Assessment of threatened status, phytochemical composition and biological properties of three Aconitum species from Kashmir Himalaya - India

Genus Aconitum (Ranunculaceae) is represented by 6–8 species from Kashmir Himalaya.  Traditionally Aconitum species are used to treat a wide array of diseases, but their ethnopharmacological validation and phytochemistry are hitherto unreported from Kashmir Himalaya. The present study was undertaken to bring insights into the traditional use and distribution pattern of three Aconitum species from the region. An ethnobotany-directed approach was employed to study the conservation status of three Aconitum species. Their phytochemical profiles and biological properties were screened under in vitro conditions. Folin–ciocalteu and Aluminium chloride assays were employed to measure their total phenolic and total flavonoid contents, respectively. Plant extracts were evaluated for antioxidant, antimicrobial and anti-inflammatory activities. Three Aconitum species, viz. Aconitum heterophyllum Wall. ex Royle, Aconitum violaceum Jacquem. ex Stapf and Aconitum chasmanthum Stapf. ex Holmes showed dwindling conservation status in Kashmir Himalaya. Aconitum extracts showed significant variations in total phenolic and flavonoid contents. Antioxidant activity of Aconitum chasmanthum methanolic extract was studied to be comparatively higher (80.115%). Aconitum chasmanthum DCM & methanolic extracts showed a good MIC value of 0.125 mg/ml against Candida albicans and Streptococus pyogenes, respectively. The percent inhibition of NLRP inflammasome was found significant in Aconitum violaceum ethyl acetate extract (74.61%). The present study revealed that Aconitum species are constantly declining at least in investigated habitats of Kashmir Himalaya and hence need strategic conservation planning. The results also emphasized the utility of Aconitum species as an antioxidant, antimicrobial and anti-inflammatory agent that could be used to manage various health problems

Isolation, identification and bioactive potential of bacterial endophytes from Coleus

Coleus (Lamiaceae) is a large and widespread genus comprising of species with diverse ethnobotanical uses. In the present study, bacterial endophytes were isolated from Coleus forskohlii and Coleus aromaticus. Endophytes are the microorganisms which reside within the plants without showing any harmful effect on its host. Diverse types of endophytes live symbiotically within almost all plants and in turn help the plant in a number of ways such as imparting resistance against biotic and abiotic stresses, producing compounds involved in attraction of pollinators, inducing the plant defense mechanisms, etc. The bacterial endophytes isolated in this study, were characterized by microscopic examination (using gram staining) and molecularly identified by sequencing the 16S rRNA. Extracts were prepared from endophytic biomass using solvents of different polarities (methanol, ethyl acetate and butanol) and were screened for their bioactive potential (in vitro cytotoxicity anti-microbial, and anti-oxidant activity). Scale-up of endophytes showing promising results is under process, which will help in isolation of pure compounds

Molecular Typing of Methicillin Resistant Staphylococcus aureus using coa Gene Polymerase Chain Reaction-Restriction Fragment Length Polymorphism: A Cross-sectional Study

Introduction: The need for fast, precise diagnostic tests to identify active tuberculosis is essential, mainly in endemic nations such as India. An automated real-time Polymerase Chain Reaction (PCR) method for pulmonary tuberculosis (TB) detection known as the Cartridge Based Nucleic Acid Amplification Test (CBNAAT) or GeneXpert assay shows great promise as a complement to the TrueNat and conventional sputum microscopy techniques. Aim: To compare sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV) of CBNAAT with TrueNat and smear microscopy in the detection of Mycobacterium tuberculosis. Materials and Methods: A cross-sectional comparative study on 175 patients with suspected pulmonary TB was conducted from June 2021 to November 2021 in a tertiary care hospital at Government Medical College, Doda District of Jammu and Kashmir, India. The sensitivity, specificity, PPV, NPV, and diagnostic accuracy for the diagnosis of tuberculosis were calculated for Acid Fast Bacilli (AFB) smear microscopy, TrueNat, and the GeneXpert and compared with each other. Statistical analysis of the data was conducted with Statistical Package for the Social Science (SPSS) version 20.0. Results: Out of the total 175, 168 (96%) patients were TB positive by CBNAAT, 162 (92.6%) by TrueNat, and 148 (84.6%) as per smear microscopy. Sensitivity, specificity, PPV, NPV, and accuracy of Ziehl-Neelsen (ZN) stain in the detection of pulmonary TB in sputum samples were 86.31%, 57.14%, 97.97%, 14.81%, and 85.14%, respectively. Whereas in the case of the TrueNat technique sensitivity, specificity, PPV, NPV, and accuracy were 94.05%, 42.86%, 97.53%, 23.08%, and 92.00%, respectively. In the case of CBNAAT sensitivity, specificity, PPV, NPV, and accuracy in the detection of pulmonary TB in sputum samples were 97.02%, 28.57%, 97.02%, 28.57%, and 94.29%, respectively. Conclusion: In respiratory samples, CBNAAT is more sensitive than ZN smear microscopy and TrueNat. Positive CBNAAT, but TrueNat and AFB microscopy negative results should be read cautiously and be well correlated with the clinical and treatment history of the patien

