Mycobacterium tuberculosis Acetyltransferase Suppresses Oxidative Stress by Inducing Peroxisome Formation in Macrophages.

Mycobacterium tuberculosis (Mtb) inhibits host oxidative stress responses facilitating its survival in macrophages; however, the underlying molecular mechanisms are poorly understood. Here, we identified a Mtb acetyltransferase (Rv3034c) as a novel counter actor of macrophage oxidative stress responses by inducing peroxisome formation. An inducible Rv3034c deletion mutant of Mtb failed to induce peroxisome biogenesis, expression of the peroxisomal ß-oxidation pathway intermediates (ACOX1, ACAA1, MFP2) in macrophages, resulting in reduced intracellular survival compared to the parental strain. This reduced virulence phenotype was rescued by repletion of Rv3034c. Peroxisome induction depended on the interaction between Rv3034c and the macrophage mannose receptor (MR). Interaction between Rv3034c and MR induced expression of the peroxisomal biogenesis proteins PEX5p, PEX13p, PEX14p, PEX11ß, PEX19p, the peroxisomal membrane lipid transporter ABCD3, and catalase. Expression of PEX14p and ABCD3 was also enhanced in lungs from Mtb aerosol-infected mice. This is the first report that peroxisome-mediated control of ROS balance is essential for innate immune responses to Mtb but can be counteracted by the mycobacterial acetyltransferase Rv3034c. Thus, peroxisomes represent interesting targets for host-directed therapeutics to tuberculosis.

Delineating FtsQ-mediated regulation of cell division in Mycobacterium tuberculosis.

Identifying and characterizing the individual contributors to bacterial cellular elongation and division will improve our understanding of their impact on cell growth and division. Here, we delineated the role of ftsQ, a terminal gene of the highly conserved division cell wall (dcw) operon, in growth, survival, and cell length maintenance in the human pathogen Mycobacterium tuberculosis (Mtb). We found that FtsQ overexpression significantly increases the cell length and number of multiseptate cells. FtsQ depletion in Mtb resulted in cells that were shorter than WT cells during the initial growth stages (4 days after FtsQ depletion) but were longer than WT cells at later stages (10 days after FtsQ depletion) and compromised the survival in vitro and in differentiated THP1 macrophages. Overexpression of N- and C-terminal FtsQ regions altered the cell length, and the C-terminal domain alone complemented the FtsQ depletion phenotype. MS analyses suggested robust FtsQ phosphorylation on Thr-24, and although phosphoablative and -mimetic mutants rescued the FtsQ depletion-associated cell viability defects, they failed to complement the cell length defects. MS and coimmunoprecipitation experiments identified 63 FtsQ-interacting partners, and we show that the interaction of FtsQ with the recently identified cell division protein SepIVA is independent of FtsQ phosphorylation and suggests a role of FtsQ in modulating cell division. FtsQ exhibited predominantly septal localization in both the presence and absence of SepIVA. Our results suggest a role for FtsQ in modulating the length, division, and survival of Mtb cells both in vitro and in the host.

Operando X-ray absorption spectroscopy study of the Fischer-Tropsch reaction with a Co catalyst.

This article describes the setting up of a facility on the energy-scanning EXAFS beamline (BL-09) at RRCAT, Indore, India, for operando studies of structure-activity correlation during a catalytic reaction. The setup was tested by operando X-ray absorption spectroscopy (XAS) studies performed on a Co-based catalyst during the Fischer-Tropsch reaction to obtain information regarding structural changes in the catalyst during the reaction. Simultaneous gas chromatography (GC) measurements during the reaction facilitate monitoring of the product gases, which in turn gives information regarding the activity of the catalyst. The combination of XAS and GC techniques was used to correlate the structural changes with the activity of the catalyst at different reaction temperatures. The oxide catalyst was reduced to the metallic phase by heating at 400°C for 5 h under H2 at ambient pressure and subsequently the catalytic reaction was studied at four different temperatures of 240, 260, 280 and 320°C. The catalyst was studied for 10 h at 320°C and an attempt has been made to understand the process of its deactivation from the XANES and EXAFS results.

