on Madagascar's past and current terrestrial and freshwater biodiversity by compiling and presenting comprehensive data on species diversity, endemism, and rates of species description and human uses, in addition to presenting an updated and simplified map of vegetation types. We report a substantial increase of records and species new to science in recent years; however, the diversity and evolution of many groups remain practically unknown (e.g., fungi and most invertebrates). Digitization efforts are increasing the resolution of species richness patterns and we highlight the crucial role of field- and collections-based research for advancing biodiversity knowledge and identifying gaps in our understanding, particularly as species richness corresponds closely to collection effort. Phylogenetic diversity patterns mirror that of species richness and endemism in most of the analyzed groups. We highlight humid forests as centers of diversity and endemism because of their role as refugia and centers of recent and rapid radiations. However, the distinct endemism of other areas, such as the grassland-woodland mosaic of the Central Highlands and the spiny forest of the southwest, is also biologically important despite lower species richness. The documented uses of Malagasy biodiversity are manifold, with much potential for the uncovering of new useful traits for food, medicine, and climate mitigation. The data presented here showcase Madagascar as a unique "living laboratory" for our understanding of evolution and the complex interactions between people and nature. The gathering and analysis of biodiversity data must continue and accelerate if we are to fully understand and safeguard this unique subset of Earth's biodiversity.

Chenopodium album L. and Chenopodium murale L. are two principal weed species, causing substantial damage to numerous winter crops across the globe. For sustainable and resource-efficient management strategies, it is important to understand weeds’ germination behaviour under diverse conditions. For the germination investigations, seeds of both species were incubated for 15 days under different temperatures (10−30 °C), salinity (0−260 mM NaCl), osmotic stress (0−1 MPa), pH (4−10), and heating magnitudes (50−200 °C). The results indicate that the germination rates of C. album and C. murale were 54−95% and 63−97%, respectively, under a temperature range of 10 to 30 °C. The salinity levels for a 50% reduction in the maximum germination (GR50) for C. album and C. murale were 139.9 and 146.3 mM NaCl, respectively. Regarding osmotic stress levels, the GR50 values for C. album and C. murale were 0.44 and 0.43 MPa, respectively. The two species showed >95% germination with exposure to an initial temperature of 75 °C for 5 min; however, seeds exposed to 100 °C and higher temperatures did not show any germination. Furthermore, a drastic reduction in germination was observed when the pH was less than 6.0 and greater than 8.0. The study generated information on the germination biology of two major weed species under diverse ecological scenarios, which may be useful in developing efficient weed management tactics for similar species in future agri-food systems.

Competing Interests: There are no conflicts of interest.

Background: Founder mutation is a heritable genetic alteration observed with high frequency in a geographically and culturally isolated population where one or more ancestors becomes the forebearer of the altered gene. The current study reports two founder mutations in the BRCA1 gene in the Nepalese people., Methods: Germline BRCA testing in all surface epithelial ovarian cancers and the selected case of breast, prostate, and pancreatic cancers has been the standard practice from 2016 to 2021. One thousand one hundred thirtythree probands were screened for germline BRCA variants by next generation sequencing. The variants were classified as per the American Society of Medical Genetics and Genomics recommendations. Pathogenic (class V) and likely pathogenic (class IV) were considered clinically relevant and utilized for cascade screening., Results: Nepalese population made up a subcohort of 5.12% (58/1,133) of probands tested for germline BRCA1/2 variants. Twenty-seven of these 58 tested harbored pathogenic genetic alterations in BRCA1/2 genes, with 23 being BRCA1 mutant. Sixteen of 23 BRCA1 mutant cases shared one common pathogenic mutation c.2214&#95;2215insT (p.Lys739Ter) (NM&#95;007294.4). Additionally, a second highly recurrent mutation in BRCA1 gene c.5068A>T (p.Lys1690Ter) (NM&#95;007294.4) was noted in six patients from this population., Conclusions: The overwhelming abundance of the above two variants in a geographically confined population confers these two genetic alterations a status of founder mutations amongst the people of Nepal. A more extensive population-based study to reaffirm these findings will help establish a dual site-specific germline testing similar to the "Multisite-3-assay" in Ashkenazi Jews as the primary screening tool, especially in a resource-constrained environment.

Background: Exon del19 and L858R mutations account for 90% of EGFR mutant non-small cell lung cancer (NSCLC). LUX lung 3 and 6 initially reported a survival difference between these two. However, other studies did not demonstrate the same. By using machine learning (ML), it is possible to discover novel patterns for cancer susceptibility, recurrence, prognostication, and therapy. We evaluate the effect of these two molecular subtypes on overall survival/progression-free survival (OS/PFS)., Methods: 413 patients with stage IV EGFR mutant NSCLC were analyzed for clinicopathologic features, treatment details, and survival outcome. PFS prediction models were built using ensemble decision trees, and random forest. Ensemble decision trees were built and validation was performed using survival analysis. Clustering regression techniques were then applied to train and test the prediction of the 1 st PFS of patients., Results: The median age of the cohort was 59 years comprising 53% males and 47% females. 275 (66.5%) patients showed a del19 mutation type and 138 (33.5%) harbored L858R. After clustering, the most important variables were age (P<0.05), ECOG performance status (PS) (P<0.04), PDL1 (P<0.09), smoking status (P<0.01) and to a lesser extent, number of extrathoracic metastasis (ETM) sites (median 1.2, P<0.06), brain metastasis (P<0.06) and gender (P<0.08). The prediction for 1 st PFS for del19 showed mean absolute error of 2.6 months and 4.72 months for L858R. The accuracy was 79.8% with 82% sensitivity, 79% specificity and AUC: 0.72. The precision was 92% with a Mathews correlation coefficient of 0.59., Conclusion: This study used machine learning modeling with fair accuracy to demonstrate that ECOG PS, age at diagnosis, and smoking status are the three main predictive factors of PFS in these patients., Competing Interests: None., (AJTR Copyright © 2022.)

