TB Subjects Research Articles

Immunomodulatory effects of cysteamine and its potential use as a host-directed therapy for tuberculosis.

Cysteamine, a drug approved to treat cystinosis, has been proposed as a host-directed therapy for M. tuberculosis (Mtb) and SARS-CoV-2. The impact of cysteamine on the immune responses has not been fully investigated. We aimed to in vitro evaluate the immunomodulatory effects of cysteamine on peripheral blood mononuclear cells (PBMCs) using the purified protein derivative (PPD) as a recall antigen, and an unspecific stimulus as staphylococcal enterotoxin B (SEB). PBMCs isolated from subjects with tuberculosis infection (TBI), those with tuberculosis disease (TB), and healthy controls (HC) were in vitro stimulated with PPD or SEB and treated or not with cysteamine at different concentrations (50 µM-400 µM) for 6 hours (h) and 24 h. We evaluated the T helper1 (Th1) and T cytotoxic1 (Tc1) cell cytokine production by flow cytometry and immune-enzymatic assays. In HC, we also evaluated apoptosis and/or necrosis by flow cytometry. We observed an immunomodulatory effect of cysteamine at 400 µM in PBMCs from TB and TBI subjects. It significantly reduced PPD-specific Th1 responses at 24 h and at 6 h (p=0.0004 and p=0.0009, respectively), and a similar non-significant trend was observed with cysteamine at 200 µM (p=0.06 at 24 h and p=0.14 at 6 h). Moreover, cysteamine at both 400 µM (p<0.0001 and p=0.0187 at 24 h, respectively, and p<0.0001 at 6 h for both) and 200 µM (p=0.0119 and p=0.0028 at 24 h and p=0.0028 and p=0.0003 at 6 h, respectively) significantly reduced SEB-induced Th1 and Tc1 responses. Furthermore, we found that cysteamine induced morphological lymphocyte changes and significantly reduced the lymphocyte percentage in a dose- and time-dependent manner. Cysteamine at 400 µM induced 8% late apoptosis and 1.6% necrosis (p<0.05) at 24 h. In contrast, despite significant differences from untreated conditions (p<0.05), cysteamine at 400 µM for 6 h induced approximately 1% late apoptosis and 0.1% necrosis in the cells. High doses of cysteamine in vitro reduce the percentages of PPD- and SEB-induced Th1 and Tc1 cells and induce late apoptosis and necrosis. Differently, cysteamine at lower doses retains the immunomodulatory effect without affecting cell viability. These findings suggest cysteamine as a potential adjunct to antimicrobial regimens as in the TB or COVID-19 field, for its ability to reduce the inflammatory status.

Interleukin-4 expression is increased in patients with tuberculosis: A systematic review and meta-analysis.

Interleukin-4 (IL-4) is an important cytokine in the Th2 differentiation of CD4+ T cells, which modulates immune responses and participates in host defense against Mycobacterium tuberculosis. The present study aimed to evaluate the significance of IL-4 concentration in patients with tuberculosis. Data from this study will be helpful in understanding the immunological mechanisms of tuberculosis and in clinical practice. A data search was conducted from January 1995 to October 2022 in electronic bibliographic databases such as China National Knowledge Infrastructure, Wan Fang, Embase, Web of Science, and PubMed. The Newcastle-Ottawa Scale was used to assess the quality of the included studies. Heterogeneity between the studies was assessed using I2 statistics. Publication bias was determined by funnel plot, and Egger's test was used to confirm the presence of publication bias. All qualified studies and statistical analyses were performed using Stata 11.0. Fifty-one eligible studies comprising 4317 subjects were included in the meta-analysis. The results depicted a considerably increased level of serum IL-4 in patients with tuberculosis than in the controls (standard mean difference [SMD] = 0.630, [95% confidence interval (CI), 0.162-1.092]). However, there was no significant difference in plasma IL-4 levels between patients with TB and controls (SMD = 0.290, [95% CI, -0.430 to 1.010]). In addition, the infection status, TB focus location, drug resistance, race, research design type, and detection method divided the subjects into different subgroups for the meta-analysis. The results of the comparison of healthy controls and TB subjects showed that in the Asian population, the serum IL-4 level in patients with TB was higher than that in controls (SMD = 0.887, [95% CI, 0.202 to -1.573]) and patients with active TB as well as people with pulmonary TB showed increased serum IL-4 levels compared to controls (SMD = 0.689, [95% CI, 0.152-1.226]). In the case of the control group with latent TB, the active TB group had higher serum IL-4 levels than the control group (SMD = 0.920, [95% CI, 0.387-1.452]). The present meta-analysis showed that serum IL-4 varied in healthy individuals and patients with TB. Patients with active TB may also exhibit higher IL-4 concentrations.

