In silico studies of alkaloids and their derivatives against N-acetyltransferase EIS protein from Mycobacterium tuberculosis.

Antibiotic resistance against Mycobacterium tuberculosis (M.tb.) has been a significant cause of death worldwide. The Enhanced intracellular survival (EIS) protein of the bacteria is an acetyltransferase that multiacetylates aminoglycoside antibiotics, preventing them from binding to the bacterial ribosome. To overcome the EIS-mediated antibiotics resistance of M.tb., we compiled 888 alkaloids and derivatives from five different databases and virtually screened them against the EIS receptor. The compound library was filtered down to 87 compounds, which underwent additional analysis and filtration. Moreover, the top 15 most prominent phytocompounds were obtained after the drug-likeness prediction and ADMET screening. Out of 15, nine compounds confirmed the maximum number of hydrogen bond interactions and reliable binding energies during molecular docking. Additionally, the Molecular dynamics (MD) simulation of nine compounds showed the three most stable complexes, further verified by re-docking with mutated protein. The density functional theory (DFT) calculation was performed to identify the HOMO-LUMO energy gaps of the selected three potential compounds. Finally, our selected top lead compounds i.e., Alkaloid AQC2 (PubChem85634496), Nobilisitine A (ChEbi68116), and N-methylcheilanthifoline (ChEbi140673) demonstrated more favourable outcomes when compared with reference compounds (i.e., 39b and 2i) in all parameters used in this study. Therefore, we anticipate that our findings will help to explore and develop natural compound therapy against multi and extensively drug-resistant strains of M.tb.Communicated by Ramaswamy H. Sarma.

Assessment of Static and Dynamic Load Tolerance of the Implant Mounts on its Particular Implant.

Objectives: We compared the loads at which the implant holders from Astra Tech (AST) (AstraOsseoSpeed) and Osseotite Certain failed under static compression after experiencing fatigue, as well as the gap that resulted from dynamic loading between the implant-holder complexes. Materials and Method: The ISO 14801 recommendation served as the foundation for the test protocol. Each brand's five implant-implant holder assemblies underwent dynamic loading. A load of 200 N was applied at a stress frequency of 12 Hz and a cycle rate of 5105. (Eden Prairie, MN, USA). Using scanning electron microscopy (S3700N, HITACHI, Japan), the gap (m) at the interface was measured post-fatigue. Static loading was then used to determine the highest load (N) after the point of failure. Controls included definitive abutment-implant complexes. Statistics were used to analyze the data. Result: The Osseotite Certain group showed a slight trend toward greater resistance, but there was no diversity among the two implant holder groups (P 0.05). AST (AstraOsseoSpeed) implants had a larger interface gap, but the difference was not statistically significant. Conclusion: With respect to greatest compression load or the interface gap following dynamic loading, there were no discernible differences between the two experimental groups.

Complete genome resource unravels the close relation of an Indian Xanthomonas oryzae pv. oryzae strain IXOBB0003 with Philippines strain causing bacterial blight of rice.

Xanthomonas oryzae pv. oryzae (Xoo) is a pathogen of concern for rice growers as it limits the production potential of rice varieties worldwide. Due to their high genomic plasticity, the pathogen continues to evolve, nullifying the deployed resistance mechanisms. It is pertinent to monitor the evolving Xoo population for the virulent novel stains, and the affordable sequencing technologies made the task feasible with an in-depth understanding of their pathogenesis arsenals. We present the complete genome of a highly virulent Indian Xoo strain IXOBB0003, predominantly found in northwestern parts of India, by employing next-generation sequencing and single-molecule sequencing in real-time technologies. The final genome assembly comprises 4,962,427 bp and has 63.96% GC content. The pan genome analysis reveals that strain IXOBB0003 houses total of 3655 core genes, 1276 accessory genes and 595 unique genes. Comparative analysis of the predicted gene clusters of coding sequences and protein count of strain IXOBB0003 depicts 3687 of almost 90% gene clusters shared by other Asian strains, 17 unique to IXOBB0003 and 139 CDSs of IXOBB0003 are shared with PXO99A. AnnoTALE-based studies revealed 16 TALEs conferred from the whole genome sequence. Prominent TALEs of our strain are found orthologous to TALEs of the Philippines strain PXO99A. The genomic features of Indian Xoo strain IXOBB0003 and in comparison with other Asian strains would certainly contribute significantly while formulating novel strategies for BB management. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03596-x.

