Genetic screening and metabolomics identify glial adenosine metabolism as a therapeutic target in Parkinson's disease.

Parkinson's disease (PD) is the second most common neurodegenerative disorder and lacks disease-modifying therapies. We developed a Drosophila model for identifying novel glial-based therapeutic targets for PD. Human alpha-synuclein is expressed in neurons and individual genes are independently knocked down in glia. We performed a forward genetic screen, knocking down the entire Drosophila kinome in glia in alpha-synuclein expressing flies. Among the top hits were five genes (Ak1, Ak6, Adk1, Adk2, and awd) involved in adenosine metabolism. Knockdown of each gene improved locomotor dysfunction, rescued neurodegeneration, and increased brain adenosine levels. We determined that the mechanism of neuroprotection involves adenosine itself, as opposed to a downstream metabolite. We dove deeper into the mechanism for one gene, Ak1, finding rescue of dopaminergic neuron loss, alpha-synuclein aggregation, and bioenergetic dysfunction after glial Ak1 knockdown. We performed metabolomics in Drosophila and in human PD patients, allowing us to comprehensively characterize changes in purine metabolism and identify potential biomarkers of dysfunctional adenosine metabolism in people. These experiments support glial adenosine as a novel therapeutic target in PD.

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.

Establishment of a 6-OHDA Induced Unilaterally Lesioned Male Wistar Rat Model of Parkinson's Disease.

Robust preclinical models of Parkinson's disease (PD) are valuable tools for understanding the biology and treatment of this complex disease. 6-Hydroxydopamine (6-OHDA) is a selective catecholaminergic drug injected into the substantia nigra pars compacta (SNc), medial forebrain bundle (MFB), or striatum, which is then metabolized to induce parkinsonism. Unilateral injection of 6-OHDA produces loss of dopaminergic (DAergic) neurons on the injected side with a marked motor asymmetry known as hemiparkinsonism, typically characterized by a rotational behavior to the impaired side. The present work describes a stable unilateral 6-OHDA-lesioned rat model of PD. 6-OHDA was administered into the MFB, leading to the consistent loss of striatal dopamine (DA) and behavioral imbalance in unilateral 6-OHDA-lesioned rats to establish the model of PD. This model of PD is a valuable tool for understanding the mechanisms underlying the generation of parkinsonian symptoms.

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.

The Trend of C-Reactive Protein After Corticosteroid Therapy in COVID-19 Patients Admitted to IGIMS, Patna.

BACKGROUND: C-reactive protein (CRP) is a routine inflammation biomarker. Increased CRP levels are correlated with COVID-19. We found a marked reduction in CRP concentration on corticosteroid therapy, which in turn led to reduced mortality and duration of hospital stay. METHODS: In this retrospective cohort study, CRP levels were measured on admission and at 72 hours and compared between two groups of patients, with and without corticosteroid therapy. The study sample consisted of 105 RT-PCR-confirmed patients admitted to the ICU of the COVID ward. Out of the total patients, 57 received one or more doses of dexamethasone in addition to usual treatment, and 48 were given only usual care. RESULT: CRP at the time of admission was comparable for both groups. Also, a significant decrease in the CRP was noted in both groups 72 hours post-admission. Moreover, the decline was more marked in the steroid-administered group (CRP-baseline: 34.3mg/dL (+/-8.44), CRP at 72 hours 18.5mg/dL(+/-7.95) (p <0.00) compared to non-steroid group (CRP_baseline: 34.04mg/dL (+/-10.06), CRP at 72. Those with comorbidities were administered steroids (n=38, 66.7%) compared to those who were not (n=08, 16.7%). The average duration of hospital stay was less (5 to 7 days) in the corticosteroid-administered group compared to the other group (7 to 10 days). CONCLUSION: Routine CRP tests can predict the outcome and treatment of severe coronavirus disease. Corticosteroid treatment in COVID-19 patients is associated with reduced CRP levels within 72 hours after therapy.

Epidemiological and microbiological profiles of microbial keratitis in a tertiary eye center in Eastern India (Bihar).

