A disposable, portable electrochemical immunosensor for rapid in situ detection of bovine tuberculosis.

This contribution describes the development of a simple, fast, cost-effective, and sensitive impedimetric immunosensor for quantifying bovine tuberculosis (TB) in bovine serum samples. The construction of the immunosensor involved immobilizing the purified protein derivative (PPD) of M. bovis onto a screen-printed electrode that was modified with gold nanoparticles (AuNPs) and a polypyrrole (pPy) film synthesized electrochemically. The immunosensor exhibited a linear range from 0.5 µg mL-1 to 100 µg mL-1 and achieved a limit of detection (LD) of 100 ng mL-1 for the detection of anti-M. bovis antibody. The recovery percentages obtained in bovine serum samples were excellent, ranging between 98 % and 103 %. This device presents several advantages over alternative methods for determining TB in bovine serum samples. These include direct, in situ measurement without the need for pre-treatment, utilization of small volumes, thus avoiding harmful solvents and expensive reagents, and portability. In addition, the immunosensor exhibits both physical and chemical stability, retaining effectiveness even after 30 days of modification. This allows simultaneous incubations and facilitates large-scale detection. Hence, this immunosensor presents itself as a promising diagnostic tool for detecting anti-M. bovis antibodies in bovine serum. It serves as a viable alternative to tuberculin and ELISA tests.

Factores relacionados con la adherencia al tratamiento de tuberculosis en el Departamento de Boyacá / Factors related with adherence to treatment of tuberculosis in the Department of Boyacá

Resumen Introducción: la tuberculosis es una enfermedad infecciosa crónica causada por Mycobacterium tuberculosis, capaz de afectar cualquier órgano o tejido siendo la forma más común la enfermedad pulmonar. Objetivo: identificar los factores que afectan la adherencia al tratamiento antituberculoso en pacientes del Departamento de Boyacá durante los años 2017-2019. Metodología: estudio descriptivo retrospectivo que utilizó como fuentes de información datos registrados en la base del programa departamental de Tuberculosis y SIVIGILA. Se realizó análisis univariado, determinando la distribución de las variables y análisis bivariado en el que se exploró la asociación entre la adherencia al tratamiento de tuberculosis y las variables independientes, se utilizó la prueba de Chi cuadrado. Resultados: se incluyeron 402 pacientes, la adherencia al tratamiento antituberculoso fue 96,5% y la no adherencia de 3,5%. El 66,7% eran de sexo femenino; 55,7% vivián en zona urbana; 66,5% pertenecían al régimen subsidiado; 89,7% mestizos; 91,2% correspondían a otros grupos poblacionales, seguido del 7,5% de población privada de la libertad y 1,3% habitantes de calle. Dentro de las comorbilidades la coinfección con VIH con 4,2% fue la de mayor presencia. Entre las causas de no adherencia al tratamiento fueron desinterés, cambio frecuente de domicilio, contrato finalizado con la EPS y suspensión del tratamiento por toxicidad hepática. El paciente con mayor probabilidad de abandonar el tratamiento pertenece al sexo masculino, al grupo poblacional de migrantes o habitantes de calle y se encuentra afiliado al régimen subsidiado, ​​​​se encontró una relación estadísticamente significativa entre estas variables y el resultado con la adherencia al tratamiento. Conclusiones: si bien el resultado obtenido en el presente trabajo no es elevado, es importante realizar vigilancia de la adherencia al tratamiento antituberculoso para disminuir el riesgo de complicaciones derivadas de su abandono como mayor mortalidad, desarrollo de resistencia bacteriana y un período de contagiosidad más prolongado.