The rice-wheat cropping system (RWCS) is the backbone of Indian farming, especially in the north-western region. But continuous adoption of the RWCS in northwest India has resulted in major challenges and stagnation in the productivity of this system. Additionally, the Indo-Gangetic Plains of Pakistan, Nepal, and Bangladesh are also facing similar challenges for sustainable production of the RWCS. Several emerging problems, such as the exhausting nutrient pool in soil, deteriorating soil health, groundwater depletion, escalating production cost, labor scarcity, environmental pollution due to crop residue burning and enhanced greenhouse gas emissions, climatic vulnerabilities, and herbicide resistance in weed species, are a few major threats to its sustainability. To address these challenges, a wide range of sustainable intensification technologies have been developed to reduce the irrigation and labor requirements, tillage intensity, and straw burning. Awareness and capacity building of the stakeholders and policy matching/advocacy need to be prioritized to adopt time- and need-based strategies at the ground level to combat these challenges. This review summarizes the current status and challenges of the RWCS in the northwest region of the country and also focuses on the precision management options for achieving high productivity, profitability, and sustainability., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Dhanda, Yadav, Yadav and Chauhan.)

In the current era of the COVID-19 pandemic, the world has never been more interested in the process of vaccine development. While researchers across the globe race to find an effective yet safe vaccine to protect populations from the newly emergent SARS-CoV-2 virus, more than one-third of the world has been subjected to either full or partial lockdown measures. With communities having felt the burden of prolonged isolation, finding a safe and efficacious vaccine will yield direct beneficial effects on protecting against COVID-19 morbidity and mortality and help relieve the psychological and economic load on communities living with COVID-19. There is hope that with the extraordinary efforts of scientists a vaccine will become available. However, given the global public health crisis, development of a COVID-19 vaccine will need to be fast tracked through the usual prelicensing development stages and introduced with limited clinical trial data compared with those vaccines that are developed conventionally over more than a decade. In this scenario, surveillance of the vaccine in the real world becomes even more paramount. This responsibility falls to observational researchers who can provide an essential safety net by continuing to monitor the effectiveness and safety of a COVID-19 vaccine after licensing. Postauthorisation observational studies for safety and effectiveness are complementary to prelaunch clinical trials and not a replacement. In this paper, we highlight the importance of postmarketing studies for future newly licensed COVID-19 vaccines and the key epidemiological considerations., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Objectives: To compare the safety profile of Seasonique, a 91-day levonorgestrel-containing combined oral contraceptive (COC LNG ), to 28-day COC LNG regarding the risk of venous thromboembolism (VTE) and arterial thromboembolism (ATE)., Study Design: A new user cohort study was conducted in a US health care database from 2006 to 2017. Each 91-day COC LNG treatment episode in females was matched to up to four 28-day COC LNG treatment episodes by propensity score. We identified VTE cases in either (1) an inpatient setting with ICD-9 and ICD-10 diagnosis codes of PE and/or DVT in the primary position, or (2) an outpatient setting with ICD-9 or ICD-10 diagnosis codes of DVT in conjunction with an anticoagulant medication dispensing or alteplase (thrombolytic) during the 30-day period following the date of DVT diagnosis. VTE was validated using medical records. We assessed the study endpoints in the two cohorts using incidence rates and Cox proportional hazards models adjusted for potential confounders., Results: Of the 25,593 treatment episodes in 91-day COC LNG and 76,586 treatment episodes in 28-day COC LNG , 35 and 68 patients had VTEs, respectively, corresponding to a hazard ratio (HR) of 1.40 (95% confidence interval [CI], 0.90-2.19). The VTE algorithm had a positive predictive value of 76.4% (95% CI, 66.2%-84.8%). ATEs were recorded in 13 and 28 episodes, respectively, with a corresponding HR of 1.21 (95% CI, 0.58-2.53)., Conclusions: These results do not indicate a significant difference between 91-day COC LNG and 28-day COC LNG in terms of VTE or ATE risk., Implications: Compared to use of 28-day COC LNG , use of 91-day extended COC LNG was not associated with a significant difference in risk of venous and arterial thromboembolism., (Copyright © 2021. Published by Elsevier Inc.)

Introduction: With the International Association for the Study of Lung Cancer (IASLC) recommendations promoting liquid biopsy as a primary detection tool, a new era of research has begun. The authors aimed to study the concordance of plasma genotyping platforms against the tissue gold standard. Methods: 184 patients with non-small cell lung cancer underwent EGFR genotyping using Cobas, droplet digital polymerase chain reaction (ddPCR) and Therascreen assays from 2019-2020. Results: Of 184 cases, 70 were positive by Cobas, 51 by ddPCR and 69 by Therascreen. The sensitivity of Cobas was 97.1% and the sensitivity of ddPCR was 71%. Receiver operating characteristic analysis showed an area under the curve of 0.977 for Cobas and 0.846 for ddPCR. Conclusion: In line with the FLAURA trial of osimertinib making its way to first-line and given the IASLC recommendations, it is important to understand the attributes of these tests to initiate appropriate treatment.