Purpose: To determine the demography, risk factors, and causative organisms of microbial keratitis (MK) in Bihar, an eastern state of India. Design: Retrospective study. Methods: We reviewed the demographic, clinical, and microbiological data of 2303 patients with MK (non-viral) presenting between January 2019 and December 2022. Results: This study revealed a predominance of males (65.0%) compared to females (34.9%), with a mean age of 48.4 ± 16.5 years. The majority of patients (63.1%) presented after 2 weeks from the onset of symptoms. The most common risk factor observed was corneal injury (58.1%), followed by ocular surface diseases (13.6%) and diabetes mellitus (13.3%). The majority of patients (73.16%) were involved in agriculture. Prior to presentation, almost all patients (92%) had received topical antibiotics. Unsupervised use of topical corticosteroids was observed in 29.2% of the patients for the median duration of 3 days (odds ratio, 0.17). At presentation, the median size of corneal ulcers was 5 mm, the best-corrected visual acuity was less than 20/400 in 51.4% of patients, and corneal perforation was in 14% of patients. The smear and culture positivity rate were 75.4% and 47.9%, respectively. The common causative organism was fungus (48.8%), followed by bacteria (17.4%). Aspergillus spp. and Staphylococcus spp. were the most commonly identified organisms; a quarter of the patients (24.5%) remained unidentified. All bacteria showed good sensitivity to vancomycin. Conclusion: MK is a significant cause of ocular morbidity in Bihar. The knowledge of epidemiology, risk factors, and microbiological profiles of MK can provide a valuable approach to disease prevention, diagnosis, and management.

A Comparative Analysis of Microscopy, Culture, and the Xpert Mycobacterium tuberculosis/Rifampicin Assay in Diagnosing Pulmonary Tuberculosis in Human Immunodeficiency-Positive Individuals.

INTRODUCTION: Individuals with human immunodeficiency virus (HIV) infection have an increased likelihood of developing tuberculosis (TB). The primary objective of this study was to compare the diagnostic accuracy of microscopy, culture, and the Xpert Mycobacterium tuberculosis/Rifampicin (MTB/RIF) assay in the diagnosis of pulmonary TB in sputum samples of HIV-infected patients. The secondary objectives were to evaluate the sensitivity and specificity of these three methods along with a comparison of diagnostic approaches for detecting drug-resistant strains. MATERIAL AND METHODS: This prospective, laboratory-based study was done in the Microbiology Department of IGIMS, Patna. The study included sputum samples of 102 individuals who were HIV-positive and exhibited symptoms indicative of tuberculosis. RESULTS: Out of 102 individuals suspected of having tuberculosis, 18 tested positive for M. tuberculosis. Male individuals between the ages of 31 and 40 were more affected by both HIV and tuberculosis, and in most of these cases, their CD4 cell count was below 200 cells/µl. Among the 102 sputum samples collected, 18% (18 samples) were found to be positive using the Mycobacterium Growth Indicator Tube (MGIT) 960 liquid culture method. Two samples were contaminated, and 14.7% (15 cases) tested positive using the cartridge-based nucleic acid amplification test (CBNAAT) method. Additionally, 3.92% (four samples) were positive using the ZN staining method. CONCLUSION: The study found that Xpert MTB/RIF outperformed other methods in identifying resistance to RIF, showed better agreement with gene sequencing results for RIF resistance, and had higher accuracy in detecting tuberculosis cases, including both smear-positive and smear-negative cases.

Characterization of Genetic Mutations in Multi-Drug-Resistant Isolates of Mycobacterium tuberculosis Bacilli Conferring Resistance to a Second-Line Anti-tuberculosis Drug.

INTRODUCTION: Multi-drug-resistant tuberculosis (MDR-TB) has become a major public health concern globally. Mutations in first- and second-line drug targets such as katG, inhA, rpoB, rrs, eis, gyrA, and gyrB have been associated with drug resistance. Monitoring predominant mutations in the MDR-TB patient population is essential to monitor and devise future therapeutic regimes. The present study is aimed to characterize genetic mutations in MDR isolates of Mycobacterium tuberculosis (MTB) bacilli conferring resistance to a second-line anti-tuberculosis drug in the Eastern Indian population. METHODS: This cross-sectional study was conducted in the Department of Microbiology, Indira Gandhi Institute of Medical Sciences, Patna, Bihar, and in the Tuberculosis Demonstration & Training Centre, Agamkuan, Patna. A total of 3270 patients suspected to have MDR-TB were recruited in the study. Two sputum samples, one on the spot, and the other in the morning were collected from each patient and the diagnosis of rifampicin-sensitive (RS)/rifampicin-resistant (RR/MDR) TB was done by Gene-Xpert test. One hundred fifty RS-TB samples and 150 RR/MDR-TB samples were considered for line probe assay (LPA). RS samples were subjected to first-line LPA using Genotype® MTBDR Plus ver 2.0 and RR/MDR samples were considered for second-line LPA using Genotype® MTBDRsl ver 2.0. All sputum samples were subjected to sputum smear microscopy using the Ziehl-Neelsen staining method. Statistical analysis was done using Statistical Package for Social Sciences (SPSS) version 26.0 (IBM Corp. Armonk, NY) and R (version 4.1; R Core Team 2021). RESULTS: In the present study, out of 3270 patients, we detected RR/MDR-TB in 235 patients (7.19%), RS-TB in 812 patients (24.83%), the rest of the patients negative for MTB (2223, 67.98%). Out of 150 RR/MDR-TB sputum samples tested, resistance to fluoroquinolone (FQ) was observed in 41 samples. The selected patients had predominantly FQ resistance due to the gyrA gene mutations (97.56%, n=40) compared to the gyrB gene mutations (2.44%, n=1). We observed >60% of the mutations in the gyrA gene in codon 94 (MUT3C (D94G), MUT3A (D94A), and MUT3D (D94H). In addition, we found the mutations MUT1 (A90V) and MUT2 (S91P) in the codons 90 and 91 of the gyrA gene in the considered MTB patient population. CONCLUSION: The identified genes can be further validated to be considered as therapeutic targets, but more therapeutics and advanced strategies should be applied in the management of MTB.