Differential expression of chemokine receptors on monocytes in TB and HIV S

In the present study, we defined multiple chemokine receptors expressed by classical, intermediate and non-classical monocyte subsets in TB, HIV and TB/HIV co-infection and associate it with the perturbation of monocyte subsets due to the diseases. Peripheral blood mononuclear cells from TB+ (n = 34), HIV+ (n = 35), TB + HIV+ (n = 12), as well as TB-HIV- healthy controls (n = 39), were tested for monocyte phenotyping by flow cytometry. Frequencies of intermediate and non-classical monocytes were significantly higher in TB and/or HIV disease relative to healthy controls. CCR2 and CX3CR1 were significantly higher on monocytes in TB disease, whereas CCR4 and CCR5 were present at higher levels in HIV disease. TB/HIV co-infected patients exhibited CCR2, CCR5 and CX3CR1 levels intermediate to TB and HIV subjects, while CCR4 was at a higher level than HIV. Despite the increase in the expression of chemokine receptors due to disease conditions, chemokine receptors maintained their original expression pattern on monocyte subsets. Our data provided new insight into the disease-specific but not monocyte subsets-specific modulation of chemokine receptors in TB and HIV.

Correlation of TB LAM Ag (Tuberculosis Lipoarabinomannan Antigen) Results from Urine with Adenosine Deaminase Levels from Pleural Fluid Patients with Pulmonary TB Accompanied by Pleural Effusion

Background: One of the newest testing methods for active TB uses TB LAM Antigen, in which lipoarabinomannan (LAM) as the examined substance serves as the main component of the cell wall of M. tuberculosis (MTB) and the degradation product of MTB by macrophages in the human body. Patients with active TB with or without HIV infection may experience a decreased antibody response, which causes LAM not to bind to antibodies so that it can pass through the normal glomerular basement membrane and can be detected in the urine. Lateral flow urine lipoarabinomannan assay (LF-LAM) can detect this material with urine samples from active TB patients. Another TB screening method uses the measurement of adenosine deaminase (ADA) activity. ADA has a very important function for the proliferation and differentiation of lymphoid cells, especially T cells, and assists in the maturation of monocytes into macrophages. Objective: This study aims to qualitatively prove the correlation between the pleural fluid ADA levels of pulmonary TB patients with pleural effusion using the LF-LAM test results and examine the potential of TB LAM Ag, which correlates with increased pleural fluid ADA levels, as a diagnostic tool for diagnosing pulmonary TB. Method: It is an observational analytic study with a crosssectional design. Materials for this study included temporary urine from active TB patients at Dr. Soetomo Regional Public Hospital collected in May-August 2022. The descriptive analysis was conducted using SPSS 25.0. The data were tested for normal distribution and with the homogeneity test of Shapiro-Wilk's data. The statistical analysis was performed using the Mann-Whitney test, while the kappa suitability test was carried out to determine the correlation and level of concordance between the ADA cut-off value of pleural fluid and the TB LAM Ag test results. Results: This study involved 50 subjects consisting of 22 (44%) subjects with ADA test results between 40-100 IU/L compared to 28 subjects (56%) with ADA test results below 40 IU/L. There were 14 (28%) subjects with positive TB LAM Ag test results, consisting of 10 (45%) active TB subjects with a high ADA test cut-off of 40-100 IU/L and 4 (14%) active TB subjects with a low ADA test cut-off below 40 IU/L, with a p-value = 0.012. Conclusions: There is a correlation between TB Lipoarabinomannan antigen test results of TB patients with pleural effusion. TB LAM Ag can be detected positive on ADA test results of < 40 IU/L and more positive in active TB patients with high ADA test results of > 40-100 IU/L. Therefore, the TB LAM Ag test is more sensitive than the ADA test. The cut-off value of the ADA test reached 76.4 U/L, and a positive TB LAM was found.