Water-assisted stability of carbene: cyclic voltammetric investigation of 1-ethyl-3-methylimidazolium ethylsulfate ionic liquid.

In this work, we report electrochemical studies on imidazolium-based ionic liquids with an objective to explore the possibility of carbene formation in their dilute aqueous solutions. Conventionally, water plays a detrimental role during investigations involving ionic liquids, and this role has been investigated via electrochemical studies in aqueous ionic liquid solutions. There are varying opinions regarding the influence of water on the physicochemical behaviour of ionic liquids that require an in-depth understanding. To eludicate the role of water, we attempted to evaluate the electrochemical performance of ionic liquids in water as a solvent, and the influence of water on ionic liquids was explored through feasibility and stability studies on carbene formed in an aqueous imidazolium-based ionic liquid solution. The electrochemical investigation of an aqueous solution of 1-ethyl-3-methylimidazolium ethylsulfate ([EMIM][EtSO4]) revealed a redox couple. Detailed investigations suggest that reduction of the imidazolium cation occurs at the C2 position, with subsequent formation of carbene. Furthermore, an anodic peak was found to be associated with the oxidation of carbene. The coulometric process associated with the anodic peaks indicated that the two-electron oxidation of carbene occurred. The stability of carbene in water was evaluated through the use of different protic and aprotic solvents. The hydrogen bond-forming ability of carbene with water seems to be responsible for its improved stability in water.

Protein kinase G confers survival advantage to Mycobacterium tuberculosis during latency-like conditions.

Protein kinase G (PknG), a thioredoxin-fold-containing eukaryotic-like serine/threonine protein kinase, is a virulence factor in Mycobacterium tuberculosis, required for inhibition of phagolysosomal fusion. Here, we unraveled novel functional facets of PknG during latency-like conditions. We found that PknG mediates persistence under stressful conditions like hypoxia and abets drug tolerance. PknG mutant displayed minimal growth in nutrient-limited conditions, suggesting its role in modulating cellular metabolism. Intracellular metabolic profiling revealed that PknG is necessary for efficient metabolic adaptation during hypoxia. Notably, the PknG mutant exhibited a reductive shift in mycothiol redox potential and compromised stress response. Exposure to antibiotics and hypoxic environment resulted in higher oxidative shift in mycothiol redox potential of PknG mutant compared with the wild type. Persistence during latency-like conditions required kinase activity and thioredoxin motifs of PknG and is mediated through phosphorylation of a central metabolic regulator GarA. Finally, using a guinea pig model of infection, we assessed the in vivo role of PknG in manifestation of disease pathology and established a role for PknG in the formation of stable granuloma, hallmark structures of latent tuberculosis. Taken together, PknG-mediated GarA phosphorylation is important for maintenance of both mycobacterial physiology and redox poise, an axis that is dispensable for survival under normoxic conditions but is critical for non-replicating persistence of mycobacteria. In conclusion, we propose that PknG probably acts as a modulator of latency-associated signals.

Antimicrobial metabolites from Saraca asoca impairs the membrane transport system and quorum-sensing system in Pseudomonas aeruginosa.

This study was conducted to explore the antimicrobial mechanism of metabolites from Saraca asoca (SA1) using differential proteomics and metabolic profile of Pseudomonas aeruginosa after treatment with effective sub-MIC dose of 312 µg/mL. SA1 fraction was found to contain antibacterial metabolites catechol, protocatechuic acid, and epigallocatechin gallate. Proteome analysis revealed 33 differentially expressed proteins after SA1 treatment. Protein network analysis showed that SA1 treatment upregulated the DNA topological and metabolic processes. Furthermore, it revealed that T2SS, cellular component biogenesis, and response to chemical stimuli were inhibited by SA1 treatment, supported by down-regulated Na+/H+ antiporter, SdeX, ompK, and trbD proteins. Statistical analysis of mass data revealed the altered level of 20 metabolites includes HSLs, PQS, rhamnolipid, and pyocyanin. Proteome and metabolome results showed that treatment impaired cell membrane functions and quorum-sensing system. It was further confirmed by increased MDA (3.95 fold), and rhamnolipids (4.3 fold) production and, therefore, oxidative stress (36.9%) after SA1 treatment.

