Neutrophil extracellular traps characterize caseating granulomas.

Tuberculosis (TB) remains one of the top 10 causes of death worldwide and still poses a serious challenge to public health. Recent attention to neutrophils has uncovered unexplored areas demanding further investigation. Therefore, the aim of this study was to determine neutrophil activation and circulatory neutrophil extracellular trap (NET) formation in various types of TB. Sera from TB patients (n = 91) and healthy controls (NHD; n = 38) were analyzed for NE-DNA and MPO-DNA complexes, cell-free DNA (cfDNA), and protease activity (elastase). We show that these NET parameters were increased in TB sera. Importantly, NET formation and NE activity were elevated in TB patients with extensive tissue damage when compared to those with minor damage and in patients with relapse, compared to new cases. We discuss the importance of balancing NET formation to prevent tissue damage or even relapse and argue to analyze circulating NET parameters to monitor the risk of disease relapse. To investigate the tissues for NETs and to find the source of the circulating NET degradation products, we collected sections of granulomas in lung and lymph node biopsies. Samples from other diseases with granulomas, including sarcoidosis (SARC) and apical periodontitis (AP), served as controls. Whereas NET formation characterizes the caseating granulomas, both caseating and non-caseating granulomas harbor DNA with unusual conformation. As TB is associated with hypercoagulation and thromboembolism, we further imaged the pulmonary vessels of TB patients and detected vascular occlusions with neutrophil aggregates. This highlights the dual role of neutrophils in the pathology of TB.

Drug-Resistant Tuberculosis, Georgia, Kazakhstan, Kyrgyzstan, Moldova, and Ukraine, 2017-2022.

In 2021, the World Health Organization recommended new extensively drug-resistant (XDR) and pre-XDR tuberculosis (TB) definitions. In a recent cohort of TB patients in Eastern Europe, we show that XDR TB as currently defined is associated with exceptionally poor treatment outcomes, considerably worse than for the former definition (31% vs. 54% treatment success).

Treatment outcomes and risk factors for an unsuccessful outcome among patients with highly drug-resistant tuberculosis in Ukraine.

OBJECTIVES: To describe demographics, clinical features, and treatment outcomes of patients with highly drug-resistant tuberculosis (TB) in Ukraine, and to evaluate risk factors for an unsuccessful outcome. METHODS: Data from patients with multi-, pre-extensively, or extensively drug-resistant TB were collected prospectively from TB dispensaries in 15 out of 24 Ukrainian oblasts (regions) from 2020 to 2021. Treatment outcomes were evaluated using WHO definitions. Risk factors for an unsuccessful outcome were identified using a multivariable logistic regression model. RESULTS: Among 1748 patients, the overall proportion of successful outcomes was 58% (95% confidence interval [95% CI] 56-60) (n = 1015/1748), ranging from 65% (95% CI: 62-69) (n = 531/814) for multidrug-resistant TB to 54% (95% CI: 49-58) (n = 301/563) for pre-extensively drug-resistant TB and 49% (95% CI: 44-55) (n = 183/371) for extensively drug-resistant TB. Results were similar across oblasts, with few exceptions. The strongest risk factors for an unsuccessful outcome were extensively drug-resistant TB (adjusted OR [aOR] 3.23; 95% CI: 1.88-5.53), total serum protein below 62 g/L in adults and below 57 g/L for children and adolescents (aOR 2.79; 95% CI: 1.93-4.04), psychiatric illness (aOR 2.79; 95% CI: 1.46-5.33), age at TB diagnosis >65 years (aOR 2.50; 95% CI: 1.42-4.42), and alcohol misuse (aOR 2.48; 95% CI: 1.89-3.26). DISCUSSION: The overall proportion of successful outcomes among Ukrainians treated for highly drug-resistant TB was 58%, notably better compared with previous years, but still low for extensively drug-resistant TB. Risk factors for unsuccessful outcomes highlight that addressing socioeconomic factors in TB management is crucial. Efforts in maintaining TB dispensaries during and following the ongoing war are highly warranted.

Chest X-ray as an alternative method of making a preliminary diagnosis in patients with susceptible or drug-resistant pulmonary tuberculosis.

