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Background: Despite multifactorial evidence, the safe and effective elimination of free-floating micro-organisms remains a significant scientific challenge. ZeBox Technology exploits microbial Zeta Potential, to extract and eliminate them from free-flowing air, using a non-ionizing electric field, in combination with a microbicidal surface.Aim: Evaluation of ZeBox Technology against aerosolized SARS-CoV-2 and Mycobacterium tuberculosis under controlled conditions.Methods: SARS-CoV-2 and M. tuberculosis H37Ra were used in this study. Individual micro-organisms were aerosolized using a Collison nebulizer inside an air-sealed test chamber. Air samples were collected from the chamber on to a Mixed Cellulose Ester membrane, at various time points, and used for enumeration. SARS-CoV-2 was enumerated using qRT-PCR, while M. tuberculosis H37Ra was quantified using standard microbiological procedures.Findings: We established a viable aerosolized microbial load of ∼10E9 and ∼10E6 for SARS-CoV-2 and M. tuberculosis H37Ra, respectively, inside the test chamber. Under ideal conditions, the floating microbial load was at a steady-state level of 10E9 for SARS-CoV-2 and 10E6 for M. tuberculosis. When the ZeBox-Technology-enabled device was operated, the microbial load reduced significantly. A reduction of ∼10E4.7 was observed for M. tuberculosis, while a reduction of ∼10E7 for SARS-CoV-2 was observed within a short duration. The reduction in airborne SARS-CoV-2 load was qualitatively and quantitatively measured using fluorescence analysis and qRT-PCR methods, respectively.Conclusion: This validation demonstrates the efficacy of the developed technology against two of the deadliest micro-organisms that claim millions of lives worldwide. In conjunction with the existing reports, the present validation proved the true broad-spectrum elimination capability of ZeBox technology. [ABSTRACT FROM AUTHOR]

Abstract Background Tuberculosis (TB) is a leading cause of morbidity and mortality worldwide. Control of TB is lingering by the lack of diagnostic tests that are simple, rapid, yet accurate. Thus, smear-negative pulmonary TB often misses the diagnosis. The study evaluated the performance of GeneXpert MTB/RIF assay for the detection of Mycobacterium tuberculosis (MTB). Methods The study was carried out from June to December 2016 in Nepal Tuberculosis Center, Bhaktapur, Nepal. A total of 173 sputum samples were collected and processed by microscopy [Auramine-O staining and Ziehl–Neelsen (ZN) staining], followed by GeneXpert MTB/RIF assay and culture in Lowenstein-Jensen (LJ) medium. Results Of 173 sputum samples, 162 (93.6%) were smear-negative. Of 162 smear-negative sputum samples, 35 (21.6%) were confirmed to have MTB by culture, and 31 (19.1%) by GeneXpert MTB/RIF assay. Of 31 GeneXpert-positive samples, 25 (80.6%) were susceptible, 4 (12.9%) were resistant, and 2 (6.45%) were intermediate to rifampicin. The sensitivity, specificity, positive predictive value, and negative predictive value of GeneXpert MTB/RIF assay for smear-negative sputum samples were 74.3%, 96.6%, 86.7%, and 92%, respectively. The GeneXpert MTB/RIF has a substantial diagnostic agreement of 90.91% with culture (Cohen’s Kappa coefficient = 0.73). Conclusion The diagnostic performance of GeneXpert MTB/RIF assay was almost on par with culture, and thus can be relied upon for MTB detection in smear-negative sputum samples.

Abstract Background Tuberculosis (TB) is a communicable disease remains a major global health problem and the leading cause of death from a single infectious agent. Even though many of the WHO recommended TB control strategies were implemented; there is still a major gap in TB case detection and treatment. This study aimed to determine the prevalence of Mycobacterium tuberculosis among presumptive TB patients in Gedeo Zone, Southern Ethiopia. Methods A cross-sectional study was conducted on 384 TB suspected patients in Gedeo Zone from February to July 2021. Data were collected using a pretested structured questionnaire. Laboratory examination was processed using Xpert-MTB/RIF assay. Data entry was made using Epi info version 7 and analyzed by SPSS version 24. Logistic regression models were used to determine the risk factors. Results Out of 384 study participants suspected with TB, M. tuberculosis was isolated from 103 giving an overall prevalence of 26.8%. Males (AOR) = 1.95; 95% CI 1.56–2.65, P = 0.01) were more likely to develop TB than females. Study participants who were illiterate (AOR 2.10; 95% CI 1.17–2.51, P = 0.014) were more likely to develop TB than the educated ones. Cigarette smokers (AOR 2.89; 95% CI 2.10–3.84, P = 0.01), khat chewers (AOR 2.86; 95% CI 1.28–3.79, P = 0.01), vaccination (AOR 0.52; 95% CI 0.21–0.88, P = 0.02), close contact (AOR 3.42; 95% CI 2.24–4.50, P = 0.01) and being positive for HIV (AOR 2.01; 95% CI 1.07–3.52, 0.01) were more likely to develop TB. Conclusion Despite implementation of national and international TB control strategies, TB still remains one of the major public health problems in the country especially in the study area. The high prevalence of MTB was reported different risk groups. Early case detection and management of TB should be given special attention to strengthen and an appropriate control and prevention methods to reduce the emergence and increasing of MTB cases.

[Display omitted] Over the past 2000 years, tuberculosis (TB) has been responsible for more deaths than any other infectious disease. In recent years, there has been a recovery of research and development (R&D) efforts focused on TB drugs. This is driven by the pressing need to combat the global spread of the disease and develop improved therapies for both drug-sensitive and drug-resistant strains. Many new TB drug candidates have recently entered clinical trials, marking the beginning of a rebirth in this area after decades of neglect. The problem is that very few of the hundreds of compounds identified each year as potential anti-TB drugs really make it to the clinical development stage. This perspective focuses on the primary obstacles and approaches involved in the development of new medications for TB. This will help medicinal chemists better understand TB drug challenges and develop novel drug candidates. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is a serious public health issue in India. Numerous molecular mechanisms and immunological responses play significant roles in the pathogenesis of tuberculosis. This study aimed to identify host immune-related biomarkers that are significantly differentially expressed in active TB and that play a vital role in disease progression. The methodology employed in this study included data collection, pre-processing, analysis, and interpretation of the results. Six microarray datasets were used to identify differentially expressed genes (DEGs), and only the common DEGs were used for further downstream analysis, such as hub gene identification, gene ontology, pathway enrichment analysis, and drug-gene interaction analysis. The study identified 1728 DEGs, including 906 upregulated and 822 downregulated genes. Five hub genes were identified that were: STAT1, GBP5, GBP1, FCGR1A, and BATF2. Gene ontology and pathway enrichment revealed that most of the genes were involved in interferon-gamma signaling. In addition, through drug-gene interactions, known drugs have been identified for STAT1, FCGR1A and GBP1. The findings of this study may contribute to early detection and treatment of active TB. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) can directly kill Gram-positive bacteria, Gram-negative bacteria, mycobacteria, fungi, enveloped viruses, and parasites. At sublethal concentrations, some AMPs and also conventional antibiotics can stimulate bacterial response increasing their resilience, also called the hormetic response. This includes stimulation of growth, mobility, and biofilm production. Here, we describe the discovery of AMPs that stimulate the growth of certain mycobacteria. Peptide 14 showed a growth stimulating effect on Mycobacteria tuberculosis (MTB), M. bovis, M. avium subsp. paratuberculosis (MAP), M. marinum, M. avium-intracellulare, M. celatum, and M. abscessus. The effect was more pronounced at low bacterial inocula. The peptides induce a faster transition from the lag phase to the log phase and keep the bacteria longer in the log phase before entering stationary phase when compared to nontreated controls. In some cases, an increase in the division rate was observed. An initial screen using MAP and a collection of 75 peptides revealed 13 peptides with a hormetic effect. For MTB, a collection of 25 artificial peptides were screened and 13 were found to reduce the time to positivity (TTP) by at least 5%, improving growth. A screen of 43 naturally occurring peptides, 11 fragments of naturally occurring peptides and 5 designed peptides, all taken from the database APD3, identified a further 44 peptides that also lowered TTP by at least 5%. Lasioglossin LL-III (Bee) and Ranacyclin E (Frog) were the most active natural peptides, and the human cathelicidin LL37 fragment GF-17 and a porcine cathelicidin protegrin-1 fragment were the most active fragments of naturally occurring peptides. Peptide 14 showed growth-stimulating activity between 10 ng/mL and 10 µg/mL, whereas the stability-optimised Peptide 14D had a narrow activity range of 0.1–1 µg/mL. Peptides identified in this study are currently in commercial use to improve recovery and culture for the diagnostics of mycobacteria in humans and animals.

