Secondary infections in COVID-19: Antemortem and postmortem culture study.

Background: Secondary bacterial infections during COVID-19 hospitalization have been reported in about 6-15% of patients. Aims: To study the secondary bacterial infections that affected the COVID-19 patients during their hospitalisation and to unearth the bacteriological profile of samples obtained after their demise. Settings and Design: This prospective study was carried out at a COVID-19 dedicated, apex tertiary care centre in North India from July 2020 to April 2021. Methods and Materials: Samples of 268 patients were considered for the study. Nasopharyngeal swab specimen, blood, and tissue (lung) were collected from the deceased body as early as possible and processed. Statistical Analysis: Statistical analyses were performed using STATA version 11.1 (Stata Corp., College Station, TX, USA). Results: A total of 170 samples were received from patients before their death, which included blood, urine, respiratory samples, pus, and cerebrospinal fluid. Forty-four pathogens were isolated, which consisted of Acinetobacter baumannii (43.1%), Klebsiella pneumoniae (36.3%), Escherichia coli (11.3%), and Pseudomonas aeruginosa (4.5%), Enterococcus faecium (4.5%). Two hundred fifty-eight samples were collected from the deceased bodies wherein the nasopharyngeal sample was highest, followed by tissue and blood. A total of 43 pathogens were isolated among them which included A. baumannii (44.1%), followed by K. pneumoniae (25.5%), E. coli (20.9%), P. aeruginosa (6.97%) and Enterobacter cloacae (2.3%). All these isolates were highly resistant to antimicrobials. Conclusions: In our study, bacterial profiles in antemortem and postmortem samples were found to be similar, suggesting that resistant pathogens may be the cause of mortality in COVID-19 infected hospitalised patients.

Evaluation of iso-quinoline alkaloids by RP-HPLC-PDA in different Berberis species collected from Western Himalayan Region.

The genus Berberis has high significance in Indian and other traditional systems of medicine due to the presence of iso-quinoline alkaloids. This study was conducted to record the metabolic variation in eight Berberis spp. collected from the Western Himalayan region. The RP-HPLC-PDA chromatogram separates the markers jatrorrhizine, palmatine and berberine at specific retention time, 11.45, 16.41 and 18.15 min respectively. The method was validated on linearity, precision and recovery indices as per International Conference on Harmonisation (ICH) guidelines. The maximum content of berberine, palmatine and jatrorrhizine was found in Berberis asiatica (1.74% ± 0.032, 1.63% ± 0.028 and 0.264% ± 0.012, respectively) on % dry weight basis. This study will help in phyto-marker-based identification and differentiation of various Berberis species, which can be used as an alternative to the official drug Daruharidra i.e. Berberis aristata.

Anti-rheumatic potential vis-à-vis aconitine and hypaconitine content analysis in different Aconitum spp. from Sikkim Himalayas (India).

Aconitum spp. are important medicinal plants mentioned in Ayurveda as Ativisa or Vatsanabha. The present study aims to evaluate anti-rheumatic potential in seven Aconitum species and correlation with aconitine and hypaconitine content. Anti-rheumatic potential was analyzed through in vitro xanthine oxidase inhibition, anti-inflammatory and ROS scavenging assays; and quantification of aconitine and hypaconitine with RP-HPLC method validated as per ICH guidelines. The findings reveal that A. palmatum possessed the most promising response (IC50 =12.68±0.15 µg/ml) followed by A. ferox (IC50 =12.912±1.87 µg/ml) for xanthin oxidase inhibition. We observed a wide variation in aconitine and hypaconitine content ranging from 0.018 %-1.37 % and 0.0051 %-0.077 % respectively on dry weight basis. Aconitine and hypaconitine showed moderate positive correlation (r=0.68 and 0.59 respectively) with anti-rheumatic potential. The study identifies potential alternative species of Aconitum that can help in sustainable availability of quality raw material.

Nutritional characteristics of Stereospermum chelonoides (L.f.) DC., an underutilized edible wild fruit of dietary interest.