The United States COVID-19 Forecast Hub dataset

Academic researchers, government agencies, industry groups, and individuals have produced forecasts at an unprecedented scale during the COVID-19 pandemic. To leverage these forecasts, the United States Centers for Disease Control and Prevention (CDC) partnered with an academic research lab at the University of Massachusetts Amherst to create the US COVID-19 Forecast Hub. Launched in April 2020, the Forecast Hub is a dataset with point and probabilistic forecasts of incident cases, incident hospitalizations, incident deaths, and cumulative deaths due to COVID-19 at county, state, and national, levels in the United States. Included forecasts represent a variety of modeling approaches, data sources, and assumptions regarding the spread of COVID-19. The goal of this dataset is to establish a standardized and comparable set of short-term forecasts from modeling teams. These data can be used to develop ensemble models, communicate forecasts to the public, create visualizations, compare models, and inform policies regarding COVID-19 mitigation. These open-source data are available via download from GitHub, through an online API, and through R packages

Breast cancer management pathways during the COVID-19 pandemic: outcomes from the UK ‘Alert Level 4’ phase of the B-MaP-C study

Abstract: Background: The B-MaP-C study aimed to determine alterations to breast cancer (BC) management during the peak transmission period of the UK COVID-19 pandemic and the potential impact of these treatment decisions. Methods: This was a national cohort study of patients with early BC undergoing multidisciplinary team (MDT)-guided treatment recommendations during the pandemic, designated ‘standard’ or ‘COVID-altered’, in the preoperative, operative and post-operative setting. Findings: Of 3776 patients (from 64 UK units) in the study, 2246 (59%) had ‘COVID-altered’ management. ‘Bridging’ endocrine therapy was used (n = 951) where theatre capacity was reduced. There was increasing access to COVID-19 low-risk theatres during the study period (59%). In line with national guidance, immediate breast reconstruction was avoided (n = 299). Where adjuvant chemotherapy was omitted (n = 81), the median benefit was only 3% (IQR 2–9%) using ‘NHS Predict’. There was the rapid adoption of new evidence-based hypofractionated radiotherapy (n = 781, from 46 units). Only 14 patients (1%) tested positive for SARS-CoV-2 during their treatment journey. Conclusions: The majority of ‘COVID-altered’ management decisions were largely in line with pre-COVID evidence-based guidelines, implying that breast cancer survival outcomes are unlikely to be negatively impacted by the pandemic. However, in this study, the potential impact of delays to BC presentation or diagnosis remains unknown

Exogenously Applied Rohitukine Inhibits Photosynthetic Processes, Growth and Induces Antioxidant Defense System in Arabidopsis thaliana