Background: Making a preliminary diagnosis using X-ray methods for the study of resistant and resistant tuberculosis (TB) will help to make a preliminary diagnosis and determine further tactics for the treatment of TB, even with limited resources for microbiological diagnosis of drug resistance of TB. The present study was aimed at identifying chest X-ray differences between susceptible and resistant TB. Methods: A prospective cohort study of data from all consecutive patients with culture-confirmed pulmonary TB admitted during the year to the Kharkiv TB Dispensary No. 1 in Kharkiv, Ukraine. Results: One hundred and sixty-eight patients with lung TB were examined. Patients were divided into two groups: 1st patients with pulmonary TB with resistance of Mycobacterium tuberculosis (MTB) to at least isoniazid and rifampicin (resistant TB) and 2nd pulmonary TB with preserved susceptibility of MTB to anti-TB drugs (susceptible-TB). Patients of 1st group often had lesions in two lobes of the lungs 31.1% and one lung 43.3% versus 15.4% and 2.6% of patients with susceptible TB (P < 0.001). In addition, more than 3 cavities in the lungs 45.5% were significantly more often observed in patients with resistant TB versus 7.9%-the 2nd group (P < 0.001). Smaller cavities were observed in patients with susceptible TB up to 1.99 cm 74% versus 35.2% in 1st group (P < 0.001). We did not observe any significant radiological features depending on the right or left lung, as well as the lobar localization of the TB process. Conclusions: For resistant forms of TB, radiologically, a more widespread TB process in the lungs with the presence of a larger number of cavities and their larger size against a background of a more pronounced clinical picture and mycobacterium excretion than with susceptible TB is characteristic.

Effectiveness of inhaled hypertonic saline application for sputum induction to improve Mycobacterium tuberculosis identification in patients with pulmonary tuberculosis.

BACKGROUND: This study assessed the effectiveness and diagnostic significance of hypertonic saline sputum induction for improving Mycobacterium tuberculosis (MTB) detection. METHODS: A prospective, randomized, open, two-arm, comparative study on MTB identification effectiveness when using inhaled sodium chloride hypertonic solution was performed in patients diagnosed with pulmonary tuberculosis (TB). Patients were randomly assigned into two groups: group 1 (inhalation group) included patients who inhaled a 7% sodium chloride solution upon admission to the hospital, and group 2 (control group) coughed up their sputum as usual. For both groups, specimens were tested by bacterioscopic, bacteriological, and molecular genetic methods. Diagnostic chest radiography was performed for all participants. RESULTS: In this study, 644 patients (mean age 42.2 years; 151 women, 23.4%) were randomly divided into two groups. Low-quality sputum samples were observed in 7.4% of patients from the inhalation group and 28.8% in the control group (p < 0.001). Acid-fast bacilli (AFB) smear was positive in 65.1% of patients from the inhalation group and 51.3% of controls (p = 0.002). A similar statistically significant situation was observed when culture methods (93.9% inhalation group and 81.9% control group, p < 0.001) and molecular genetic tests (92.2% inhalation group and 79.4% control group, p < 0.001) were used. Thus, active pulmonary TB was not verified microbiologically in 6.1% of patients from the inhalation group and in 18.1% of controls (p < 0.001). CONCLUSIONS: Hypertonic saline sputum induction improves the quality of collected samples. This method may be appropriate to increase the rate of MTB detection in sputum using microscopic, bacteriological, and molecular genetic methods for diagnosing TB on the day of specimen collection. Hypertonic saline sputum induction is suitable for middle- and low-income countries with limited resources and causes no severe adverse effects in TB patients.

Treatment effectiveness and outcome in patients with a relapse and newly diagnosed multidrug-resistant pulmonary tuberculosis.

