Unlocking the versatility of nitric oxide in plants and insights into its molecular interplays under biotic and abiotic stress.

In plants, nitric oxide (NO) has become a versatile signaling molecule essential for mediating a wide range of physiological processes under various biotic and abiotic stress conditions. The fundamental function of NO under various stress scenarios has led to a paradigm shift in which NO is now seen as both a free radical liberated from the toxic product of oxidative metabolism and an agent that aids in plant sustenance. Numerous studies on NO biology have shown that NO is an important signal for germination, leaf senescence, photosynthesis, plant growth, pollen growth, and other processes. It is implicated in defense responses against pathogensas well as adaptation of plants in response to environmental cues like salinity, drought, and temperature extremes which demonstrates its multifaceted role. NO can carry out its biological action in a variety of ways, including interaction with protein kinases, modifying gene expression, and releasing secondary messengers. In addition to these signaling events, NO may also be in charge of the chromatin modifications, nitration, and S-nitrosylation-induced posttranslational modifications (PTM) of target proteins. Deciphering the molecular mechanism behind its essential function is essential to unravel the regulatory networks controlling the responses of plants to various environmental stimuli. Taking into consideration the versatile role of NO, an effort has been made to interpret its mode of action based on the post-translational modifications and to cover shreds of evidence for increased growth parameters along with an altered gene expression.

Microbial CO2 fixation and biotechnology in reducing industrial CO2 emissions.

The rapid acceleration in emissions of inevitably generated CO2 due to numerous activities mainly anthropogenic have devastating environmental effects leading to climatic concerns. Hence, significant, sustainable approaches should be developed for reduction of CO2 emission targets, balancing the existing needs of the current population. Biological carbon acquisition, storage and usage are considered crucial alternative strategies in assimilating inorganic carbon, manifested by diverse variety of microorganisms. Furthermore, central biochemical routes along with associated enzymes serve as considerable factors for understanding molecular microbial CO2 assimilation. Microorganisms exhibit an impeccable capability to facilitate evolved mechanisms in sequestering inorganic carbon at higher pace to produce biomaterials like biofuels, bioplastics etc. This review endorses the importance of microorganisms in reducing the concomitant release of CO2 by providing supervision in biotechnological applications (such as genetic engineering, microbial electrosynthesis, gas fermentation and protein engineering) to mitigate CO2 at an industrial scale.

Design, development and characterization of a chimeric protein with disulfide reductase and protease domain showing keratinase activity.

Keratin is one of the major components of solid waste, and the degradation products have extensive applications in various commercial industries. Due to the complexity of the structure of keratin, especially the disulfide bonds between keratin polypeptides, keratinolytic activity is efficient with a mixture of proteins with proteases, peptidases, and oxidoreductase activity. The present work aimed to create an engineered chimeric protein with a disulfide reductase domain and a protease domain connected with a flexible linker. The structure, stability, and substrate interaction were analyzed using the protein modeling tools and codon-optimized synthetic gene cloned, expressed, and purified using Ni2+-NTA chromatography. The keratinolytic activity of the protein was at its maximum at 70 °C. The suitable pH for the enzyme activity was pH 8. While Ni2+, Mg2+, and Na+ inhibited the keratinolytic activity, Cu2+, Ca2+, and Mn2+ enhanced it significantly. Biochemical characterization of the protease domain indicated significant keratinolytic activity at 70 °C at pH 10.0 but was less efficient than the chimeric protein. Experiments using feathers as the substrate showed a clear degradation pattern in the SEM analysis. The samples collected from the degradation experiments indicated the release of proteins (2-fold) and amino acids (8.4-fold) in a time-dependent manner. Thus, the protease with an added disulfide reductase domain showed excellent keratin degradation activity and has the potential to be utilized in the commercial industries.

Ayush [Indian System of Medicines] Prophylaxis Against COVID-19: A Living Systematic Review and Meta-Analysis (Second Update).