Oral non-steroidal anti-inflammatory drugs (NSAIDs) are known to be associated with an increased risk of bleeding. The NSAID, flurbiprofen, in the form of 8.75 mg lozenge or oromucosal spray is indicated for the symptomatic relief of sore throat. Despite the low dose as compared to alternative flurbiprofen preparations, concerns have been raised regarding its safety in terms of haemorrhagic events. This systematic review was conducted to identify existing evidence on the risk of haemorrhagic events with flurbiprofen 8.75 mg dose (any formulation), particularly where this may be due to potential interactions with other medicinal products. The systematic review examined studies reporting haemorrhagic events in patients receiving flurbiprofen 8.75 mg dose. Six individual electronic databases were searched up to 28th April 2020. Records were initially screened for relevance followed by further review of potentially eligible studies. Data extraction was performed for eligible studies and risk of bias in studies was assessed. The search strategy identified 1093 individual records. Of these, 1038 records were excluded after initial review; the majority of these records related to flurbiprofen in alternative formulations with alternative doses (e.g., eye drops, skin patches, oral tablets) thus were not considered relevant for further review. The 55 remaining records related to flurbiprofen 8.75 mg dose (any formulation) or flurbiprofen lozenge/oromucosal spray where the dose was not specified. After further review, 52 of these records were not considered eligible. Thus, only three records were included in this systematic review. The three studies reported a total of five haemorrhagic events in patients taking flurbiprofen 8.75 mg lozenge; the corresponding risk in each of the studies was 8.33, 1.98 and 1.96%. Where possible, comparison of flurbiprofen 8.75 mg lozenge to placebo produced risk ratios of 0.96 (95% CI 0.07, 13.25) and 2.00 (95% CI 0.10, 118.0). This systematic review found limited evidence on the risk of haemorrhagic events with flurbiprofen when used at a dose of 8.75 mg. Counts were low across all studies and results comparing flurbiprofen and placebo treatment arms were non-significant. However, scarcity of studies and low certainty of evidence for the outcome of haemorrhagic events limits the conclusions of this systematic review., Competing Interests: AT was employed by Reckitt Benckiser Healthcare (UK) Ltd. GC and AK are employed by Reckitt Benckiser Healthcare (UK) Ltd. This study received funding from the marketing authorisation holder of flurbiprofen 8.75 mg lozenge (Strefen Honey and Lemon), Reckitt Benckiser Healthcare (UK) Ltd. The funder had the following involvement with the study; study design, decision to publish, and preparation of the manuscript. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Dhanda, Evans, Roy, Osborne, Townsley, Coutinho, Kulasekaran and Shakir.)

A 77-year-old female patient with Marfan syndrome presented with a 1-week history of worsening malaise and poor appetite. This was associated with a dramatic unintentional loss of weight as well as shortness of breath worsened by exertion. She has significant medical histories of a hiatal hernia and chronic type B aortic dissection. CT scan of the chest was able to confirm a type IV paraoesophageal hiatal hernia compressing on the lower lobe of her left lung without any progression of her aortic dissection. As surgical intervention was contraindicated in light of her advanced age and comorbidities, she was managed conservatively with a clinically satisfied outcome in the short term. Her long-term prognosis, however, is still poor with a high mortality of 1 and 5 years., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2021. No commercial re-use. See rights and permissions. Published by BMJ.)

Objective: As part of the risk management plan in Europe, this study was conducted to characterize drug utilization patterns of Seasonique, a 91 day extended levonorgestrel-containing combined oral contraceptive (COC LNG )., Methods: A retrospective observational study was conducted in France, Italy and Belgium using electronic medical record databases obtained from general practitioners (GPs) in all participating countries and gynecologists in France from 2015 to 2018. The study population included women receiving ≥1 prescription of 91 day COC LNG during the study period. Prescribing patterns of 91 day COC LNG were examined including: (1) treatment duration; (2) indication; (3) use of combined oral contraceptive (COC) before 91 day COC LNG initiation; and (4) switch from and to combined hormonal contraceptives (CHCs) or other contraceptives., Results: Totals of 235, 220, 207 and 659 women using 91 day COC LNG were identified in French, Italian and Belgian GP, and French gynecologist databases, respectively. Across databases, 46-76% of women were prescribed a single 91 day COC LNG prescription and median treatment duration ranged from 3 to 6 months. The most common indication was contraception (42-81%), followed by menstrual migraines (2-14%). Use of COC during the 6 months prior to 91 day COC LNG initiation was 14% across GP databases, but was lower (8%) in the French gynecologist database. The frequency of switching from 91 day COC LNG to CHCs or other contraceptives was generally low (5-12%), with the highest proportion being among patients of French gynecologists., Conclusions: Findings indicate that 91 day COC LNG was prescribed for relatively short durations and predominantly as indicated for contraception. Most results were comparable across all participating countries. KEY POINTSFindings from this drug utilization study in European databases across general practitioners and French gynecologists confirmed that 91 day extended levonorgestrel-containing combined oral contraceptive (COC LNG ) was prescribed for relatively short durations (median 3-6 months); predominantly for the intended indication of contraception.Combined oral contraceptive use during the 6 months prior to 91 day COC LNG initiation, and switching from 91 day COC LNG to combined hormonal contraceptives or other contraceptives, were generally low (14% or less).These findings were mostly consistent across participating countries.

Outbreak of COVID-19 by SARS-CoV-2 infection caused severe acute respiratory syndrome that has been declared a public health emergency of international concern. To control infections, there is urgent need to develop an effective therapeutic strategy. COVID-19 viral spike glycoprotein and proteases play major role in viral entry and mediating virus replication and spread and thus can serve as potential antiviral drug target. Being highly specific, efficacious and safe, peptides hold their place in therapeutics. In present study, molecular docking of 21 pharmacologically active non ribosomal peptides (NRPs) from marine microbes with SARS-CoV-2 spike glycoprotein and papain such as protease was done. Dactinomycin, Tyrocidine A and Gramicidin S showed highest binding interaction with target proteins. The binding affinity of Dactinomycin and Gramicidin S docked with SARS-CoV-2 spike glycoprotein was - 12.4 kcal/mol and - 11.4 kcal/mol, respectively. This suggested their potential to destabilize SARS spike protein binding with human host ACE2 receptor and thus hindering viral entry to the cells. Binding affinity of Tyrocidine A and Gramicidin S with SARS-CoV-2 papain-like protease was - 13.1 kcal/mol and - 11.4 kcal/mol, respectively which might be inhibited COVID-19 by acting on the protease. Gramicidin S showed same binding affinity for both target proteins and thus expected to be most potent. Based on the binding energy score, it was suggested that these pharmacologically active NRPs are potential molecules to be tested against SARS-CoV-2 and used to develop effective antiviral drugs., Competing Interests: Conflict of interestAuthors Poonam Bansal, Raman Kumar, Jasbir Singh and Suman Dhanda declare that they have no conflict of interest., (© The Author(s), under exclusive licence to Springer Nature Singapore Pte Ltd. 2021.)