Anti-Tuberculosis Therapy in The Drug Resistant Tuberculosis Patients of Kashmir Valley

Drug-resistant tuberculosis (DR-TB) strains are a significant threat to global TB control. Due to a lack of resources and inadequate healthcare infrastructure, it is difficult to identify and monitor drug resistance situations in low and middle-income nations. The study aimed to evaluate the anti-tuberculosis therapy regimen in drug-resistant tuberculosis patients of the mountainous valley of Kashmir. The current study was carried out at Government Medical College, Department of Pulmonary Medicine, Chest Diseases Hospital (CDH), Srinagar, Kashmir. A total of 1100 suspected tuberculosis subjects were included in the study, and out of 1100 TB subjects, 195 were documented medication resistance cases. Of the total 1100 Line Probe Assay (LPA) tests done, 69 (6.2%) such samples in the Tuberculosis band were absent. Of the real samples tested by LPA, Tuberculosis band (TUB) was present in 1031 samples (93.7%), of which 24 (2.3%) were multidrugresistant (MDR), 121 (11.7%) were rifampicin mono-resistant, 42 (4.0%) were isoniazid mono-resistant, 8 (0.77%) were extensively drugresistant (XDR), and 836 (81%) samples were pan-sensitive. Smoking remained an independent risk factor for poor treatment outcomes in the multivariate analysis (p-value = 0.014, OR = 4.356, 95% CI [1.425�12.45]). This study found a low incidence of Drug-Resistant Tuberculosis (DR-TB). Most of the cases were resistant to first-line anti-tuberculosis drugs. Special attention should be paid to high-risk DR-TB patients to improve treatment results even further. Further studies are recommended with a large sample size.

Effect of age and gender on high - Sensitivity C - Reactive protein levels serum on health worker with latent tuberculosis and healthy control

BackgroundLatent tuberculosis is defined as a state of persistent immune response stimulated by Mycobacterium tuberculosis antigens with no evidence and signs of active TB . Health workers have a high risk of developing latent TB disease due to occupational exposure from patients. High sensitivity CRP (hs-CRP) assays have been developed for special values that may indicate low-grade inflammatory lesions as is true in measurement of latent tuberculosis infection. Factors that affect CRP levels are gender and age. Our study is conducted to asses effect of age and gender on Hs- CReactive protein leves serum on health worker with latent tuberculosis and healthy control. MethodThis research is a cross sectional study using primary data. The research was conducted at Wahidin Sudirohusodo Makassa Hospital and Community Center For Lung Health In South Sulawesi. Studied subject were recruited by consecutivesampling, in which the patient who met the inclusion criteria and then the serum HsCRP test was measured. Data analysis was performed using SPSS version 25. ResultDuring the study period , 80 subjects met the inclusion criteria. At age ≤ 32 years, the mean HsCRP was found to be lower in latent TB than in healthy controls, but not statistically significant (p>0.370). At age >32 years, the mean HsCRP was found to be higher in latent TB than in healthy controls, but not statistically significant (p>2.49). In males, the mean HsCRP was found to be higher in latent TB than in healthy controls, but not statistically significant (P =0.584). In women, the mean HsCRP was found to be lower in latent TB than in healthy controls, but not statistically significant (P =0.712). ConclusionSerum HsCRP levels were found to be higher in latent TB subjects with increasing age and male gender but not statistically significant.

Exploring the value of Mycobacterium tuberculosis modified lipoprotein as a potential biomarker for TB detection in children

BackgroundCurrent TB diagnostic methods available have been developed for adults and development efforts have neglected the differences in disease and sampling that occur between adults and children. Diagnostic challenges are even greater in HIV co-infected children and infants.Methods and resultsWe established a sandwich ELISA assay to detect Mycobacterium tuberculosis modified lipoprotein (TLP) ex vivo in plasma. The study population contains plasma samples from 21 patients with active TB and 24 control samples with no TB, collected in the International Maternal Pediatric Adolescent AIDS Clinical Trails (IMPAACT) P1041 study. Retrospective analysis was performed and the results demonstrate that the median plasma levels of TLP in control subjects are 2.7 fold higher than the median plasma values in active TB subjects (p < 0.001).ConclusionsPlasma levels of TLP are elevated with active TB disease in HIV positive subjects and deserves further exploration as an indicator for TB detection in children.

PDL1 expression on monocytes is associated with plasma cytokines in Tuberculosis and HIV.