Background: Ayush (Ayurveda, Yoga and Naturopathy, Unani, Siddha, and Homeopathy) system of medicine has been extensively used in India for the prevention and management of coronavirus disease 2019 (COVID-19). The present report is the second update of a living systematic review and meta-analysis and has been intended to assess the preventive potential and safety of Ayush drugs against COVID-19. Methods: A search of databases such as PubMed, the Cochrane central register of controlled trials, the World Health Organization (WHO) COVID-19 database, the clinical trial registry-India, the AYUSH research portal, and preprint repositories was performed till March 1, 2023. Randomized controlled trials or analytical observational studies were included. Incidences of COVID-19 infections and mortality were taken as primary outcome measures; and symptomatic severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) infection, incidences of hospital admissions, and adverse events were taken as secondary outcomes. The risk of bias was evaluated by version 2 of the Cochrane risk-of-bias tool for randomized trials (RoB-2) and the Risk of Bias in Non-Randomized Studies-of Interventions (ROBINS-I) tool. Data were synthesized through the RevMan 5.4 tool, and the certainty of the evidence was ranked through the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach. Results: Out of 2350 articles, 15 studies were included in the review. The pooled estimate of Guduchi [Tinospora cordifolia (Thunb.) Miers], Chyawanprasha, and Ayuraksha kit was obtained. Meta-analysis of the data suggested that Guduchi may reduce the incidences of COVID-19 infections (risk ratio [RR]: 0.67 [95% confidence interval, CI: 0.48-0.95]) and the symptom development among the infected (RR: 0.88 [95% CI: 0.73-1.08]), however, no change was observed in hospital admissions (RR: 0.96 [95% CI: 0.20-4.53]). It is uncertain that Chyawanprasha reduces the incidence of COVID-19 infection (RR: 0.28; [95% CI: 0.07-1.08]). Ayuraksha kit as chemoprophylaxis may reduce the incidences of COVID-19 infections (RR: 0.49; [95% CI: 0.32-0.77]). The certainty of the evidence was low to moderate mainly due to the serious risk of bias. Conclusion: Prophylactic use of Ayush medicines for populations at risk may help to prevent COVID-19 infection by reducing incidence and severity. However, considering the certainty and fewer studies, high-quality studies are still desired to confirm the efficacy and safety of Ayush chemoprophylaxis for COVID-19. Nevertheless, this update will serve as a torchbearer for policymakers, physicians, and other stakeholders for their decision-making through evidence-based medicine.

Effectiveness of high flow nasal cannula (HFNC) versus bilevel positive airway pressure (BiPAP) in preventing tracheal reintubation in patients with high risk of extubation failure in intensive care unit - A randomised comparative trial.

Background and Aims: The incidence of tracheal extubation failure in high-risk patients is higher, and non-invasive ventilation is suggested to avoid tracheal reintubation. This study compares the effectiveness of bilevel positive airway pressure (BiPAP) and high flow nasal cannula (HFNC) to reduce the rate of reintubation in intensive care unit (ICU) patients with increased risk of extubation failure. Methods: This randomised comparative trial was conducted on 60 high-risk patients on mechanical ventilators admitted to the ICU, ready for weaning after a spontaneous breathing trial. They were randomised to Group H for HFNC and Group B for BiPAP therapy. Designated therapy was administered in these high-risk patients for up to 48 hours after tracheal extubation. Haemodynamic parameters [mean arterial pressure (MAP), heart rate (HR), respiratory rate (RR), a saturation of peripheral oxygen (SpO2), electrocardiogram (ECG)], arterial blood gas analysis (ABG) parameter [potential of hydrogen (pH), partial pressure of carbon dioxide (pCO2), partial pressure of oxygen/fraction of inspired oxygen (paO2/FiO2) ratio], the effectiveness of cough, comfort level was recorded and continuous monitoring for signs of respiratory distress and failure was done. Results: Most of the patients were obese and had more than two risk factors for extubation failure. Several patients in Group B have significantly higher successful extubation than in Group H (P = 0.044). Most of the reintubation took place within 24 hours. The HFNC therapy was more comfortable and acceptable to patients. Conclusion: BiPAP therapy was more efficient than HFNC in preventing tracheal reintubation among patients with a high risk of extubation failure.

Diagnosis of Paediatric Extrapulmonary Tuberculosis by the MPT64 Antigen at a Tertiary Care Hospital.