Hepatic encephalopathy (HE) is a debilitating neuropsychiatric complication associated with acute and chronic liver failure. It is characterized by diverse symptoms with variable severity that includes cognitive and motor deficits. The aim of the study is to assess metabolic alterations in the brain and liver using nuclear magnetic resonance (NMR) spectroscopy and subsequent multivariate analyses to characterize metabolic signatures associated with HE. HE was developed by bile duct ligation (BDL) that resulted in hepatic dysfunctions and cirrhosis as shown by liver function tests. Metabolic profiles from control and BDL rats indicated increased levels of lactate, branched-chain amino acids (BCAAs), glutamate, and choline in the liver, whereas levels of glucose, phenylalanine, and pyridoxine were decreased. In brain, the levels of lactate, acetate, succinate, citrate, and malate were increased, while glucose, creatine, isoleucine, leucine, and proline levels were decreased. Furthermore, neurotransmitters such as glutamate and GABA were increased, whereas choline and myo-inositol were decreased. The alterations in neurotransmitter levels resulted in cognitive and motor defects in BDL rats. A significant correlation was found among alterations in NAA/choline, choline/creatine, and NAA/creatine with behavioral deficits. Thus, the data suggests impairment in metabolic pathways such as the tricarboxylic acid (TCA) cycle, glycolysis, and ketogenesis in the liver and brain of animals with HE. The study highlights that metabolic signatures could be potential markers to monitor HE progression and to assess therapeutic interventions.

Introduction and Objective: COVID-19 is an ongoing, global public health crisis for which safe and effective treatments need to be identified. The benefit-risk balance for the use of lopinavir-ritonavir in COVID-19 needs to be monitored on an ongoing basis, therefore a systematic benefit-risk assessment was designed and conducted. A key objective of this study was to provide a platform for a dynamic systematic benefit-risk evaluation; although initially this evaluation is likely to contain limited information, it is required because of the urgent unmet public need. Importantly, it allows additional data to be incorporated as they become available, and re-evaluation of the benefit-risk profile., Methods: A systematic benefit-risk assessment was conducted using the Benefit-Risk Action Team (BRAT) framework. The exposure of interest was lopinavir-ritonavir treatment in severe COVID-19 compared to standard of care, placebo or other treatments. A literature search was conducted in PubMed and Embase to identify peer-reviewed papers reporting clinical outcomes. Two clinicians constructed a value tree and ranked key benefits and risks in order of considered clinical importance., Results: We screened 143 papers from PubMed and 264 papers from Embase for lopinavir-ritonavir; seven papers were included in the final benefit-risk assessment. In comparison to standard of care, data for several key benefits and risks were identified for lopinavir-ritonavir. Time to clinical improvement was not significantly different for lopinavir-ritonavir in comparison to standard of care (hazard ratio 1.31, 95% confidence interval 0.95-1.80). From one study, there were fewer cases of acute respiratory distress syndrome with lopinavir-ritonavir compared with standard of care (13% vs 27%). There also appeared to be fewer serious adverse events with lopinavir-ritonavir (20%) vs standard of care (32%). Limited data were available for comparison of lopinavir-ritonavir to other treatments., Conclusions: Based on currently available data, there was no clear benefit for the use of lopinavir-ritonavir compared to standard of care in severe COVID-19. Risk data suggested a possible decrease in serious adverse events. There was a reduction in acute respiratory distress syndrome with lopinavir-ritonavir in one study. Overall, the benefit-risk profile for lopinavir-ritonavir in severe COVID-19 cannot be considered positive until further efficacy and effectiveness data become available.

Introduction: There is a need to identify effective, safe treatments for COVID-19 (coronavirus disease) rapidly, given the current, ongoing pandemic. A systematic benefit-risk assessment was designed and conducted to examine the benefit-risk profile of remdesivir in COVID-19 patients compared with standard of care, placebo or other treatments. A key objective of this study was to provide a platform for a dynamic systematic benefit-risk evaluation, which starts with inevitably limited information (to meet the urgent unmet public health need worldwide), then update the benefit-risk evaluation as more data become available., Methods: The Benefit-Risk Action Team (BRAT) framework was used to assess the overall benefit-risk of the use of remdesivir as a treatment for COVID-19 compared with standard of care, placebo or other treatments. We searched PubMed, Google Scholar and government agency websites to identify literature reporting clinical outcomes in patients taking remdesivir for COVID-19. A value tree was constructed and key benefits and risks were ranked by two clinicians in order of considered importance., Results: Using the BRAT method, several key benefits and risks for use of remdesivir in COVID-19 compared with placebo have been identified. In one trial, the benefit of time to clinical improvement was not statistically significant (21 vs 23 days, HR 1.23, 95% CI 0.87-1.75), although the study was underpowered. In another trial, a shorter time to recovery in patients treated with remdesivir was observed (11 vs 15 days), with non-significant reduced mortality risk (8% vs 12%). Risk data were only available from one trial. This trial reported fewer serious adverse events in patients taking remdesivir (18%) compared with the placebo group (26%); however, more patients in the remdesivir group discontinued treatment as a result of an adverse event compared with those patients receiving placebo (12% vs 5%)., Conclusions: Preliminary clinical trial results suggest that there may be a favourable benefit-risk profile for remdesivir compared with placebo in severe COVID-19 infection and further data on benefits would strengthen this evaluation. There is limited safety data for remdesivir, which should be obtained in further studies. The current framework summarises the key anticipated benefits and risks for which further data are needed. Ongoing clinical trial data can be incorporated into the framework when available to provide an updated benefit-risk assessment.