Aim To investigate the treatment effectiveness and outcome in patients with pulmonary tuberculosis relapse and newly diagnosed multidrug resistant pulmonary tuberculosis (MDR-TB). Methods A total of 240 pulmonary MDR-TB patients, including 114 ones with tuberculosis relapse and 126 cases of newly diagnosed pulmonary tuberculosis, were examined. Effectiveness of the basic antimycobacterial therapy course was evaluated based on the time to normalization of tuberculosis clinical manifestation, sputum culture and acid-fast bacilli stain conversion, cavity closure, disappearance of infiltrative and focal changes in the pulmonary tissue. Treatment outcomes were evaluated as cured, treatment completed, treatment failed, died and lost to follow-up according to the World Health Organization guidelines. Results When assessing the treatment effectiveness in patients with MDR-TB, a worse clinical and chest radiograph dynamics was observed in tuberculosis relapse against the background of high parameters of treatment failure (18.4 %) and low cured (34.2 %) compared with newly diagnosed pulmonary tuberculosis (7.1% and 58.7 %, respectively) (p=0.008 and p<0.001, respectively). Conclusion Standard treatment effectiveness in patients with newly diagnosed MDR-TB manifested by faster improvement and stabilization of health, earlier sputum culture and smear conversion, higher frequency of cavity closure and achievement of certain clinical and radiographic improvement against the background of fewer cases of treatment failure and a higher number of cured patients compared with MDR-TB relapse.

Effectiveness of Intravenous Isoniazid and Ethambutol Administration in Patients with Tuberculosis Meningoencephalitis and HIV Infection.

BACKGROUND: The aim of this study was to investigate the effectiveness of intravenous isoniazid (H) and ethambutol (E) administered in patients with new sputum positive drug-susceptible pulmonary tuberculosis (TB) with tuberculous meningoencephalitis (TM) and human immunodeficiency virus (HIV) co-infection in the intensive phase of treatment. METHODS: Fifty-four patients with TB/TM and HIV co-infection were enrolled for this study. Group 1 comprised of 23 patients treated with E and H intravenously, while rifampicin and pyrazinamide were prescribed orally. Group 2 consisted of 31 patients treated with the first-line anti-TB drugs orally. The concentrations of H and E in blood serum were detected using a chromatographic method. RESULTS: A significant improvement in the clinical symptoms and X-ray signs in patients treated intravenously with H and E was observed and compared to group 2. The sputum Mycobacterium tuberculosis positivity was observed during the second month of the treatment in 25.0% of patients from group 1 and 76.1% of the patients from the control group (p=0.003). In addition, nine patients (39.1%) died up to 6 months when H and E were prescribed intravenously compared with 22 (70.9%) in group 2 (p=0.023). CONCLUSION: In TB/TM with HIV, the intravenous H and E treatment was more effective than oral H and E treatment at 2 months of intensive treatment in sputum conversion as well as in clinical improvement, accompanied by significantly higher mean serum concentrations. In addition, the mortality rate was lower in intravenous H and E treatment compared to oral treatment.

Effectiveness of anti-tuberculosis chemotherapy in patients with tuberculosis relapse compared with newly diagnosed patients.

Background: To study the effectiveness of antituberculosis chemotherapy in patients with relapse pulmonary tuberculosis (RTB) compared with patients with the newly diagnosed process. Methods: We examined 285 TB patients, including 126 individuals with RTB (Group 1) and 159 patients with newly diagnosed pulmonary tuberculosis (NDPTB) (Group 2). All patients were diagnosed with infiltrative PTB. Effectiveness of the basic course of antimycobacterial treatment was assessed in accordance with the following data: time required for the normalization of clinical manifestations, smear conversion, cavity healing, disappearance of infiltrative and focal changes in the pulmonary tissue, as well as the final clinical effectiveness of therapy. Results: Disappearance of clinical symptoms was statistically significantly faster in Group 2 compared with RTB patients in 2.25 ± 0.11 and 3.40 ± 0.15 months, respectively (P < 0.001). Sputum culture conversion was observed after 6 months of treatment in 138 (86.79%) patients with NDPTB and 89 (72.22%) patients from Group 1 (P = 0.0023). Closure of cavities and disappearance of infiltrative and focal changes in the lungs occurred within 6 months of chemotherapy only in 55 (43.65%) patients with RTB and 93 (58.49%) patients with NDPTB (P = 0.0133). Conclusions: Standard treatment for patients with NDPTB is considered successful in case of faster health improvement and stabilization, less pronounced rates of toxic adverse reactions to antiTB drugs, faster sputum smear and culture conversion and cavity healing, signs of clinical and radiological convalescence, and the reduced number of large residual changes after the treatment compared with RTB.