Enthalpic interactions in aqueous strong electrolytes upon addition of ionic liquids.

The present study deals with the inter-ionic interactions between strong electrolytes and ionic liquids based on the thermodynamic properties such as excess partial molar enthalpy, HEIL, relative apparent molar enthalpy, φL, and the enthalpic interaction parameters. The thermodynamic properties of the systems are the key indicators to understand the interionic interactions. We have conducted a systematic investigation of the enthalpic behavior of aqueous solution of salts and ionic liquids and their mixtures. The present study also emphasizes how the HEIL values for the mixture of aqueous solution of ionic liquids and salts deviate from linearity as compared to those of the constituent aqueous ionic liquid or salt. This deviation from linearity for the HEIL values has been discussed here.

Concentration-dependent apparent partition coefficients of ionic liquids possessing ethyl- and bi-sulphate anions.

This study deals with the concentration dependent apparent partition coefficients log P of the ethyl and bisulfate-based ionic liquids. It is observed that the bisulfate-based ionic liquids show different behaviour with respect to concentration as compared to ethyl sulphate-based ionic liquids. It is significant and useful analysis for the further implementation of alkyl sulfate based ionic liquids as solvents in extraction processes. The log P values of the ethyl sulphate-based ionic liquids were noted to vary linearly with the concentration of the ionic liquid, whereas a flip-flop trend with the concentration for the log P values of the bisulphate-based ionic liquids was observed due to the difference in hydrogen bond accepting basicity and possibility of aggregate formation of these anions. The π-π interactions between the imidazolium and pyridinium rings were seen to affect the log P values. The alkyl chain length of anions was also observed to influence the log P values. The hydrophobicity of ionic liquid changes with the alkyl chain in the anion in the order; [HSO4](-) < [EtSO4](-) < [BuSO4](-).

Studies on Aspirin Crystals Generated by a Modified Vapor Diffusion Method.

The objectives of the current investigation were (1) to study the influence of selected two different non-solvents (diethylether and dichloromethane) on the drug crystal formation of a model drug, aspirin (ASP-I) by the modified vapor diffusion method and (2) to characterize and compare the generated crystals (ASP-II and ASP-III) using different analytical techniques with that of unprocessed ASP-I. When compared to the classical vapor diffusion method which consumes about 15 days to generate drug crystals, the modified method needs only 12 h to get the same. Fourier transform-infrared spectroscopy (FT-IR) reveals that the internal structures of ASP-II and ASP-III crystals were identical when compared with ASP-I. Although the drug crystals showed a close similarity in X-ray diffraction patterns, the difference in the relative intensities of some of the diffraction peaks (especially at 2θ values of around 7.7 and 15.5) could be attributed to the crystal habit or crystal size modification. Similarly, the differential scanning calorimetry (DSC) study speculates that only the crystal habit modifications might occur but without involving any change in internal structure of the generated drug polymorphic form I. This is further substantiated from the scanning electron microscopy (SEM) pictures that indicated the formation of platy shape for the ASP-II crystals and needle shape for the ASP-III crystals. In addition, the observed slow dissolution of ASP crystals should indicate polymorph form I formation. Thus, the modified vapor diffusion method could routinely be used to screen and legally secure all possible forms of other drug entities too.

Angiotensin-converting enzyme inhibitor prevents oxidative stress, inflammation, and fibrosis in carbon tetrachloride-treated rat liver.