IntroductionPDL1 and its interaction with PD1 is implicated in immune dysfunction in TB and HIV. The expression of PDL1 on multiple subsets of monocytes as well as their associations with cytokines and microbial products have not been well studied.MethodHIV (TB-HIV+), TB (TB+HIV-) and TB/HIV co-infected (TB+HIV+) patients as well as apparently healthy controls (TB-HIV-) were recruited. TB and HIV patients were treatment naïve while TB/HIV patients were both ART naïve and experienced but not yet started TB therapy. Monocyte subsets were evaluated for PDL1 expression by flow cytometry; plasma TNFα, IL6, IP10, IFNγ and IL10 were measured by Luminex; and cytokine mRNA from purified monocytes quantitated by qPCR. The association of PDL1 with cytokines, clinical and microbial indices, including HIV viral load, TB smear microscopy and TB urinary lipoarabinomannan (LAM) were assessed.ResultsMonocyte expression of PDL1 was significantly higher in TB, HIV and TB/HIV co-infected patients compared with healthy controls (p = 0.0001), with the highest levels in TB/HIV co-infected patients. The highest expression of PDL1 was on intermediate (CD14+CD16+) monocytes in all participant groups. PDL1 strongly correlated with HIV viral load in TB/HIV while weakly correlated in HIV. PDL1 levels moderately correlated with plasma TNFα, IL6, IP10, IFNγ and IL10 level in TB subjects whereas weakly correlated with TNFα and IP10 in HIV patients. However, cytokine mRNA from purified monocytes showed no association with either plasma cytokines or monocyte PDL1 expression, implying that if cytokines modulate PDL1, they are likely not originating from circulating monocytes themselves. These results underscore the importance of further characterization of multiple monocyte subsets and their phenotypic and functional differences in different disease states.

Mycobacterial heparin-binding hemagglutinin (HBHA)-induced interferon-γ release assay (IGRA) for discrimination of latent and active tuberculosis: A systematic review and meta-analysis.

The Mycobacterial heparin-binding hemagglutinin (HBHA) is an important latency-associated antigen that can be used to distinguish between latent tuberculosis infection (LTBI) and active tuberculosis (ATB). Although many studies were explored the efficiency of the HBHA-induced interferon-γ release assay (IGRA) in different populations, the clinical differential value of HBHA-IGRA is still controversial. Therefore, the aim of this study was to determine whether the HBHA-IGRA can be used as an efficient test for the discrimination of LTBI and ATB by a systematic review and meta-analysis. Relevant articles were retrieved from PubMed, Embase, Web of Science, and the Cochrane Library on Oct 18, 2020, with no start date limitation. The quality of each study was evaluated using Review Manager 5.4. The Stata MP v.14.0 software was used to combine sensitivity, specificity, likelihood ratio (LR), diagnostic odds ratio (DOR), summary receiver operating characteristic (SROC) curve, and area under SROC (AUC) to evaluate the diagnostic value of HBHA-IGRA for discrimination of LTBI and ATB. Meta-regression and subgroup analysis were performed for the sources of heterogeneity based on the selection criteria for active TB, the population, the TB burden, the type of antigen, the type of sample, and the time of antigen stimulation. A total of 13 studies (14 results) were included in this meta-analysis, including 603 ATB patients and 514 LTBI individuals. The pooled sensitivity and specificity of the HBHA-IGRA for discrimination of the LTBI and ATB were 0.70 (95% CI, 0.57~0.80) and 0.78 (95% CI, 0.71~0.84), respectively. The pooled positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR) were 3.15 (95%CI, 2.43~4.09), 0.39 (95% CI, 0.27~0.56), and 8.11 (95% CI, 4.81~13.67), respectively. The AUC was 0.81 (95% CI, 0.77~0.84). The subgroup analysis showed that the main source of heterogeneity was due to the HIV-infected population incorporated, and the different selection criteria of active TB subjects would also lead to the variation of the pooled sensitivity and specificity. Different TB burdens, HBHA antigen types, sample types, antigen stimulation time and BCG vaccination did not affect the heterogeneity in this analysis. The HBHA-IGRA is a promising immunodiagnostic test for discrimination of latent and active TB, which can be added in commercial IGRAs to enhance the differential diagnostic performance.

Influence of NOD-like receptor 2 gene polymorphisms on muramyl dipeptide induced pro-inflammatory response in patients with active pulmonary tuberculosis and household contacts

PurposeThe immune response induced by nucleotide-binding oligomerization domain-2(NOD2) is associated with the production of cytokines affected by the host's genetic background. The present study aimed to examine the effects of NOD2; 802C > T, 2105G > A polymorphisms associated with altered cytokine levels in patients with active pulmonary tuberculosis disease, Latent TB subjects (household contacts(HHC) and healthy controls(HC). MethodsGenetic polymorphisms were analyzed by Restriction Fragment Length Polymorphism(RFLP) in 102-PTB patients, 102-HHC, and 132-HC. QuantiFERON-TB Gold In-Tube test was performed to identify latent TB infection in 60-HHC. Estimated their cytokine levels by ELISA in MDP (muramyl dipeptide) stimulated culture supernatants of all the groups. Further, we studied pre-mRNA structures by insilico analysis and relative gene expression by RT-PCR. ResultsRecessive genetic models of NOD2 802C > T SNP with TT genotype and AA genotype of NOD2 2105G > A SNP were significantly associated with increased TB risk in PTB patients and HHC compared with HC. In vitro stimulations were performed with NOD2 ligand MDP in PTB patients and latent TB subjects: QuantiFERON positive household contacts (QFT + ve HHC)and QuantiFERON negative household contacts(QFT-ve HHC). The results showed that reduced TNF-α and enhanced IL-12, IL-1β indicate that these cytokines may play an essential role in the initial maintenance of cell-mediated immunity. Our study demonstrated the correlation between NOD2 polymorphism with IL-1β, TNF-α, IL-12 levels. Insilico analysis represents the pre-mRNA secondary structures affected by NOD2 SNPs. We also observed the difference in m RNA levels in variant and wild genotypes. ConclusionThis finding may lead to the forthcoming development of immunotherapy and may be used as predictive markers to identify high-risk individuals for TB disease.