Background Tuberculosis (TB) remains a global health concern, with India bearing a substantial burden. Paediatric TB, especially extrapulmonary TB (EPTB), presents unique diagnostic challenges due to its paucibacillary nature and the difficulty in obtaining suitable samples in children. Accurate and timely diagnosis is crucial to initiate appropriate treatment and mitigate disease spread. The MPT64 antigen test has shown promise in diagnosing TB, but its performance in paediatric EPTB remains underexplored. This study aimed to evaluate the diagnostic utility of the MPT64 antigen test in paediatric EPTB cases at a tertiary care hospital in India. Methods We conducted a prospective cross-sectional study at the Indira Gandhi Institute of Medical Sciences (IGIMS), a tertiary care hospital in India. A total of 250 paediatric participants, aged 0-18 years, with clinical suspicion of extrapulmonary tuberculosis (EPTB) were included. Diagnostic samples (e.g., tissue biopsies, pus, cerebrospinal fluid (CSF), and lymph node aspirates) were obtained, and tests including microscopy for acid-fast bacilli (AFB), mycobacterial cultures, GeneXpert MTB/RIF assay, and the TB Antigen MPT64 Rapid ICT Kit were performed. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of the MPT64 antigen test were calculated using culture and GeneXpert as reference standards. Results Among the 250 participants, 34 (13.6%) were confirmed to have EPTB. The MPT64 antigen test demonstrated a sensitivity of 70.6% and specificity of 92.1% in detecting EPTB cases. Mycobacterial cultures had the highest sensitivity (91.2%) and specificity (97.7%). GeneXpert showed a sensitivity of 70.6% and specificity of 93.9%. Overall diagnostic accuracy ranged from 88.7% for acid-fast bacteria (AFB) staining to 96.9% for mycobacterial cultures. The MPT64 antigen test had an area under the curve (AUC) of 0.814, indicating a good diagnostic accuracy. Conclusion The MPT64 antigen test demonstrates promising sensitivity and specificity for diagnosing paediatric EPTB, making it a valuable diagnostic tool, especially in resource-limited settings. However, mycobacterial cultures maintain the highest accuracy. Combining the MPT64 antigen test with other methods may enhance diagnostic capabilities.

Comparison of Cytokines Profiles and Monocyte Response Among Tuberculosis Patients Versus Patients Coinfected With Intestinal Helminth and Tuberculosis.

Background Tuberculosis (TB) and intestinal helminth infections often coexist, posing a significant health challenge. TB, caused by Mycobacterium tuberculosis, and helminths elicit distinct immune responses - Th1 for TB and Th2 for helminths. Co-infection introduces a complex immunological challenge, potentially compromising TB control. This study addresses the research gap by comparing cytokine profiles and monocyte responses in TB patients, helminth-infected individuals, and those with both. Insights gained may enhance diagnosis, treatment, and disease control strategies where TB and helminths prevail. Methods A cross-sectional observational study conducted at Indira Gandhi Institute of Medical Sciences, Patna, Bihar, aimed to compare cytokine profiles and monocyte responses in TB patients and those coinfected with TB and helminths. The study included 150 newly diagnosed active TB individuals aged 18 to 65 years. TB diagnosis was confirmed through clinical assessment, sputum microscopy, and GeneXpert (Cepheid, Sunnyvale, CA, USA) testing. Stool examination employed various methods, including the Kato-Katz technique and formalin-ether concentration. Blood samples were collected for hematological analysis, cytokine profiling, and monocyte isolation. Statistical analysis, using SPSS version 20.0 (IBM Corp., Armonk, NY, USA), included descriptive statistics, and t-test analyses. Results In our study of 150 participants, half (50.0%) showed positive helminth status. The sociodemographic analysis revealed no significant differences in age, gender, education, occupation, marital status, smoking, alcohol, BMI, diabetes, and hypertension between TB patients (n=75) and TB+Helminth patients (n=75), ensuring baseline matching. The prevalence of specific helminth infections in TB+Helminth patients included Ascaris lumbricoides (24.0%), Trichuris trichiura (18.7%), and others. Hematological parameters showed significant differences, with TB+Helminth patients exhibiting higher RBC count, hemoglobin, hematocrit, neutrophil count, and monocyte count; also eosinophil count was more raised in TB+Helminth patients (0.36 x 103/µL) when compared to TB patients (0.25 x 103/µL). Cytokine profiles and monocyte responses varied significantly between the groups, with TB patients having higher IL-4, IL-6, IFN-γ, TNF-α, and IL-1ß levels, while TB+Helminth patients had elevated IL-10. Monocyte response time did not differ significantly. Conclusion The observed differences in hematological parameters and cytokine profiles emphasize the need for tailored approaches to diagnosis and treatment in co-infected individuals. These findings suggest that the management of TB patients should consider the potential influence of helminth co-infections.

Case Report: An Atypical Case of Post-Kala-Azar Dermal Leishmaniasis with Ulcers and Verrucous Lesions: Clinical and Therapeutic Implications.