Vision centre helps reduce blindness and vision impairment. Proof of concept in a four-year longitudinal study in an urban area in India.

Purpose: To estimate the prevalence of blindness and severe visual impairment (SVI) by using a door-to-door screening and vision center (VC) examination strategy in an urban area in western Maharashtra (Pune), India and repeat the exercise after 4 years to study its impact. Methods: Four trained community health workers measured the visual acuity and performed an external ocular examination in patients' homes. People with vision <6/18 were requested to visit the VC for a comprehensive eye examination by an optometrist. An ophthalmologist examined people whose vision did not improve to 6/12. A home examination was done for people who did not visit the VC despite two requests. The same population was examined twice in an interval of 4 years. Results: In the study, 44,535 people in 2015-16 and 98.14% (n = 43,708) of them in 2018-19 were examined. Blindness (vision < 3/60 in better eye), and moderate-to-severe visual impairment (MSVI, vision 6/18-6/60 in better eye) were 0.26% and 1.3%, respectively, in the first cohort, and 0.16% and 1.1%, respectively, in the second cohort (P < 0.001). When the worse eye was considered, the prevalence of blindness reduced from 0.72% to 0.44%, SVI reduced from 0.1% to 0.07%, and MVI decreased from 1.7% to 1.49% between 2015 and 2019 (P < 0.001). Females (P < 0.001) and older individuals (P < 0.001) were more likely to have blindness or SVI. In the VC, 8211 people were examined in 4 years. Conclusion: The reduction of blindness and MSVI in the urban area of Pune can be partly ascribed to the presence of a VC and attendant screening in this locality.

Flavanones: A potential natural inhibitor of the ATP binding site of PknG of Mycobacterium tuberculosis.

Over the years, Mycobacterium tuberculosis has been one of the major causes of death worldwide. As several clinical isolates of the bacteria have developed drug resistance against the target sites of the current therapeutic agents, the development of a novel drug is the pressing priority. According to recent studies on Mycobacterium tuberculosis, ATP binding sites of Mycobacterium tuberculosis serine/threonine protein kinases (MTPKs) have been identified as the new promising drug target. Among the several other protein kinases (PKs), Protein kinase G (PknG) was selected for the study because of its crucial role in modulating bacterium's metabolism to survive in host macrophages. In this work, we have focused on the H37Rv strain of Mycobacterium tuberculosis. A list of 477 flavanones obtained from the PubChem database was docked one by one against the crystallized and refined structure of PknG by in-silico techniques. Initially, potential inhibitors were narrowed down by preliminary docking. Flavanones were then selected using binding energies ranging from -7.9 kcal.mol-1 to -10.8 kcal.mol-1. This was followed by drug-likeness prediction, redocking analysis, and molecular dynamics simulations. Here, we have used experimentally confirmed drug AX20017 as a reference to determine candidate compounds that can act as potential inhibitors for PknG. PubChem165506, PubChem242065, PubChem688859, PubChem101367767, PubChem3534982, and PubChem42607933 were identified as possible target site inhibitors for PknG with a desirable negative binding energy of -8.1, -8.3, -8.4, -8.8, -8.6 and -7.9 kcal.mol-1 respectively. Communicated by Ramaswamy H. Sarma.

Production of Plant Beneficial and Antioxidants Metabolites by Klebsiellavariicola under Salinity Stress.