Tuberculosis (TB) is a fatal infectious disease which affected millions of people worldwide for many decades and now with mutating drug resistant strains, it poses bigger challenges in treatment of the patients. Computational techniques might play a crucial role in rapidly developing new or modified anti-tuberculosis drugs which can tackle these mutating strains of TB. This research work applied a computational approach to generate a unique recommendation list of possible TB drugs as an alternate to a popular drug, EMB, by first securing an initial list of drugs from a popular online database, PubChem, and thereafter applying an ensemble of ranking mechanisms. As a novelty, both the pharmacokinetic properties and some network based attributes of the chemical structure of the drugs are considered for generating separate recommendation lists. The work also provides customized modifications on a popular and traditional ensemble ranking technique to cater to the specific dataset and requirements. The final recommendation list provides established chemical structures along with their ranks, which could be used as alternatives to EMB. It is believed that the incorporation of both pharmacokinetic and network based properties in the ensemble ranking process added to the effectiveness and relevance of the final recommendation.

Tuberculosis (T.B.) is an infectious disease caused primarily by the bacterial pathogen Mycobacterium tuberculosis (Mtb) in humans. The emergence of various drug-resistant Mtb strains threatens to disrupt worldwide attempts to control the infection. The state of disease can be attributed to the host–pathogen protein interaction network. Hence in this study, we have analyzed the protein interaction network at the intra-species level, i.e., within Mtb using the Louvain community detection and at the inter-species level, i.e., between human and Mtb proteins. We observed a higher inter-connectedness in the intra-species protein network as compared to the inter-species network. This reflects a critical role of protein interaction network modulation during host–pathogenesis conditions. After analyzing 269 Mtb proteins and 2287 human proteins using the interlog approach, we observed a total of 640 host–pathogen protein interaction pairing involving 120 humans and 90 Mtb proteins. The H.P.I.s were filtered using functional protein annotations and publicly available experimental results. In the Mtb PPI network, Rv1286, Rv0462, Rv2215, Rv3559c, Rv3504, and Rv3523 are associated as a motif (a governing unit in a network) and identified as key proteins. This interaction network will help researchers better understand host–pathogen protein–protein interactions, as well as shed light on how Mtb interacts with its host.

Background: Molecular tumor boards (MTBs) match molecular alterations with targeted anticancer drugs upon failure of the available therapeutic options. Special and local needs are most likely to emerge through the comparative analysis of MTB networks, but these are rarely reported. This manuscript summarizes the state-of-art of 16 active Italian MTBs, as it emerges from an online survey curated by Alliance Against Cancer (ACC). Main text: Most MTBs (13/16) are exclusively supported through local Institutional grants and meet regularly. All but one adopts a fully virtual or a mixed face-to-face/virtual calling/attendance meeting model. It appears that the ACC MTB initiative is shaping a hub-and-spoke virtual MTB network reminiscent of non-redundant, cost-effective healthcare organization models. Unfortunately, public awareness of MTB opportunities presently remains insufficient. Only one center has a website. Dedicated e-mail addresses are for the exclusive use of the MTB staff. More than half of ACC members consider a miscellanea of most or all solid and hematological malignancies, and more than one-third consider neoplasms arising at any anatomical location. The average number of Staff Members in MTBs is 9. More than 10 staff members simultaneously attend MTB meetings in 13 MTBs. A medical oncologist is invariably present and is in charge of introducing the clinical case either with (45%) or without previous discussion in organ-specific multidisciplinary Boards. All but two MTBs take charge of not only patients with no standard-of-care (SoC) therapy option, but also cases receiving NGS profiling in SoC settings, implying a larger number of yearly cases. All MTBs run targeted NGS panels. Three run whole-exome and/or RNAseq approaches. ESCAT-ESMO and/or Onco-KB levels of evidence are similarly used for diagnostic reporting. Most MTBs (11) provide a written diagnostic report within 15 days. Conclusions are invariably communicated to the patient by the medical oncologist. Conclusions: MTB networking is crucial not only for molecular diagnosis and therapy assignment, but also for healthcare governance. Survey results show that MTBs review therapeutic opportunities at the crossover between standard-of-care with off-label, the former task being much beyond their scope. Societal and scientific implications of this beyond-the-scope MTB function may be relevant for healthcare in Italy and abroad. [ABSTRACT FROM AUTHOR]

Simple Summary: In Molecular Tumor Boards (MTBs), clinicians and researchers discuss the biology of tumor samples from individual patients to find suitable therapies. MTBs have therefore become key elements of precision oncology programs. Patients living in urban areas with specialized medical centers can easily access MTBs. Dedicated efforts are necessary to also grant equal access for patients from rural areas. To address this challenge, the four German cancer centers in Würzburg, Erlangen, Regensburg and Augsburg collectively measured the regional efficacy of their MTBs. By jointly analyzing the residences of all MTB patients, we uncovered regional differences in our mostly rural catchment area. Mapping and further understanding these local differences—especially the underrepresented white spots—will help resolving inequalities in patient access to precision oncology. Our study represents a hands-on approach to assessing the regional efficacy of a precision oncology program. Moreover, this approach is transferable to other regions and clinical applications. (1) Background: molecular tumor boards (MTBs) are crucial instruments for discussing and allocating targeted therapies to suitable cancer patients based on genetic findings. Currently, limited evidence is available regarding the regional impact and the outreach component of MTBs; (2) Methods: we analyzed MTB patient data from four neighboring Bavarian tertiary care oncology centers in Würzburg, Erlangen, Regensburg, and Augsburg, together constituting the WERA Alliance. Absolute patient numbers and regional distribution across the WERA-wide catchment area were weighted with local population densities; (3) Results: the highest MTB patient numbers were found close to the four cancer centers. However, peaks in absolute patient numbers were also detected in more distant and rural areas. Moreover, weighting absolute numbers with local population density allowed for identifying so-called white spots—regions within our catchment that were relatively underrepresented in WERA MTBs; (4) Conclusions: investigating patient data from four neighboring cancer centers, we comprehensively assessed the regional impact of our MTBs. The results confirmed the success of existing collaborative structures with our regional partners. Additionally, our results help identifying potential white spots in providing precision oncology and help establishing a joint WERA-wide outreach strategy. [ABSTRACT FROM AUTHOR]

• TB patients have lower IFN-γR1 and IFN-γR2 expression. • There is sex-dependent modulation of IFN-γR1 and IFN-γR2 expression in TB patients. • IFN-γR1 mRNA and its surface expression shows significant positive correlation in HCs. Interferon-γ (IFN-γ) is an essential pro-inflammatory cytokine against tuberculosis (TB). To initiate immune response, IFN-γ binds to its receptor complex which consists of two subunits IFN-γ receptor 1 (IFN-γR1) and IFN-γ receptor 2 (IFN-γR2). The deficiency in either receptor subunit can alter IFN-γ signalling thus influencing host susceptibility to TB. In the present study IFN-γ receptor expression at transcriptional and translational level was analysed in pulmonary TB patients from North India. A total of 46 pulmonary TB patients (at 0 day of anti-tuberculosis therapy) and 48 healthy controls (HCs) were recruited. It was found that the mRNA expression of IFN-γR1 was decreased in male TB patients (p = 0.003). The surface expression of IFN-γR1 (p = 0.0005) and IFN-γR2 (p = 0.024) was also found to be decreased in male TB patients. In conclusion, we found sex-dependent regulation of IFN-γR1 and IFN-γR2 expression in pulmonary TB patients of studied population. [ABSTRACT FROM AUTHOR]

In eukaryotes, RNA N 6 -methyladenosine (m 6 A) modification and microRNA (miRNA)-mediated RNA silencing represent two critical epigenetic regulatory mechanisms. The m 6 A methyltransferase complex (MTC) and the microprocessor complex both undergo liquid-liquid phase separation to form nuclear membraneless organelles. Although m 6 A methyltransferase has been shown to positively regulate miRNA biogenesis, a mechanism of reciprocal regulation between the MTC and the microprocessor complex has remained elusive. Here, we demonstrate that the MTC and the microprocessor complex associate with each other through the METHYLTRANSFERASE B (MTB)-SERRATE (SE) interacting module. Knockdown of MTB impaired miRNA biogenesis by diminishing microprocessor complex binding to primary miRNAs (pri-miRNAs) and their respective MIRNA loci. Additionally, loss of SE function led to disruptions in transcriptome-wide m 6 A modification. Further biochemical assays and fluorescence recovery after photobleaching (FRAP) assay indicated that SE enhances the liquid-liquid phase separation and solubility of the MTC. Moreover, the MTC exhibited enhanced retention on chromatin and diminished binding to its RNA substrates in the se mutant background. Collectively, our results reveal the substantial regulatory interplay between RNA m 6 A modification and miRNA biogenesis., (© 2024 The Author(s). Journal of Integrative Plant Biology published by John Wiley & Sons Australia, Ltd on behalf of Institute of Botany, Chinese Academy of Sciences.)