About 75% cases of post-kala-azar dermal leishmaniasis (PKDL) occur in India. Although the classic description of PKDL is the progression from initial hypopigmented macular lesions to papules to plaques and nodular lesions, atypical morphologies are also seen and are easily missed or misdiagnosed. We report a case of a 27-year-old man who presented to us with multiple acral ulcers and verrucous lesions for 5 years. A diagnosis of PKDL was made based on slit skin smear, histopathology, and quantitative polymerase chain reaction. The patient was given combination therapy with four doses of liposomal amphotericin B and miltefosine 50 mg twice daily for 45 days. In this report, we discuss unusual morphologies of PKDL, the pathway to the diagnosis, and the therapeutic options available along with their efficacy.

Comparative Evaluation of Broth Microdilution With Disc Diffusion and VITEK 2 for Susceptibility Testing of Colistin on Multidrug-Resistant Gram-Negative Bacteria.

Background The rise of antibiotic resistance, particularly in Gram-negative bacteria, poses a significant global health threat. Colistin, a last-resort antibiotic, has witnessed renewed use. However, accurate susceptibility testing for colistin is challenging, with various methods available, leading to potential discrepancies. Ensuring reliable testing is crucial for effective patient treatment and antimicrobial stewardship. This study addresses the need to compare different colistin susceptibility testing methods, providing insights into their accuracy and relevance in clinical settings. Methods In this one-year prospective observational cross-sectional study conducted at Indira Gandhi Institute of Medical Sciences (IGIMS), Bihar, India, a tertiary care hospital from July 2021 to June 2022, we aimed to evaluate the concordance between two widely used methods, VITEK 2 and Disc Diffusion, for antibiotic susceptibility testing in clinical multidrug-resistant Gram-negative bacterial isolates. These isolates, including species like Klebsiella pneumoniae, Acinetobacter baumannii, Klebsiella oxytoca, Pseudomonas aeruginosa, Citrobacter freundii, and Escherichia coli, were isolated from various clinical specimens. After rigorous species-level identification and quality control measures, antibiotic susceptibility testing was performed using both methods, and their agreement was assessed through Percentage Agreement analysis. Results In our study, we isolated and identified bacterial isolates from 105 patients, with a mean age of 47.30 years, demonstrating a wide age range. Pus samples were the most common type (25.7%), and K. pneumoniae was the most prevalent organism (45.7%). Antibiotic resistance patterns revealed significant challenges in treating infections caused by K. pneumoniae and A. baumannii, with resistance rates exceeding 70% for certain antibiotics. Among the 48 isolates of K. pneumoniae, the agreement was 93.8%, with 89.6% being sensitive and 6.3% being resistant by Disc Diffusion, while VITEK 2 indicated 0% resistance. E. coli isolates (n=21) had an agreement of 90.5%, with 90.5% sensitivity and 9.5% resistance by Disc Diffusion, and no resistance by VITEK 2. Conclusion The comparative analysis of antibiotic susceptibility testing methods reveals the superior performance of the VITEK 2 system, particularly in sensitivity and negative predictive value, emphasizing its potential as a reliable tool for guiding antibiotic therapy decisions.

AYUSH (Indian System of Medicines) Therapeutics for COVID-19: A Living Systematic Review and Meta-Analysis (First Update).

Background: In India, alternative and complementary therapies (Ayurveda, Yoga and Naturopathy, Unani, Siddha, and Homeopathy [AYUSH] medicines) are extensively utilized in COVID-19 management, and some were investigated clinically. This study assessed the effectiveness of AYUSH therapeutic on COVID-19 through a living systematic review and meta-analysis approach. Methods: Databases like PubMed; the Cochrane central register of controlled trials; WHO COVID-19 database; the central trial registry-India; Digital Helpline for Ayurveda Research Articles and AYUSH research portal, and preprint repositories were searched till August 1, 2021. Randomized controlled trials or analytical observational studies were included only. Primary outcomes selected were clinical improvement, WHO ordinal scale, viral clearance, and mortality, whereas secondary outcomes were the use of O2 therapy or mechanical ventilator, admission to high dependency unit or emergency unit, duration of hospitalization, the time to symptom resolution, and adverse events. The risk of bias was evaluated by Version 2 of the Cochrane risk-of-bias tool for randomized trials (RoB-2) and Risk of Bias in Nonrandomized Studies-of Interventions (ROBINS-I) tools; data were synthesized through RevMan 5.4 tool, and the certainty of the evidence was ranked through the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach. Results: Of 3609 studies retrieved, 17 were included in the systematic review, and 3 AYUSH therapeutics were meta-analyzed. Meta-analysis suggested that add-on AYUSH-64 likely provides therapeutic benefits by reducing time to symptom resolution (mean difference [MD] 2.35 days lower [95% confidence interval, CI; 4.05 lower to 0.65 lower]) and hastening clinical improvement (365 more per 1000 [95% CI; 4 more to 1000 more]) in mild-to-moderate COVID-19 patients. Kabasura Kudineer adjuvant to standard care is likely to reduce symptom resolution (MD; 1.93 days lower [95% CI; 2.28 lower to 1.58 lower]) and hospital stay (MD; 4.2 days lower [95% CI; 4.97 lower to 3.43 lower]) in mild-to-moderate COVID-19 patients. Co-administration of Guduchi (Tinospora cordifolia [Willd.] Miers.) to standard care may reduce the duration of hospitalization (MD; 3.93 days, lower [95% CI; 8.83 lower to 0.97 higher]) in mild-to-moderate COVID-19 patients. Furthermore, all three agents seemed safe in adjunct usage to standard care. The certainty of evidence for most outcomes was moderate to low, primarily due to the high risk of bias or imprecision owing to the small sample size. Conclusion: Rational use of integrated or standalone AYUSH interventions in mild-to-moderate COVID-19 patients is safe and may provide therapeutic benefits. The effect estimates may be changed with additional evidence in upcoming updates.