Hepatic fibrosis is a common feature of chronic liver injury, and the involvement of angiotensin II in such process has been studied earlier. We hypothesized that anti-angiotensin II agents may be effective in preventing hepatic fibrosis. In this study, Long Evans female rats were used and divided into four groups such as Group-I, Control; Group-II, Control + ramipril; Group-III, CCl4; and Group-IV, CCl4 + ramipril. Group II and IV are treated with ramipril for 14 d. At the end of treatment, the livers were removed, and the level of hepatic marker enzymes (aspartate aminotransferase, Alanine aminotransferase, and alkaline phosphatase), nitric oxide, advanced protein oxidation product , catalase activity, and lipid peroxidation were determined. The degree of fibrosis was evaluated through histopathological staining with Sirius red and trichrome milligan staining. Carbon-tetrachloride (CCl4) administration in rats developed hepatic dysfunction and raised the hepatic marker enzymes activities significantly. CCl4 administration in rats also produced oxidative stress, inflammation, and fibrosis in liver. Furthermore, angiotensinogen-inhibitor ramipril normalized the hepatic enzymes activities and improved the antioxidant enzyme catalase activity. Moreover, ramipril treatment ameliorated lipid peroxidation and hepatic inflammation in CCl4-treated rats. Ramipril treatment also significantly reduced hepatic fibrosis in CCl4-administered rats. In conclusion, our investigation suggests that the antifibrotic effect of ramipril may be attributed to inhibition of angiotensin-II mediated oxidative stress and inflammation in liver CCl4-administered rats.

An atypical Trypanosoma lewisi infection in a 22-day-old neonate from India: An emergent zoonosis.

Reports on atypical human trypanosomiasis, caused by Trypanosoma lewisi, are rare and so far a total of 19 reports on human infection with animal trypanosomes, which includes nine cases from Trypanosoma lewisi exist. Trypanosoma lewisi, a Stercorarian trypanosoma of rats, is transmitted by the fecal contamination of the wound or the bite caused by rat flea Ceratophyllus fasciatus. We report here an atypical neonatal infection of T. lewisi in a 22-day-old infant from Agra. The infant presented with a history of high fever, poor appetite, and lethargy for 3 days. The hematological parameters were normal except for a low platelet count. A high C-reactive protein (CRP) concentration of 70.49 mg/L indicated marked inflammation. The Leishman-stained thin blood smears were microscopically positive for the hemoflagellate. Based on the morphological features and further confirmed by polymerase chain reaction (PCR) assay, the hemoflagellate was identified as T. lewisi. Symptomatic treatment and antibiotic therapy helped in an uneventful recovery of the patient.

Maxillary Mucinous Adenocarcinoma Mimicking a Lesion of Endodontic Origin: A Rare Case Report.

Periapical lesions of endodontic origin are fairly common in the oral cavity in association with tooth pulp infection. Most of these lesions will resolve with adequate root canal treatment and rarely cause suspicion of more insidious disease. Most clinicians tend to skip histopathological examination in cases where the lesion is excised or curetted. We present a rare case of mucinous adenocarcinoma in association an endodontically treated maxillary discoloured central incisor in a 38 year old patient with a history of root canal treatment about 15 years ago. Root canal re-treatment and wide excision was performed. Histology showed epithelial islands suggestive of a neoplasm. Immunohistochemistry was positive for CK7 and S100. Metastasis was ruled out and no evidence of recurrence has been noted in the 12-month follow up period. It is emphasized that any tissue removed from the surgical site should be analysed microscopically. (EEJ-2022-01-013).

Amelioration of CCl4-induced oxidative stress and hepatotoxicity by Ganoderma lucidum in long evans rats.

