We present an open Molecular Crystal (MC) database of Machine-Learned Interatomic Potentials (MLIPs) called MolCryst-MLIPs. The first release comprises fine-tuned MACE models for nine molecular crystal systems─Benzamide, Benzoic acid, Coumarin, Durene, Isonicotinamide, Nicotinic acid, Niacinamide, Pyrazinamide, and Resorcinol─developed using the Automated Machine Learning Pipeline (AMLP), which streamlines the entire MLIP development workflow, from reference data generation to model training and validation, into a reproducible and user-friendly pipeline. Models are fine-tuned from the MACE-MH-1 foundation model (omol head), yielding a mean energy MAE of 0.141 kJ·mol-1·atom-1 and a mean force MAE of 0.648 kJ·mol-1·Å-1 across all systems. Benchmarked against three state-of-the-art foundation models on the DFT-labeled polymorph set, only the fine-tuned models reliably identify the most stable polymorph, with a mean Kendall τ = 0.397 indicating a moderate rank correlation with the full DFT landscape. Dynamical stability and structural integrity, as assessed through energy conservation, P2 orientational order parameters, and radial distribution functions, are evaluated using molecular dynamics simulations. The released models and data sets constitute a growing open database of validated MLIPs, ready for production MD simulations of molecular crystal polymorphism across the polymorphic landscape of each target compound under different thermodynamic conditions.

Whole-genome sequencing (WGS) is an increasingly adopted platform for predicting drug resistance in Mycobacterium tuberculosis; however, diagnostic accuracy varies substantially across bioinformatic tools and analytical frameworks, generating considerable uncertainty for clinical laboratory implementation. We conducted a prospectively registered (PROSPERO: CRD420261342739), PRISMA-DTA-compliant systematic review and meta-analysis of diagnostic accuracy studies. PubMed (MEDLINE), Embase, Web of Science, and Cochrane CENTRAL were searched from 1 January 2000 through 28 January 2026. Primary overall sensitivity and specificity were estimated using a tool-level bivariate random-effects model. Exploratory subgroup analyses and meta-regression examined the association between algorithm category and diagnostic-performance heterogeneity. Twenty-eight drug-level evaluations from seven tools (rifampicin, isoniazid, ethambutol, and pyrazinamide for each tool) were compiled from the extracted 2 × 2 data. For the primary tool-level composite analysis, pooled sensitivity was 0.930 (95% CI: 0.907-0.948) and pooled specificity was 0.962 (95% CI: 0.929-0.981). In secondary drug-specific analyses, sensitivity was highest for rifampicin (0.960, 95% CI: 0.934-0.976) and isoniazid (0.933, 95% CI: 0.906-0.953), and lowest for pyrazinamide (0.860, 95% CI: 0.800-0.904). Exploratory tool-level comparisons produced pooled sensitivity estimates of 0.920 for rule-based tools, 0.899 for machine learning tools, and 0.951 for hybrid tools. These comparisons involved only seven tool-level analytic units and cannot disentangle algorithm type from individual tool identity, training data, mutation catalogue version, or validation population. WGS-based bioinformatic tools provide highly specific and generally sensitive predictions of Mycobacterium tuberculosis resistance for first-line drugs across diverse clinical settings. Exploratory differences between tool categories should not be interpreted as causal effects of algorithmic architecture. Future studies should use prospective head-to-head evaluations on shared, geographically diverse isolate collections, alongside continued improvement of resistance catalogues and external validation.

The development of dual-acting hybrid antibiotics is a promising strategy to combat the ongoing spread of drug-resistant tuberculosis. Inspired by the structure of the natural dithiolopyrrolone hybrid antibiotic thiomarinol and based on the synergistic effects confirmed by checkerboard assays, we employed a molecular hybridization strategy. The dithiolopyrrolone natural product thiolutin was covalently linked to the pharmacophores of four clinical antitubercular drugs-cycloserine, linezolid, isoniazid, and pyrazinamide-through alkyl linkers of 7-10 carbon atoms via amide condensation, leading to the design and synthesis of 15 novel hybrids. In vitro antitubercular activity evaluation revealed that the cycloserine series exhibited the best activity, with MIC values as low as 1 μg/mL, followed by the isoniazid series. Cytotoxicity assays showed that all cycloserine hybrids had IC50 values > 40 μg/mL against RAW 264.7 mouse macrophages, markedly lower than that of thiolutin alone. Among them, T1-CS and T4-CS displayed the best selectivity indices, achieving an effective reduction in cytotoxicity. This study successfully constructed a class of thiolutin-cycloserine hybrids with low cytotoxicity and high selectivity, providing a valuable molecular template for the discovery of novel antitubercular lead compounds.