Abstract Introduction: Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis, capable of affecting any organ or tissue, the most common form being lung disease. Objective: Identify the factors that affect adherence to tuberculosis treatment in patients from the Department of Boyacá during the years 2017-2019. Methodology: Retrospective descriptive study that used as information sources data registered in the database of the departmental Tuberculosis program and SIVIGILA. A univariate analysis was carried out, determining the distribution of the variables and a bivariate analysis in which the association between adherence to tuberculosis treatment and the independent variables was explored, using the Chi-square test. Results: 402 patients were included, adherence to anti-tuberculosis treatment was 96.5% and non-adherence 3.5%. 66.7% were female; 55.7% lived in an urban area; 66.5% belonged to the subsidized regime; 89.7% mestizo; 91.2% corresponded to other population groups, followed by 7.5% of the population deprived of liberty and 1.3% homeless. Within the comorbidities, coinfection with HIV 4.2% was the one with the greatest presence. Among the causes of non-adherence to treatment were lack of interest; frequent change of address; Terminated contract with the EPS and suspension of treatment due to liver toxicity. The patient with the highest probability of abandoning treatment belongs to the male sex, to the population group of migrants or street dwellers and is affiliated with the subsidized regime, a statistically significant relationship was found between these variables and the result with adherence to treatment. Conclusions: Although the result obtained in this study is not high, it is important to monitor adherence to tuberculosis treatment to reduce the risk of complications derived from its abandonment, such as increased mortality, development of bacterial resistance, and a longer period of contagiousness.

[Pancreatic tuberculosis]. / Tuberculosis pancreática.

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis. It is currently considered a public health problem due to the socioeconomic conditions of the world population and the increase in other infections such as that due to Human Immunodeficiency Virus (HIV). Pulmonary involvement is the most common form of clinical presentation, although in recent decades extrapulmonary involvement has increased. Among these, digestive disorders represent less than 10%. We present a case of exclusive pancreatic tuberculosis, in the context of an immunocompromised patient. A 43-year-old man with a history of HIV with a low CD4 count and high viral load, without antiretroviral treatment, with one-month history of symptoms characterized by low-grade fever associated with weight loss and nonspecific abdominal pain. An abdominal tomography was performed which showed a collection in the tail of the pancreas; the puncture revealed material with positive culture for M. tuberculosis. Antifimic and antiretroviral treatment was started with a good response. Primary pancreatic tuberculosis is an uncommon extrapulmonary form, given that it is a gland that would be biologically protected by the enzymes it produces. We highlight the rarity of the case and although the prognosis is good with anti-tuberculosis treatment, it could be fatal without correct diagnosis and treatment. The high index of suspicion of pancreatic tuberculosis by the physician and the performance of fine needle aspiration puncture to obtain histopathological evidence are important for a correct diagnosis, especially in HIV patients.

La tuberculosis es una enfermedad infectocontagiosa producida por el Mycobacterium tuberculosis. Actualmente se considera un problema de salud pública debido a las condiciones socioeconómicas de la población mundial y al incremento de otras infecciones causantes de inmunosupresión, como el virus de la inmunodeficiencia humana (HIV). La afección pulmonar es la forma de presentación clínica más frecuente aunque en las últimas décadas el compromiso extrapulmonar se ha visto incrementado. Dentro de este la afección digestiva representa menos del 10%. Presentamos un caso de localización exclusivamente pancreática en un paciente inmunocomprometido. Varón de 43 años HIV positivo, con bajo recuento de CD4 y alta carga viral, sin tratamiento antirretroviral, consultó por cuadro de un mes de evolución caracterizado por registros subfebriles asociado a pérdida de peso y dolor abdominal inespecífico. Se realizó tomografía de abdomen la cual arrojó colección en cola de páncreas, y posteriormente punción del material con rescate en cultivo de M. tuberculosis. Se inició tratamiento antifímico y antirretroviral con buena respuesta al mismo. La tuberculosis pancreática primaria es una forma extrapulmonar infrecuente, dado que es una glándula que estaría biológicamente protegida por las enzimas que origina. Destacamos la infrecuencia del caso y aunque el pronóstico es bueno con el tratamiento antifímico, podría ser fatal sin un diagnóstico y tratamiento correctos. El alto índice de sospecha de tuberculosis pancreática por parte del médico y la realización de punción aspiración con aguja fina (PAAF) para obtener evidencia histopatológica son importantes para un diagnóstico correcto, especialmente en pacientes HIV.

Systematic review of tuberculous meningitis in high-risk populations: mortality and diagnostic disparities.

Aim: Tuberculous meningitis (TBM) is one of the most severe clinical forms of tuberculosis (TB). Since epidemiological studies can contribute to TB control, we conducted a review and meta-analysis of epidemiological publications of adults TBM cases in countries with high incidence of TB.Materials & methods: The search resulted in 11,855 articles, in which 21 ultimately were included in our review and 15 in our meta-analysis.Results: TBM mortality was 25% with death rates of 70% in Africa. The review showed different and non-concordant diagnostic techniques and treatment schemes.Conclusion: Adults living in the African region are at high risk of death from TBM, highlighting an urgent need of guidelines to support diagnosis and treatment, and ultimately, to reduce mortality.