The secondary metabolite rohitukine has been reported in only a few plant species, including Schumanniophyton magnificum, S. problematicum, Amoora rohituka, Dysoxylum acutangulum and D. gotadhora. It has several biological activities, such as anticancer, anti-inflammatory, antiadipogenic, immunomodulatory, gastroprotective, anti-implantation, antidyslipidemic, anti-arthritic and anti-fertility properties. However, the ecological and physiological roles of rohitukine in parent plants have yet to be explored. Here for the first time, we tried to decipher the physiological effect of rohitukine isolated from D. gotadhora on the model system Arabidopsis thaliana. Application of 0.25 mM and 0.5 mM rohitukine concentrations moderately affected the growth of A. thaliana, whereas a remarkable decrease in growth and the alteration of various morphological, physiological and biochemical mechanisms were observed in plants that received 1.0 mM of rohitukine as compared to the untreated control. A. thaliana showed considerable dose-dependent decreases in leaf area, fresh weight and dry weight when sprayed with 0.25 mM, 0.5 mM and 1.0 mM of rohitukine. Rohitukine exposure resulted in the disruption of photosynthesis, photosystem II (PSII) activity and degradation of chlorophyll content in A. thaliana. It also triggered oxidative stress in visualized tissues through antioxidant enzyme activity and the expression levels of key genes involved in the antioxidant system, such as superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX). Rohitukine-induced changes in levels of metabolites (amino acids, sugars, organic acids, etc.) were also assessed. In light of these results, we discuss (i) the likely ecological importance of rohitukine in parent plants as well as (ii) the comparison of responses to rohitukine treatment in plants and mammals

Mannitol Stress Directs Flavonoid Metabolism toward Synthesis of Flavones via Differential Regulation of Two Cytochrome P450 Monooxygenases in Coleus forskohlii

Cytochrome P450 monooxygenases (CYP450s) are known to play important roles in biosynthesis of all secondary metabolites, including flavonoids. Despite this, few CYP450s have been functionally characterized in model plants and roles of fewer CYP450s are known in non-model, medicinal, and aromatic plants. Our study in Coleus forskohlii indicates that flavone synthase (CYP93B) and flavonoid 3′ monooxygenase (CYP706C) are key enzymes positioned at a metabolic junction, to execute the biosynthesis of different sub-classes of flavonoids (flavones, flavonol, anthocynanin, isoflavones etc.) from a common precursor. Such branch points are favored targets for artificially modulating the metabolic flux toward specific metabolites, through genetic manipulation or use of elicitors that differentially impact the expression of branch point genes. Genkwanin, the only flavone reported from C. forskohlii, is known to possess anti-inflammatory activity. It is biosynthesized from the general flavonoid precursor: naringenin. Two differentially expressed cytochrome P450 genes (CfCYP93B, CfCYP706C), exhibiting maximum expression in leaf tissues, were isolated from C. forskohlii. Mannitol treatment resulted in increased expression of CfCYP93B and decrease in expression of CfCYP706C. Metabolite quantification data showed that genkwanin content increased and anthocyanin levels decreased in response to mannitol treatment. Alignment, phylogenetic analysis, modeling, and molecular docking analysis of protein sequences suggested that CfCYP93B may be involved in conversion of naringenin to flavones (possibly genkwanin via apigenin), while CfCYP706C may act on common precursors of flavonoid metabolism and channel the substrate toward production of flavonols or anthocynanins. Decrease in expression of CfCYP706C and increase in accumulation of genkwanin suggested that mannitol treatment may possibly lead to accumulation of genkwanin via suppression of a competitive branch of flavonoids in C. forskohlii

The effect of pre-etching with 37% orthophosphoric acid on the shear bond strength of orthodontic brackets bonded using self-etching primer-adhesive system

Objectives: The purpose of this study was to compare the effect of pre-etching with 37% orthophosphoric acid on the shear bond strength (SBS) of orthodontic brackets bonded using self-etching primer (SEP). Materials and Methods: Two hundred freshly extracted human premolars were divided randomly into two groups based on pre-etching with 37% orthophosphoric acid: Group 1 (control, n = 50) without pre-etching and Group 2 (experimental, n = 150) with pre-etching. Group 2 was further divided into three sub groups 2a, 2b, and 2c with 50 teeth each and was first pre-etched with 37% orthophosphoric acid for 10, 30, and 60 s, respectively. Brackets were bonded on the teeth in both the groups using SEP and light cure adhesive. The SBS were determined using universal testing machine. The comparison of SBS was done using one-way analysis of variance and Tukey′s honest significant difference test. Results: The SBS of experimental Group 2 was significantly higher than the control Group 1 (p < 0.0001). Further, the SBS of Group 2a was highest, followed by Group 2b and 2c (p < 0.0001). Conclusion: It was found that pre-etching for 10 s prior to application of SEP-adhesive system increases the SBS of orthodontic brackets