A green and efficient one-pot synthetic method has been developed to prepare novel 5-amino-3-(aryl)-1-(aryl)-1H-pyrazole-4-carbonitrile analogues, employing ammonium acetate as a mild, green catalyst. The proposed methodology provides a rapid, operationally simple, and sustainable approach for the construction of biologically relevant heterocyclic scaffolds with good yields. Confirmation of the synthesized compounds was achieved using IR, 1H NMR, 13C NMR, and HRMS. All synthesized derivatives were subjected to in vitro biological assessment for their anti-inflammatory, antitubercular, antibacterial, and antifungal activities. Several compounds demonstrated promising pharmacological profiles. In particular, compounds 4d, and 4f exhibited significant COX-2 inhibitory activity, with IC50 value of 77.62 and 65.95 μg/mL. Antitubercular screening revealed compounds 4d and 4e showed enhanced potency, displaying sensitivity at the MIC value of 3.12 μg/mL, compared to the standard reference drug pyrazinamide. Furthermore, the synthesized derivatives exhibited notable antibacterial and antifungal activities comparable to reference drugs, indicating their broad-spectrum antimicrobial potential. Computational studies, including molecular docking, MD simulations, and MM-GBSA, provided further support for the observed COX-2 inhibition and indicated favorable pharmacokinetic profiles. These findings highlight the synthetic efficiency of the developed one-pot protocol and the promising biological potential of the synthesized 5-Amino-4-cyanopyrazole derivatives. Further studies involving structural optimisation, in vivo assessment, and mechanistic investigations are warranted to advance these compounds toward pharmaceutical development.

Extrapulmonary tuberculosis (EPTB) is characterized by low bacterial load and complex matrix in lesion samples. Therefore, current tuberculosis (TB) diagnostic methods often lose sensitivity to detect Mycobacterium tuberculosis (Mtb), leading to missed diagnosis and delayed treatment. Herein, we presented a single-cell metabolic labeling probe for tuberculosis (SCMLP-TB), which exhibited supporting investigative evidence for diagnosis of renal TB case, and enabled investigative monitoring of therapeutic efficacy of anti-TB regimens. A 51-year-old patient presented with fever, generalized weakness and muscle soreness on admission. Chest computed tomography (CT) revealed calcified mediastinal lymph nodes, and relevant renal function indicators were abnormal. Conventional TB diagnostic methods, including Xpert MTB/RIF, TB-DNA testing, and culture, yielded negative results using sputum, blood, and urine samples. Notably, SCMLP-TB assay detected Mtb in urine samples, suspecting renal TB. SCMLP-TB assay further detected gradually decreased bacterial viability along with anti-TB treatment (isoniazid, rifapentine, ethambutol, and pyrazinamide), revealing the efficacy of current regimen. The patient's symptom resolution and anemia/renal function improvement provided auxiliary evidence of antimicrobial efficacy. Ultimately, SCMLP-TB showed great potential for non-invasive, sensitive detection of suspected renal TB, and enabled investigative monitoring of the therapeutic efficacy.

Objective: To analyze the drug resistance spectrum of clinical isolates of Mycobacterium (M.) tuberculosis and the mutations in drug resistance-related genes in the genomic sequences of rifampicin-resistant isolates from elderly pulmonary tuberculosis (TB) patients in Beijing in 2024. Methods: A total of 474 M. tuberculosis isolates from elderly pulmonary TB patients were included. Drug susceptibility testing against 10 anti-TB drugs was performed with microbroth dilution method. Whole-genome sequencing (WGS) was performed for 27 rifampicin-resistant isolates to systematically characterize resistance-associated gene mutations and genotype-phenotype consistency. Results: The total drug resistance rate was 24.26% in the 474 M. tuberculosis isolates. The resistance rate was significantly higher in retreatment patients (44.00%) than in new patients (23.16%), and a higher multidrug resistance rate was observed in men (6.75% vs. 2.03%). WGS revealed that all the rifampicin-resistant strains belonged to lineage 2 (East Asian lineage), with rpoB p.Ser450Leu (55.56%) being the most prevalent mutation. Consistency between WGS and phenotypic drug susceptibility was 96.30% for isoniazid, pyrazinamide, and rifampicin, but was low for ethambutol (22.22%). Conclusions: The drug resistance of M. tuberculosis was severe in elderly pulmonary TB patients in Beijing, which was associated with treatment history and gender. Rifampicin-resistant strains showed lineage 2 clustering, indicating a risk for local transmission. The resistance genotypes detected by WGS were highly consistent with the phenotypic drug susceptibility of isoniazid and pyrazinamide, but showed low consistency with ethambutol.