Tuberculosis (TB) is a disease that mostly affects the lungs. It can also affect other organs. When TB affects the brain and spinal cord, it is called tuberculous meningitis (TBM). We looked to analyze the traits of the adults with TBM that live in countries with a high number of cases of TB. We searched scientific publications that studied these populations to find information that may help to control the disease. The death rate of TBM was 25%, reaching up to 70% in Africa. We found some disparities regarding diagnosis and treatment. Adults living in Africa have a higher risk of dying from TBM. We need guidelines about the diagnosis and treatment of TBM to help reduce TBM deaths in these countries.

Bedaquiline versus injectable containing regimens for rifampicin-resistant and multidrug-resistant tuberculosis in a reference center in Brazil - a real-world evidence study using a retrospective design.

BACKGROUND: Drug resistance (DR) is one of the several challenges to global tuberculosis (TB) control. The implementation of bedaquiline (BED) for DR-TB after more than 40 years was expected to improve treatment outcomes as well as microbiologic conversion and adverse events (AE) occurrence. METHODS: Retrospective cohort study based on secondary data of patients with rifampicin-resistant (RR) or multidrug-resistant (MDR) TB reported to the Outpatient Clinic of Mycobacterial Diseases of the Thorax Diseases Institute - Federal University of Rio de Janeiro - Brazil, between 2016 and 2023. We aimed to evaluate microbiologic conversion, AE and TB treatment outcomes and compare them according to the treatment regimen used for RR/MDR-TB patients under routine conditions [Injectable Containing Regimens (ICR) versus BED Containing Regimens (BCR)]. Logistic regression and survival analysis using Cox regression and Kaplan Meier curve were used for statistical analysis. RESULTS: Of the 463 DR-TB patients notified during the study period, 297 (64.1%) were included for analysis (ICR = 197 and BCR = 100). Overall AEs were more frequent (83.7 vs. 16.3%, p < 0.001) and occurred earlier in the ICR group (15 days vs. 65 days, p = 0.003). There were no cases of cardiotoxicity requiring interruption of BED treatment. None of the regimens of treatment tested were associated with smear or culture conversion on Cox regression analysis (p = 0.60 and 0.88, respectively). BED-containing regimens were also associated with favorable outcomes in multivariable logistic regression [adjusted odds ratio (aOR) = 2.63, 95% confidence interval (CI)1.36-5.07, p = 0.004], as higher years of schooling, primary drug resistance, and no previous TB treatment. In the survival analysis, BCR was inversely associated with the occurrence of AE during treatment follow-up (aHR 0.24, 95% CI 0.14-0.41, p < 0.001). In addition, TB treatment regimens with BED were also associated with favorable outcomes (aHR 2.41, 95% CI 1.62-3.57, p < 0.001), along with no illicit drug use and primary drug resistance. CONCLUSIONS: The implementation of a fully oral treatment for RR/MDR-TB in a reference center in Brazil was safe and associated with favorable outcomes under routine conditions, despite social, demographic, and behavioral factors that may influence TB treatment completion.

Case Report: Spontaneous acute hemopericardium.

Acute hemopericardium is generally produced by complications of interventional procedures or traumatisms to the chest wall. In absence of those antecedents, clinicians face an arduous process of etiological diagnosis and treatment. We present the case of a male patient with history of Hodgkin's lymphoma and aortic endovascular treatment years ago, who develop an episode of fever, chest pain and dyspnea that was complicated with cardiac tamponade diagnosed with echocardiogram and angio- tomography. In the operating room hemopericardium was diagnosed and drained with resolution of symptoms. Diagnosis work out was carried out with suspicion of tuberculous or neoplastic pericarditis with negative results. Patient was discharged with the diagnoses of viral or idiopathic pericarditis. The case highlights the use of multimodality images and laboratories procedures to lead to a correct diagnoses and treatment.

Macrophages, Metabolism, Mitochondria, Circadian Rhythmicity and the Pathogen: The Multidimensional Nature of Tuberculosis.