Bacteria that surround plant roots and exert beneficial effects on plant growth are known as plant growth-promoting rhizobacteria (PGPR). In addition to the plant growth-promotion, PGPR also imparts resistance against salinity and oxidative stress and needs to be studied. Such PGPR can function as dynamic bioinoculants under salinity conditions. The present study reports the isolation of phytase positive multifarious Klebsiella variicola SURYA6 isolated from wheat rhizosphere in Kolhapur, India. The isolate produced various plant growth-promoting (PGP), salinity ameliorating, and antioxidant traits. It produced organic acid, yielded a higher phosphorous solubilization index (9.3), maximum phytase activity (376.67 ± 2.77 U/mL), and copious amounts of siderophore (79.0%). The isolate also produced salt ameliorating traits such as indole acetic acid (78.45 ± 1.9 µg/mL), 1 aminocyclopropane-1-carboxylate deaminase (0.991 M/mg/h), and exopolysaccharides (32.2 ± 1.2 g/L). In addition to these, the isolate also produced higher activities of antioxidant enzymes like superoxide dismutase (13.86 IU/mg protein), catalase (0.053 IU/mg protein), and glutathione oxidase (22.12 µg/mg protein) at various salt levels. The isolate exhibited optimum growth and maximum secretion of these metabolites during the log-phase growth. It exhibited sensitivity to a wide range of antibiotics and did not produce hemolysis on blood agar, indicative of its non-pathogenic nature. The potential of K. variicola to produce copious amounts of various PGP, salt ameliorating, and antioxidant metabolites make it a potential bioinoculant for salinity stress management.

Clinicoepidemiological study of skin manifestations in patients of chronic renal failure on hemodialysis.

BACKGROUND: Chronic renal failure (CRF) is associated with a variety of cutaneous manifestations as a result of underlying etiology as well as the various treatment modalities. AIM: To evaluate the prevalence of various dermatoses in patients with CRF on hemodialysis and to study the effect of hemodialysis on the intensity of pruritus. MATERIALS AND METHODS: A total of 35 patients of CRF on hemodialysis having at least one cutaneous manifestation were included in the study. RESULTS: Twenty-four (68.71%) cases in our study belonged to the age group of 50-69 years, out of which 16 cases were in the sixth decade. Xerosis and pruritus occurred in 80% and 65.71% of cases, respectively. Other common findings included pallor (68.57%), dyspigmentation (34.29%), cutaneous infections (34.39%), acquired perforating dermatosis (17.4%), and nail changes (60%). Hemodialysis failed to improve pruritus in 17 (73.9%) of our patients. Twenty-six patients (74.28%) suffered from hypertension, 13 of them also were known cases of type II diabetes mellitus. Five patients suffered exclusively from type II diabetes mellitus. CONCLUSIONS: In our small study, xerosis was the commonest finding and pruritus, the commonest symptom. The intensity of pruritus was largely unaffected by hemodialysis.

Differential migration of human monocyte-derived dendritic cells after infection with prevalent clinical strains of Mycobacterium tuberculosis.

Dendritic cells (DCs) play a key role in the host immune response to infections. Mycobacterium tuberculosis (MTB) can inhibit the maturation of DCs and impair their ability to stimulate T cell proliferation. Here, we assessed in vitro migratory behavior of human monocyte-derived DCs (MoDC) when infected with various MTB strains (H37Rv and prevalent clinical strains S7 and S10 from South India). The migration of Rv and S7 infected MoDC towards secondary lymphoid chemokine (CCL21) was 50% lower after 1 day of infection compared to LPS stimulation. This reduced cell migration may be due to a block in the chemokine receptor switch from CCR5 to CCR7 expression on MoDC. Only clinical strain S10 infected MoDC showed an up-regulation of CCR7 and down-regulation of CCR5 expression, similar to LPS stimulated MoDC. While Rv and S7 infected MoDC did not display any alteration in expression of these receptors. Similarly, Rv and S7 infected MoDC did not induce IL-8, IP-10 and MCP-1 chemokine production. This reduction in chemokine levels was reflected in the reduced chemoattraction of CD4(+) T cells also. These findings suggest that there is variation in the stimulation of MoDC with different clinical strains of MTB and this variation may be dependent upon the virulence of the strain.