Multidrug-resistant TB (MDR TB) disease is caused by TB bacteria that are resistant to at least isoniazid and rifampicin, the most effective first-line TB treatment drugs. MDR-TB poses a significant global health challenge due to its resistance to multiple antibiotics. World Health Organization (WHO) and other global health agencies have initiated targeted efforts to control its spread. The recent initiative taken by WHO is ENDTB Universal access to TB prevention and care. However, the eradication of the TB epidemic remains a challenging problem due to the emergence of specialized and modified drug resistance mechanisms. Among these mechanisms, the major facilitator superfamily (MFS) transporters play a crucial role in mediating drug resistance. The Rv0191 efflux transporter protein, a putative member of the MFS, has recently emerged as a potential target for investigating the molecular mechanisms behind the multidrug resistance (MDR) mechanism in Mtb, as it is involved in the efflux of PZA. To investigate the detailed molecular mechanism of Rv0191-mediated drug efflux, two conformations were studied: an outward-open (protonated) conformation for protonation analysis and an inward-open (unprotonated) conformation to examine substrate efflux. Pyrazinamide (PZA), a first-line anti-tuberculosis drug, was selected as the model substrate. Both conformations revealed a central tunnel with a single, distinct binding pocket. The efflux process was explored through a protonation-driven mechanism, highlighting the functional distinctions between the two conformational states. The systems were embedded in a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine lipid bilayer, and 500 ns of molecular dynamics simulations in triplicate were conducted for the PZA-docked structure. Trajectory-based structural analyses revealed specific interactions between PZA and the binding pocket, accompanied by conformational changes in Rv0191 that MD simulations indicate substrate movement. The observed structural dynamics suggest a plausible efflux pathway, allowing translocation of PZA from the cytoplasmic to the periplasmic side through the central tunnel, thereby suggesting a plausible mechanism of MFS-mediated drug resistance.

Miliary tuberculosis (TB) represents a potentially fatal form of disseminated TB resulting from the lymphohematogenous dissemination of Mycobacterium tuberculosis. Although accounting for approximately 1-2% of all TB cases in immunocompetent adults, its mortality rate exceeds 30%. This review screened peer-reviewed, original research studies via four databases (PubMed, EMBASE, MEDLINE, and CENTRAL) to synthesize up-to-date evidence. Key risk factors include human immunodeficiency virus (HIV) co-infection, anti-tumor necrosis factor-α (TNF-α) agents and chronic malnutrition. The clinical presentation of miliary TB is notoriously nonspecific, with fever, night sweats, weight loss, and constitutional symptoms predominating. Over 60% of patients have extrapulmonary complications, with central nervous system (CNS) involvement in 10-30%, abdominal involvement in 15-30%, and skeletal involvement in up to 17%. Laboratory abnormalities are common, including elevated erythrocyte sedimentation rate (ESR), anemia, lymphopenia, and nearly universal C-reactive protein (CRP) elevation. Diagnoses remain difficult due to paucibacillary lesions; novel molecular assays and a validated CNS prediction scoring system greatly improve diagnostic efficiency, while multiple microbiological and invasive tissue sampling modalities are briefly summarized for comprehensive clinical reference. Standard 6-month isoniazid, rifampicin, pyrazinamide ethambutol and isoniazid, rifampicin (HRZE-HR) regimens apply to isolated pulmonary miliary TB, whereas 9-12-month extended therapy plus adjunctive dexamethasone is required for CNS complications. Drug resistant cases demand 18-24-month individualized regimens with high blood-brain barrier penetration. Severe complications including tuberculous meningitis and high mortality acute respiratory distress syndrome (ARDS) are fully discussed, and TNF-α antagonists act as salvage therapy for steroid-refractory tuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS). Bacillus Calmette-Guérin (BCG) vaccination provides substantial protection against miliary TB in children, with efficacy of approximately 77% and favorable cost-effectiveness. Existing reviews lack integrated post-pandemic epidemiological data, cutting-edge molecular diagnostics and modern extrapulmonary treatment algorithms; this paper fills such gaps to offer balanced clinical guidance for frontline clinicians.

Pulmonary tuberculosis (TB) remains a major global health concern and may present with radiographic and laboratory findings that complicate timely diagnosis. Interferon-gamma release assays (IGRAs), including QuantiFERON-TB Gold, are blood tests that measure T-cell immune sensitization to Mycobacterium tuberculosis antigens; they do not distinguish latent infection from active disease and cannot exclude active TB. We report a 23-year-old immunocompetent Venezuelan-born man who presented with fever, night sweats, productive cough, and intermittent hemoptysis after approximately seven months of respiratory symptoms. Chest imaging demonstrated multilobar cavitary disease involving both upper lobes and the left lower lobe. Despite a negative QuantiFERON-TB Gold result, two sputum acid-fast bacilli smears were positive, M. tuberculosis nucleic acid amplification testing was positive, and mycobacterial culture confirmed the M. tuberculosis complex. The patient was placed in airborne isolation and treated with rifampin, isoniazid, pyrazinamide, and ethambutol. Public health authorities coordinated directly observed therapy, contact investigation, and outpatient sputum monitoring. This case demonstrates that a negative IGRA cannot rule out active pulmonary TB and that atypical multilobar cavitation should not lower clinical suspicion when epidemiologic, clinical, and microbiologic findings support the diagnosis.