Bronchoalveolar lavage (BAL) is the saline wash of the bronchial tree, which aids in diagnosing various pulmonary pathologies. The present study was contemplated with an aim to know the clinical, microbiological profile of BAL samples along with its sensitivity pattern and to assess its utility as a diagnostic tool. This was a prospective observational study, carried over 90 patients presenting with lower respiratory tract infections. The total microbial recovery rate from BAL was 39 (43.3%). The sensitivity, specificity and positive predictive value of BAL were found to be 76.4%, 89.7% and 90.6% respectively. Maximum isolates were bacteria (25.5%) followed by Mycobacterium tuberculosis (MTB) (16.6%) and fungi (1.1%). Predominant bacterial isolates were Gram-negative (81.5%) compared to Gram-positive (18.5%). Multidrug resistance (MDR) in bacteria was seen in 59.2% of isolates. BAL is a valuable diagnostic tool to find not only bacterial but mycobacterial and fungal infections in patients with lower respiratory tract infection (LRTI). A trend towards LRTI with Gram-negative infections is on the rise and they tend to be multidrug-resistant. Hence checking susceptibility patterns is crucial to start evidence-based treatment.

Kuma Diriba,1 Ephrem Awulachew,1 Gemechu Churiso2 1Department of Medical Laboratory Sciences, Health Science and Medical College, Dilla University, Dilla, Ethiopia; 2Department of Medical Laboratory Sciences, Immunology Unit, Health Science and Medical College, Dilla University, Dilla, EthiopiaCorrespondence: Kuma Diriba Tel +251913384550Email kumadiriba47@gmail.comBackground: Tuberculosis (TB) remains a major global health problem causing death among millions of people each year. The new barrier that challenges the control of tuberculosis is the emerging and the increasing number of drug-resistant TB that becomes a world concern. This study aimed to determine the magnitude of rifampicin-resistant Mycobacterium tuberculosis (RR-MTB) among presumptive TB patients attending Dilla University Referral Hospital, Gedeo Zone, Ethiopia.Methods: A retrospective cross-sectional study was conducted at Dilla University Referral Hospital from January 2014 to December 2020. Sputum results were done using Xpert MTB/RIF assay and other necessary data were collected from the registration logbooks using a standardized data extraction format and analyzed using SPSS version 23 statistical software.Results: A total of 17,745 presumptive TB patients were included, of which 62.2% were males. The overall prevalence of Mycobacterium tuberculosis (MTB) was 11.8%, of which 5.1% were confirmed to have RR-MTB. Extra-pulmonary TB was reported in 1.5% of the study participants. The highest prevalence of MTB and RR-MTB was recorded in 2017 with a prevalence of 20.1% and 8.5%, respectively. All age groups were significantly associated with a higher prevalence of MTB (p < 0.036). TB patients with a history of previous treatment and HIV positive were significantly associated with MTB (P < 0.021), while RR-MTB was only significantly associated with patients with a history of previous treatment (P < 0.018).Conclusion: A high magnitude of MTB and RR-MTB was reported among TB patients with HIV and a history of previous treatment. Therefore, coordinated efforts should be applied to the improvement of treatment adherence of known TB cases, and appropriate control and prevention methods to reduce the emergence and increase of MTB and RR-MTB cases.Keywords: presumptive TB, MTB, RR-MTB, Xpert MTB/RIF assay, previous treatment, HIV/TB

Pulmonary tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (MTB). Whole-genome sequencing (WGS) holds great promise as an advanced technology for accurately predicting anti-TB drug resistance. The development of a reliable method for detecting drug resistance is crucial in order to standardize anti-TB treatments, enhance patient prognosis, and effectively reduce the risk of transmission. In this study, our primary objective was to explore and determine the potential of WGS for assessing drug resistance based on genetic variants recommended by the World Health Organization (WHO). A total of 1105 MTB strains were selected from samples collected from 2014–2018 in Zhejiang Province, China. Phenotypic drug sensitivity tests (DST) of the anti-TB drugs were conducted for isoniazid (INH), rifampicin (RFP), streptomycin, ethambutol, fluoroquinolones (levofloxacin and moxifloxacin), amikacin, kanamycin, and capreomycin, and the drug-resistance rates were calculated. The clean WGS data of the 1105 strains were acquired and analyzed. The predictive performance of WGS was evaluated by the comparison between genotypic and phenotypic DST results. For all anti-TB drugs, WGS achieved good specificity values (>90%). The sensitivity values for INH and RFP were 91.78% and 82.26%, respectively; however, they were ≤60% for other drugs. The positive predictive values for anti-TB drugs were >80%, except for ethambutol and moxifloxacin, and the negative predictive values were >90% for all drugs. In light of the findings from our study, we draw the conclusion that WGS is a valuable tool for identifying genome-wide variants. Leveraging the genetic variants recommended by the WHO, WGS proves to be effective in detecting resistance to RFP and INH, enabling the identification of multi-drug resistant TB patients. However, it is evident that the genetic variants recommended for predicting resistance to other anti-TB drugs require further optimization and improvement.

The BCG vaccine is prepared from a weakened strain of Mycobacterium bovis (M. bovis), a bacterium closely related to Mycobacterium tuberculosis (MTB), which causes tuberculosis (TB). The vaccine was developed over 13 years, from 1908 to 1921, in the French Institut Pasteur by Léon Charles Albert Calmette and Jean-Marie Camille Guérin, who named the product Bacillus Calmette–Guérin (BCG). BCG, the only licensed vaccine currently available to prevent TB, is given to infants at high risk of TB shortly after birth to protect infants and young children from pulmonary, meningeal, and disseminated TB. The BCG vaccine, one of the safest and most widely used live attenuated vaccines in the world, recently celebrated its 100th anniversary (from 1921 to 2021); its record of use in preventing TB in China is also approaching 100 years. In 2022, a new century of BCG vaccine immunization will begin. In this article, we briefly review the history of BCG vaccine use in China, describe its current status, and offer a preliminary outlook on the future of the vaccine, to provide BCG researchers with a clearer understanding of its use in China. [ABSTRACT FROM AUTHOR]

Background: Hormone-sensitive lipase (HSL) is a neutral lipase capable of hydrolysing various kinds of lipids. In comparison to single human Hormone Sensitive Lipase (hHSL), that is induced under nutritional stress, twelve serine hydrolases are annotated as HSL in Mycobacterium tuberculosis (mHSL). Mycobacterium is exposed to multiple stresses inside the host. Therefore, the present study was carried out to investigate if mHSL are also expressed under stress condition and if there is any correlation between various stress conditions and expression pattern of mHSL. Methods and results: The expression pattern of mHSL under different environmental conditions (in-vitro and ex-vivo) were studied using qRT-PCR in M. tuberculosis H37Ra strain with 16 S rRNA as internal control. Out of 12, only two genes (lipU and lipY) were expressed at very low level in mid log phase culture under aerobic conditions, while 9 genes were expressed at stationary phase of growth. Ten mHSLs were expressed post-infection under ex-vivo conditions in time dependent manner. LipH and lipQ did not express at any time point under ex-vivo condition. The relative expression of most of the genes under individual stress was much higher than observed in ex-vivo conditions. The expression pattern of genes varied with change in stress condition. Conclusions: Different sets of mHSL genes were expressed under different individual stress conditions pointing towards the requirement of different mHSL to combat different stress conditions. Overall, most of the mHSLs have demonstrated stress dependent expression pointing towards their role in intracellular survival of mycobacteria. [ABSTRACT FROM AUTHOR]