Identification of Fungal Isolates in Patients With Pulmonary Tuberculosis Treated at a Tertiary Care Hospital.

Introduction Pulmonary tuberculosis (TB) has long been associated with opportunistic fungal infections and could prove lethal if these fungal infections are not detected in the early stages of tuberculosis. TB patients are mostly immunocompromised, and an association with a fungal infection fuels each other, thus decreasing host immunity and making it difficult to treat. Extensive use of antibiotics and steroids has resulted in increasing trends of these fungal infections globally. Material and methods This observational, retrospective hospital-based medical record review study was conducted in the Department of Microbiology at the Indira Gandhi Institute of Medical Sciences (IGIMS), Patna, Bihar, India. A total of 200 medical records of pulmonary tuberculosis patients diagnosed by using sputum as clinical samples were evaluated and analysed for two years, from January 2020 to December 2021. This study was started after approval from the institutional ethical committee. Data were obtained from the mycology test records from the Department of Microbiology and from the data files of the medical records section over a period of two years. Results Our study included the medical records of 200 pulmonary tuberculosis patients who underwent treatment at IGIMS Patna. Out of 200 patient records, 124 (62%) were males and 76 (38%) were females. The male-to-female ratio was 1.6:1. After analysis and evaluation of 200 medical records of pulmonary tuberculosis patients, fungal species were detected in 16 (8%) sputum samples. Among 16 culture-positive sputum samples, 10 (8.06%) and six (7.1%) were diagnosed in male and female patients, respectively. Fisher's exact test showed a non-significant two-sided p-value of 1.000 with a relative risk of 0.9982. The prevalence, or positivity rate, was 8% in two years. The age group of 31-45 years had the most fungal co-infection at 37.5%. Among the fungal isolates, 5/16 (31.25%) were yeasts, and the remaining 11/16 (68.75%) isolates were mycelial fungi. Conclusion According to the findings of the present study, pulmonary fungal infections co-exist in tuberculosis patients, although the prevalence rates of all the coinfections are low and statistically not significant. Being chronic in nature and with confusing clinical and radiological findings, these fungal infections are misdiagnosed as reactivation of tuberculosis. Hence, the increasing rate of morbidity and mortality can decrease if adequate measures are taken for the diagnosis at an early stage and appropriate treatment of these fungal mycoses with antifungal therapy is instituted.

Covalent organic frameworks (COFs) and metal-organic frameworks (MOFs) as electrochemical sensors for the efficient detection of pharmaceutical residues.

Pharmaceutical residues are the undecomposed remains from drugs used in the medical and food industries. Due to their potential adverse effects on human health and natural ecosystems, they are of increasing worldwide concern. The acute detection of pharmaceutical residues can give a rapid examination of their quantity and then prevent them from further contamination. Herein, this study summarizes and discusses the most recent porous covalent-organic frameworks (COFs) and metal-organic frameworks (MOFs) for the electrochemical detection of various pharmaceutical residues. The review first introduces a brief overview of drug toxicity and its effects on living organisms. Subsequently, different porous materials and drug detection techniques are discussed with materials' properties and applications. Then the development of COFs and MOFs has been addressed with their structural properties and sensing applications. Further, the stability, reusability, and sustainability of MOFs/COFs are reviewed and discussed. Besides, COFs and MOFs' detection limits, linear ranges, the role of functionalities, and immobilized nanoparticles are analyzed and discussed. Lastly, this review summarized and discussed the MOF@COF composite as sensors, the fabrication strategies to enhance detection potential, and the current challenges in this area.