A Comparative Study of Bacterial Infections Between COVID-19 and Non-COVID-19 Patients With Respect to Different Isolates and Their Antibiotic Sensitivity Pattern.

Background Following the pandemic caused by SARS-CoV-2, the emergence of and following the pandemic has required major modifications to healthcare systems and frameworks. Antimicrobials have more than a few potential roles in managing COVID-19. Experimental cures for the treatment of SARS-CoV-2 are being explored. The availability of limited data suggests that nosocomial infections are associated with a higher risk of death and severity of COVID-19. To fill this knowledge gap, we conducted a study to assess the spectrum of bacteriological isolates in different samples from COVID-19 patients. Our study aimed to evaluate the antibiotic resistance pattern of these bacterial isolates and compare the spectrum of bacteriological isolates in different samples and their antibiotic sensitivity pattern between COVID-19 and non-COVID-19 patients. Methodology An observational cross-sectional, partly retrospective, and partly prospective study was carried out in the bacteriology section of the Department of Microbiology, Indira Gandhi Institute of Medical Sciences (IGIMS), Patna, Bihar, for a total duration of six months from February 2021 to July 2021. The profile of pathogens isolated from 105 clinical samples from COVID-19 patients was studied. To detect COVID-19, RT-PCR was performed. All clinical specimens (urine, blood, pus, respiratory sample, etc.) were processed and cultured on different media to support the growth of the bacteria as per our standard operating procedures (SOP) for bacteriology samples. Antimicrobial susceptibility testing was carried out based on the Clinical and Laboratory Standards Institute. Results A total of 105 clinical samples were received in the bacteriology section of the Department of Microbiology, IGIMS, Patna, Bihar, from admitted COVID-19 patients. The mean age of study participants was 51.57 ± 14.76 years, and males (66.7%, 70/105) were more than females (33.3%, 35/105). The majority of the patients were 91 out of 105 (86.67%) from the ward and 14 from the ICU (13.33%). Of the total samples tested, 62 (59%) were urine samples, 26 (24.8%) were respiratory specimens, 13 (12.4%) were pus samples, 3 (2.9%) were body fluids, and 1 (1%) were tissue samples. Among the total pathogen isolates (n=57) obtained from patients with SARS-CoV-2 admitted to the ward and ICU, 56.14% (32) were gram-negative, 26.31% (15) were Candida, and 17.54% (10) were gram-positive pathogens. The most isolated pathogen was Escherichia coli (39.02%, 16/41) followed by Klebsiella pneumoniae (29.26%, 12/41), Acinetobacter baumannii (7.31%, 3/41), and Enterobacter cloacae (2.43%, 1/41). Enterococcus spp. as gram-positive bacteria were isolated in 21.95% (9/41) of patients. Among the gram-negative bacteria (Enterobacterales), the highest resistance was seen in ampicillin (100%,29/29). For non-Enterobacterales, the highest resistance was seen in ceftriaxone (66.66%,2/3). Enterococcus spp. showed maximum ciprofloxacin, and gentamicin (high level) resistance was 100% (9/9). Conclusion Secondary infections with resistant pathogens in COVID-19 patients highlight the importance of antimicrobial management programs focused on the optimal selection of empirical treatments based on culture reports.