Efficacy and safety of intravenous chemotherapy during intensive treatment phase in patients with newly diagnosed pulmonary tuberculosis.

The purpose of our study was to examine the efficacy and safety of intravenous chemotherapy during intensive treatment phase for patients with newly diagnosed pulmonary tuberculosis (pulmonary TB). MATERIALS AND METHODS: The study involved 92 patients with newly diagnosed pulmonary TB aged between 20 years and 68 years. All patient with newly diagnosed pulmonary TB and chemosensitive tuberculosis were enrolled in this study. The patients were allocated to two groups. The first (control) group of 46 patients received standard chemotherapy orally. The second (main) group consisted of 46 patients who were prescribed isoniazid, rifampin, ethambutol by i / v transfusion, and pyrazinamide orally as a part of the standard treatment. RESULTS: Symptoms of intoxication in pulmonary TB patients from the second group were eliminated faster (1.42±0.35) of a month than the same symptoms of the group 1-(2.96±0.24) of the months,p < 0.05; disappearance of respiratory symptoms of the group 2-(1.34±0.29) of a month, group 1-(2.65±0.43) of the months,p < 0.05. In the group 2, the bacterioexcretion time was reducing faster and up to 2 months it reached 37(80.43±5.85%) while the time for the control group reached 25(54.35±7.34%),p < 0.05. Destruction healing and healing frequency of destruction cavities up to 4 months amounted to 38(82.61±5.59%) (in control group - 28(60.87±7.20%),p < 0.05) and residual changes were reducing (small changes or absence of even minimal radiological changes were found in 29(63.04 ±7.12%) patients versus 18(39.13±7.20%) of the group 1, and large residual changes accordingly in 17(36.96±7.12%) and 28(60.87±7.20%),p < 0.05. CONCLUSIONS: Thanks to i/v chemotherapy clinical manifestations of the in patients with pulmonary TB were eliminated faster, severe side effects to anti-TB drugs were not noticed, time of bacterial excretion and healing destruction reduced, healing frequency of destruction cavities increased and the residual changes decreased.

Association between effectiveness of tuberculosis treatment and cytochrome P-4502E1 polymorphism of the patients.

CONTEXT: The risk of antituberculosis (TB) drug-induced liver injury could be determined by patients' genotype polymorphism of the xenobiotic-metabolizing enzymes. To find the meaning of cytochrome P-4502E1 (CYP2E1) polymorphism in TB patients. Corresponding of CYP2E1 polymorphism in TB patients with the level of isoniazid and rifampicin as well as for the outcome and toxicity development during inpatient TB treatment. METHODS: CYP2E1 genotype was detected with the help of polymerase chain reaction and endonuclease analysis. The level of rifampicin, isoniazid, diene conjugates (DC), and catalase activity in the blood was determined spectrophotometrically. We have considered medical records at the beginning and at the end of inpatient treatment. STATISTICAL ANALYSIS USED: Kruskal-Wallis, ANOVA, and Chi-square tests were used in this study. RESULTS: The concentration of rifampicin 6 h after its intake was 17.6% higher in carriers of slow metabolizer (SM) CYP2E1 genotype than in patients with rapid metabolizer (RM) genotype that proved a participation of hepatic enzyme CYP2E1 in metabolism of rifampicin. According to obtained results in TB patients with RM genotype, the indexes of cytolysis (alanine aminotransferase, aspartate aminotransferase) and bile stasis (gamma-glutathione transferase) were higher comparatively to SM genotype both before and after inpatient treatment. This correlated with a higher concentration of DC in the blood (+8.6%) and lower plasma catalase activity (-50.0%) in the patients with RM genotype comparatively with the patients with SM genotypes. CONCLUSION: Polymorphism of CYP2E1 genotype is an important criterion for the development of hepatotoxicity before and during TB treatment while increased rifampicin level has no influence on it.