Motivation: Despite conservation in general architecture of promoters and protein-DNA interaction interface of RNA polymerases among various prokaryotes, identification of promoter regions in the whole genome sequences remains a daunting challenge. The available tools for promoter prediction do not seem to address the problem satisfactorily, apparently because the biochemical nature of promoter signals is yet to be understood fully. Using 28 structural and 3 energetic parameters, we found that prokaryotic promoter regions have a unique structural and energy state, quite distinct from that of coding regions and the information for this signature state is in-built in their sequences. We developed a novel promoter prediction tool from these 31 parameters using various statistical techniques., Results: Here, we introduce SEProm, a novel tool that is developed by studying and utilizing the in-built structural and energy information of DNA sequences, which is applicable to all prokaryotes including archaea. Compared to five most recent, diverged and current best available tools, SEProm performs much better, predicting promoters with an 'F-value' of 82.04 and 'Precision' of 81.08. The next best 'F-value' was obtained with PromPredict (72.14) followed by BProm (68.37). On the basis of 'Precision' value, the next best 'Precision' was observed for Pepper (75.39) followed by PromPredict (72.01). SEProm maintained the lead even when comparison was done on two test organisms (not involved in training for SEProm)., Availability and Implementation: The software is freely available with easy to follow instructions (www.scfbio-iitd.res.in/software/TSS&#95;Predict.jsp)., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Aggregation and adhesion capability and survival efficacy of candidate probiotic strain Pediococcus acidilactici NCDC 252 under simulated gastric, intestinal and vaginal conditions was studied. The strain exhibited strong autoaggregation phenotype and coaggregation with other Lactic acid bacteria (LAB) and E. coli. The adhesion studies of NCDC 252 to pig's intestinal epithelial cells showed its adhesive ability. Aggregation and adhesiveness were related through cell surface proteins as removal/extraction of surface proteins resulted in altered aggregation and no adhesiveness. Cell surface proteins were analysed by SDS-PAGE and also in silico analysed from its genome. SDS-PAGE analysis of cell surface proteins of NCDC 252 revealed two potential proteins of approximately 74.3 and 53.6 kDa to be involved in host-probiotic interaction. Removal of cell surface proteins by LiCl-treatment (5 mol l -1 ) resulted in loss of aggregation and adhesiveness. Further survival of NCDC 252 under simulated gastrointestinal and vaginal conditions in terms of high viable counts confirmed its efficacy for its survival under gut and urogenital conditions. These observations suggest that it can be used further in functional foods, nutraceuticals and in combating urogenital infections. As NCDC 252 was able to survive in intestinal conditions, interaction of its cell surface proteins with intestinal mucins was studied in silico by docking. Highest affinity of adhesion was observed for MUC3B. In conclucion, NCDC 252, exhibited aggregation phenotype and adhesion capability. Survivability of NCDC 252 under simulated conditions and its interaction with human mucins confirms its efficacy to be used as probiotic.

An annotated checklist of the 271 strict-endemic taxa (235 species) and 387 near-endemic taxa (337 species) of vascular plants in Mozambique is provided. Together, these taxa constitute c. 9.3% of the total currently known flora of Mozambique and include five strict-endemic genera ( Baptorhachis , Emicocarpus , Gyrodoma , Icuria and Micklethwaitia ) and two near-endemic genera ( Triceratella and Oligophyton ). The mean year of first publication of these taxa is 1959, with a marked increase in description noted following the onset of the two major regional floristic programmes, the "Flora of Tropical East Africa" and "Flora Zambesiaca", and an associated increase in botanical collecting effort. New taxa from Mozambique continue to be described at a significant rate, with 20 novelties described in 2018. Important plant families for endemic and near-endemic taxa include Fabaceae, Rubiaceae and Euphorbiaceae s.s. There is a high congruence between species-rich plant families and endemism with the notable exceptions of the Poaceae, which is the second-most species rich plant family, but outside of the top ten families in terms of endemism, and the Euphorbiaceae, which is the seventh-most species rich plant family, but third in terms of endemism. A wide range of life-forms are represented in the endemic and near-endemic flora, with 49% being herbaceous or having herbaceous forms and 55% being woody or having woody forms. Manica Province is by far the richest locality for near-endemic taxa, highlighting the importance of the cross-border Chimanimani-Nyanga (Manica) Highlands shared with Zimbabwe. A total of 69% of taxa can be assigned to one of four cross-border Centres of Endemism: the Rovuma Centre, the Maputaland Centre sensu lato, and the two mountain blocks, Chimanimani-Nyanga and Mulanje-Namuli-Ribaue. Approximately 50% of taxa have been assessed for their extinction risk and, of these, just over half are globally threatened (57% for strict-endemics), with a further 10% (17% for strict-endemics) currently considered to be Data Deficient, highlighting the urgent need for targeted conservation of Mozambique's unique flora. This dataset will be a key resource for ongoing efforts to identify "Important Plant Areas - IPAs" in Mozambique, and to promote the conservation and sustainable management of these critical sites and species, thus enabling Mozambique to meet its commitments under the Convention on Biological Diversity (CBD)., (Iain Darbyshire, Jonathan Timberlake, Jo Osborne, Saba Rokni, Hermenegildo Matimele, Clayton Langa, Castigo Datizua, Camila de Sousa, Tereza Alves, Alice Massingue, Jeneen Hadj-Hammou, Sonia Dhanda, Toral Shah, Bart Wursten.)

Purpose: Retinal hemangioblastomas (RHs) are characteristic of von Hippel-Lindau (VHL) disease. Early diagnosis of retinal lesions may aid in systemic diagnosis. Early identification of VHL is life-saving and also prevents vision loss. Fundus fluorescein angiography (FFA) is a useful tool in the diagnosis and management of RHs. The aim of this study is to report FFA features of RH using ultra-widefield (UWF) imaging., Methods: A retrospective cross-sectional study of consecutive patients of RH who underwent UWF FFA at a tertiary eye care center. Images were analyzed and assessed by authors. The main outcome measures were (a) the number and size of RH in each eye and (b) vascular characteristics of the retina. UWF-FFA characteristics in each eye were tabulated. The number of clock hours involved by these characteristics and their correlation with the number and size of RH were analyzed., Results: The study evaluated 24 eyes of 13 patients. The mean age was 28.4 years. The median number of RHs in an eye was 3.5 (range 1-16), and the size of RHs varied from 0.1 to 4 disc diameters. Novel UWF-FFA findings noted in this study were the presence of abnormal capillary network in 22 of 24 eyes (91.7%), capillary leakage in 15 of 24 eyes (62.5%), and capillary telangiectasia in 7 of 24 eyes (29.2%). In addition, feeder arterioles and venules showed bulbous projections in 8 of 24 eyes (33.3%)., Conclusion: The UWF-FFA characteristics of RH, which have not been described before, were identified. These add to our understanding of the pathogenesis of the disease and may pave the way for future therapeutic targets., Competing Interests: None