Malnutrition and hunger is a serious global issue, however, wild fruits possess the potential of combatting it being rich in nutrients. Stereospermum chelonoides (L.f.) DC., commonly known as "Patala" in Ayurvedic text, is a large wild tree bearing edible, yet, underutilized fruits consumed by the locals in Western parts of India and neighboring countries. The present study focuses on the nutritional profile of S. chelonoides fruit along with quantification of bioactive constituents using RP-HPLC-PDA and evaluation of in-vitro anti-oxidant and, anti-microbial activity. The fruit was found rich in nutritional composition having protein (2.41 % ± 0.007), fibre (3.46 % ± 0.02) and carbohydrate (90.19 % ± 1.73) with energy value of 368.2 ± 3.94 Kcal/100g. The elemental analysis of fruit resulted in macronutrients Ca, Mg and Na and micronutrients Fe, Mn, Zn, and Cu in amounts comparable to common marketed fruits. The RP-HPLC-PDA analysis revealed the presence of six phenolic compounds in all 3 extracts made from the fruit in which highest amount are present in hydro-alcoholic extract. All the extracts exhibited potent antioxidant activity evaluated through DPPH assay and oxygen radical absorbing capacity (ORAC), with highest activity in hydro-alcoholic extract. All the analyzed extracts also exhibited potent inhibition, against four human pathogens namely Pseudomonas aeruginosa, Vibrio cholerae, Escherichia coli, and Shigella flexneri. Therefore, it is evident from the study that the fruit of S. chelonoides has immense potential as a nutraceutical supplement and may help in the management of nutrient deficiency and malnutrition among rural and tribal communities.

Nutritional characterization of an underutilized legume Entada rheedii Spreng. seeds and validation of its folklore uses.

Entada rheedii seeds are a rich source of protein (23.99% ± 0.07), starch (42.04% ± 0.05) and potassium (1670.15 ppm ± 116.732). HPTLC-UV analysis (seeds) reveals galactose in considerable amount, that is, 2.60% ± 0.008. Additionally, the species is low in fat and anti-nutrient metabolites like tannin. Interestingly, the proximates in Entada seeds was found comparable with the commonly consumed legumes like cowpea, green gram, and so on. The species exhibits promising anti-radical, anti-inflammatory and anti-diabetic activity. Data advocates the folklore use of E. rheedii seeds and can be a promising alternative source of dietary nutrition, fortified with medicinal value. Standardization and validation of scientific knowledge will bring such underutilized, low-cost legumes into daily dietary intake and are a promising source for Protein-Energy-Malnutrition.

Renal vein thrombosis in pancreatitis: a rare vascular complication.

Extrasplanchnic venous thrombosis is a rare complication of chronic pancreatitis. Thrombosis of inferior vena cava and renal vein, in particular, is extremely rare. We present our recent experience of managing two patients of chronic pancreatitis who presented with renal vein thrombosis. We also highlight the treatment dilemmas facing a clinician managing patients with this atypical and rare vascular complication.

In silico guided in vitro study of traditionally used medicinal plants reveal the alleviation of post-menopausal symptoms through ERß binding and MAO-A inhibition.

The slumping level of estrogen and serotonin in menopausal women is directly associated with the occurrence of menopausal symptoms where, estrogen receptor-ß (ERß) and monoamine oxidase-A (MAO-A) are directly involved. The present investigation aimed for validation of promising plants traditionally used to alleviate menopausal symptoms with ERß mediated MAO-A inhibition potential through in silico disease-target network construction using Cytoscape plugins followed by molecular docking of phytomolecules through AutoDock vina. ADMET parameters of identified bioactive phytomolecules were analysed through swissADME and ProTox II. The efficacy of promising plant leads was further established through in vitro ERß competitive binding, MAO-A inhibition, enzyme kinetics and free radical quenching assays. In silico analysis suggested glabrene (ΔG = -9.7 Kcal/mol) as most promising against ERß in comparison to 17ß-estradiol (ΔG = -11.4 Kcal/mol) whereas liquiritigenin (ΔG = -9.4 Kcal/mol) showed potential binding with MAO-A in comparison to standard harmine (ΔG = -8.8 Kcal/mol). In vitro analysis of promising plants segregated Glycyrrhiza glabra (IC50 = 0.052 ± 0.007 µg/ml) as most promising, followed by Hypericum perforatum (IC50 = 0.084 ± 0.01 µg/ml), Trifolium pratense (IC50 = 0.514 ± 0.01 µg/ml) and Rumex nepalensis (IC50 = 2.568 ± 0.11 µg/ml). The enzyme kinetics of promising plant leads showed reversible and competitive nature of inhibition against MAO-A. The potency of plant extracts in quenching free radicals was at par with ascorbic acid. The identified four potent medicinal plants with ERß selective, MAO-A inhibitory and free radical quenching abilities could be used against menopausal symptoms however, finding needs to be validated further for menopausal symptoms in in vivo conditions for drug development.Communicated by Ramaswamy H. Sarma.