Neural interfacing architecture enables enhanced motor control and residual limb functionality postamputation

Despite advancements in prosthetic technologies, patients with amputation today suffer great diminution in mobility and quality of life. We have developed a modified below-knee amputation (BKA) procedure that incorporates agonist-antagonist myoneural interfaces (AMIs), which surgically preserve and couple agonist-antagonist muscle pairs for the subtalar and ankle joints. AMIs are designed to restore physiological neuromuscular dynamics, enable bidirectional neural signaling, and offer greater neuroprosthetic controllability compared to traditional amputation techniques. In this prospective, nonrandomized, unmasked study design, 15 subjects with AMI below-knee amputation (AB) were matched with 7 subjects who underwent a traditional below-knee amputation (TB). AB subjects demonstrated significantly greater control of their residual limb musculature, production of more differentiable efferent control signals, and greater precision of movement compared to TB subjects (P < 0.008). This may be due to the presence of greater proprioceptive inputs facilitated by the significantly higher fascicle strains resulting from coordinated muscle excursion in AB subjects (P < 0.05). AB subjects reported significantly greater phantom range of motion postamputation (AB: 12.47 ± 2.41, TB: 10.14 ± 1.45 degrees) when compared to TB subjects (P < 0.05). Furthermore, AB subjects also reported less pain (12.25 ± 5.37) than TB subjects (17.29 ± 10.22) and a significant reduction when compared to their preoperative baseline (P < 0.05). Compared with traditional amputation, the construction of AMIs during amputation confers the benefits of enhanced physiological neuromuscular dynamics, proprioception, and phantom limb perception. Subjects' activation of the AMIs produces more differentiable electromyography (EMG) for myoelectric prosthesis control and demonstrates more positive clinical outcomes.

Biochemical changes in tuberculosis.

Introductiontuberculosis (TB) remains a global health issue with high morbidity and mortality rates especially in the developing countries. It is a multi-organ disease and can influence biochemical changes. This study sought to determine the influence of tuberculosis and its drug treatment on serum biochemical parameters in patients in Nigeria.Methodsit was a descriptive observational cohort study on 150 subjects whose blood samples were analyzed for serum albumin, serum sodium, and serum potassium. The subjects were grouped into 3: TB group= 50 new TB subjects not on treatment, F group= 50 TB subjects on treatment for 2/12 or more and C group= 50 non-TB control subjects. These biochemical variables were compared between the 3 groups.Resultsmale/female ratio was 1: 1.5, mean age 37.1±0.92 years, and range 18-65 years. The differences in mean values of serum albumin, calcium and sodium between the three groups were significant (p<0.001), whereas that of serum potassium was not significant (p=0.056). Those patients with new case TB had a significantly lower serum sodium, serum albumin and serum calcium than the control group and those on treatment, p<0.001. There was significant positive correlation between serum albumin and serum calcium (r=0.0.420, p<0.001) as well as serum sodium (r=0.310, p<0.001) in the study population. Similarly, the correlation between serum calcium and serum sodium was positive and significant (r=0.200, p=0.014). In contrast, the correlation between serum potassium and serum albumin and that between serum potassium and serum calcium was not significant.Conclusiontuberculosis with or without anti-tuberculous medications was associated with significant reduction in serum albumin, serum sodium and serum calcium in this study.

Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

Indonesia is one of the countries with the highest multidrug-resistant tuberculosis cases in the world. Rapid molecular test using the Xpert MTB/RIF assay is one of the detection methods for MDR-TB. Early detection of MDR-TB is crucial for early initiation of treatment. However, Xpert MTB/RIF assay only detects the rpoB gene mutations associated with Rifampicin resistance. Recently, WHO recommends the use of Pyrosequencing, a DNA sequencing method that can detect not only the rpoB gene but also katG and/or inhA gene mutations associated with Isoniazid resistance. The aims of this study were to compare the interpretation between the two methods and to determine the differences in codon mutation position detection of the rpoB gene and mutation detection of the katG and/or inhA gene. This was a cross-sectional comparative observational study on patients ≥18 years old interpreted as RR-TB patients based on Xpert MTB/RIF assay results who had not received MDR-TB drugs at Dr. Hasan Sadikin General Hospital, Bandung, Indonesia. Results showed there were 40 Rifampicin-resistant TB subjects interpreted by Xpert MTB/RIF assay while Pyrosequencing interpreted 30 MDR-TB, 9 RR-TB and one Isoniazid-resistant TB subjects in January - February 2020. The detection of rpoB gene codon mutation position between Xpert MTB/RIF assay and Pyrosequencing methods was not significantly different (p=0.389). Pyrosequencing had detected 27 katG gene mutations, 3 inhA gene mutations, one katG and inhA gene mutation. To conclude, Pyrosequencing can be used for accurate detection of Rifampicin and Isoniazid resistance in MDR-TB.

1478-P: Having Prediabetes before the Detection of Tuberculosis: Does It Have a Negative Influence on Treatment?

Aim: Coexistence of DM and TB influences TB treatment outcomes. TB increases hyperglycaemia due to inflammatory response. Effect of prediabetes (PDM) on TB treatment outcomes is unknown. The aim was to assess the effect of PDM on treatment outcomes after TB treatment. Methods: This is a prospective observational cohort study (EDOTS - part of RePORT India Consortium) of 569 eligible New smear positive cases screened for DM between 2014 to 2018 in TB units in North Chennai, South India. Out of 569 subjects, 179 with a known history of DM were excluded.The remaining 390 screened TB subjects, 120 were excluded based on culture results and unwillingness to participate in the study. HbA1c was assessed at baseline and categorised into two groups (Gp): normoglycaemia (Gp1) (&amp;lt;5.7%) and PDM (Gp 2) (5.7 to 6.4%) and followed them at 3rd and 6th month and treatment outcomes were assessed at the end of the TB treatment. Results: Prevalence of Prediabetes was 41.1%. There was no significant difference in the treatment outcomes between the groups(p=0.54). Total cure rate was 72.7% with no significant difference between the groups. There was a higher proportion of treatment failures (7.9%) amongGp1 as compared to Gp 2 (4.5%) (p = 0.33). There were 15.8 vs. 17.1% treatment defaulters. Higher proportion of deaths occurred during the course of DOTS in Gp 2 (7.2%) compared to Gp 1 (1.3%) (p = 0.06). At the end of intensive phase of DOTS treatment, about 23.8 % were observed to have positive sputum smear in Gp 2 compared to 8.6% in Gp 1 (p=0.019). The estimated relative risk to remain as sputum smear positive among prediabetes at the end of intensive phase was 3.0 (95% CI: 1.2-7.6). Conclusion: Higher prevalence of prediabetes was evident among TB patients. Among those who had prediabetes, death rate was higher during the course of the treatment and higher proportion remained sputum smear positive at the end of the intensive phase of TB treatment. Disclosure V. Viswanathan: None. A. Devarajan: None. S. Kumpatla: None. M. Dhanasekaran: None. S. Babu: None. H. Kornfeld: None. Funding Government of India (BT/MB/INDO-US/REPORT/03/2013-140)

Determination of Malondialdehyde and Superoxide Dismutase in Pulmonary Tuberculosis Subjects

Pulmonary tuberculosis (PTB) is a dangerous bacterial infection that attacks the lungs. It has long been documented that there is an increase circulating levels of free radicals and oxidative stress markers in TB subjects. Malnutrition and deficient antioxidant capabilities further complicate the patient’s situation. The aim of this study is to determine the serum levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in pulmonary tuberculosis patients and normal volunteers at Rasheed Shekoni Teaching Hospital Dutse, Jigawa State, Nigeria. The present study consists of one hundred and thirty-four (134) subjects, Ninety (90) among which are diagnosed TB patients and forty-four (44) apparently healthy controls attending Rasheed Shekoni Teaching Hospital Dutse. Venous samples were collected from ninety consecutive, consenting PTB on drugs and those that have not started medications. MDA levels were analysed using the method of Nadigar et al (1986). The mean values of MDA in the 24 tuberculosis infected males who are yet to commence drug was 4.0±0.32, 2.8±0.53 in those that are on drugs and 2.0±0.23 in the male control subjects. It was 4.1±0.35, 2.9±0.29 and 1.9±0.32 in female that are TB infected and yet to commence drugs, those on drugs and the control subjects respectively. The total mean MDA value for those on tuberculosis drugs was 4.0±0.33, 2.8±0.46 for those that are yet to commence the treatment and 2.0±0.28 for the control subjects, the serum levels of SOD was found significantly lower (p&lt;0.005) in PTB subjects on treatment compared to those without treatment and control subjects. Conclusively, the findings of the current study showed that pulmonary tuberculosis patients are predisposed to oxidative stress leading to an increased MDA and consequent decreased SOD levels as compared to the control subjects.