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB) was first identified in 1882 by Robert Koch, and it is estimated that this pathogen has been around for as long as 3 million years.The World Health Organization (WHO) reported that in 2022 alone an estimated 10.6 million people developed TB worldwide, making TB the world's second leading cause of death from a single infectious agent, just after coronavirus disease (COVID-19), despite TB being a preventable and usually curable disease.Moreover, epidemiological studies suggest that approximately a quarter of the global population has been infected with TB bacteria, of which 5-10% will eventually develop symptoms and TB disease. Poverty, obesity, diabetes, and alcohol use contribute to the burden of TB.Alveolar macrophages play a pivotal role in the clearance of airborne pathogenic microorganisms and are the primary target of M. tuberculosis.Macrophage activity depend on metabolism and circadian rhythmicity, and mitochondria are a central hub that coordinates the communication between metabolism, circadian rhythmicity, and the immune system.Recent evidence has thrown light on how M. tuberculosis metabolism may regulate macrophage activity and the overall host responses to M. tuberculosis infection.This chapter explores how all these biological domains relate to each other, highlighting the multidimensional nature of TB, and positioning macrophages at center stage.

Case report: granulomatous hepatitis due to Mycobacterium avium in an Atlantic yellow-nosed albatross (Thalassarche chlororhynchos) from Southern Brazilian coast.

This study reports a granulomatous hepatitis caused by Mycobacterium avium in an Atlantic yellow-nosed albatross (Thalassarche chlororhynchos) found dead on Brazil southern coast. At necropsy, the albatross was cachectic and the liver was severely enlarged with multifocal to coalescing white nodules. Histopathological evaluation revealed multifocal to coalescing granulomas with caseous necrosis, surrounded by an infiltrate of macrophages and multinucleated giant cells, and a thin capsule of fibrous connective tissue. The Fite-Faraco staining technique revealed multiple acid-fast bacilli (AFB) stained in magenta, predominantly in the areas of necrosis. Bacterial culture and polymerase chain reaction (PCR) analysis confirmed the presence of Mycobacterium avium in liver samples. This case underscores the importance of wildlife surveillance in coastal regions. Pelagic birds like the Atlantic yellow-nosed albatross can harbor pathogenic agents that represent a threat to wildlife and domestic animals. Enhanced monitoring and research are essential to understand the epidemiology and potential risks associated with such infections in coastal ecosystems.

Repurposing antiparasitic N,N'-aliphatic diamine derivatives as promising antimycobacterial agents.

In previous studies, we demonstrated the potent activity of a library of 25 N,N'-disubstituted diamines (NNDDA) toward Trypanosomatid and Apicomplexa parasites. Considering the structure similarity between this collection and SQ109, an antituberculosis compound, and its compelling antiparasitic properties, we aimed to repurpose this library for tuberculosis treatment. We assayed this collection against Mycobacterium tuberculosis H37Rv and M. avium, obtaining several compounds with MIC values below 10 µM. The most active analogs were also evaluated against M. smegmatis, a non-pathogenic species, and the non-tuberculosis mycobacteria M. abscessus, M. kansasii, and M. fortuitum. 3c stands out as the lead mycobacterial compound of the collection, with potent activity against M. tuberculosis (minimal inhibitory concentration [MIC] = 3.4 µM) and moderate activity against M. smegmatis, M. kansasii, and M. fortuitum (all with MIC values of 26.8 µM). To unravel the mechanism of action, we employed the web-based platform Polypharmacology Browser 2 (PPB2), obtaining carbonic anhydrases as potential drug targets. Nevertheless, none of the compounds displayed experimental inhibition. In summary, our study confirms the validity of the repurposing approach and underscores the antimycobacterial potential of NNDDA compounds, especially the analog 3c, setting a stepping stone for further studies.

[Telephone surveys for the study of catastrophic costs due to tuberculosis in Colombia: a novel toolPesquisas telefônicas para um estudo de custos catastróficos da tuberculose na Colômbia: uma ferramenta inovadora]. / Encuestas telefónicas para el estudio de costos catastróficos por tuberculosis en Colombia: una herramienta innovadora.