Pediococcus acidilactici NCDC 252 is a facultative anaerobe of dairy origin that possessed all studied in vitro probiotic attributes and several useful enzyme activities. Its whole genome was sequenced and analysed for its evolutionary relationship with other lactic acid bacteria (LAB). This is a novel sequence and first report of genome sequence of P. acidilactici of dairy origin. Its genome is relatively larger than other studied genomes of P. acidilactici and is comprised of 40 scaffolds that totals to 3,243,337 bases and 44.5% GC content. A total of 3054 coding sequences (CDS) were identified by RAST and DIAMOND servers. The genome also encoded different enzyme activities required for utilization of various carbohydrates. This was also confirmed by carbohydrate utilization studies. The genome also encoded genes for probiotics properties. The phylogenetic analysis of P. acidilactici NCDC 252 genome was done using Maximum Parsimony and Maximum Likelihood methods to study its evolution and relatedness to other LABs based upon their 16S rDNA sequences. The strain exhibited highest resemblance to Lactobacillus plantarum WCFS1 and is also much close to P. acidilactici based on similarity of ribosomal protein. The strain seems to have acquired some genes for its adaptation in dairy/environmental niche. This genome sequence is novel with genome more similar to L. plantarum and biochemical and phenotypic characteristics of P. acidilactici.

The presence of minimal residual disease (MRD) is one of the strong predictors of disease outcome in various hematological malignancies including B-ALL and T-ALL, independent of pre-therapeutic risk factors. There is scant Indian data on MRD by flowcytometry in T-ALL including gating strategies, clinical correlation etc. The primary aim of this retrospective observational study was to define the clinico-hematologic characteristics and prognostic significance of patients with ETP/near-ETP versus non-ETP immunophenotype, especially in terms of minimal residual disease at different time points as well as event-free survival (1 year). Baseline hematologic characteristics along with post-induction (Day-35) and post-consolidation (Day-78) MRD in bone marrow samples from newly diagnosed T-ALL patients were studied. 14.3% patients had ETP-ALL immunophenotype, 11.4% were near-ETP ALL patients and the remaining 74.5% were of non-ETP subtype. The ETP/near ETP patients was significantly associated with higher risk of MRD positivity ( > 0.01%) at the end of induction in comparison to the non-ETP patients ( p  = 0.033). Also, these patients showed a trend towards proclivity to anemia ( p  = 0.06) and higher rates of induction failure ( p  = 0.07) However, no difference was observed between the two subgroups in terms of age, high TLC, thrombocytopenia, adverse cytogenetics, steroid responsiveness on Day + 8 of induction, MRD-positivity > 0.01% at the end of consolidation and EFS-1 year. Through this preliminary study, it can be stated clearly that ETP status is associated with MRD > 0.01% post-induction but has no significant impact on long-term survival of these patients., Competing Interests: Conflict of interestAll authors declare that they have no conflict of interest., (© Indian Society of Hematology and Blood Transfusion 2019.)

Competing Interests: None

Competing Interests: There are no conflicts of interest

Pediococcus acidilactici is a probiotic lactic acid bacteria possessing studied in-vitro probiotic properties. Study of membrane proteins is crucial in developing technological and health applications of probiotic bacteria. Genome analysis of Pediococcus acidilactici revealed about more than 60 proteases/peptidases which need characterization. Dipeptidyl peptidase-III (DPP-III) is studied for first time in prokaryotes and it is a membrane protein in P. acidilactici that has been purified to apparent homogeneity. The enzyme was purified 81.66 fold with 36.75% yield. The specific activity of purified DPP-III was 202.67 U/mg. The protein moved as single band on native PAGE. The purity was also confirmed by in-situ gel assay. However SDS-PAGE analysis revealed it as high molecular weight heterotetramer with molecular weight of 108 kDa. The enzyme was maximally active at pH 8.5 and at 37 C. Purified DPP-III specifically hydrolyzed Arg-Arg-4-βNA with micromolar affinity (K m = 9.0 µM) and none of studied endopeptidase and monopeptidase substrate was hydrolyzed. Inhibition study revealed purified DPP-III to be a serine protease with involvement of metal ion at active site. The significance of this enzyme as membrane protein is yet to be studied.

The present study was designed to investigate the mechanisms involved in blood-brain barrier (BBB) permeability in bile duct ligation (BDL) model of chronic hepatic encephalopathy (HE). Four weeks after BDL surgery, a significant increase was observed in serum bilirubin levels. Masson trichrome staining revealed severe hepatic fibrosis in the BDL rats. 99m Tc-mebrofenin retention was increased in the liver of BDL rats suggesting impaired hepatobiliary transport. An increase in permeability to sodium fluorescein, Evans blue, and fluorescein isothiocyanate (FITC)-dextran along with increase in water and electrolyte content was observed in brain regions of BDL rats suggesting disrupted BBB. Increased brain water content can be attributed to increase in aquaporin-4 mRNA and protein expression in BDL rats. Matrix metalloproteinase-9 (MMP-9) mRNA and protein expression was increased in brain regions of BDL rats. Additionally, mRNA and protein expression of tissue inhibitor of matrix metalloproteinases (TIMPs) was also increased in different regions of brain. A significant decrease in mRNA expression and protein levels of tight junction proteins, viz., occludin, claudin-5, and zona occluden-1 (ZO-1) was observed in different brain regions of BDL rats. VCAM-1 mRNA and protein expression was also found to be significantly upregulated in different brain regions of BDL animals. The findings from the study suggest that increased BBB permeability in HE involves activation of MMP-9 and loss of tight junction proteins.

