Estimation of country-specific tuberculosis resistance antibiograms using pathogen genomics and machine learning.

BACKGROUND: Global tuberculosis (TB) drug resistance (DR) surveillance focuses on rifampicin. We examined the potential of public and surveillance Mycobacterium tuberculosis (Mtb) whole-genome sequencing (WGS) data, to generate expanded country-level resistance prevalence estimates (antibiograms) using in silico resistance prediction. METHODS: We curated and quality-controlled Mtb WGS data. We used a validated random forest model to predict phenotypic resistance to 12 drugs and bias-corrected for model performance, outbreak sampling and rifampicin resistance oversampling. Validation leveraged a national DR survey conducted in South Africa. RESULTS: Mtb isolates from 29 countries (n=19 149) met sequence quality criteria. Global marginal genotypic resistance among mono-resistant TB estimates overlapped with the South African DR survey, except for isoniazid, ethionamide and second-line injectables, which were underestimated (n=3134). Among multidrug resistant (MDR) TB (n=268), estimates overlapped for the fluoroquinolones but overestimated other drugs. Globally pooled mono-resistance to isoniazid was 10.9% (95% CI: 10.2-11.7%, n=14 012). Mono-levofloxacin resistance rates were highest in South Asia (Pakistan 3.4% (0.1-11%), n=111 and India 2.8% (0.08-9.4%), n=114). Given the recent interest in drugs enhancing ethionamide activity and their expected activity against isolates with resistance discordance between isoniazid and ethionamide, we measured this rate and found it to be high at 74.4% (IQR: 64.5-79.7%) of isoniazid-resistant isolates predicted to be ethionamide susceptible. The global susceptibility rate to pyrazinamide and levofloxacin among MDR was 15.1% (95% CI: 10.2-19.9%, n=3964). CONCLUSIONS: This is the first attempt at global Mtb antibiogram estimation. DR prevalence in Mtb can be reliably estimated using public WGS and phenotypic resistance prediction for key antibiotics, but public WGS data demonstrates oversampling of isolates with higher resistance levels than MDR. Nevertheless, our results raise concerns about the empiric use of short-course fluoroquinolone regimens for drug-susceptible TB in South Asia and indicate underutilisation of ethionamide in MDR treatment.

Intra-regional variations and contextual effects on facility-based delivery in Bangladesh: A multi-level analysis.

We examined intra-regional variations and contextual influences on institutional delivery of women using the nationally representative 2014 Bangladesh Demographic and Health Survey data. Due to the hierarchical structure of the data, we employed multi-level logistic regression analysis. Of the women who had had a live birth in the last three years preceding the survey, only 38% availed the opportunity of institutional delivery. From the findings of this study, we observed that women of the Eastern region were less likely and those of the Western region were more likely to use FBD compared to the women of the Central region. Both individual- and community-level factors influence women to use facility-based delivery. Community-level programs aimed at improving availability and easy accessibility to economically deprived and geographically disadvantaged areas may increase safe motherhood practices among women.

Feature Importance Score-Based Functional Link Artificial Neural Networks for Breast Cancer Classification.

Growth of malignant tumors in the breast results in breast cancer. It is a cause of death of many women across the world. As a part of treatment, a woman might have to go through painful surgery and chemotherapy that may further lead to severe side effects. However, it is possible to cure it if it is diagnosed in the initial stage. Recently, many researchers have leveraged machine learning (ML) techniques to classify breast cancer. However, these methods are computationally expensive and prone to the overfitting problem. A simple single-layer neural network, i.e., functional link artificial neural network (FLANN), is proposed to overcome this problem. Further, the F-score is used to reduce the issue of overfitting by selecting features having a higher significance level. In this paper, FLANN is proposed to classify breast cancer using Wisconsin Breast Cancer Dataset (WBCD) (with 699 samples) and Wisconsin Diagnostic Breast Cancer (WDBC) (with 569 samples) datasets. Experimental results reveal that the proposed models can diagnose breast cancer with higher performance. The proposed model can be used in the early breast cancer diagnosis with 99.41% accuracy.

Child marriage and its association with Maternal Health Care Services utilisation among women aged 20-29: a multi-country study in the South Asia region.