The study of catastrophic costs incurred by people affected by tuberculosis (TB), conducted in Colombia during the COVID-19 pandemic, provided the opportunity to implement telephone surveys for data collection. This constitutes a methodological innovation regarding the standards established by the World Health Organization (WHO) which, for this type of study, usually rely on face-to-face surveys of patients attending health facilities. The study design, objectives, and methodology were adapted from the WHO publication Tuberculosis patient cost surveys: a handbook. A total of 1065 people affected by tuberculosis were selected as study participants and, by telephone, were administered a standard questionnaire adapted to the Colombian context. This allowed the collection of structured data on the direct and indirect costs faced by TB patients and their families. Greater than 80% completeness was achieved for all variables of interest, with an average survey duration of 40 minutes and a rejection rate of 8%. The described survey method to determine the baseline for further study of catastrophic costs in Colombia was novel because of its telephone-based format, which adheres to the information standards required to allow internationally comparable estimates. It is a useful means of generating standardized results in contexts in which the ability to conduct face-to-face surveys is limited.

O estudo dos custos catastróficos incorridos pelas pessoas afetadas pela tuberculose realizado na Colômbia durante a pandemia de COVID-19 representou uma oportunidade de implementar pesquisas telefônicas como forma de coleta de dados. Constitui-se uma inovação metodológica dos padrões estabelecidos pela Organização Mundial da Saúde (OMS), que, para esse tipo de estudo, geralmente se baseiam no uso de pesquisas presenciais com os pacientes que frequentam estabelecimentos de saúde. O delineamento, os objetivos e a metodologia do estudo foram adaptados do manual prático da OMS para a realização de pesquisas de custos da tuberculose. Um total de 1065 pessoas afetadas pela tuberculose foram selecionadas para participar do estudo. O questionário padrão, adaptado ao contexto colombiano, foi aplicado pelo telefone. Foi possível obter dados estruturados sobre os custos diretos e indiretos enfrentados pelos pacientes com tuberculose e suas famílias. Em geral, observou-se que todas as variáveis de coleta atingiram uma completude de mais de 80%, com um tempo médio de pesquisa de 40 minutos e uma taxa de recusa de 8%. A metodologia de pesquisa telefônica desenvolvida para determinar a linha de base do estudo de custos catastróficos na Colômbia foi inovadora devido ao formato telefônico, que mantém os padrões de informação necessários para permitir estimativas comparáveis internacionalmente e é uma forma útil de gerar resultados padronizados em circunstâncias em que há limitações para a realização de pesquisas presenciais.

Tuberculosis Diagnosis: Current, Ongoing, and Future Approaches.

Tuberculosis (TB) remains an impactful infectious disease, leading to millions of deaths every year. Mycobacterium tuberculosis causes the formation of granulomas, which will determine, through the host-pathogen relationship, if the infection will remain latent or evolve into active disease. Early TB diagnosis is life-saving, especially among immunocompromised individuals, and leads to proper treatment, preventing transmission. This review addresses different approaches to diagnosing TB, from traditional methods such as sputum smear microscopy to more advanced molecular techniques. Integrating these techniques, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP), has significantly improved the sensitivity and specificity of M. tuberculosis identification. Additionally, exploring novel biomarkers and applying artificial intelligence in radiological imaging contribute to more accurate and rapid diagnosis. Furthermore, we discuss the challenges of existing diagnostic methods, including limitations in resource-limited settings and the emergence of drug-resistant strains. While the primary focus of this review is on TB diagnosis, we also briefly explore the challenges and strategies for diagnosing non-tuberculous mycobacteria (NTM). In conclusion, this review provides an overview of the current landscape of TB diagnostics, emphasizing the need for ongoing research and innovation. As the field evolves, it is crucial to ensure that these advancements are accessible and applicable in diverse healthcare settings to effectively combat tuberculosis worldwide.

Comparative Genomic Analysis of Mycobacterium tuberculosis Isolates Circulating in North Santander, Colombia.

Tuberculosis (TB) is an important infectious disease in relation to global public health and is caused species of the Mycobacterium tuberculosis complex (MTBC). In this study, we used whole-genome sequencing (WGS) and comparative genomics to investigate the genetic diversity of M. tuberculosis (Mtb) isolates circulating in North Santander (NS), Colombia. WGS was used for the phylogenetic and lineage characterization of 18 isolates of Mtb typed with orphan genotypes from 11 municipalities of NS between 2015 and 2018. The isolates studied were included in six sublineages from L4; the most frequent were 4.1.2.1, 4.3.3, and 4.3.4.2, corresponding to a proportion of 22.2%. The genome analysis conducted allowed the identification of a set of genetic variants mainly associated with determinants of virulence and evasion of the immune system (PPE34 and PE_PGRS2); adaptation and survival (PGL/p-HBAD); stress response (sigJ and sigM); geographic variability (PPE34); and carbohydrate and lipid metabolism (aldA, rocA, and cyp144). This is the first description of the molecular epidemiology of Mtb isolates circulating in NS achieved through WGS. It was possible to perform comparative genomics analyses between Mtb isolates against the universal reference H37Rv and Colombian UT205 genome, which can help us to understand the local genetic diversity and is relevant for epidemiological studies, providing insight into TB transmission dynamics in NS.