Liver disease is a serious health problem affecting people worldwide at an alarming rate. The present study aimed to investigate the protective effects of Ganoderma lucidum against CCl4-induced liver toxicity in rats. The experimental Long Evans rats were divided into five groups, of which four groups were treated with carbon tetrachloride (CCl4). Among the CCl4 treated groups, one of the groups was treated with silymarin and two of them with ethanolic extract of G. lucidum at 100 and 200 mg/Kg body weight. The oxidative stress parameters and endogenous antioxidant enzyme concentrations were assessed by biochemical tests. Liver enzymes ALT, AST, and ALP were determined spectrophotometrically. Histopathological examinations were carried out to assess hepatic tissue damage and fibrosis. Reverse transcription PCR (RT-PCR) was performed to determine the expression of IL-1ß, IL-6, IL-10, TNF-α, and TGF-ß genes. Gas Chromatography-Mass Spectroscopy (GC-MS) analysis revealed that G. lucidum is rich in several phytochemicals including 6-Octadecanoic acid (55.81%), l-( +)-Ascorbic acid 2,6-dihexadecanoate (18.72%), Cis-11-Eicosenamide (5.76%), and Octadecanoic acid (5.26%). Treatment with the G. lucidum extract reduced the elevated ALT, AST, ALP levels, and cellular oxidative stress markers and increased the endogenous antioxidant levels. Histopathology observations revealed that the inflammation, infiltration of immune cells, and aberration of collagen fibers in the hepatocytes were altered by the G. lucidum treatment. The increased expression of inflammatory cytokines TNF-α, TGF-ß, IL-1 ß, and IL-6 were markedly suppressed by G. lucidum extract treatment. G. lucidum also prevented the suppression of protective IL-10 expression by CCl4. This study strongly suggests that G. lucidum extract possesses significant hepatoprotective activity as evidenced by reduced oxidative stress and inflammation mediated by suppression in inflammatory cytokine expression and increased protective IL-10 cytokine expression.

Nanostructured chitosan-polyphenolic patch for remote NIR-photothermal controlled dermal drug delivery.

We describe the synthesis of a nanostructured dermal patch composed of chitosan-tannic acid (CT) that can carry near-infrared (NIR) active Indocyanine green (ICG) dye for performing photothermal heat conversion activity. The NIR-responsive CT-I dermal patch can deliver topical antibiotic drugs (Neomycin). The CT-I and drug-loaded CT-I/N patches have been demonstrated by FTIR, SEM/EDX, TGA, and DSC analysis. The in vitro drug release from the CT-I/N patch are favorable in the dermal environment (pH = 5.5) and significantly increases 25 % more at higher temperatures of 40 to 45 °C. The CT-I/N showed increasing photothermal heat in response to NIR (808 nm) light. The in vivo thermograph demonstrated that the CT-I/N patch can generate >45 °C within 5 min NIR irradiation. As a result, sustained wound healing was shown in H&E (hematoxylin and eosin) staining dermal tissue. Such NIR-active nanostructure film/patch is promising for the future of any sustained on-demand drug delivery system.

Arsenic causing gallbladder cancer disease in Bihar.

In recent times Gallbladder cancer (GBC) incidences increased many folds in India and are being reported from arsenic hotspots identified in Bihar. The study aims to establish association between arsenic exposure and gallbladder carcinogenesis. In the present study, n = 200 were control volunteers and n = 152 confirmed gallbladder cancer cases. The studied GBC patient's biological samples-gallbladder tissue, gallbladder stone, bile, blood and hair samples were collected for arsenic estimation. Moreover, n = 512 gallbladder cancer patients blood samples were also evaluated for the presence of arsenic to understand exposure level in the population. A significantly high arsenic concentration (p < 0.05) was detected in the blood samples with maximum concentration 389 µg/L in GBC cases in comparison to control. Similarly, in the gallbladder cancer patients, there was significantly high arsenic concentration observed in gallbladder tissue with highest concentration of 2166 µg/kg, in gallbladder stones 635 µg/kg, in bile samples 483 µg/L and in hair samples 6980 µg/kg respectively. Moreover, the n = 512 gallbladder cancer patient's blood samples study revealed very significant arsenic concentration in the population of Bihar with maximum arsenic concentration as 746 µg/L. The raised arsenic concentration in the gallbladder cancer patients' biological samples-gallbladder tissue, gallbladder stone, bile, blood, and hair samples was significantly very high in the arsenic exposed area. The study denotes that the gallbladder disease burden is very high in the arsenic exposed area of Bihar. The findings do provide a strong link between arsenic contamination and increased gallbladder carcinogenesis.