The Compositive Passive Control method (CPC method) of "interlayer seismic isolation" + "shock absorption between adjacent towers" is applied to Multi-Tower Building (MTB) with a large podium. Taking the total vibration energy of the structure as the optimal control objective, the response expressions of a multi-DOF layer shear model and an equivalent single-DOF layer shear model are derived. The rationality of the equivalent model is demonstrated from the perspective of mode characteristics and time history response. Based on the Kanai–Tajimi spectral seismic motion model, the control effect and the optimal control parameters of the CPC method are studied. Compared with the conventional seismic scheme and the interlayer seismic isolation scheme, the influence of the CPC method on the structure natural vibration characteristics and dynamic response under different parameters of the connecting control device is discussed and analyzed. The numerical analysis proves that the CPC method provides a significant damping effect compared with the interlayer seismic isolation scheme. It maintains the advantages of the interlayer seismic isolation scheme and reduces the defect of the response amplification caused by the interlayer isolation. This analysis constitutes a good reference for the follow-up study of the CPC method of MTB. [ABSTRACT FROM AUTHOR]

Targeting enzymes that play a role in the biosynthesis of the bacterial cell wall has long been a strategy for antibacterial discovery. In particular, the cell wall of Mycobacterium tuberculosis (Mtb) is a complex of three layers, one of which is Peptidoglycan, an essential component providing rigidity and strength. UDP-GlcNAc, a precursor for the synthesis of peptidoglycan, is formed by GlmU, a bi-functional enzyme. Inhibiting GlmU Uridyltransferase activity has been proven to be an effective anti-bacterial, but its similarity with human enzymes has been a deterrent to drug development. To develop Mtb selective hits, the Mtb GlmU substrate binding pocket was compared with structurally similar human enzymes to identify selectivity determining factors. Substrate binding pockets and conformational changes upon substrate binding were analyzed and MD simulations with substrates were performed to quantify crucial interactions to develop critical pharmacophore features. Thereafter, two strategies were applied to propose potent and selective bacterial GlmU Uridyltransferase domain inhibitors: (i) optimization of existing inhibitors, and (ii) identification by virtual screening. The binding modes of hits identified from virtual screening and ligand growing approaches were evaluated further for their ability to retain stable contacts within the pocket during 20 ns MD simulations. Hits that are predicted to be more potent than existing inhibitors and selective against human homologues could be of great interest for rejuvenating drug discovery efforts towards targeting the Mtb cell wall for antibacterial discovery. [ABSTRACT FROM AUTHOR]

Rbab Taha,1 Shahd Abureesh,2 Shuruq Alghamdi,2 Rola Y Hassan,3 Mohamed M Cheikh,4,5 Rania A Bagabir,6 Hani Almoallim,2,7 Altaf Abdulkhaliq8 1Department of Transplant Infectious Disease, University Health Network, Toronto, Ontario, Canada; 2Department of Medicine, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia; 3Department of Internal Medicine, King Fahad Armed Forces Hospital, Jeddah, Saudi Arabia; 4Department of Internal Medicine, Doctor Soliman Fakeeh Hospital, Jeddah, Saudi Arabia; 5Department of Medicine, Fakeeh College for Medical Sciences, Jeddah, Saudi Arabia; 6Department of Immunology and Hematology, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia; 7Alzaidi Chair of Research in Rheumatic Diseases, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia; 8Department of Clinical Biochemistry, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi ArabiaCorrespondence: Rola Y Hassan Email r.toufiq@hotmail.comAbstract: Vitamin D is proposed to have a potential role in the pathogenicity, clinical presentation, prognosis, complications, and treatment of several diseases. In addition to its well-known role in calcium metabolism, vitamin D regulates both innate and adaptive immunity, and subsequently modulates the antiviral and antibacterial inflammatory immune responses. In view of the emerging coronavirus disease 2019 (COVID-19) pandemic, searching for potential therapeutic and protective strategies is of urgent interest, and vitamin D is one of the promising agents in this field. In this review, we present data from literature that supports the promising role of vitamin D in treatment and/or prevention of several infections including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This review summarizes vitamin D metabolism and its role in inflammation, thrombosis and immune regulation. It also reviews, in short, the role of vitamin D and the impact of its deficiency in several infections namely tuberculosis, influenza, human immunodeficiency virus (HIV), and SARS-CoV-2. Considering the roles of vitamin D on immune modulation, controlling of thrombosis, and attacking several microorganisms, the current review will elaborate on the association between these salient roles of vitamin D and the pathogenicity of various infectious agents including COVID-19. Consequently, the comprehensive finding of the current review shows a possible significant impact of vitamin D supplement as a hope in preventing, treating, and/or improving the progression of certain infections, specifically during the worldwide attempts to fight against the COVID-19 pandemic and minimize the severity of health complications encountered accordingly. In addition, avoiding a status of vitamin D deficiency to obtain its positive effects on the immune system and its protective mechanism during infections will be a general benefit overall.Keywords: 1,25(OH)2D, 1,25-dihydroxyvitamin D, 25(OH)D, 25-hydroxyvitamin D, HIV, human immunodeficiency virus, SARS, COVID-19, severe acute respiratory syndrome, TB, tuberculosis, MTB, Mycobacterium tuberculosis, influenza

ABSTRACT Individuals co-infected with HIV and Mycobacterium tuberculosis (Mtb) are more likely to develop severe tuberculosis (TB) disease than HIV-naive individuals. To understand how a chronic pre-existing Simian immunodeficiency virus (SIV) infection impairs the early immune response to Mtb, we used the Mauritian cynomolgus macaque (MCM) model of SIV/Mtb co-infection. We examined the relationship between peripheral viral control and Mtb burden, Mtb dissemination, and T cell function between SIV+ spontaneous controllers, SIV+ non-controllers, and SIV-naive MCM who were challenged with a barcoded Mtb Erdman strain 6 months post-SIV infection and necropsied 6 weeks post-Mtb infection. Mycobacterial burden was highest in the SIV+ non-controllers in all assessed tissues. In lung granulomas, the frequency of TNF-α-producing CD4+ T cells was reduced in all SIV+ MCM, but IFNγ-producing CD4+ T cells were only lower in the SIV+ non-controllers. Further, while all SIV+ MCM had more PD1+ and TIGIT+ T cells in the lung granulomas relative to SIV-naive MCM, SIV+ controllers exhibited the highest frequency of cells expressing these markers. To measure the effect of SIV infection on within-host bacterial dissemination, we sequenced the molecular barcodes of Mtb present in each tissue and characterized the Mtb population complexity. While Mtb population complexity was not associated with SIV infection group, lymph nodes had increased complexity when compared with lung granulomas across all groups. These results provide evidence that SIV+ animals, independent of viral control, exhibit a dysregulated T cell immune response and enhanced dissemination of Mtb, likely contributing to the poor TB disease course across all SIV/Mtb co-infected animals. IMPORTANCE HIV and TB remain significant global health issues, despite the availability of treatments. Individuals with HIV, including those who are virally suppressed, are at an increased risk to develop and succumb to severe TB disease when compared with HIV-naive individuals. Our study aims to understand the relationship between the extent of SIV replication, mycobacterial growth, and T cell function in the tissues of co-infected Mauritian cynomolgus macaques during the first 6 weeks of Mtb infection. Here we demonstrate that increased viral replication is associated with increased bacterial burden in the tissues and impaired T cell responses, and that the immunological damage attributed to virus infection is not fully eliminated when animals spontaneously control virus replication.

Abstract Background Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium tuberculosis (MTB). An estimated 1.7 billion people worldwide are infected with Mycobacterium tuberculosis (LTBI) during the incubation period without any obvious symptoms. Because of MTB’s high infection and mortality rates, there is an urgent need to develop a fast, portable, and sensitive diagnostic technology for its detection. Methods We included research from PubMed, Cochrane Library, Web of Science, and Embase and extracted the data. MetaDisc and STATA were used to build forest plots, Deek’s funnel plot, Fagan plot, and bivariate boxplot for analysis. Results Forty-six articles were analyzed, the results of which are as follows: sensitivity and specificity were 0.92 (0.91–0.93) and 0.95 (0.94–0.95) respectively. The NLR and PLR were 0.04 (95% CI 0.03–0.07) and 25.32 (95% CI 12.38–51.78) respectively. DOR was 639.60 (243.04–1683.18). The area under the SROC curve (AUC) was 0.99. Conclusions MPT64 exhibits good diagnostic efficiency for MTB. There is no obvious heterogeneity between the three commercial kits.