Post-COVID-19 cavitary lung lesion due to Aspergillus flavus and Enterobacter cloacae in a patient suffering from COVID-19 pneumonia - a case report.

The coronavirus disease 2019 (COVID-19) pandemic has manifested as a multifaceted paradigm but has primarily affected the respiratory system. Though a rare sequela after-COVID-19, we present a case of cavitary lung lesion in an adult patient, which manifested with common symptoms such as fever, cough and dyspnoea during the post-COVID-19 recovery period. Aspergillus flavus and Enterobacter cloacae were found to be the main causative organisms. Fungal and bacterial coinfection may be thought of in similar situations and appropriate treatment may be given to prevent further morbidity and mortality.

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.

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.

Design and synthesis of benzo[d]thiazol-2-yl-methyl-4-(substituted)-piperazine-1-carbothioamide as novel neuronal nitric oxide inhibitors and evaluation of their neuroprotecting effect in 6-OHDA-induced unilateral lesioned rat model of Parkinson's disease.

Neuronal nitric oxide synthase (nNOS) is an enzyme constitutively expressed in the mammalian brain and skeletal muscles. The excessive activation of nNOS in the neurons results in oxidative and nitrosative stress associated with neuronal loss in various neurological disorders. Several nNOS inhibitors have been reported to limit the excessive activation of nNOS. In the present work, we have designed and carried the synthesis of benzo[d]thiazol-2-yl-methyl-4-(substituted)-piperazine-1-carbothioamide as novel neuronal nitric oxide inhibitors (5-28, twenty-four compounds). Stably transfected HEK 293 cells expressing NOS isoforms treated with the compounds (5-28) showed that the eight compounds exhibited > 95% cell survival in the MTT assay. nNOS inhibition assay of the eight compounds illustrated that the compound 18 was most selective for nNOS (nNOS=66.73 ± 1.51; eNOS=28.70 ± 1.39; iNOS =13.26 ± 1.01) in HEK 293 cells expressing NOS isoforms. 6-OHDA-induced unilaterally lesioned rats treated with the compound 18 showed the improvement in motor and non-motor functions. Furthermore, the compound 18 showed the increased levels of dopamine and decreased levels of glutamate and nitrite ions in the isolated rat brain. In the docking analysis, the compound 18 showed the significant binding affinity with the nNOS binding site (the ∆G value = - 9.0 kcal/mol). Overall results demonstrated that the N-(benzo[d]thiazol-2-ylmethyl)-4-(4-nitrophenyl) piperazine-1-carbothioamide (the compound 18) possessed significant nNOS inhibiting activity and neuroprotecting potential in 6-OHDA-induced unilaterally lesioned rat model of PD and more work will be required to establish the role of the compound 18 in the therapy of PD and other neurodegenerative disorders.

Candidemia Among ICU Patients: Species Characterisation, Resistance Pattern and Association With Candida Score: A Prospective Study.

Introduction Candidiasis is a significant cause of morbidity and mortality in immunocompromised patients admitted in intensive care units. Identification of Candida species is essential for effective treatment. However, in absence of proven fungemia, guidelines to initiate therapy are yet to be defined. Materials and methods During the study (16 months: September 2018 to December 2019), samples (urine, sputum, blood, tracheal aspirate, urinary catheter) were collected from ICU patients and prospectively evaluated. Microscopy, culture, and antifungal susceptibility testing were performed as per standard laboratory protocol. Demographic details and risk factors were noted from case records and correlated with Candida score. Results One hundred twenty-five non-duplicate samples (120 patients) positive on culture were included in the study. The most common co-morbid condition associated with fungemia was diabetes mellitus. The most common risk factor was total parenteral nutrition. Non-albicansCandida(C. tropicalis) was predominant. Candida species showed good sensitivity to voriconazole (80%) followed by fluconazole (67.78%) and amphotericin (62.22%). Twenty-nine patients had a Candida score of more than three. Conclusion Fluconazole available in both oral and parenteral formulations is an effective antifungal agent against the candida spp. Voriconazole should be reserved for non-responders. Rising resistance to common antifungals among Candida albicans is a matter of concern.

Rationalizing the Use of Polyphenol Nano-formulations in the Therapy of Neurodegenerative Diseases.