Effectiveness of midurethral slings in recurrent stress urinary incontinence: a systematic review and meta-analysis.

INTRODUCTION AND HYPOTHESIS: Midurethral slings (MUS) are the gold standard primary procedure for the surgical treatment of stress urinary incontinence (SUI). There is no robust evidence on the success with MUS in the treatment of recurrent SUI. Our objective was to evaluate the effectiveness and complications of MUS in women with recurrent SUI by systematic review and meta-analysis of the literature. METHODS: A systematic literature search was carried out (up to August 2011) using relevant search terms in MEDLINE, EMBASE, CENTRAL and Google Scholar. Relevant randomised controlled trials (RCT) and prospective studies were selected and then analysed by two independent reviewers. Meta-analysis of cure stated in prospective cohort studies was performed with a random effects model using Stata 8. RESULTS: There was 1 randomised trial and 11 good quality prospective studies included in this systematic review. The overall subjective cure rate per meta-analysis of prospective cohort studies following MUS for recurrent SUI after any previous surgery was found to be 78.5 % [95 % confidence interval (CI) 69-88] at the follow-up of 29.72 ± 29.49 months. The subjective cure rate following MUS after previous failed MUS was 73.3 % (95 % CI 55-97) at the follow-up of 15.7 ± 7.7 months. CONCLUSIONS: The studies report good cure rates of SUI after MUS surgery following previous incontinence surgery (62-100 %). There seems to be a lower cure rate with transobturator compared to the retropubic tape for recurrent SUI after previous surgery.

Cytokines Expression and Nitric Oxide Production under Induced Infection to Salmonella Typhimurium in Chicken Lines Divergently Selected for Cutaneous Hypersensitivity.

In the present study, the impact of Salmonella Typhimurium on cell-mediated immunity (CMI) was investigated in 5 week-old immuno divergent broiler lines selected for the high and low response to phytohemagglutinin-P. The immune response was assessed in peripheral-blood mononuclear cells (PBMCs) induced with Salmonella Typhimurium at different time intervals (0 h, 0.5 h, 2 h, 4 h, 6 h, 12 h and 24 h). The differential mRNA expression patterns of IFN-γ, IL-2 and iNOS were evaluated by quantitative real time PCR. In-vitro production of nitric oxide (NO) was also estimated in the culture supernatant and correlated with iNOS mRNA expression. Present study showed higher production of NO in the high cell-mediated line (HCMI) as compared to the low cell-mediated line (LCMI) upon stimulation with Salmonella Typhimurium. Correspondingly, higher mRNA expression of iNOS and IFN-γ were observed in high response birds (HCMI); but IL-2 was down regulated in this line compared to the low response birds (LCMI). Significantly (p<0.05) higher expression of iNOS, IFN-γ and higher production of NO in high line indicated that the selection for PHA-P response might be employed for increasing the immune competence against Salmonella Typhimurium in chicken flocks.

The predictive value of depth of invasion and tumor size on risk of neck node metastasis in squamous cell carcinoma of the oral cavity: A prospective study.