Can Interferon-γ Release Assays Be Useful for Monitoring the Response to Anti-tuberculosis Treatment?: A Systematic Review and Meta-analysis.

The number of studies which evaluated interferon-gamma release assays (IGRAs) results after anti-tuberculosis (TB) treatment has been rapidly increasing. The aim of this study was to investigate the potential use of IGRAs (QFT-GIT, T-SPOT.TB, QFT-Plus) in assessing the response to anti-TB treatment. We searched all studies in English language published from 1 October 2011 to 18 November 2018 in PubMed, Web of Science, and Scopus. Our search included the term "tuberculosis treatment AND interferon-γ release assay". We included studies evaluating the performance of commercial IGRAs (including QFT-GIT, T-SPOT.TB and QFT-Plus) before and after the anti-TB treatment. We performed subgroup analysis based on the age (children, adults), type of TB (active, latent, active and latent, and contacts exposed to MDR defined as MDR LTBI), type of IGRAs (QFT-GIT and T-SPOT.TB), and follow-up interval (2, 3, 4, 6, 9months). Of the 18 included studies, 12 used QFT-GIT for assessment of IGRA performance after therapy, 1 used T-SPOT.TB, and 3 used both QFT-GIT and T-SPOT.TB. Since then, only two studies have assessed the QFT-Plus performance during therapy. According to the results of the meta-analysis, the pooled rate of positive IGRAs (QFT-GIT and T-SPOT.TB) following anti-TB therapy was estimated at 76% [95% CI 70-81%] and no difference was found compared to the pooled positive rate of IGRAs before initiation of therapy which was 76% [95% CI 60-89%]. The subgroup analysis showed that the pooled rate of positive IGRAs (QFT-GIT and T-SPOT.TB) after anti-TB therapy was significantly higher in monitoring active TB subjects [80% (95% CI 74-88%)] than LTBI [71% (95% CI 70-81%)]. Available data are now sufficient to suggest that monitoring changes in the IGRAs (QFT-GIT and T-SPOT.TB) response during anti-TB treatment may have limited use in evaluating the effectiveness of treatment, while the monitoring changes in QFT-Plus during anti-tubercular treatment are recommended to determine treatment efficacy or for treatment monitoring. Further research is needed to establish the efficacy of this new assay as marker on a larger scale for treatment monitoring.

Screening and identification of plasma lncRNAs uc.48+ and NR_105053 as potential novel biomarkers for cured pulmonary tuberculosis

BackgroundTuberculosis (TB) treatment takes a long time, and a gold standard test to define TB cure is lacking. This may lead to early discharge of TB patients, resulting in an increased risk of disease transmission and drug resistance. Plasma lncRNAs might act as potential biomarkers to evaluate TB cure in an efficient and precise manner. MethodsA lncRNA microarray assay was used to screen differentially expressed plasma lncRNAs in untreated TB and cured TB subjects. The expression levels of lncRNAs were verified by qPCR. Target genes of lncRNAs were predicted using a coding–non-coding gene co-expression network and mRNA–lncRNA–miRNA interaction network analysis. ResultsThe expression levels of lncRNAs uc.48+ (p < 0.001) and NR_105053 (p = 0.03) were found to differ significantly between the untreated TB group and the cured TB group. The predicted target genes of uc.48+ were EP300, BAI1 and NR_105053 were TLR9, MYD88, BAI1, respectively. A predictive model for cured TB was established by the combination of uc.48+ and NR_105053 expression, with a sensitivity of 90.00% and specificity of 86.36%, and an area under the curve (AUC) value of 0.945. ConclusionslncRNAs uc.48+ and NR_105053 may serve as potential biomarkers to distinguish between untreated TB patients and cured TB subjects. This study provides an experimental basis to evaluate the effect of TB treatment and may also provide new clues to the pathological mechanisms of TB.