Leptin response in patients with tuberculous pleuritis.

BACKGROUND & OBJECTIVES: Tuberculous pleuritis is used as a model to understand the protective immune response in tuberculosis. It is predominated by Th1 response at the site of infection, where a possible role for the leptin, a known enhancer of Th1 response, could be speculated. Hence, we investigated leptin levels in pleural effusions in patients with both tuberculous (TP) and non-tuberculous (NTP) pleural effusion. METHODS: Leptin and cytokine levels were assessed in serum and pleural fluid of TP and NTP patients (N = 20 each) by ELISA. Multivariate regression analysis were performed to find the possible determinants of leptin taking leptin as the dependent and body mass index (BMI), gender, source of leptin [i.e., serum or pleural fluid (PF)], age and disease status as independent variables. RESULTS: PF leptin levels were significantly higher than serum leptin levels in both the groups however the PF leptin levels were significantly lower in TP subjects compared to NTP. The results showed that the leptin was found to be dependent on BMI but not on the other parameters. However, regression analysis based on the source of leptin showed males to be a better predictor of leptin. No correlation was observed between leptin and measured immune parameters. INTERPRETATION & CONCLUSION: Our findings demonstrated that the decreased leptin levels were associated with reduction in BMI but not with the disease status in tuberculous pleuritis.

Plasmid distribution & antimicrobial susceptibility patterns of Acinetobacter genospecies from healthy skin of a tribal population in western India.

BACKGROUND & OBJECTIVE: Acinetobacter spp., the only Gram negative commensal found on human skin are emerging nosocomial pathogens, and are rapidly developing multiple drug resistance. Most studies in India and over the world are on clinical and environmental Acinetobacter isolates. The present investigation was undertaken to study plasmid distribution and antibiotic resistance profile of Acinetobacter genospecies isolated from healthy human skin of a tribal population from western India. METHODS: One hundred and eighteen strains of Acinetobacter spp. were isolated from healthy human skin and biochemically classified into nine genospecies. Plasmid profiles were determined using four standard methods with minor modifications and by a commercial kit. Molecular weights were determined by comparison with standard plasmids pUC19, pBR322, RP4 and molecular weight marker, V517. Antimicrobial susceptibility patterns of Acinetobacter genospecies to 30 antibiotics belonging to different groups was determined using Kirby Bauer method. RESULTS: All isolates, except two showed a maximum of three plasmids with molecular size 1.5 to 40 kb. Five different plasmid types were observed. Only eight strains had one or two stably maintained low molecular weight plasmids in addition to 40 kb plasmid seen in most of the strains. Quinolones and aminoglycosides were most active group followed by, cephalosporins and beta lactams. INTERPRETATION & CONCLUSION: Acinetobacter spp. isolated from the tribal skin flora had low number of plasmids as compared to clinical and environmental isolates. Acinetobacter spp. harbouring low molecular weight plasmids were found to be sensitive to most of the antibiotics tested suggesting these might be coding for some other novel properties or may be cryptic, whereas, those having 40 kb plasmid showed intermediate to low level resistance to some antibiotics.

Detection of rare beta-thalassemia mutations by denaturing gradient gel electrophoresis among Indians.