Docking experiments suggest that gloriosine has microtubule-targeting properties similar to colchicine.

Gloriosine, the predominant metabolite of Gloriosa superba L., shares chemical properties with colchicine. We analyze the microtubule-binding affinity of gloriosine at the colchicine binding site (CBS) using an in silico-in vivo approach. The In silico docking of gloriosine showed a binding score of (-) 7.5 kcal/Mol towards ß-tubulin at CBS and was validated by overlapping the coupling pose of the docked ligand with co-crystallized colchicine. 2D plots (Ligplot +) showed > 85% overlap between gloriosine and colchicine. The ADMET profile of gloriosine was in accordance with Lipinski's rule of five. Gloriosine belongs to class II toxicity with anLD50 value of 6 mg/kg. In vivo and transmission electron microscopy studies revealed that gloriosine induces abnormalities in cell division such as condensed chromosomes in C-metaphase and enlarged nucleus with increased nuclear material. Gloriosine treated cells exhibited mitotic index of about 14% compared to control of 24% and high anti-proliferative activity i.e. 63.94% cell viability at a low concentration (0.0004 mg/ml). We conclude that gloriosine has a strong affinity for ß-tubulin at CBS and thus can be used as a colchicine alternative in cytology and other clinical conditions.

Nutritional potential of an edible terrestrial orchid Eulophia nuda LINDL and validation of its traditional claim in arthritis.

ETHNO PHARMACOLOGICAL RELEVANCE: Eulophia nuda, locally known as "Amarkand" is an edible orchid, traditionally used as food and ethnomedicine in arthritis, as a blood purifier, vermifuge, in bronchitis, scrofulous glands etc. AIM: The present study focuses on the proximate-nutrient analysis, metabolic profiling of bioactive phenolic acids (PA's) and validation of anti-arthritic activity in E. nuda. MATERIALS: The proximate, nutrition and element (macro-micro) content were evaluated as per standard protocols. The anti-arthritic activity was evaluated via different Invitro models and bioactive phenolics were quantified through calibrated HPLC-UV (PDA) method, as per ICH guidelines. RESULTS: The species contains a considerable amount of proximate i.e. ash, fiber, crude alkaloid, total phenolics, and flavonoid. It is a rich source of macro-micro nutrients, carbohydrates and energy, at par with conventional cereals and super-foods like finger millet, foxtail millet etc. It also contains seven PA's viz. gallic acid, protocatechuic acid, caffeic acid, syringic acid, vanillin acid, ferulic acid and quercetin. The PA's content varies from 4.00 to 83.50 µg/ml. The anti-arthritic potential of the plant extract based on several in-vitro-models showed a promising inhibitory effect on inflammation and uric acid synthesis. CONCLUSION: The study scientifically validates the traditional claims of this traditional orchid as food and ethnomedicine. The species can be commercially explored as a supplement to combat nutritional deficiency among rural communities.

Impact of seasonal variation on four labdane-type diterpenoids in Coleus forskholii Briq.