Chronic liver disease per se induces neuroinflammation that contributes to cognitive deficits in hepatic encephalopathy (HE). However, the processes by which pro-inflammatory molecules result in cognitive impairment still remains unclear. In the present study, a significant increase in the activity of liver function enzymes viz. alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) was observed along with increase in plasma ammonia levels after four weeks of bile duct ligation (BDL) in rats suggesting hepatocellular damage. A significant increase was observed in mRNA expression of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) in brain regions and liver of BDL rats. Concomitantly, IL-6, TNF-α and MCP-1 protein levels were also increased in brain regions, liver and serum of BDL rats suggesting the involvement of blood-brain-axis in inflammatory response. However, a significant decrease was observed in glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule-1 (Iba-1) expression at transcriptional and translation level in brain of BDL rats. Immunohistochemical and flowcytometric analysis revealed reduced number of GFAP-immunopositive astrocytes and Iba1-immunopositive microglia in the brain regions of BDL rats. Further, a significant decline was observed in cognitive functions in BDL rats assessed using Morris water maze and novel object recognition tests. Expression of pro and mature form of brain derived neurotrophic factor (BDNF) and its upstream transcription element showed significant reduction in brain of BDL rats. Taken together, the results of the present study suggest that systemic inflammation and reduced expression of BDNF and its upstream transcription factor plays a key role in cognitive decline in HE., (Copyright © 2018 Elsevier Inc. All rights reserved.)

β-galactosidase is a commercially important enzyme that was purified from probiotic Pediococcus acidilactici. The enzyme was extracted from cells using sonication and subsequently purified using ammonium sulphate fractionation and successive chromatographies on Sephadex G-100 and Q-Sepharose. The enzyme was purified 3.06-fold up to electrophoretic homogeneity with specific activity of 0.883 U/mg and yield of 28.26%. Molecular mass of β-galactosidase as estimated by SDS-PAGE and MALDI-TOF was 39.07 kDa. The enzyme is a heterodimer with subunit mass of 15.55 and 19.58 kDa. The purified enzyme was optimally active at pH 6.0 and stable in a pH range of 5.8-7.0 with more than 97% activity. Purified β-galactosidase was optimally active at 50 °C. Kinetic parameters K m and V max for purified enzyme were 400 µM and 1.22 × 10 -1 U respectively. Its inactivation by PMSF confirmed the presence of serine at the active site. The metal ions had different effects on enzyme. Ca 2+ , Mg 2+ and Mn 2+ slightly activated the enzyme whereas NH 4 + , Co 2+ and Fe 3+ slightly decreased the enzyme activity. Thermodynamic parameters were calculated that suggested that β-galactosidase is less stable at higher temperature (60 °C). Purified enzyme effectively hydrolysed milk lactose with lactose hydrolysing rate of 0.047 min -1 and t 1/2 of 14.74 min. This is better than other studied β-galactosidases. Both sonicated Pediococcus acidilactici cells and purified β-galactosidase synthesized galactooligosaccharides (GOSs) as studied by TLC at 30% and 50% of lactose concentration at 47.5 °C. These findings indicate the use of β-galactosidase from probiotic bacteria for producing delactosed milk for lactose intolerant population and prebiotic synthesis. pH and temperature optima and its activation by Ca 2+ shows that it is suitable for milk processing., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Membrane bound proline iminopeptidase (PIP) from lactic acid bacteria (LAB) L. plantarum was extracted and purified using CM-sephadex, Sephadex G-100 and Q-sepharose column chromatography. PIP was purified with purification fold 7.13 and 33.5% yield. SDS-PAGE and MALDI-TOF revealed it as homodimer with molecular weight of 37.9 kDa and subunit of mass 18.9 kDa. Purified enzyme exhibited maximum activity at 45 °C and pH 7.0. K m and V max of purified PIP were 65 μM and 25.9 nm/min/ml respectively. Inhibition by PMSF confirmed it a serine protease. Metal ions and EDTA showed no effect on enzyme activity. The enzyme mainly hydrolysed Pro-4mβNA. The effectiveness of enzyme in purified form, membrane bound form and in combination with other enzymes to degrade collagen resulting in pharmaceutically significant collagen hydrolysates and in meat tenderization marks its industrial importance. There are very few PIPs are characterized from LAB, and therefore this study is industrially significant and brings some new knowledge into this area., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Perturbations in the cerebral energy metabolism are anticipated to be an important factor by which ammonia may exert its toxic effects on the central nervous system. The present study was designed to investigate the role of impaired mitochondrial functions and cerebral energy metabolism in the development hepatic encephalopathy (HE) induced by of bile duct ligation (BDL). After four weeks of BDL, a significant increase in hepatic hydroxyproline and collagen content was observed which confirmed biliary fibrosis. Brain regions viz. cortex, hippocampus, striatum and cerebellum of BDL rats had impaired activity of mitochondrial respiratory chain enzymes. This was accompanied by increase in mitochondrial reactive oxygen species (ROS), malondialdehyde (MDA) and protein carbonyl levels in the brain. Mitochondrial redox ratio was significantly reduced in the brain of BDL rats. In addition, mitochondria from brain of BDL rats were depolarized and swollen compared to the sham controls. Ultrastructure analysis of mitochondria from cortex and hippocampus of BDL animals revealed aberrant cristae, ruptured membranes and non-dense matrix. Further, a significant decrease was observed in creatine kinase activity, glucose uptake and CO 2 production in the brain regions of BDL rats. ATP/ADP ratio, a critical parameter of cellular energy status, was also significantly reduced in brain regions of rats with HE. Overall, the findings clearly demonstrate that BDL induced HE involves mitochondrial respiratory chain dysfunctions, mitochondrial depolarization and swelling that accentuates oxidative stress which in turn leads to compromise in cerebral energy metabolism thereby contributing to the pathophysiology of chronic HE.