We report four rare beta-thalassemia (thal) mutations, viz. AATAAA --> AACAAA [polyadenylation (poly A) site mutation]. IVS-II-745 (C --> G), codon 121 (G --> T) and IVS-II-1 (G --> A), detected by denaturing gradient gel electrophoresis (DGGE) among Indians. Of these, the poly A site mutation has been found in combination with deletional delta(beta)-thal in one case, and with the IVS-1-5 (G --> C) mutation in another. Two DGGE patterns, corresponding to the same IVS-II-1 (G --> A) mutation, were seen in one family. Framework (FW) analyses in family studies have shown that the poly A site mutation is associated with FW-1, while both the codon 121 (G --> T) and IVS-II-1 (G --> A) mutations are associated with FW-2. Denaturing gradient gel electrophoresis facilitates the screening of rare beta-thal mutations in the diverse Indian population with its many ethnic groups, covering a vast geographic territory.

Prenatal diagnosis in a family at risk for beta-thalassemia and hemophilia A: an uncommon association.

beta-Thalassemia (thal) is an autosomal recessive disorder with a prevalence of 2-3% in Indians, while hemophilia A is X-linked with a prevalence of 1 in 5,000-10,000 male births. The chances of both these disorders being present together is extremely rare (1 in 250,000). We report an interesting consanguineous family from Western India with a combination of these two disorders, which was referred to us for prenatal diagnosis.

Usefulness of automated chromatography for rapid fetal blood analysis for second trimester prenatal diagnosis of beta-thalassemia.

Prenatal diagnosis of beta-thalassemia is now ideally done in the first trimester of pregnancy by chorionic villus tissue DNA analysis. Nevertheless, fetal blood analysis in the second trimester is required either when the mutation in both parents cannot be characterised or when the couple comes late for investigations. We evaluated the usefulness of analysis of fetal blood on the Biorad Variant Hemoglobin Testing System using the beta-thalassemia short programme in comparison with the conventional globin biosynthesis in 58 pregnancies. The beta/alpha biosynthesis ratios in 13 homozygous fetuses ranged from 0 to 0.03 and the adult hemoglobin (HbA) levels by automated chromatography varied from 0% to 0.4%. The normal or heterozygous fetuses had beta/alpha ratios of >0.04 and HbA levels ranging from 2.1% to 10.6%. In 17 fetuses we also correlated the beta gene mutations with the predicted genotypes using automated high-performance liquid chromatography (HPLC). Follow-up of 18 unaffected fetuses using the Variant System at birth showed a significant increase in HbA levels.

Growth inhibition of human promonocytic leukaemic U937 cells by interferon gamma is irreversible and not cell cycle phase-specific.

Growth of human promonocytic leukaemic U937 cells was found arrested within 24 h upon exposure to interferon gamma (IFN-gamma). Removal of the interferon did not result in the resumption of growth, as is evident from the absence of doubling of viable cell count and(3)H-thymidine incorporation. 5-Bromo-2'-deoxyuridine-based flow cytometric analysis of the growth-arrested cells, 24 h subsequent to the removal of IFN-gamma, showed absence of DNA synthesis, confirming the irreversible nature of the growth inhibition. Propidium iodide-based flow cytometric analysis of the growth-arrested cells showed a distribution which is typical of a growth inhibition without resulting in the accumulation of cells in any specific phase of the cell cycle. These results indicated that IFN-gamma arrested growth of U937 cells in an irreversible and cell cycle phase-independent manner. These observations were in contrast to our earlier report on the reversible and cell cycle phase-specific growth inhibition of human amniotic (fetal epithelial) WISH cells by the interferon.

IFN-gamma inhibits growth of WISH cells in a cell cycle phase-specific manner.

Treatment of WISH (human amnion) cells with interferon-gamma (IFN-gamma) inhibits their growth. Release of the cells from IFN-gamma-mediated growth inhibition led to a rapid and significant increase in DNA synthesis, followed by doubling of cell numbers. The DNA synthesis profile was strikingly similar to that shown by WISH cells released from growth arrest by the G1/S phase inhibitor, aphidicolin. This strongly suggested that IFN-gamma treatment leads to growth inhibition of WISH cells at the G1/S boundary of the cell cycle. In contrast, IFN-alpha blocked growth of these cells at the G0/G1 boundary.