Neurodegenerative diseases are a heterogeneous group of disorders among aging populations worldwide characterized by the progressive degeneration of the structure and function of brain cells and the nervous system. Alzheimer's disease and Parkinson's disease are common neurodegenerative diseases (NDs). Classic pathological features of AD are the accumulation of the amyloid betaprotein and aggregates of hyperphosphorylated tau protein around the brain cells. Dopaminergic neuronal death in the midbrain and accumulation of α- synuclein in the neurons are the hallmark of Parkinson's disease. The pathogenesis is multifactorial, and both neurodegenerative disorders have complex etiology. Oxidative stress closely linked with mitochondrial dysfunction, excitotoxicity, nitric oxide toxicity, and neuro-inflammation, is anticipated to trigger neuronal death. Ample evidence has implicated that oxidative stress and inflammation contribute to the pathology of neurodegeneration in AD and PD. Currently, acetylcholinesterase inhibitors are the main treatment option for AD, while L-DOPA is the gold standard therapy for PD. Along with the main therapy, many endogenous antioxidants, like vitamin E, selenium, etc., are also given to the patients to combat oxidative stress. Current treatment for these NDs is limited due to the blood-brain barrier (BBB) that hinders drug targeting towards neurons. In this review, we emphasize adjunct treatment with anti-inflammatory agents that act at the site of the disease and can halt the disease progression by attenuating the effect of ROS triggering neuro-inflammatory response. Polyphenols, either as purified compounds or extracts from various natural plant sources, have been well studied and documented for anti-inflammatory effects, but their use for ND is limited due to their physicochemical attributes. Nanoparticle-mediated drug delivery system exhibits immense potential to overcome these hurdles in drug delivery to the CNS, enabling nanoparticle-based therapies to directly target the inflammation and release bioactive compounds with anti-inflammatory properties to the site of action.

Genome-wide identification of carbapenem-resistant Gram-negative bacterial (CR-GNB) isolates retrieved from hospitalized patients in Bihar, India.

Carbapenemase-producing clinical isolates are becoming more common over the world, posing a severe public health danger, particularly in developing nations like India. Carbapenem-resistant Gram-negative bacterial (CR-GNB) infection has become a fast-expanding global threat with limited antibiotic choice and significant mortality. This study aimed to highlight the carbapenem-resistance among clinical isolates of hospital admitted patients in Bihar, India. A cross-sectional study was conducted with 101 clinical isolates of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa. All GNB isolates were tested for their antimicrobial susceptibility using Kirby-Bauer disc diffusion method. Double disc synergy test / modified Hodge test (DDST/MHT) were used to detect carbapenemase production by these isolates. Subsequently, these isolates were evaluated for carbapenem-resistance genes using whole-genome sequencing method. The overall percentage of carbapenem-resistance among GNB was (17/101) 16.8%. The genomic analysis of antimicrobial-resistance (AMR) demonstrates a significantly high prevalence of blaCTX-M followed by blaSHV, blaTEM, blaOXA, and blaNDM ß-lactam or carbapenem resistance genes among clinical isolates of GNB. Co-occurrence of blaNDM with other beta-lactamase-encoding genes was found in 70.6% of carbapenemase-producing isolates. Our study highlights the mechanism of carbapenem-resistance to curb the overwhelming threat posed by the emergence of drug-resistance in India.

Development of novel carbazole derivatives with effective multifunctional action against Alzheimer's diseases: Design, synthesis, in silico, in vitro and in vivo investigation.

Carbazole based novel multifunctional agents has been rationally designed and synthesized as potential anti-Alzheimer agents. Multi-functional activity of these derivatives have been assessed by performing various in-vitro assays and these compounds appeared to be potent AChE inhibitors, Aß aggregation inhibitors, anti-oxidant and neuroprotective agents. Among the entire series, MT-1 and MT-6 were most potent multifunctional agents which displayed effective and selective AChE inhibition, Aß disaggregation, anti-oxidant and metal chelation action. Neuroprotective activity of MT-6 has been examined against H2O2 induced toxicity in SHSY-5Y cells and they have shown effective neuroprotection. Additionally, MT-6 did not display any significant toxicity in SHSY-5Y cells, indicating its non-toxic nature. Molecular docking and MD simulation studies have been also performed to explore molecular level interaction with AChE and Aß. Finally, MT-6 was evaluated against scopolamine induced dementia model of mice and this compound actively improved memory deficit and cognition impairment in scopolamine treated mice. Thus, novel carbazole derivative MT-6 has been explored as an effective and safe multifunctional agent against AD and this molecule may be used as a suitable lead for development of effective anti-Alzheimer agents in future.

Biofuel and co-products from algae solvent extraction.