This study aimed to assess the consistency and replicability of treatment recommendations provided by ChatGPT 3.5 compared to gastrointestinal tumor cases presented at multidisciplinary tumor boards (MTBs). It also aimed to distinguish between general and case-specific responses and investigated the precision of ChatGPT's recommendations in replicating exact treatment plans, particularly regarding chemotherapy regimens and follow-up protocols. A retrospective study was carried out on 115 cases of gastrointestinal malignancies, selected from 448 patients reviewed in MTB meetings. A senior resident fed patient data into ChatGPT 3.5 to produce treatment recommendations, which were then evaluated against the tumor board's decisions by senior oncology fellows. Among the examined cases, ChatGPT 3.5 provided general information about the malignancy without considering individual patient characteristics in 19% of cases. However, only in 81% of cases, ChatGPT generated responses that were specific to the individual clinical scenarios. In the subset of case-specific responses, 83% of recommendations exhibited overall treatment strategy concordance between ChatGPT and MTB. However, the exact treatment concordance dropped to 65%, notably lower in recommending specific chemotherapy regimens. Cases recommended for surgery showed the highest concordance rates, while those involving chemotherapy recommendations faced challenges in precision. ChatGPT 3.5 demonstrates potential in aligning conceptual approaches to treatment strategies with MTB guidelines. However, it falls short in accurately duplicating specific treatment plans, especially concerning chemotherapy regimens and follow-up procedures. Ethical concerns and challenges in achieving exact replication necessitate prudence when considering ChatGPT 3.5 for direct clinical decision-making in MTBs. • ChatGPT 3.5 shows promise as a valuable tool in clinical decision-making. • ChatGPT 3.5 provides case-specific recommendations when provided with sufficient information. • In 83% of cases, ChatGPT 3.5 generates conceptual treatment recommendations that align with the MTB. • ChatGPT 3.5 struggles to generate case-specific recommendations, achieving a 65% concordance rate. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is one of the leading causes of infectious mortality from a single infectious agent, Mycobacterium tuberculosis (MTB). This study evaluated the performance of the newly developed BZ TB/NTM NALF assay, which integrated loop-mediated isothermal amplification and lateral flow immunochromatographic assay technologies, for the detection of MTB. A total of 80 MTB-positive samples and 115 MTB-negative samples were collected, all of which were confirmed by TB real-time PCR (RT-PCR) using either AdvanSureTM TB/NTM RT-PCR Kit or Xpert® MTB/RIF Assay. The performance of the BZ TB/NTM NALF assay was evaluated by calculating its sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) in comparison to those of the RT-PCR methods. Compared to the RT-PCR, the sensitivity, specificity, PPV, and NPV of BZ TB/NTM NALF assay were 98.7%, 99.1%, 98.7%, and 99.1%, respectively. The concordance rate between BZ TB/NTM NALF and RT-PCR was 99.0%. Rapid and simple detection of MTB is essential for global case detection and further elimination of TB. The performance of the BZ TB/NTM NALF Assay is acceptable with a high concordance with RT-PCR, indicating that it is reliable for use in a low-resource environment.

Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium tuberculosis (MTB) and a significant health concern worldwide. The main threat to the elimination of TB is the development of resistance by MTB to the currently used antibiotics and more extended treatment methods, which is a massive burden on the health care system. As a result, there is an urgent need to identify new, effective therapeutic strategies with fewer adverse effects. The traditional medicines found in South Asia and Africa have a reservoir of medicinal plants and plant-based compounds that are considered another reliable option for human beings to treat various diseases. Abundant research is available for the biotherapeutic potential of naturally occurring compounds in various diseases but has been lagging in the area of TB. Plant-based compounds, or phytoproducts, are being investigated as potential anti-mycobacterial agents by reducing bacterial burden or modulating the immune system, thereby minimizing adverse effects. The efficacy of these phytochemicals has been evaluated through drug delivery using nanoformulations. This review aims to emphasize the value of anti-TB compounds derived from plants and provide a summary of current research on phytochemicals with potential anti-mycobacterial activity against MTB. This article aims to inform readers about the numerous potential herbal treatment options available for combatting TB.

For most vaccination studies, the assessment of vaccine-induced CD4+ and CD8+ T cells has relied upon the measurement of antigen-specific polyfunctional cells, typically using recombinant antigen or peptide pools. However, this approach leaves open the question as to whether or not these cells are responsive to the Mtb-infected cell within the context of Mtb infection and hence leaves open the possibility that a key parameter of vaccine immunogenicity may be overlooked. In this review, we discuss the case that these measurements almost certainly over-estimate the capacity of both CD4+ and CD8+ T cells to recognize the Mtb-infected cell. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) is a leading cause of morbidity and mortality worldwide. Control of TB is lingering by the lack of diagnostic tests that are simple, rapid, yet accurate. Thus, smear-negative pulmonary TB often misses the diagnosis. The study evaluated the performance of GeneXpert MTB/RIF assay for the detection of Mycobacterium tuberculosis (MTB).Methods: The study was carried out from June to December 2016 in Nepal Tuberculosis Center, Bhaktapur, Nepal. A total of 173 sputum samples were collected and processed by microscopy [Auramine-O staining and Ziehl-Neelsen (ZN) staining], followed by GeneXpert MTB/RIF assay and culture in Lowenstein-Jensen (LJ) medium.Results: Of 173 sputum samples, 162 (93.6%) were smear-negative. Of 162 smear-negative sputum samples, 35 (21.6%) were confirmed to have MTB by culture, and 31 (19.1%) by GeneXpert MTB/RIF assay. Of 31 GeneXpert-positive samples, 25 (80.6%) were susceptible, 4 (12.9%) were resistant, and 2 (6.45%) were intermediate to rifampicin. The sensitivity, specificity, positive predictive value, and negative predictive value of GeneXpert MTB/RIF assay for smear-negative sputum samples were 74.3%, 96.6%, 86.7%, and 92%, respectively. The GeneXpert MTB/RIF has a substantial diagnostic agreement of 90.91% with culture (Cohen's Kappa coefficient = 0.73).Conclusion: The diagnostic performance of GeneXpert MTB/RIF assay was almost on par with culture, and thus can be relied upon for MTB detection in smear-negative sputum samples. [ABSTRACT FROM AUTHOR]

Fused‐azepinones are interesting heterocyclic scaffolds present in various natural products and synthetic derivatives with potent kinase inhibition, anti‐cancer, anti‐inflammatory, anti‐HIV, neuroprotective, anti‐fouling and other biological activities. These fused‐azepinones are also known for their antimicrobial activity against the Plasmodium falciparum, Leishmania, Escherichia coli, and Streptococcus pneumonia. In the present work, we synthesized a library of 3,4‐dihydro‐1H‐benzo[b]azepine‐2,5‐dione derivatives and evaluated them for their antibacterial potential against a panel of bacterial pathogens. The Structure‐activity relationship studies revealed the essential structural features for the promising antibacterial properties against Staphylococcus aureus and Mycobacterium tuberculosis with MIC 4–64 μg/mL. In addition, these compounds exhibited favourable selectivity index (SI≥10) in cytotoxicity studies. With the interesting antibacterial properties exhibited and good selectivity index, these compounds have emerged as promising candidates for further development. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) is a communicable disease remains a major global health problem and the leading cause of death from a single infectious agent. Even though many of the WHO recommended TB control strategies were implemented; there is still a major gap in TB case detection and treatment. This study aimed to determine the prevalence of Mycobacterium tuberculosis among presumptive TB patients in Gedeo Zone, Southern Ethiopia. Methods: A cross-sectional study was conducted on 384 TB suspected patients in Gedeo Zone from February to July 2021. Data were collected using a pretested structured questionnaire. Laboratory examination was processed using Xpert-MTB/RIF assay. Data entry was made using Epi info version 7 and analyzed by SPSS version 24. Logistic regression models were used to determine the risk factors. Results: Out of 384 study participants suspected with TB, M. tuberculosis was isolated from 103 giving an overall prevalence of 26.8%. Males (AOR) = 1.95; 95% CI 1.56–2.65, P = 0.01) were more likely to develop TB than females. Study participants who were illiterate (AOR 2.10; 95% CI 1.17–2.51, P = 0.014) were more likely to develop TB than the educated ones. Cigarette smokers (AOR 2.89; 95% CI 2.10–3.84, P = 0.01), khat chewers (AOR 2.86; 95% CI 1.28–3.79, P = 0.01), vaccination (AOR 0.52; 95% CI 0.21–0.88, P = 0.02), close contact (AOR 3.42; 95% CI 2.24–4.50, P = 0.01) and being positive for HIV (AOR 2.01; 95% CI 1.07–3.52, 0.01) were more likely to develop TB. Conclusion: Despite implementation of national and international TB control strategies, TB still remains one of the major public health problems in the country especially in the study area. The high prevalence of MTB was reported different risk groups. Early case detection and management of TB should be given special attention to strengthen and an appropriate control and prevention methods to reduce the emergence and increasing of MTB cases. [ABSTRACT FROM AUTHOR]

For most vaccination studies, the assessment of vaccine-induced CD4+ and CD8+ T cells has relied upon the measurement of antigen-specific polyfunctional cells, typically using recombinant antigen or peptide pools. However, this approach leaves open the question as to whether or not these cells are responsive to the Mtb-infected cell within the context of Mtb infection and hence leaves open the possibility that a key parameter of vaccine immunogenicity may be overlooked. In this review, we discuss the case that these measurements almost certainly over-estimate the capacity of both CD4+ and CD8+ T cells to recognize the Mtb-infected cell.