Amiodarone-induced acute respiratory distress syndrome masquerading as acute heart failure.

BACKGROUND: Amiodarone use has been rarely associated with the development of acute respiratory distress syndrome (ARDS), usually in association with surgery or pulmonary angiography. In patients with preexisting left ventricular dysfunction, the diagnosis may be overlooked. CASE REPORT: A 92-year-old woman with a history of atrial fibrillation who was on low-dose amiodarone presented to the Emergency Department with sudden onset of shortness of breath. The patient was started on treatment for acute heart failure based on the physical examination and the elevated brain natriuretic peptide level. Despite adequate diuresis, the patient showed no improvement. A chest computed tomography scan revealed acute interstitial pneumonitis. The patient received corticosteroids due to suspected amiodarone-induced acute interstitial pneumonitis resulting in ARDS. She returned to her baseline activity within 2 weeks of the therapy. CONCLUSION: Although rare, clinicians should be vigilant for amiodarone-induced acute interstitial pneumonitis resulting in ARDS, as delay in treatment may result in a high risk of mortality. In addition, the development of ARDS occurred in our patient in the absence of precipitating factors such as surgery or pulmonary angiography.

Role of Ayurveda treatment in the management of hepatitis B (Ubhayapatha Ashrita Swatantra Kamala)- A case report.

Hepatitis B is one of the most common causes of liver disease, and due to unawareness of the safety measures, more prone to spread. As per the World Health Organization, for the South East Asia region, its prevalence is 2%. Yellowish discoloration of the eyes, body, and urine, abdominal discomfort, and vomiting is its cardinal symptoms. In Ayurveda, this set of symptoms is known as Kamala. This case report illustrates the effectiveness of the Ayurveda treatment modality in a patient with a viral load of 3705.71 IU/ml, and the values of AST (aspartate aminotransferase) and alanine transaminase (ALT) were 140 IU/ml and 173 IU/ml, respectively. A 40-year-old female patient with a complaint of yellowish discoloration of urine, eyes, and skin with fatigue and irritability was diagnosed with Ubhayapatha Ashrita Swatantra Kamala. The patient was treated with Ayurveda drugs and Virechana Karma (therapeutic purgation). After the treatment for 13 months, the disease was cured. Improvement was observed based on hepatitis B virus deoxyribonucleic acid (DNA) polymerase chain reaction (PCR) (hepatitis B DNA PCR) (decreased from 3705.71 IU/ml to <50.0 IU/ml) and the values of AST (decreased from 140 IU/ml to 19.0 IU/ml) and ALT (decreased from 173 IU/ml to 28 IU/ml). The patient was stable and asymptomatic during the follow-up period of 4 months.

Bacterial profile and antibiotic susceptibility pattern of uropathogens causing urinary tract infection in the eastern part of Northern India.

Urinary tract infection (UTI) is a common infectious disease that affects men and women. It is a significant health concern due to multidrug-resistant (MDR) organisms. Therefore, it is necessary to have a current understanding of the antibiotic susceptibility (AS) pattern of uropathogens to manage UTI effectively. Since the bacterial pathogen causing UTI and its AS vary with time and place, the prevailing AS pattern of the causative agents are essential for empirical antibiotic therapy. This study aims to determine the prevalence and AS of uropathogens isolated from UTI patients in the eastern part of Northern India. The study was carried out between November 2018 and December 2019. Clean catch midstream urine samples were collected and processed using standard guidelines for microbiological procedures. Positive microbiological cultures were found in 333 of the 427 patients, where 287 were gram-negative bacteria (GNB), and 46 were gram-positive bacteria (GPB). Females had a higher prevalence of UTI (60.7%) than males (39.3%) (p = 0.00024). The most susceptible age group in females was 18-50 years as compared to males, whereas at the age of 51-80 years and >80 years males were more susceptible than females (p = 0.053). The most prevalent pathogen identified were Escherichia coli (55.0%), followed by Proteus sp. (6.9%), Klebsiella pneumoniae (6.6%), Pseudomonas aeruginosa (6.3%), of which 96.0% were MDR bacteria. The susceptibility pattern of our study also revealed that amikacin, gentamycin and imipenem were the most effective drugs against GNB. In contrast, nitrofurantoin, vancomycin, and chloramphenicol were the most effective drugs against GPB. According tothe findings, MDR pathogens are very much prevalent. Since UTI is one of the most frequent bacterial diseases, proper management necessitates extensive investigation and implementation of antibiotic policy based on AS patterns for a particular region.