The present study has been planned to evaluate the impact of seasonal variation in labdane-type diterpenoids namely isoforskolin, forskolin, 1,9-dideoxyforskolin and 1-deoxyforskolin in Coleus forskholii (roots). The plant samples were harvested in different seasons from our experimental field located at CSIR-NBRI garden, Lucknow (India) and metabolite contents were estimated through validated high performance thin layer chromatography (HPTLC) method. The HPTLC plate was developed in tertiary mobile phase of toluene-ethyl acetate-methanol (8.5-1-0.05 v/v) for separation of all the four metabolites. The metabolite content viz. isoforskolin, forskolin, 1,9-dideoxyforskolin and 1-deoxyforskolin varies from 0.0247% to 0.198%, 0.238 to 0.730%, 0.056 to 0.161% and 0.0401 to 0.332% on dry weight basis respectively. The maximum content of metabolites was recorded in winter season and was found optimum for harvesting of C. forskholii roots. Optimization of harvesting season for this industrially valuable medicinal plant will lead to sustainable sources of good quality raw material to herbal drug industry.

Evaluation of Coscinium fenestratum (Goetgh.) Colebr. stem extracts for urolithiasis and quantification of bioactive alkaloids to validate the traditional claims.

Coscinium fenestratum (Goetgh.) Colebr. is widely used for urinary disorders and kidney stones by ethnic communities in southern India. The species is documented in various ancient Indian Ayurvedic literatures having therapeutic use in 'Ashmari' i.e., urolithiasis. The present study aims at validation of in-vitro anti-urolithiatic potential of various extracts of C. fenestratum stem along with identification and quantification of major bioactive alkaloids, i.e., berberine and palmatine through HPTLC and LC-MS/MS. Water extract showed maximum anti-urolithiatic activity which on further kinetic analysis, showed concentration dependent inhibitory delay in nucleation and aggregation of calcium oxalate crystals. Berberine and palmatine were quantified with maximum content in methanolic extract (0.478 ± 0.003 and 0.0358 ± 0.001) followed by chloroform and petroleum ether extracts. The study validates ethnobotanical use of C. fenestratum as anti-urolithiatic agent. Further, species can also be explored as a substitute for Berberis spp. for the alkaloid metabolites i.e., berberine and palmatine.

Study on Chemotypic Variability of Coleus forskohlii Briq., Samples Collected from Different Phytogeographical Locations of India and Evaluation of Its Inhibitory Potential.

Coleus forskohlii Briq. is an important medicinal herb, endowed with a wide range of medicinal properties against the variety of ailments. Seven germplasm of C. forskohlii collected from different phyto-geographical locations and identification of elite chemotype was performed with the help of high performance thin layer chromatography. Data of soil analysis correlated with the bioactive compounds and inhibitory potential of the species. Quantification of forskolin and its isomer (iso-forskolin) content were done in all the collected samples of C. forskohlii, which revealed a wide range of variations, varying from 1.15-0.004% and 0.0091 to 0.1077% per dry weights basic, respectively. Variation in the bioactive content may be due to the soil nature and environmental factors. Soil analysis of collected samples demonstrated that there is significant variation in available NPK and micronutrient content and may be reasoned for existing chemotypic variability. In vitro biological activity (antioxidant and antidiabetic) analyses were performed, which reveals that germplasms have a high amount of forskolin and iso-forskolin, both show more activity. The aim of this study was to elucidate the effect of elicitors and precursors on the production of bioactive compounds and identification of best elite germplasm among the populations, to provide basic lead to the industry for commercial exploitability including its location-specific commercial cultivation.

Liver function tests in COVID 19: A retrospective record-based study from a tertiary care centre in urban Maharashtra, India.

Background: COVID-19 is a multi system disorder and causes various abnormalities in liver function tests. The aim of this study was to estimate the prevalence of abnormal liver function tests in patients of COVID-19 and to describe the association of liver function tests with clinical features and disease severity in these patients. Methods: We retrospectively evaluated and analyzed the liver function tests of all real-time polymerase chain reaction (RT-PCR) positive COVID-19 patients admitted to a tertiary care hospital in Western Maharashtra. The Institutional Ethics Committee of our hospital approved the study. Results: Of the 533 patients included in our study, 50% had abnormal albumin levels while 40.1%, 43.5%, 9.3%, and 6.3% patients had deranged alanine transaminase (ALT) aspartate transaminase (AST), total protein and bilirubin levels, respectively. Hepatocellular injury was observed in 21 (3.9%) patients, and cholestatic liver injury was observed in seven (1.3%) patients. Abnormal liver function test (LFT) was significantly associated with disease severity but not with mortality. Conclusion: Abnormal LFT in patients of COVID-19 is associated with severe disease but not mortality. Liver injury is common in patients of COVID-19.