Mangroves are trees or large shrubs which have special adaptations to survive in this environment and grow within the intertidal zone in tropical and subtropical regions. Sesuvium portulacastrum (seapurslane) is, herbaceous, perennial, dichotomous, halophyte fast growing plant that belongs to family Aizoaceae. Salicornia brachiata Roxb, is a halophyte, commonly known as sea asparagus widely used as food item in several countries. Then, In vitro studies like, antioxidant, anti-inflammatory and antimycobacterial assay was carried out. The IC50 values of Salicornia brachiata and Sesuvium portulacastru was found to be 288.166, 369.451 and 369.764 µg/ml for DPPH scavenging activity. The presence of various primary and secondary metabolites was revealed in the present study. It also showed the better in vitro activities and was the most active plants against MDRTB, MTB and H37RV strain. This study validates the medicinal use of plants in the treatment of TB which have promising effect against MDRTB, MTB and H37RV. [ABSTRACT FROM AUTHOR]

This study analyses the challenges and implications of technological development in mountain-bike sports for Alpine tourism destinations. We investigate the motivations of e-mountain bikers in tourism destinations and derive implications for tourism managers to handle dynamic technological developments in a sustainable manner. A mixed-method design including qualitative in-depth interviews with relevant actors and a quantitative survey with 175 e-mountain bikers was applied. The e-bike boom offers potential and challenges for sustainable tourism management in Alpine destinations. Tourism managers must develop diverse, participatory, and nature-friendly concepts as well as collaborations with nearby destinations and recreational areas to control the flow of tourists within ecologically sensitive areas. [ABSTRACT FROM AUTHOR]

Purpose: This study was aimed at characterising Mycobacterium tuberculosis (Mtb) H37Rv Rv0292 protein peptides. As this protein forms a structural part of the cell wall and Mtb type VII secretion system, it is believed that Rv0292 has important functions regarding the disease's pathogenesis and could represent a good candidate when designing a synthetic anti-tuberculosis vaccine. Methods: Receptor-ligand binding assays were used for identifying peptides from the Rv0292 protein sequence having high specific binding activity (HABP) to infection target cells; some HABPs were able to inhibit Mtb H37Rv entry to such cell lines in in vitro assays. Fluorescent microsphere internalisation assays were carried out for corroborating such peptides' role in mycobacterial entry. The protein's predicted alfa helical structure was confirmed by circular dichroism of the peptides forming its sequence. Results: It was found that peptide 31050 (101 LTLIARYLDRYGIRADTIRI 120) had high binding capacity for the cell lines used here and inhibited mycobacterial entry to such cell lines; it was also found that this peptide was not recognised by sera from individuals who had been in contact with the mycobacteria. Conclusions: The aforementioned characteristics led to suggesting that peptide 31050 sequence plays an important role in Mtb H37Rv Rv0292 protein interaction with infection target cells and could be considered a candidate when designing a synthetic anti-tuberculosis vaccine. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is a serious lung infection caused by the bacterium Mycobacterium tuberculosis (M. tuberculosis) is highly contagious, and can be spread through the air from an infected person to others. The aim of this article was to evaluate the immunological role of CD4 and CD20 in patients infected with M. tuberculosis and multi-drug resistant tuberculosis (MDRTB). A total of 182 individuals suspected with M. tuberculosis admitted to the tuberculosis center in AL-Najaf City, Iraq. A sputum acid-fast stain was performed for each individual and GeneXpert® heminested real time PCR has been performed for MDR-M. tuberculosis detection. CD4 and CD20 have been measurement in serum of infected individual using ELISA technique. Serum CD4 and CD20 levels were significantly elevated (P

Tuberculosis (TB) is a lung infection caused by the bacterium Mycobacterium tuberculosis (MTB). TB is a leading cause of death globally, accounting for approximately 1.5 million deaths per year. The aim of this article was to evaluate the immunological role of Interleukin (IL) 18 and Interleukin 22 in patients infected with MTB and multi-drug resistant tuberculosis MDRTB. A total of 182 individuals suspected with MTB admitted to the tuberculosis center in AL-Najaf City, Iraq. A sputum acid-fast stain was performed for each individual and GeneXpert® heminested real time PCR has been performed for MDRTB detection. The two Interleukins was measurement in serum of infected individual using ELISA technique. Out of 182 individuals, there were 20 infected with MTB and 15 infected with MDRTB. Serum IL18 and IL22 levels were significantly elevated (P

N6-adenosine methylation (m6A) is a prevalent form of RNA modification found in the expressed transcripts of many eukaryotic organisms. Moreover, m6A methylation is a dynamic and reversible process that requires the functioning of various proteins and their complexes that are evolutionarily conserved between species and include methylases, demethylases, and m6A-binding proteins. Over the past decade, the m6A methylation process in plants has been extensively studied and the understanding thereof has drastically increased, although the regulatory function of some components relies on information derived from animal systems. Notably, m6A has been found to be involved in a variety of factors in RNA processing, such as RNA stability, alternative polyadenylation, and miRNA regulation. The circadian clock in plants is a molecular timekeeping system that regulates the daily and rhythmic activity of many cellular and physiological processes in response to environmental changes such as the day-night cycle. The circadian clock regulates the rhythmic expression of genes through post-transcriptional regulation of mRNA. Recently, m6A methylation has emerged as an additional layer of post-transcriptional regulation that is necessary for the proper functioning of the plant circadian clock. In this review, we have compiled and summarized recent insights into the molecular mechanisms behind m6A modification and its various roles in the regulation of RNA. We discuss the potential role of m6A modification in regulating the plant circadian clock and outline potential future directions for the study of mRNA methylation in plants. A deeper understanding of the mechanism of m6A RNA regulation and its role in plant circadian clocks will contribute to a greater understanding of the plant circadian clock.

ATP synthase subunit c (AtpE) is an enzyme that catalyzes the production of ATP from ADP in the presence of sodium or proton gradient from Mycobacterium tuberculosis (MTB). This enzyme considered an essential target for drug design and shares the same pathway with the target of Isoniazid. Thus, this enzyme would serve as an alternative target of the Isoniazid. The three dimensional (3D) model structure of the AtpE was constructed based on the principle of homology modeling using the Modeller9.16. The developed model was subjected to energy minimization and refinement using molecular dynamic (MD) simulation. The minimized model structure was searched against Zinc and PubChem database to determine ligands that bind to the enzyme with minimum binding energy using RASPD and PyRx tool. A total of 4776 compounds capable of bindings to AtpE with minimum binding energy were selected. These compounds further screened for physicochemical properties (Lipinski rule of five). All the compounds that possessed the desirable property selected and used for molecular docking analysis. Five (5) compounds with minimum binding energies ranged between ─8.69, and ─8.44 kcal/mol, less than the free binding energy of ATP were selected. These compounds further screened for the absorption, distribution, metabolism, excretion, and toxicity (ADME and toxicity) properties. Of the five compounds, three (ZINC14732869, ZINC14742188, and ZINC12205447) fitted all the ADME and toxicity properties and subjected to MD simulation and Molecular Mechanics Generalized Born and Surface Area (MM-GBSA) analyses. The results indicated that the ligands formed relatively stable complexes and had free binding energies, less than the binding energy of the ATP. Therefore, these ligands considered as prospective inhibitors of MTB after successful experimental validation.