Downregulation of PTEN Promotes Autophagy via Concurrent Reduction in Apoptosis in Cardiac Hypertrophy in PPAR α-/- Mice.

Cardiac hypertrophy is characterized by an increase in the size of the cardiomyocytes which is initially triggered as an adaptive response but ultimately becomes maladaptive with chronic exposure to different hypertrophic stimuli. Prolonged cardiac hypertrophy is often associated with mitochondrial dysfunctions and cardiomyocyte cell death. Peroxisome proliferator activated receptor alpha (PPAR α), which is critical for mitochondrial biogenesis and fatty acid oxidation, is down regulated in hypertrophied cardiomyocytes. Yet, the role of PPAR α in cardiomyocyte death is largely unknown. To assess the role of PPAR α in chronic hypertrophy, isoproterenol, a ß-adrenergic receptor agonist was administered in PPAR α knock out (PPAR α-/-) mice for 2 weeks and hypertrophy associated changes in cardiac tissues were observed. Echocardiographic analysis ensured the development of cardiac hypertrophy and compromised hemodynamics in PPAR α-/- mice. Proteomic analysis using high resolution mass spectrometer identified about 1,200 proteins enriched in heart tissue. Proteins were classified according to biological pathway and molecular functions. We observed an unexpected down regulation of apoptotic markers, Annexin V and p53 in hypertrophied heart tissue. Further validation revealed a significant down regulation of apoptosis regulator, PTEN, along with other apoptosis markers like p53, Caspase 9 and c-PARP. The autophagy markers Atg3, Atg5, Atg7, p62, Beclin1 and LC3 A/B were up regulated in PPAR α-/- mice indicating an increase in autophagy. Similar observations were made in a high cholesterol diet fed PPAR α-/-mice. The results were further validated in vitro using NRVMs and H9C2 cell line by blocking PPAR α that resulted in enhanced autophagosome formation upon hypertrophic stimulation. The results demonstrate that in the absence of PPAR α apoptotic pathway is inhibited while autophagy is enhanced. The data suggest that PPAR α signaling might act as a molecular switch between apoptosis and autophagy thereby playing a critical role in adaptive process in cardiac hypertrophy.

Possible role of CYP2C9 & CYP2C19 single nucleotide polymorphisms in drug refractory epilepsy.

BACKGROUND & OBJECTIVES: Multiple drug resistance in epilepsy is a common problem and one third of epilepsy patients remain non responsive to antiepileptic drug (AED) therapy. In this study we aimed to investigate the relationship between the genetic polymorphism of cytochrome P450 genes, namely CYP2C9 and CYP2C19 with multiple drug resistance in epilepsy patients. METHODS: A total of 402 patients with epilepsy were enrolled in this study; 128 were drug resistant and 274 were drug responsive. The peripheral blood samples of the patients with epilepsy were collected. Drug compliance was confirmed in 20 per cent patient population using HPLC. Genotyping of CYP2C9 (FNx012 and FNx013), and CYP2C19 (FNx012 andFNx013) was carried out by PCR-RFLP. RESULTS: The genotype frequencies of CYP2C9 430 C>T (FNx012 variant) and CYP2C9 1075 A>C (FNx013 variant) did not differ significantly in drug resistant versus responsive patients. After combining CYP2C9 FNx012 and CYP2C9 FNx013, the frequency of CYP2C9FNx011/FNx013 was significantly lower in drug resistant as compared to drug responsive epilepsy patients (P=0.03, OR=0.53, 95%CI=0.30-0.95). Similarly, combined frequency of all the slow and poor metabolizer variants (2C9 FNx011/FNx012, FNx011/FNx013 and FNx012/FNx013) was also lower as compared to drug resistant group (P=0.03, OR=0.60, 95% CI 0.38-0.96). There was no significant differences in genotypic or allelic distribution of CYP2C190FNx012 while CYP2C19FNx013 was monomorphic in northern Indian population. INTERPRETATION & CONCLUSIONS: Our results demonstrated significant involvement of CYP2C9 genetic variants in the modulation of epilepsy pharmacotherapy confirming the important role of CYP2C9 mutants preventing epilepsy patients from developing drug resistance.