The BCG vaccine and SARS-CoV-2: Could there be a beneficial relationship?

The COVID-19 disease continues to cause complications and deaths worldwide. Identifying effective immune protection strategies remains crucial to address this ongoing challenge. The Bacillus Calmette-Guérin (BCG) vaccine, developed initially to prevent pulmonary tuberculosis, has gained relevance due to its ability to induce cross-protection against other pathogens of the airways. This review summarizes research on the immunological protection provided by BCG, along with its primary clinical and therapeutic uses. It also explores the immunological features of COVID-19, the mechanisms implicated in host cell death, and its association with chronic pulmonary illnesses such as tuberculosis, which has led to complications in diagnosis and management. While vaccines against COVID-19 have been administered globally, uncertainty still exists about its effectiveness. Additionally, it is uncertain whether the utilization of BCG can regulate the immune response to pathogens such as SARS-CoV-2.

Massive cerebral tuberculomas, Pott's disease and hypercalcaemia secondary to Mycobacterium bovis in a patient with chronic kidney disease on peritoneal dialysis.

Tuberculosis (TB) is still a health problem in developing countries. Pulmonary involvement remains the most common clinical presentation. However, multiorgan involvement can be life-threatening. We present the case of a young woman on peritoneal dialysis who was admitted to hospitalisation for hypercalcaemia and low back pain. In his biochemical evaluation, suppressed intact parthyroid hormone (iPTH) and elevated 1,25-hydroxyvitamin D were detected. On a lumbar CT scan, a hypodense lesion in vertebral bodies compatible with Pott's disease was found. Positive cultures for Mycobacterium bovis were obtained in bronchoalveolar lavage and peritoneal fluid, for which specific treatment was initiated. Due to neurological deterioration, a CT scan was performed showing the presence of multiple tuberculomas. Retrospectively, the lack of an etiological diagnosis of chronic kidney disease, the initiation of dialysis 8 months before and the clear evidence of long-standing TB strongly suggest mycobacterium infection as the cause or trigger for the rapid decline in kidney function.

Embryonic and larval zebrafish models for the discovery of new bioactive compounds against tuberculosis.

Tuberculosis (TB) is a world health challenge the treatment of which is impacted by the rise of drug-resistant strains. Thus, there is an urgent need for new antitubercular compounds and novel approaches to improve current TB therapy. The zebrafish animal model has become increasingly relevant as an experimental system. It has proven particularly useful during early development for aiding TB drug discovery, supporting both the discovery of new insights into mycobacterial pathogenesis and the evaluation of therapeutical toxicity and efficacy in vivo. In this review, we summarize the past two decades of zebrafish-Mycobacterium marinum research and discuss its contribution to the field of bioactive antituberculosis therapy development.

Genome sequences of Mycobacterium sp. Elmwood and accompanying phage, isolated from a public swimming pool in Nebraska.

Genome sequencing of a non-tuberculosis Mycobacterium species, isolated from a public pool, shows that the genome contains several genes for antibiotic resistance and anti-phage defense, which are absent from other related Mycobacteria. Metagenomic binning also provided the genome of the accompanying phage, which is distinct from other mycobacterial phages.

A Genome-Focused Investigation Reveals the Emergence of a Mycobacterium tuberculosis Strain Related to Multidrug-Resistant Tuberculosis in the Amazon Region of Brazil.