Objective: Mycobacterium tuberculosis (Mtb) infection is quickly countered by the host immune system; but, the pathogen is never eradicated. In this infection, 10% of infections are considered as open tuberculosis, while the remaining 90% are becoming latent, a state that can persist in the host for decades until immune control is lost. To date, more than 350 plant species have been studied in mycobacterial research. Methods: In this research, we used a mixture of software and database explorations to find the anti-mycobacterial possibilities of phytoconstituents. Maestro v12.7 and Cytoscape v3.8.0 were applied as software. In addition, the String v11.0, KEGG, Reactome and Pass Server databases assisted us in understanding TB cellular processes and phytoconstituent drug possibilities. Results: The racemosol and pacharin have registered higher docking scores of 7.823 and -7.715 respectively. Furthermore, these compounds have substantial binding affinities in the active pocket of Mtb, and their docking metrics have consisted close to that of an approved drug (Rifampin). The network pharmacological exploration is also revealed possible connections with all of the TB-associated genes and other important signaling pathways. Conclusions: This computational study found that racemosol and pacharin were the safest anti-mycobacterial drugs for tuberculosis. Following comprehensive research of the pharmacological properties of racemosol and pacharin, we decided that this would be the first study to focus on anti-mycobacterial activity utilizing those compounds. Also, we believe that racemsosol and pacharin compounds will reveal potential actions when used in mycobacterial research. Furthermore, the derivatives of these compounds will be acted as possible drugs for tuberculosis and associated diseases. More pharmacological and toxicological explorations are required to have a better understanding of these compounds.

Tuberculosis (TB) is one of the three leading causes of death from a single infectious agent, Mycobacterium tuberculosis (MTB), together with COVID-19 and HIV/AIDS. This disease places a heavy burden on countries with low socio-economic development and aggravates existing inequalities. For the year 2021, estimations for Ecuador were 8500 TB cases, of which 370 were associated to multiple drug resistance (TB-MDR), and 1160 deaths. In the same year, Ecuador notified 5973 total cases, 401 of them were TB-MDR, pointing out an under diagnosis problem. The few molecular epidemiology studies available conclude that L4 is the most prevalent MTB lineage in Ecuador (with LAM as the main L4 sublineage), but L2-Beijing family is also present at low prevalence. Nevertheless, with less than 1 % MTB isolates genetically characterized by either MIRU-VNTR, spolygotyping or WGS to date, molecular epidemiology research must me improved to assist the TB surveillance and control program in Ecuador., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Author(s).)

N 6 -methyladenosine (m 6 A), a well-characterized RNA modification, is involved in regulating multiple biological processes; however, genome-wide identification and functional characterization of the m 6 A modification in legume plants, including soybean ( Glycine max (L.) Merr.), remains lacking. In this study, we utilized bioinformatics tools to perform comprehensive analyses of molecular writer candidates associated with the RNA m 6 A modification in soybean, characterizing their conserved domains, motifs, gene structures, promoters, and spatial expression patterns. Thirteen m 6 A writer complex genes in soybean were identified, which were assigned to four families: MT-A70, WTAP, VIR, and HAKAI. It also can be identified that multiple cis elements in the promoters of these genes, which were classified into five distinct groups, including elements responsive to light, phytohormone regulation, environmental stress, development, and others, suggesting that these genes may modulate various cellular and physiological processes in plants. Importantly, the enzymatic activities of two identified m 6 A writers, GmMTA1 and GmMTA2, were confirmed in vitro . Furthermore, we analyzed the expression patterns of the GmMTA s and GmMTB s under different abiotic stresses, revealing their potential involvement in stress tolerance, especially in the response to alkalinity or darkness. Overexpressing GmMTA2 and GmMTB1 in soybean altered the tolerance of the plants to alkalinity and long-term darkness, further confirming their effect on the stress response. Collectively, our findings identified the RNA m 6 A writer candidates in leguminous plants and highlighted the potential roles of GmMTA s and GmMTB s in the response to abiotic stress in soybean., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Liu, Liu, Zhao, Yang, Guo, Chang, Xiao, Xu, Liu, Zhu, Gan and Chen.)

Mycobacterium tuberculosis (Mtb) is an intracellular pathogen capable of adapting and surviving within macrophages, utilizing host nutrients for its growth and replication. Cholesterol is the main carbon source during the infection process of Mtb. Cholesterol metabolism in macrophages is tightly associated with cell functions such as phagocytosis of pathogens, antigen presentation, inflammatory responses, and tissue repair. Research has shown that Mtb infection increases the uptake of low-density lipoprotein (LDL) and cholesterol by macrophages, and enhances de novo cholesterol synthesis in macrophages. Excessive cholesterol is converted into cholesterol esters, while the degradation of cholesterol esters in macrophages is inhibited by Mtb. Furthermore, Mtb infection suppresses the expression of ATP-binding cassette (ABC) transporters in macrophages, impeding cholesterol efflux. These alterations result in the massive accumulation of cholesterol in macrophages, promoting the formation of lipid droplets and foam cells, which ultimately facilitates the persistent survival of Mtb and the progression of tuberculosis (TB), including granuloma formation, tissue cavitation, and systemic dissemination. Mtb infection may also promote the conversion of cholesterol into oxidized cholesterol within macrophages, with the oxidized cholesterol exhibiting anti-Mtb activity. Recent drug development has discovered that reducing cholesterol levels in macrophages can inhibit the invasion of Mtb into macrophages and increase the permeability of anti-tuberculosis drugs. The development of drugs targeting cholesterol metabolic pathways in macrophages, as well as the modification of existing drugs, holds promise for the development of more efficient anti-tuberculosis medications., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Chen, Kong, Ma, Chen, Zeng, Liu, Wang and Lu.)

Background: Co-infection of Mycobacterium tuberculosis (MTB) and the Human Immunodeficiency Virus (HIV) is a major global public health issue, particularly in border areas of resource-limited nations, including Ethiopia., Objective: To explore the prevalence and associated variables of MTB/HIV co-infection among PTB presumptive patients in Northwest Ethiopia., Methods: From February to August 2021, a cross-sectional institutional investigation was conducted at the Metema and Abrehajira hospitals. Semi-structured questionnaires were used to collect socio-demographic and clinical data. The MTB/RIF Xpert assay was used to process sputum, and 3 ml of veins blood was collected for HIV rapid test (STAT-PAK, ABON, and SD BIOLINE HIV test algorithm) following the Ethiopian National HIV test algorithm. The Gene Xpert assay's sample processing control was checked to ensure data quality. Data entered into Epi-Data were exported to SPSS version 20 for analysis. Statistically significant variables (p-value ≤0.2) from bivariable analysis were included in multivariable analysis. A p-value ≤ 0.05 was judged statistically significant., Results: This study included 314 PTB presumptive patients with a median age of 35.0 years, of which 178 (56.69 %) were males. Among all patients, 40(12.7 %) and 51(16.2 %) were PTB, and HIV seropositivity, respectively. Of the PTB patients, 14/40 (35 %) (95 % CI: 24.4-45.6) were co-infected with HIV/AIDS. Married patients were 70 % less likely than unmarried individuals (AOR = 0.3 CI; 0.07-0.98) to have MTB/HIV co-infection. Patients who had contact history with MDR-TB patients (AOR = 5 CI; 1.37-18.00), and those who had a history of alcohol use (AOR = 12.2 CI; 2.56-57.8) were more likely to have MTB-HIV co-infection than their peers., Conclusion: Our findings showed that MTB-HIV co-infection is one of the most important community health concerns in the study area. Therefore, MTB/HIV cooperation activities should be fully in place to prevent co-infection and its impact on the population., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Mucheye Gizachew reports equipment, drugs, or supplies was provided by Department of Medical Microbiology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, P. O. Box 196, Gondar, Ethiopia. Mucheye Gizachew reports a relationship with Department of Medical Microbiology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, P. O. Box 196, Gondar, Ethiopia that includes: employment. Not applicable has patent pending to Not applicable. Not applicable If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

Background: Rapid and reliable diagnosis of tuberculosis (TB) represents a diagnostic challenge in compartmentalized extrapulmonary TB infection because of the small number of mycobacteria (MTB) and the frequent lack of fresh samples to perform culture. Here, we estimate the performances of homemade droplet digital PCR (ddPCR)-based assays against culture in 89 biopsies, for those fresh and formalin-fixed and paraffin-embedded (FFPE) subsamples were available. Methods: MTB diagnosis in fresh subsamples was performed by culture. Fresh subsamples were also analyzed for acid-fast bacilli smear-microscopy (AFB) and Xpert® MTB/RIF (Xpert). MTB examination was repeated in blind in the 89 FFPE subsamples by in-house ddPCR assays targeting the IS6110 and rpoB. Analytical sensitivity of ddPCR assays was evaluated using serial dilution of H37Rv strain. Limit of detection (LOD) was calculated by probit analysis. Results were expressed in copies/106 cells. Results: IS6110 and rpoB ddPCR assays showed a good linear correlation between expected and observed values (R 2: 0.9907 and 0.9743, respectively). Probit analyses predicted a LOD of 17 and 40 copies/106 cells of MTB DNA for IS6110 and rpoB, respectively. Of the 89 biopsies, 68 were culture positive and 21 were culture negative. Considering mycobacterial culture as reference method, IS6110 assay yielded positive results in 67/68 culture-positive samples with a median interquartile range (IQR) of 1,680 (550–8,444) copies/106 cells (sensitivity: 98.5%; accuracy: 98.9). These performances were superior to those reported by the rpoB assay in FFPE subsamples (sensitivity: 66.20%; accuracy: 74.1) and even superior to those reported by Xpert and AFB in fresh subsamples (sensitivity: 79.4 and 33.8%, respectively; accuracy: 84.3 and 49.4, respectively). When Xpert and AFB results were stratified according to mycobacterial load detected by rpoB and IS6110 ddPCR, bacterial load was lower in Xpert and AFB negative with respect to Xpert and AFB-positive samples (p = 0.003 and 0.01 for rpoB and p = 0.01 and 0.11 for IS6110), confirming the poor sensitivity of these methods in paucibacillary disease. Conclusion: ddPCR provides highly sensitive, accurate, and rapid MTB diagnosis in FFPE samples, as defined by the high concordance between IS6110 assay and culture results. This approach can be safely introduced in clinical routine to accelerate MTB diagnosis mainly when culture results remain unavailable. [ABSTRACT FROM AUTHOR]