Prognostic value of some serum protein fractions as Early Index of Clinical Recovery in Pulmonary Tuberculosis subjects

BackgroundMalnutrition is an important risk factor for pulmonary tuberculosis and can result to adverse treatment outcomes. AimThis was a cross-sectional study designed to assess prognostic value of some serum protein fractions in pulmonary tuberculosis (PTB) subjects at Oron, Akwa Ibom State, Nigeria. Thirty (30) TB subjects on Anti-tuberculosis therapy, thirty (30) drug naive TB subjects and thirty (30) apparently healthy control subjects aged 21–52 (35 ± 16) years were conveniently recruited. MethodsTotal protein and albumin were measured colourimetrically, Albumin-globulin ratio was calculated while demographic data was obtained using questionnaire. ResultsBMI (kg/m2), Albumin (g/dl) and AGR were significantly lower in TB subjects with or without ATT when compared with control subjects (p < 0.000 respectively), but higher in PTB subjects on ATT when compared with drug naive PTB subjects (p = 0.000 respectively). Serum Total protein (g/dl) level in PTB subjects with or without ATT was significantly higher when compared with controls (p = 0.004) while globulin (g/dl) level was lower in PTB subjects on ATT when compared with drug naive PTB subjects (p = 0.000). ConclusionDecreased BMI in TB subjects signifies reduction in muscle mass and wasting precipitated by PTB infection, while depleted albumin and AGR suggests high degree of malnutrition. Increased albumin and AGR in PTB subjects on ATT suggests improvement with ATT. Assessment of serum albumin and AGR may serve as affordable and early index of clinical recovery in PTB subjects especially in resource limited settings, and may be more reliable than the traditionally used BMI.

Blastocystis subtype 1 (allele 4); Predominant subtype among tuberculosis patients in Iran

BackgroundBlastocystis and tuberculosis are two public health issues that are frequently reported in regions with low level of hygiene. Therefore, the current study aimed to investigate Blastocystis subtype and allele distribution in TB patients. MethodsTotally, 161 stool samples were taken from TB patients who were undergoing anti-MTB treatment. Stool samples were concentrated using conventional formalin-ether technique and examined using Lugol's iodine staining under light microscopy. DNA extraction was carried out and discriminative fragment was amplified and sequenced. With comparison in GenBank database, relevant subtypes and alleles were characterized and phylogenetically analyzed using MEGA v.7 and Tamura 3-parameter model. ResultsIn total, from 161 stool samples, 19 samples were suspected to be Blastocystis-positive. The expected fragment was amplified in 13 (8.07%) of samples. Accordingly, 11/13 (84.62%) of Blastocystis cases settled in urban and 2/13 (15.38%) were villagers. Close-contact with animals was also seen among 7/13 (53.84%) of samples. Subtype 1 (7/13; 53.84%) was the most prevalent followed by subtype 2 (5/13; 38.46%) and subtype 3 (1/13, 7.69%). All ST1 were allele 4, while alleles 9, 11 and 12 were seen in ST2 and allele 34 was the only allele observed in ST3. All three subtypes were clearly separated, while there was no separation between sequences from TB and non-TB patients. ConclusionBlastocystis ST1 was the most prevalent subtype in TB patients and there was no difference between Blastocystis isolates from TB and non-TB human subjects.

Toxoplasma gondii and Tuberculosis Co-infection among Tuberculosis patients at the Bamenda Regional Hospital, North West Region, Cameroon

Aim: To determine the prevalence of toxoplasmosis and tuberculosis co-infection among (TB) patients. &#x0D; Study Design: A cross-sectional study was employed to gather data on 147 sputum positive TB patients.&#x0D; Place and Duration of Study: Bamenda Regional Hospital (BRH), Tuberculosis Reference Laboratory and Treatment Centre from October 2015 to April, 2016. &#x0D; Method: An epidemiological questionnaire was applied to gather data on positive-sputum pulmonary TB patients of both sexes on whom some risk factors for Toxoplasma were explored, followed by blood sampling. T. gondii IgM and IgG antibodies were researched in sera samples using indirect ELISA.&#x0D; Results: T. gondii antibodies prevalence amongst the TB population was 83% (122/147, 95% CI 112-130). 85.25% of the Toxoplasma infection in the study population were recent infections with T.gondii parasite while up to 53.28% of patients were reactivating, no significant difference in prevalence between HIV negative or HIV positive TB patients was established.&#x0D; Conclusion: The high seroprevalence of toxoplasmosis among TB subjects attending treatment at the Bamenda Regional Hospital is suggestive of co-infection. Toxoplasmosis and tuberculosis co-infection are opportunistic infections for HIV/AIDS, and life threatening to TB patients and could be the reason for unexplained deaths among these patients on treatment.