Despite pervasiveness of child marriage in South Asian countries, its association with the usage of Maternal Health Care Services (MHCS) is poorly explored. In this study, we examined the impact of child marriage on MHCS utilisation among the prime reproductive aged women of five South Asian countries Afghanistan, Bangladesh, India, Nepal and Pakistan. Data for this study are extracted from the most recent Demographic and Health Surveys. The effect of child marriage on the use of MHCS was assessed by the multivariate logistic regression analysis. Findings reveal that in the South Asian region, the prevalence of child marriage is more common in Bangladesh, followed by Afghanistan, Nepal, India and Pakistan. Child marriage significantly (p< .01) deters the usage of MHCS - at least once antenatal care (ANC) visit, at least four ANC visit, Institutional delivery and seeking assistance from skilled birth assistance (SBA), though the effect size was somewhat different across the countries. Findings stipulate to remain girls in schooling more and the marriage acts of the countries should be enacted properly to diminish child marriage not only for increasing MHCS use, also for individual wellbeing and overall development of each country.Impact StatementWhat is already known on this subject? Child marriage in South Asian countries is commonplace despite substantial improvements of socioeconomic status. Although child marriage have many adverse reproductive and health outcomes, empirical evidence and systematic studies, particularly the association of child marriage with the usage of Maternal Health Care Services (MHCS) in the South Asian women are few.What do the results of this study add? Findings of this study affirm that prevalence of child marriage in South Asian region is still common. Although every country has set out minimum age at first marriage by acts, the acts are poorly followed, which adversely impacts the usage of MHCS among married women.What are the implications of these findings for clinical practice and/or future research? The study findings provide important insights of the consequence of child marriage on the skilled MHCS utilisation. The marriage acts of the countries should be enacted properly to restrict child marriage not only for increasing MHCS, also for individual wellbeing and overall development of each country. Study has important implications for planners, organisation managers and policy makers.

FVC-NET: An Automated Diagnosis of Pulmonary Fibrosis Progression Prediction Using Honeycombing and Deep Learning.

Pulmonary fibrosis is a severe chronic lung disease that causes irreversible scarring in the tissues of the lungs, which results in the loss of lung capacity. The Forced Vital Capacity (FVC) of the patient is an interesting measure to investigate this disease to have the prognosis of the disease. This paper proposes a deep learning-based FVC-Net architecture to predict the progression of the disease from the patient's computed tomography (CT) scan and the patient's metadata. The input to the model combines the image score generated based on the degree of honeycombing for a patient identified based on segmented lung images and the metadata. This input is then fed to a 3-layer net to obtain the final output. The performance of the proposed FVC-Net model is compared with various contemporary state-of-the-art deep learning-based models, which are available on a cohort from the pulmonary fibrosis progression dataset. The model showcased significant improvement in the performance over other models for modified Laplace Log-Likelihood (-6.64). Finally, the paper concludes with some prospects to be explored in the proposed study.

Beneficial Effect of Low Dose Aspirin in Adult Patients with COVID-19: A Retrospective Observational Study in Bangladesh.

Presently Coronavirus disease 2019 is a huge human misery that is almost unstoppable with little remedy on board. The disease is predominantly associated with inflammation and thrombosis. Although aspirin exhibits excellent anti-thrombotic and anti-inflammatory properties, it is yet to be explored in treating Covid-19 patients. In this article we want to bring forth its beneficial effect in Covid-19 case management. It is a retrospective observational study. Adult Covid-19 patients either diagnosed with RT-PCR (reverse transcriptase polymerized chain reaction), or categorized as probable cases (as per World Health Organization case definition protocol) by medical doctors were enrolled as participants. Data were collected from doctors' office records for the period ranging from May 2020 to September 2020 in six districts of Bangladesh. Out of 44 participants 42 were eligible (2 children excluded) for the study. Among them 11 participants took low dose aspirin (75mg daily) during the disease process. All participants in aspirin group became cured without complication or death (RR 2.2, 95% CI 1.5-3.2, p<0.001). On the other hand, no aspirin group suffered complications and deaths. The study findings revealed that complications were significantly low among aspirin users. It documented that low dose aspirin is beneficial for Covid-19 patients. Further study is warranted with larger sample size.

The association between marital violence and reproductive and sexual health outcomes of women: A multi-country study of South Asia.

In this study, we examined the relationship between marital violence(MV) and reproductive and sexual health outcomes of women of Bangladesh, India, Nepal and Pakistan. The lifetime MV inflicted by husbands on their wives was highest at 53% in Bangladesh, India(37%), Pakistan(28%) and Nepal(28%) respectively. Overall, our results in this study suggest that MV perpetrated by husbands on their wife is significantly associated with unintended pregnancy, pregnancy termination, current use of any modern contraceptive methods and sexually transmitted infections, but inconsistent associations are appeared across countries. Lifetime sexual violence and physical-sexual force were significantly and positively associated with unintended pregnancy and pregnancy termination respectively, whereas, all types of MV were significantly and positively related with the symptom of sexually transmitted diseases in the women. Appropriate measures should be undertaken to combat Violence against women(VAW) and necessary services should be provided to the victimized women to uphold their reproductive and sexual rights.

Population structure, biogeography and transmissibility of Mycobacterium tuberculosis.