Bronchoalveolar lavage (BAL) is the saline wash of the bronchial tree, which aids in diagnosing various pulmonary pathologies. The present study was contemplated with an aim to know the clinical, microbiological profile of BAL samples along with its sensitivity pattern and to assess its utility as a diagnostic tool. This was a prospective observational study, carried over 90 patients presenting with lower respiratory tract infections. The total microbial recovery rate from BAL was 39 (43.3%). The sensitivity, specificity and positive predictive value of BAL were found to be 76.4%, 89.7% and 90.6% respectively. Maximum isolates were bacteria (25.5%) followed by Mycobacterium tuberculosis (MTB) (16.6%) and fungi (1.1%). Predominant bacterial isolates were Gram-negative (81.5%) compared to Gram-positive (18.5%). Multidrug-resistance (MDR) in bacteria was seen in 59.2% of isolates. BAL is a valuable diagnostic tool to find not only bacterial but mycobacterial and fungal infections in patients with lower respiratory tract infection (LRTI). A trend towards LRTI with Gram negative infections is on the rise and they tend to be multi drug resistant. Hence checking susceptibility patterns is crucial to start evidence-based treatment. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is a chronic infectious disease mainly caused by Mycobacterium tuberculosis (MTB), but other members of the Mycobacterium tuberculosis complex (MTBC), especially Mycobacterium bovis (pyrazinamide-resistant organisms), may also be involved. Thus, the ability to rapidly detect and identify MTB from other MTBC members (e.g., M. bovis , Mycobacterium microti , Mycobacterium africanum) is essential for the prevention and treatment of TB. A novel diagnostic method for the rapid detection and differentiation of MTB, which employs multiplex loop-mediated isothermal amplification (mLAMP) combined with a nanoparticle-based lateral flow biosensor (LFB), was established (mLAMP-LFB). Two sets of specific primers that target the IS6110 and mtp40 genes were designed according to the principle of LAMP. Various pathogens were used to optimize and evaluate the mLAMP-LFB assay. The optimal conditions for mLAMP-LFB were determined to be 66°C and 40 min, and the amplicons were directly verified by observing the test lines on the biosensor. The LAMP assay limit of detection (LoD) was 125 fg per vessel for the pure genomic DNA of MTB and 4.8 × 103 CFU/ml for the sputum samples, and the analytical specificity was 100%. In addition, the whole process, including the clinical specimen processing (35 min), isothermal amplification (40 min), and result confirmation (1–2 min), could be completed in approximately 80 min. Thus, mLAMP-LFB is a rapid, reliable, and sensitive method that is able to detect representative members of MTBC and simultaneously differentiate MTB from other MTBC members, and it can be used as a potential screening tool for TB in clinical, field, and basic laboratory settings. [ABSTRACT FROM AUTHOR]

ABSTRACT Magnetosomes are complex membrane organelles synthesized by magnetotactic bacteria (MTB) for navigation in the Earth’s magnetic field. In the alphaproteobacterium Magnetospirillum gryphiswaldense, all steps of magnetosome formation are tightly controlled by >30 specific genes arranged in several gene clusters. However, the transcriptional organization of the magnetosome gene clusters has remained poorly understood. Here, by applying Cappable-seq and whole-transcriptome shotgun RNA sequencing, we show that mamGFDCop and feoAB1op are transcribed as single transcriptional units, whereas multiple transcription start sites (TSS) are present in mms6op, mamXYop, and the long (>16 kb) mamABop. Using a bioluminescence reporter assay and promoter knockouts, we demonstrate that most of the identified TSS originate from biologically meaningful promoters which mediate production of multiple transcripts and are functionally relevant for proper magnetosome biosynthesis. In addition, we identified a strong promoter in a large intergenic region within mamXYop, which likely drives transcription of a noncoding RNA important for gene expression in this operon. In summary, our data suggest a more complex transcriptional architecture of the magnetosome operons than previously recognized, which is largely conserved in other magnetotactic Magnetospirillum species and, thus, is likely fundamental for magnetosome biosynthesis in these organisms. IMPORTANCE Magnetosomes have emerged as a model system to study prokaryotic organelles and a source of biocompatible magnetic nanoparticles for various biomedical applications. However, the lack of knowledge about the transcriptional organization of magnetosome gene clusters has severely impeded the engineering, manipulation, and transfer of this highly complex biosynthetic pathway into other organisms. Here, we provide a high-resolution image of the previously unappreciated transcriptional landscape of the magnetosome operons. Our findings are important for further unraveling the complex genetic framework of magnetosome biosynthesis. In addition, they will facilitate the rational reengineering of magnetic bacteria for improved bioproduction of tunable magnetic nanoparticles, as well as transplantation of magnetosome biosynthesis into foreign hosts by synthetic biology approaches. Overall, our study exemplifies how a genetically complex pathway is orchestrated at the transcriptional level to ensure the balanced expression of the numerous constituents required for the proper assembly of one of the most intricate prokaryotic organelles.

Background: Rapid and reliable diagnosis of tuberculosis (TB) represents a diagnostic challenge in compartmentalized extrapulmonary TB infection because of the small number of mycobacteria (MTB) and the frequent lack of fresh samples to perform culture. Here, we estimate the performances of homemade droplet digital PCR (ddPCR)-based assays against culture in 89 biopsies, for those fresh and formalin-fixed and paraffin-embedded (FFPE) subsamples were available.Methods: MTB diagnosis in fresh subsamples was performed by culture. Fresh subsamples were also analyzed for acid-fast bacilli smear-microscopy (AFB) and Xpert® MTB/RIF (Xpert). MTB examination was repeated in blind in the 89 FFPE subsamples by in-house ddPCR assays targeting the IS6110 and rpoB. Analytical sensitivity of ddPCR assays was evaluated using serial dilution of H37Rv strain. Limit of detection (LOD) was calculated by probit analysis. Results were expressed in copies/106 cells.Results: IS6110 and rpoB ddPCR assays showed a good linear correlation between expected and observed values (R2: 0.9907 and 0.9743, respectively). Probit analyses predicted a LOD of 17 and 40 copies/106 cells of MTB DNA for IS6110 and rpoB, respectively. Of the 89 biopsies, 68 were culture positive and 21 were culture negative. Considering mycobacterial culture as reference method, IS6110 assay yielded positive results in 67/68 culture-positive samples with a median interquartile range (IQR) of 1,680 (550–8,444) copies/106 cells (sensitivity: 98.5%; accuracy: 98.9). These performances were superior to those reported by the rpoB assay in FFPE subsamples (sensitivity: 66.20%; accuracy: 74.1) and even superior to those reported by Xpert and AFB in fresh subsamples (sensitivity: 79.4 and 33.8%, respectively; accuracy: 84.3 and 49.4, respectively). When Xpert and AFB results were stratified according to mycobacterial load detected by rpoB and IS6110 ddPCR, bacterial load was lower in Xpert and AFB negative with respect to Xpert and AFB-positive samples (p = 0.003 and 0.01 for rpoB and p = 0.01 and 0.11 for IS6110), confirming the poor sensitivity of these methods in paucibacillary disease.Conclusion: ddPCR provides highly sensitive, accurate, and rapid MTB diagnosis in FFPE samples, as defined by the high concordance between IS6110 assay and culture results. This approach can be safely introduced in clinical routine to accelerate MTB diagnosis mainly when culture results remain unavailable.