The aim of the present study was to evaluate the effect of N-acetyl-l-cysteine (NAC) on bile duct ligation (BDL) induced oxidative stress in kidneys. Male Wistar rats were randomly segregated into four groups; sham control (SC), SC + NAC, BDL and BDL + NAC group. Liver damage was induced following BDL and renal injury was assessed by kidney function tests along with lipid peroxidation, nitrite levels, thiols and antioxidant enzymes. Three weeks after BDL, rats developed renal dysfunction in terms of elevated serum creatinine levels. BDL animals exhibited an increase in lipid peroxidation, reduction in thiols and redox ratio in liver and kidney tissue along with altered antioxidant enzymes in kidneys. BDL animals that were orally administered NAC at a daily dose 100 mg/kg for duration of two weeks, showed significant reduction in serum creatinine levels. NAC was effective in lowering lipid peroxidation and was able to restore thiol levels along with GSH/GSSG ratio in both liver and kidneys along with the activity of antioxidant enzymes in the kidneys of BDL animals. The results clearly demonstrate the efficacy of NAC in attenuating oxidative stress in kidneys, suggesting a therapeutic role for NAC in individuals with renal dysfunction following BDL.

Ovarian torsion is a surgical emergency characterized by a partial or complete rotation of the ovarian vascular pedicle, causing obstruction to venous outflow followed by arterial inflow. Clinically, ovarian torsion frequently mimics other causes of acute pelvic pain such as appendicitis, diverticulitis, renal colic etc. Ultrasonography is the first-line imaging modality of choice for evaluation of ovarian torsion. MRI is useful as a problem-solving tool in patients with equivocal or non-diagnostic ultrasonography studies. CT is ordinarily not utilized in a young female with suspected ovarian torsion due to the radiation dose. However, the significant expansion in use of CT imaging in emergency departments for female patients presenting with acute abdominal pain has increased the likelihood that ovarian torsion may be first seen on CT. In addition, a non-specific clinical presentation may lead to an initial imaging with CT rather than ultrasonography. Ultrasound features of the ovarian torsion are well known and sufficiently described across literature as compared with the CT scan findings. In view of the increasing usage of CT as the modality of choice in emergency settings, it is imperative for the radiologist to familiarize with the CT features of ovarian torsion. An early correct diagnosis by the radiologist in clinically unsuspected cases, facilitating a prompt surgery to restore the ovarian blood flow can prevent permanent irreversible damage. There is limited published data available on the CT features of ovarian torsion. This pictorial essay illustrates CT findings with histological correlation of surgically proven ovarian torsion in our institution. These patients were primarily investigated with CT scan for acute pelvic pain ascribed to non-gynaecological causes such as bowel or urinary tract lesions.

Probiotics are living organisms that confer health benefits when administered in adequate amounts. Probiotics are continuously being explored for their different health beneficiary activities. Anticancer activity is one of the most important benefits both from a preventive and therapeutic point of view. Though not many studies have been conducted to date in this area, a number suggest using laboratory animal models and different cell lines that there may be a mechanistic basis for the anticancer effects of probiotics and require more scientific justification and clinical trials. Most studies of probiotics are conducted for colon cancer associated with inflammatory bowel disease. Studies are also being extended to other types of cancer in different cell lines. This review summarizes studied probiotics considered for treatment of colon cancer and some other cancers (in cancer cell lines) and also proposed mechanism how probiotics are inhibiting cancer growth along with some challenges and future perspectives.

Dipeptidylpeptidase-II (DPP-II, E.C. 3.4.14.2), an exopeptidase was purified 15.4 fold with specific activity and yield of 15.4U/mg/mL and 14.68% respectively by a simple two step procedure from a probiotic Pediococcus acidilactici. DPP-II is 38.7KDa homodimeric serine peptidase with involvement of His and subunit mass of 18.9KDa. The enzyme exhibited optimal activity at pH 7.0 and 37°C with activation energy of 24.97kJ/mol. The enzyme retained more than 90% activity upto 50°C thus adding industrial importance. DPP-II hydrolysed Lys-Ala-4mβNA with K M of 50μM and V max of 30.8nmol/mL/min. In-silico characterization studies of DPP-II on the basis of peptide fragments obtained by MALDI-TOF revealed an evolutionary relationship between DPP-II of prokaryotes and phosphate binding proteins. Secondary and three-dimensional structure of enzyme was also deduced by in-silico approach. Functional studies of DPP-II by TLC and HPLC-analysis of collagen degraded products revealed that enzyme action released free amino acids and other metabolites. Microscopic and SDS-PAGE analysis of enzyme treated analysis of chicken's chest muscle (meat) hydrolysis revealed change and hydrolysis of myofibrils. This may affect the flavor and texture of meat thereby suggesting its role in meat tenderization. Being a protein of LAB (Lactic acid bacteria), it is also expected to be safe., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Competing Interests: There are no conflicts of interest.

In vitro environments like heat stress usually increase the production of reactive oxygen species in bubaline oocytes which have been implicated as one of the major causes for reduced developmental competence. Oocytes during meiotic maturation are sensitive to oxidative stress, and heat stress accelerates cellular metabolism, resulting in the higher production of free radicals. Therefore, the aim of present work was to assess the impact of heat stress during meiotic maturation on bubaline cumulus-oocyte complexes (COC), denuded oocytes (DO), and cumulus cell mass in terms of their oxidative status. Accordingly, for control group, COC were matured at 38.5 °C for complete 24 h of meiotic maturation and heat stress of 40.5 and 41.5 °C was applied to COC during the first 12 h of maturation and then moved to 38.5 °C for rest of the 12 h. In another group, COC after maturation were denuded from the surrounding cumulus cells by manual pipetting. Results indicated that the production of reactive oxygen species (ROS), lipid peroxides, and nitric oxide (NO) was significantly (P < 0.05) higher in the oocytes subjected to heat stress (40.5 and 41.5 °C) during meiotic maturation compared to the oocytes matured under standard in vitro culture conditions (38.5 °C). Also, the antioxidant enzymatic activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase were significantly (P < 0.05) increased in all the treatment groups compared to the control group. Therefore, the present study clearly establishes that heat stress ensues oxidative stress in bubaline oocytes which triggers the induction of antioxidant enzymatic defense system for scavenging the ROS.