Establishment of antitick efficacy of a phytoformulation prepared from Annona squamosa leaf extracts for the management of acaricide resistant tick infestations on cattle.

To combat the problem of acaricide resistance in ticks, efforts have been made to develop eco-friendly herbal acaricides which are safe for animal use with no possibilities of evoking environment and residual toxicity. In the present study, the acaricidal properties of a commonly available plant, Annona squamosa have been established to develop a novel antitick phytoformulation using leaf extract, suitable solvents and emulsifier for managing resistant tick infestations in animals. The quality control of the safe and stable phytoformulation using Rutin as a major constituent was maintained by HPTLC profiling. Several phytformulations (A-E) were prepared and formulation 'C' is identifried as the most effective showing 52.5-75% antitick activity against in vitro treated resistant strains of R. microplus (IVRI-IV and V) with 33.8-40.2% inhibition of oviposition while 65-85% mortality against multiacaricide resistant field populations. The phytoformulation provided 70.4% efficacy after first larval challenge in experimentally infested animals. The field efficacy of the formulation in different multilocational field trials was 68.53-77.8%. The detail antitick efficacy of the formulation along with safety and stability are explained as a component of antitck technology to manage resistant tick problem in livestock.

Anemia in cirrhosis: An underestimated entity.

Anemia in a patient with cirrhosis is a clinically pertinent but often overlooked clinical entity. Relevant guidelines highlight the algorithmic approach of managing a patient of cirrhosis presenting with acute variceal hemorrhage but day-to-day management in hospital and out-patient raises multiple dilemmas: Whether anemia is a disease complication or a part of the disease spectrum? Should iron, folic acid, and vitamin B complex supplementation and nutritional advice, suffice in those who can perform tasks of daily living but have persistently low hemoglobin. How does one investigate and manage anemia due to multifactorial etiologies in the same patient: Acute or chronic blood loss because of portal hypertension and bone marrow aplasia secondary to hepatitis B or C viremia? To add to the clinician's woes the prevalence of anemia increases with increasing disease severity. We thus aim to critically analyze the various pathophysiological mechanisms complicating anemia in a patient with cirrhosis with an emphasis on the diagnostic flowchart in such patients and proposed management protocols thereafter.

Current and emerging therapeutic strategies in pancreatic cancer: Challenges and opportunities.

Pancreatic carcinoma (PC) is one of the leading causes of cancer-related deaths worldwide. Despite early detection and advances in therapeutics, the prognosis remains dismal. The outcome and therapeutic approach are dependent on the stage of PC at the time of diagnosis. The standard of care is surgery, followed by adjuvant chemotherapy. The advent of newer drugs has changed the landscape of adjuvant therapy. Moreover, recent trials have highlighted the role of neoadjuvant therapy and chemoradiotherapy for resectable and borderline resectable PC. As we progress towards a better understanding of tumor biology, genetics, and microenvironment, novel therapeutic strategies and targeted agents are now on the horizon. We have described the current and emerging therapeutic strategies in PC.

Characterization of deltamethrin, cypermethrin, coumaphos and ivermectin resistance in populations of Rhipicephalus microplus in India and efficacy of an antitick natural formulation prepared from Ageratum conyzoides.

Rhipicephalus microplus is posing a serious threat to productive animal husbandry. Excessive use of synthetic chemicals in tick management has led to the development of resistant tick populations. Characterization of resistance to deltamethrin, cypermethrin, coumaphos and ivermectin in ticks is necessary to develop a suitable and sustainable control strategy. Based on adult immersion test and larval packet test, the resistance ratios (RR50) for adults and larvae of R. microplus populations from two Indian states ranged from 3.8 to 19.4 and 1.35-25.0 against deltamethrin, 0.061-26.3 and 0.22-19.2 against cypermethrin, and 0.2-9.5 and 0.01-3.1 against coumaphos, respectively, were recorded. Moreover, the RR50 for adults ranged from 0.212 to 3.87 against ivermectin. The RR50 for different acaricides was significantly (p<0.01) correlated with esterases, Glutathione S-transferase and monooxygenase activity. A point mutation at the 190th position of the domain II S4-5 linker region of the sodium channel gene in synthetic pyrethroids (SP) resistant populations was also detected. An antitick natural formulation prepared from the plant Azeratum conyzoides and containing two major compounds, Precocene-I (7­methoxy-2, 2-dimethyl 2H-chromene) and Precocene II (6, 7-dimethoxy-2, 2-dimethyl- 3-chromene), was developed and tested against the resistant ticks. The LC50 values of the natural formulation against the resistant populations were in the range of 4.31-5.33% irrespective of their RR50 values. Multi-acaricide resistant populations of R. microplus are established in India and the A. conyzoides based natural formulation can be used for its management.

