Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis, continues to be a major global health threat despite being preventable and curable. This burden is particularly high in low and middle income countries. Microscopy remains essential for diagnosing TB by enabling direct visualization of Mycobacterium tuberculosis in sputum smear samples, offering a cost-effective approach for early detection and effective treatment. Given the labour-intensive nature of microscopy, automating the detection of bacilli in microscopic images is crucial to improve both the expediency and reliability of TB diagnosis. The current methodologies for detecting tuberculosis bacilli in bright field microscopic sputum smear images are hindered by limited automation capabilities, inconsistent segmentation quality, and constrained classification precision. This paper proposes a two-stage deep learning methodology for tuberculosis bacilli detection, comprising bacilli segmentation followed by classification. In the initial phase, an advanced U-Net model employing attention blocks and residual connections is proposed to segment microscopic sputum smear images, enabling the extraction of Regions of Interest (ROIs). The extracted ROIs are then classified using a Vision Transformer, which we specifically customized as TBViT to enhance the precise detection of bacilli within the images. For the experiments, a newly developed dataset of microscopic sputum smear images derived from Ziehl-Neelsen-stained slides is used in conjunction with existing public datasets. The qualitative and quantitative evaluation of the experiments using various metrics demonstrates that the proposed model achieves significantly improved segmentation performance, higher classification accuracy, and a greater level of automation, surpassing existing methods.

Thiosemicarbazones belong to the most versatile classes of physiologically active ligands in medicinal and bioinorganic chemistry due to their structural tunability, diverse coordination behaviour, and wide spectrum of pharmacological activity. They are rich in donor atoms and thione-thiol tautomerism and can coordinate a large number of transition and post-transition metal ions, usually leading to a marked increase in biological activity compared to the parent ligands. Thiosemicarbazone-based metal complexes have attracted a great deal of attention as potential therapeutic agents for the treatment of cancer, microbial infections, tuberculosis, parasitic diseases, and other pathological conditions. The therapeutic potentials are derived from a variety of processes, including metal chelation, disturbance of cellular metal homeostasis, production of reactive oxygen species, enzyme inhibition, and interactions with nucleic acids and proteins. Of note, the differences in ligand architecture, substituent pattern, donor atom environment, and metal center strongly affect biological activity, selectivity, and mechanism of action, emphasizing the role of structure-activity relationships in the rational design of effective metallodrugs. The present review critically summarizes recent progress in the coordination chemistry, biological properties, and therapeutic applications of metal complexes derived from thiosemicarbazones, focusing on anticancer, antimicrobial, antitubercular, antiparasitic, antioxidant, and diagnostic applications. Particular attention is devoted to ruthenium-thiosemicarbazone complexes owing to their unique coordination chemistry, favourable pharmacological properties, and significant therapeutic potential. Representative systems based on ruthenium, copper, silver, gold, gallium, zinc, nickel, palladium, platinum, and other biometals are described together with developing mechanistic insights and design techniques. Current issues in toxicity, selectivity, stability, pharmacokinetics, and clinical translation are also discussed. Collectively, the available evidence highlights the growing importance of thiosemicarbazone metal complexes as multifunctional therapeutic platforms and underscores their potential for the development of next-generation metallodrugs.

Effective control of tuberculosis, nontuberculous mycobacterial (NTM) disease, and melioidosis depends on rapid and accurate laboratory diagnosis. Mycobacterium tuberculosis complex (MTBC) remains a health priority, whereas NTM infections, particularly Mycobacterium avium complex (MAC), are increasingly recognized and may be misdiagnosed as tuberculosis. Melioidosis, caused by Burkholderia pseudomallei, is underdiagnosed in endemic regions. These infections have overlapping clinical presentations, and conventional diagnostics methods may not accurately differentiate among these diseases. We developed and evaluated a quadruplex real-time PCR assay (TAM-mPCR) for simultaneous detection of MTBC, MAC, and B. pseudomallei, targeting IS6110, rrs, and BPSS1187. A total of 607 pulmonary and extrapulmonary specimens from patients with suspected tuberculosis were collected at two sites in Thailand between 2024 and 2025. The limits of detection were 2 pg/reaction for MTBC, 0.1 ng/reaction for MAC, and 50 fg/reaction for B. pseudomallei, with 100% analytical specificity. For MTBC detection, TAM-mPCR demonstrated 100% sensitivity (330/330) and 99% (189/191) specificity. All three culture-confirmed MAC specimens were detected. For B. pseudomallei, the assay demonstrated 73.2% sensitivity (30/41) and 100% specificity (79/79). For MTBC, TAM-mPCR showed high concordance with NAAT and culture results, but differed significantly from AFB microscopy (P < 0.001). The assay also detected additional MTBC, MAC, and B. pseudomallei infections that were missed by routine diagnostics. TAM-mPCR enables rapid simultaneous pathogen detection and differentiation and may improve case detection and patient management in setting where tuberculosis and melioidosis co-endemic.

Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) is the leading infectious cause of death globally. Despite wide use of antiretroviral therapy (ART) by people living with HIV, the risk of TB remains increased. To understand immune interactions within lung granulomas, we compared spatial transcriptomics of Mtb and simian immunodeficiency virus (SIV) in co-infected macaques with or without ART as a model for HIV/Mtb co-infection. Spatially differentiated transcriptional profiles were observed in Mtb-only granulomas, with myeloid cells enriched in metabolic/antimicrobial pathways within the inner ring and T cell co-stimulatory/activation pathways enriched in the outer ring. These spatially distinct patterns were lost in SIV/Mtb granulomas with higher enrichment in type I IFN pathways compared to Mtb-only granulomas. SIV/ART/Mtb granulomas had an intermediate transcriptional pattern without restoration to Mtb-only granulomas, despite a lack of viral replication. Cell-cell communication was reduced among SIV/Mtb co-infected groups. These data suggest that HIV disrupts spatially organized immune functions of granulomas, which are not fully restored by ART.

Non-marneffei Penicillium species are ubiquitous molds usually dismissed as contaminants in respiratory cultures; invasive disease is rare and may mimic malignancy, tuberculosis, or sarcoidosis. A 39-year-old apparently immunocompetent man with asthma, a smoking history, and tuberculosis exposure presented with chronic cough, fever, night sweats, and weight loss. Chest computed tomography (CT) showed right lower-lobe subpleural consolidation, bilateral nodules, and mediastinal lymphadenopathy. Sputum and bronchoalveolar lavage (BAL) studies for tuberculosis were repeatedly negative; CT-guided lung biopsy showed a non-necrotizing granuloma, suggesting sarcoidosis. Five months later, the original lesions had regressed, and new contralateral pleural-based nodules appeared, establishing a migratory pattern. Repeat BAL grew a Penicillium species, a second recovery after an earlier isolate was dismissed as contamination. Surgical mediastinal lymph-node biopsy showed necrotizing granulomatous inflammation containing PAS- and GMS-positive septate hyphae, establishing proven invasive fungal disease, and fungal culture of the same tissue grew a Penicillium species; HIV and hepatitis serologies were negative. Liposomal amphotericin B followed by oral voriconazole produced near-complete radiological resolution. Attribution to Penicillium rests on recovery of the mold from the surgical tissue itself; identification nevertheless remained at genus level, because molecular identification, susceptibility testing, and fungal biomarkers were unavailable, and invasion of lung parenchyma was never demonstrated histologically. Migratory infiltrates should prompt reconsideration of fixed diagnoses; repeated mold recovery with tissue-invasive hyphae and treatment response warrants reassessment rather than dismissal as contamination.