Pharmacogenomic association study on the role of drug metabolizing, drug transporters and drug target gene polymorphisms in drug-resistant epilepsy in a north Indian population.

BACKGROUND: In epilepsy, in spite of the best possible medications and treatment protocols, approximately one-third of the patients do not respond adequately to anti-epileptic drugs. Such interindividual variations in drug response are believed to result from genetic variations in candidate genes belonging to multiple pathways. MATERIALS AND METHODS: In the present pharmacogenetic analysis, a total of 402 epilepsy patients were enrolled. Of them, 128 were diagnosed as multiple drug-resistant epilepsy and 274 patients were diagnosed as having drug-responsive epilepsy. We selected a total of 10 candidate gene polymorphisms belonging to three major classes, namely drug transporters, drug metabolizers and drug targets. These genetic polymorphism included CYP2C9 c.430C>T (*2 variant), CYP2C9 c.1075 A>C (*3 variant), ABCB1 c.3435C>T, ABCB1c.1236C>T, ABCB1c.2677G>T/A, SCN1A c.3184 A> G, SCN2A c.56G>A (p.R19K), GABRA1c.IVS11 + 15 A>G and GABRG2 c.588C>T. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods, and each genotype was confirmed via direct DNA sequencing. The relationship between various genetic polymorphisms and responsiveness was examined using binary logistic regression by SPSS statistical analysis software. RESULTS: CYP2C9 c.1075 A>C polymorphism showed a marginal significant difference between drug resistance and drug-responsive patients for the AC genotype (Odds ratio [OR] = 0.57, 95% confidence interval [CI] = 0.32-1.00; P = 0.05). In drug transporter, ABCB1c.2677G>T/A polymorphism, allele A was associated with drug-resistant phenotype in epilepsy patients (P = 0.03, OR = 0.31, 95% CI = 0.10-0.93). Similarly, the variant allele frequency of SCN2A c.56 G>A single nucleotide polymorphism was significantly higher in drug-resistant patients (P = 0.03; OR = 1.62, 95% CI = 1.03, 2.56). We also observed a significant difference at the genotype as well as allele frequencies of GABRA1c.IVS11 + 15 A > G polymorphism in drug-resistant patients for homozygous GG genotype (P = 0.03, OR = 1.84, 95% CI = 1.05-3.23) and G allele (P = 0.02, OR = 1.43, 95% CI = 1.05-1.95). CONCLUSIONS: Our results showed that pharmacogenetic variants have important roles in epilepsy at different levels. It may be noted that multi-factorial diseases like epilepsy are also regulated by various other factors that may also be considered in the future.

Impact of AYUSH interventions on COVID-19: a protocol for a living systematic review and meta-analysis.

Background: The coronavirus disease 2019 (COVID-19) pandemic has created a great burden on governments and the medical fraternity globally. Many clinical studies from the Indian system of Traditional Medicines [Ayurveda, Yoga and Naturopathy, Unani, Siddha, and Homoeopathy (AYUSH)] have been carried out to find appropriate solutions. Through a living systematic review and meta-analysis, this study aims to determine the effectiveness of the Traditional System of Indian Medicine (AYUSH system) in lowering the incidence, duration, and severity of COVID-19. Methods: We will search the following databases: Pubmed; the Cochrane central register of controlled trials (CENTRAL); the Clinical Trials Registry - India (CTRI); Digital Helpline for Ayurveda Research Articles (DHARA); AYUSH research portal; Google scholar and World Health Organization (WHO) COVID-19 database. Clinical improvement, WHO ordinal scale, viral clearance, incidences of COVID-19 infection, and mortality will be considered as primary outcomes. Secondary outcomes will be use of O2 therapy or mechanical ventilator, admission to high dependency unit or emergency unit, duration of hospitalization, the time to symptom resolution, and adverse events. Data will be synthesized, with RevMan 5.4 tool and the risk of bias will be assessed with RoB 2 (for RCTs) and ROBINS I (for NRSIs). Certainty of evidence will be assessed through the GRADE (Grading of Recommendations, Assessment, Development and Evaluations) tool. The review will be updated bi-monthly with two updates. Conclusion: This living systematic review will be the first to address AYUSH interventions in COVID-19, synthesizing the full spectrum of Indian Traditional System of Medicine against COVID-19. It will facilitate professionals, guideline developers, and authorities with up to date synthesis on interventions periodically to make health-care decisions on AYUSH therapies in the management of COVID-19.