This study reports a novel method of using algae biomass as a source of lipid and various other co-products. Solvent-based extraction techniques could yield a number of products simultaneously. Further, the study focuses on all possible characterization and utilization of the three layers obtained from chloroform-methanol extraction of lipids. The lipid from the chloroform layer was transesterified for Fatty acid methyl esters (FAME) production. The fatty acid methyl esters derived by oleic acid, palmitic acid, linoleic acid and phytol were majorly analysed by GC-MS. The methanol layer was analysed with HPLC and stachyose, maltotriose, glucose, fructose, acetic acid, butyric acid, DMSO, glycerol were identified. The cell debris was further physically activated, and physiochemical properties of raw algae, residual algae and algae bio-char were compared. Spectrum peaks of FTIR study identified many alkyl‒halide stretches. Similarly, EDX analysed the presence of carbon, nitrogen, oxygen, potassium, chlorine, calcium, iron, magnesium and phosphorus. The SEM reveals that residual algae was comparatively crystalline and hence could not be utilized directly as an adsorbent. Therefore, further physical treatment was applied, and methylene blue dye adsorption study was also conducted to know the time and capacity of biochar as an adsorbent. However, organic and mineral enriched biomass could be used directly as fertilizer for agricultural purposes.

Neuroprotective effect of IDPU (1-(7-imino-3-propyl-2,3-dihydrothiazolo [4,5-d]pyrimidin-6(7H)-yl)urea) in 6-OHDA induced rodent model of hemiparkinson's disease.

Parkinson's disease (PD) is a progressive neurodegenerative motor disorder characterized by the degeneration of dopaminergic nigrostriatal neurons. Levodopa (l-DOPA) is the most effective therapy for PD, however, PD progression continues with significant side effects in long term, thus necessitating the search for effective therapy that impedes PD progression. PD therapy through non-dopaminergic pathways offers treatment without the risk of extrapyramidal effects. In this regard, earlier, we had reported, a novel compound IDPU with potential adenosine A2A receptor antagonist effect in haloperidol (chronic treatment) induced Parkinson model. In the present study, we extended our investigation towards i) evaluation of IDPU in well-established 6-OHDA induced Parkinson rat model to establish its role in the therapy of PD ii) its function in alleviating the neuronal loss. We carried the IDPU administration (i.p.) in rats for two weeks after establishing 6-OHDA induced unilateral lesions. The behavioral activity, neurochemical alteration, oxidative stress marker and tyrosine hydroxylase positive neurons in substantia nigra were analyzed. The results showed that IDPU significantly reduced motor and non-motor deficits induced by 6-OHDA in the behavioral tasks such as apomorphine, rota rod and force swim test. Furthermore, the results of oxidative stress biomarkers revealed that IDPU successfully modulated oxidative stress associated biomarkers such as MDA, catalase, superoxide dismutase, nitric oxide and reduced glutathione level. Additionally, IDPU significantly elevated intracellular dopamine, decreased glutamate and calcium levels in brain as compared to 6-OHDA alone treated animals which is evocative of its neuroprotective behavior. Thus, the investigations clearly validated IDPU as a potent anti-parkinsonian agent which showed immense capability to protect neurodegeneration.

Pharmacological assessments of potent A2A receptor antagonist IDPU (1-(7-imino-3-propyl-2,3-dihydrothiazolo[4,5-d]pyrimidin-6(7H)-yl)urea) in rodent model of haloperidol induced Parkinson like symptoms.

A2A receptor antagonists emerged as potential candidate for management of Parkinson's disease. Earlier we had reported the therapeutic potential of 1-(7-imino-3-propyl-2,3-dihydrothiazolo[4,5-d]pyrimidin-6(7H)-yl) urea (IDPU) as A2A receptor antagonist. Herein, we have investigated the effect of IDPU in attenuation of haloperidol induced Parkinson like symptoms in rats. It has successfully restored hypo-locomotion induced by haloperidol and NECA. IDPU also displayed protective effect against oxidative stress induced by chronic haloperidol treatment in rats. The antidepressant activity of IDPU was determined in mice showed that it imperatively reduced depression like symptoms in well-established depression models viz. TST and FST. Additionally, IDPU was found to be a safe and non-toxic chemical entity in acute, sub-acute and neurotoxicity studies. In silico study of IDPU showed acceptable physicochemical parameters and in vitro screening exhibited satisfactory metabolic stability. This study clearly indicates that A2A receptor antagonist IDPU is able to ameliorate Parkinsonian symptoms without exerting any significant toxicity.