A previous study in Pará, Northern Brazil, described a strain of Mycobacterium tuberculosis with a unique genotype (SIT2517/T1) associated with multidrug-resistant tuberculosis (MDR-TB). To improve our understanding of MDR-TB transmission dynamics of these strains within this region, we performed phenotypic and genotypic drug susceptibility testing (pDST/gDST), 24-loci mycobacterial interspersed repetitive units (MIRU-VNTR) genotyping, whole-genome sequencing (WGS) and geo-epidemiology analysis. Of the 28 SIT2517/T1 isolates, 19 (67.9%) could be genotyped by 24-loci MIRU-VNTR and 15 by WGS. All belonged to sublineage 4.1.1.3, distinct from other representative Lineage 4 isolates identified in Brazil. The MDR phenotype determined by pDST was confirmed by gDST, the latter also demonstrating the presence of additional mutations conferring pre-extensively drug-resistance (pre-XDR). Discrepancies between gDST and pDST were observed for pyrazinamide and fluoroquinolones. Thirteen out of 15 isolates analyzed by WGS were clustered when applying a 12 single nucleotide polymorphisms (SNPs) cutoff. The SIT2517/T1 isolates were distributed across the metropolitan regions of Belém and Collares municipalities, showing no geographic clustering. WGS-transmission network analysis revealed a high likelihood of direct transmission and the formation of two closely linked transmission chains. This study highlights the need to implement TB genomic surveillance in the Brazilian Amazon region.

Tuberculin skin test repetition after TNF-α inhibitors in patients with chronic inflammatory arthritis: a long-term retrospective cohort in endemic area.

OBJECTIVES: To evaluate the tuberculin skin test (TST) conversion in chronic inflammatory arthropathies (CIA) patients on TNFα inhibitors (TNFi) and without previous latent tuberculosis infection (LTBI) treatment. METHODS: Patients with rheumatoid arthritis (RA), ankylosing spondylitis (AS) and psoriatic arthritis (PsA) with negative LTBI were retrospectively evaluated for TST conversion and active tuberculosis (TB) after six months of exposition to TNFi. Two groups were compared: patients who repeated TST (TST-repetition) during the follow-up and patients who did not (non-TST-repetition). RESULTS: A total of 355 CIA patients on TNFi were screened and 138 (38.9%) did not fulfill the inclusion criteria. Of the remaining 217 CIA patients, 81 (37.3%) repeated TST during TNFi treatment. TST conversion rate was observed in 18 (22.2%) patients without significant differences among CIA (p = 0.578). The number of TB cases was low (n = 10; 4.6%) and was similar in TST-repetition and non-TST-repetition groups [2 (2.5%) vs. 8 (5.9%), p = 0.328]. Of note, 30% of active TB occurred early (6-12 months of TNFi exposure) and the median (full range) time to incident TB was 1.3 (0.6-10.6) years, whereas the median (full range) time to TST repetition was later [3.3 (0.5-13.4) years]. The incidence of active TB was lower among RA patients than AS patients [342 (95% CI 41 - 1446) vs. 1.454 (95% CI 594-2993)/100,000 patient-years, p = 0.049]. CONCLUSION: These results indicate that TST repetition is associated with a high conversion rate, suggesting the need for recommended treatment. The delayed repetition of TST and low number of active TB cases hampered the evaluation of this strategy effectiveness to prevent active infection. Larger studies with systematic repetition patterns are necessary. In addition, the study highlights the need for a greater surveillance for TB in AS patients.

Development of inhaled moxifloxacin-metformin formulation as an alternative for pulmonary tuberculosis treatment.

Resistant M. tuberculosis strains threaten pulmonary tuberculosis (P-TB) control since they limit drug options. Drug repositioning and new development strategies are urgently required to overcome resistance. Studies have already shown the beneficial role of the oral antidiabetic metformin as an anti-tuberculosis adjuvant drug. This work aimed to develop an inhalatory dry powder co-formulation of metformin and moxifloxacin to figure out a future option for P-TB treatment. Pre-formulation evaluations indicated the physicochemical compatibility of constituents, demonstrating powder crystallinity and acceptable drug content. Eight moxifloxacin-metformin dry powder formulations were produced by spray drying, and solid-state characterizations showed partial amorphization, ascribed to moxifloxacin. Four formulations containing L-leucine exhibited micromeritic and in vitro deposition profiles indicating pulmonary delivery suitability, like spherical and corrugated particle surface, geometric diameters < 5 µm, high emitted doses (>85 %), and mass median aerodynamic diameters between 1-5 µm. The use of a second spray dryer model further optimized the aerodynamic properties and yield of the best formulation, demonstrating the influence of the equipment used on the product obtained. Moreover, the final formulation showed high in vitro cell tolerability and characteristics in permeability studies indicative of good drug retention in the lungs.