Rodenticide-induced acute liver failure - Uncommon presentation of commonly available poison.

Rodenticide or 'rat poison' is easily available in a predominantly agrarian economy such as India. Metal phosphides or yellow phosphorous are two common rodenticides. Acute liver failure caused by accidental or suicidal poisoning with rodenticides has been infrequently reported in literature. Liver transplantation offers the best chances of survival in severe intoxication. However, the availability of liver transplantation in resource-limited settings presents a challenge. N-acetyl cysteine has been successfully used in paracetamol poisoning. Its use in rodenticide-induced acute liver failure is not so well known. We report three cases of rodenticide-related acute liver failure, one of them being a pregnant lady. All three patients were given N-acetyl cysteine and two patients improved. It is possible that the administration of N-acetyl cysteine contributed to the improvement in these two.

Hepatitis B vaccination in Indian children: Seroprotection and age-related change in antibody titres.

BACKGROUND: Childhood immunization against hepatitis B is one of the most effective strategies for reducing the global burden of chronic hepatitis B infection and its sequelae. There are limited data from India on both the anti-Hep B antibody titres in children after vaccination and the age-related decline in the titres. This study was planned to estimate the proportion of children in the age group of 1-10 years who develop protective levels of anti-hepatitis B antibodies after childhood vaccination and to examine the change in antibody titres with age in these children. METHODS: A hospital-based cross-sectional study was carried out in children admitted to the hospital for various ailments. Basic demographic data, vaccination history and HBsAg status of the mother were recorded. All the enrolled children were evaluated for HBsAg and anti hepatitis B surface antibody (anti-HBS) titres. Institutional ethical clearance was obtained, and informed consent from the parents of the children was taken before drawing samples. RESULTS: We found that 68.86% Confidence Interval ((CI): 59.8-76.8%) of the children showed protective antibody titres after vaccination, while 31.14% (CI: 23.1-40.2%) of the children had titres less than 10 IU/L. Although 100% of children in the age group from birth to three years had titres more than 10 IU/L, this percentage showed a decline across the age groups, and 60% of children aged 9-10 years had titres less than 10 IU/L. CONCLUSION: Childhood vaccination against hepatitis B is effective in 68% children, and the antibody levels showed a steady decline with increasing age.

Influence of Soil Variation on Diosgenin Content Profile in Costus speciosus from Indo-Gangetic Plains.

Costus speciosus is a rich source of commercially important compound Diosgenin, distributed in different regions of India. The present investigation was aimed to quantify diosgenin through High Performance Thin Layer Chromatography in 34 germplasms of Costus speciosus and also to identify the superior sources and to correlate the macronutrients of rhizospheric soil. The starch content varied in microscopic examination and correlated inversely (r=-0.266) with diosgenin content. Findings revealed that the extraction process with acid hydrolysis yielded higher diosgenin content (0.15-1.88 %) as compared to non-hydrolysis (0.009-0.368 %) procedure. Germplasms from Uttar Pradesh (NBCS-4), Jharkhand (NBCS-39) and Bihar (NBCS-2) were identified as elite chemotypes based on hierarchical clustering analysis. The phosphorous content of respective rhizospheric soil correlated positively (r=0.742) with diosgenin content. Findings of present study are useful to identify the new agrotechniques. The elite germplasms can also be used as quality planting material for large scale cultivation in order to assure a sustained supply to the herbal drug industry.