Background: Tumor depth is a reliable parameter to predict nodal metastasis in oral cancers; therefore, the authors embarked upon a prospective observational study to define the relationship between the tumor depth and the risk of cervical lymph node involvement as well to determine the optimal tumor depth cutoff point for nodal metastasis. Aims: The aim was to study the predictive value of depth of invasion (DOI) and tumor size on risk of cervical node metastasis in squamous cell carcinoma of the oral cavity. Materials and Methods: Biopsy-proven Stage I-Stage III oral cavity squamous cell carcinoma patients were included in this prospective, observational study. Various histopathological characteristics (DOI, tumor size, lympho-vascular invasion [LVI], perineural spread, and grade of differentiation) were analyzed to predict the cervical node metastasis. Statistical Analysis Used: The impact of the clinical and histopathological parameters of primary tumor on cervical lymph node metastasis was analyzed by univariate as well as multivariate logistic regression analyses using NCSS 12 version 12.0.5 statistical software. Results: The independent predictors of cervical lymph node metastasis were DOI (P = 0.0014) and LVI (P = 0.0414). The incidence of cervical metastasis increased markedly when the DOI was over 5 mm, and it was a statistically significant (P < 0001) association. Conclusions: DOI is a significant predictor of cervical nodal metastasis and tumor depth 5 mm can be considered as a cutoff value in staging and management of early oral squamous cell carcinoma.

Seroprevalence of Anti-SARS-CoV-2 IgG Antibodies among HIV Infected Individuals Attending ART Centre at Pune: A Cross-Sectional Study.

Background: We aimed to determine the anti-SARS-CoV-2 IgG antibodies among people living with HIV (PLHIV) in Pune, India. Methods: This cross-sectional study was conducted between March 2021 and June 2021. Demographic and clinical information related to coronavirus disease 2019 (COVID-19) were recorded on structured questionnaires. Blood samples were collected and tested for anti-SARS-CoV-2 IgG antibodies using commercial ELISA. Results: Of the 405 HIV infected individuals enrolled in the study, 223(55.1%) were females. Mean age and CD4 count of participants were 42 years (SD: 10) and 626 cells/mm3 (SD: 284) respectively. A total of 382 (95%) PLHIV were virologically suppressed. The seropositivity against SARS-CoV-2 was found in 221 PLHIV (54.6%, 95% CI: 49.7-59.4). No significant association was found between demographic or clinical factors and seropositivity. Conclusion: A high prevalence of anti-SARS-CoV-2 IgG antibodies was found among PLHIV attending ART centre indicating an exposure to the virus among them.

Development of potential proteasome inhibitors against Mycobacterium tuberculosis.

Tuberculosis (TB) has been recently declared as a health emergency because of sporadic increase in Multidrug-resistant Tuberculosis (MDR-TB) problem throughout the world. TB causing bacteria, Mycobacterium tuberculosis has become resistant to the first line of treatment along with second line of treatment and drugs, which are accessible to us. Thus, there is an urgent need of identification of key targets and development of potential therapeutic approach(s), which can overcome the Mycobacterium tuberculosis complications. In the present study, Mycobacterium tuberculosis proteasome has been taken as a potential target as it is one of the key regulatory proteins in Mycobacterium tuberculosis propagation. Further, a library of 400 compounds (small molecule) from Medicines for Malaria Venture (MMV) were screened against the target (proteasome) using molecular docking and simulation approach, and selected lead compounds were validated in in vitro model. In this study, we have identified two potent small molecules from the MMV Pathogen Box library, MMV019838 and MMV687146 with -9.8 kcal/mol and -8.7 kcal/mol binding energy respectively, which actively interact with the catalytic domain/active domain of Mycobacterium tuberculosis proteasome and inhibit the Mycobacterium tuberculosis growth in in vitro culture. Furthermore, the molecular docking and simulation study of MMV019838 and MMV687146 with proteasome show strong and stable interaction with Mycobacterium tuberculosis compared to human proteasome and show no cytotoxicity effect. A better understanding of proteasome inhibition in Mycobacterium tuberculosis in in vitro and in vivo model would eventually allow us to understand the molecular mechanism(s) and discover a novel and potent therapeutic agent against Tuberculosis. Active efflux of drugs mediated by efflux pumps that confer drug resistance is one of the mechanisms developed by bacteria to counter the adverse effects of antibiotics and chemicals. Efflux pump activity was tested for a specific compound MMV019838 which was showing good in silico results than MIC values.Communicated by Ramaswamy H. Sarma.