Mycobacterium tuberculosis is a clonal pathogen proposed to have co-evolved with its human host for millennia, yet our understanding of its genomic diversity and biogeography remains incomplete. Here we use a combination of phylogenetics and dimensionality reduction to reevaluate the population structure of M. tuberculosis, providing an in-depth analysis of the ancient Indo-Oceanic Lineage 1 and the modern Central Asian Lineage 3, and expanding our understanding of Lineages 2 and 4. We assess sub-lineages using genomic sequences from 4939 pan-susceptible strains, and find 30 new genetically distinct clades that we validate in a dataset of 4645 independent isolates. We find a consistent geographically restricted or unrestricted pattern for 20 groups, including three groups of Lineage 1. The distribution of terminal branch lengths across the M. tuberculosis phylogeny supports the hypothesis of a higher transmissibility of Lineages 2 and 4, in comparison with Lineages 3 and 1, on a global scale. We define an expanded barcode of 95 single nucleotide substitutions that allows rapid identification of 69 M. tuberculosis sub-lineages and 26 additional internal groups. Our results paint a higher resolution picture of the M. tuberculosis phylogeny and biogeography.

Individual- and community-level factors associated with underweight and overweight among women of reproductive age in Bangladesh: a multilevel analysis.

The co-existence of under- and overweight at population level around the globe is well documented. However, this has yet to be explored using suitable statistical techniques in the context of Bangladesh. This study aimed to examine the prevalence and risk factors for being underweight and overweight or obese compared with normal weight in ever-married non-pregnant women aged 15-49 years in Bangladesh using data from the most recent Bangladesh Demographic and Health Survey conducted in 2014. Multilevel multinomial logistic regression (MLMLR) and quantile regression models were fitted to examine the associations of socioeconomic and individual-, household- and community-level factors on the nutritional status of women as measured by BMI. Overall, the prevalences of underweight, normal weight, overweight and obese women were 19%, 58%, 19% and 4%, respectively, in 2014. The MLMLR analysis revealed that women of young age, widowed/divorced/separated, having a larger family size and children aged ≤5 years in the household, currently amenorrhoeic and members of non-government organizations were at significantly increased risk of being underweight; those of older age, having higher parity, more educated, frequently watched TV and non-poor were more likely to be overweight or obese relative to normal BMI. Women from more affluent communities and urban areas were more likely to be overweight or obese relative to normal BMI than their counterparts from less-affluent and rural communities. Women's nutritional status was found to be heterogeneous across the regions of the country. The findings indicate that, along with individual-level factors, community-level characteristics are also important in explaining women's BMI in Bangladesh. The issue of under- and overweight or obesity among women in Bangladesh requires the immediate adoption of a public health policy for its mitigation. When developing intervention programmes, important determinants and uniform development of regions should be taken into consideration to combat the dual burden of under- and overweight among women in Bangladesh.

Slow radiological improvement and persistent low-grade inflammation after chemotherapy in tuberculosis patients with type 2 diabetes.

BACKGROUND: Diabetes mellitus type 2 (DM) may impede immune responses in tuberculosis (TB) and thus contribute to enhanced disease severity. In this study, we aimed to evaluate DM-mediated alterations in clinical, radiological and immunological outcomes in TB disease. METHODS: Newly diagnosed pulmonary TB patients with or without DM (TB n = 40; TB-DM n = 40) were recruited in Dhaka, Bangladesh. Clinical symptoms, sputum smear and culture conversion as well as chest radiography were assessed. Peripheral blood and sputum samples were collected at the time of diagnosis (baseline) and after 1, 2 and 6 months of standard anti-TB treatment. Blood samples were also obtained from healthy controls (n = 20). mRNA expression of inflammatory markers in blood and sputum samples were quantified using real-time PCR. RESULTS: The majority of TB-DM patients had poor glycemic control (HbA1c > 8%) and displayed elevated pulmonary pathology (P = 0.039) particularly in the middle (P < 0.004) and lower lung zones (P < 0.02) throughout the treatment period. However, reduction of clinical symptoms and time to sputum smear and culture conversion did not differ between the groups. Transcripts levels of the pro-inflammatory cytokines IL-1ß (P = 0.003 at month-1 and P = 0.045 at month-2) and TNF-α (P = 0.005 at month-1) and the anti-inflammatory cytokine IL-10 (P = 0.005 at month-2) were higher in peripheral blood after anti-TB treatment in TB-DM compared to TB patients. Conversely in sputum, TB-DM patients had reduced CD4 (P < 0.009 at month-1) and IL-10 (P = 0.005 at month-1 and P = 0.006 at month-2) transcripts, whereas CD8 was elevated (P = 0.016 at month-2). At 1- and 2-month post-treatment, sputum IL-10 transcripts were inversely correlated with fasting blood glucose and HbA1c levels in all patients. CONCLUSION: Insufficient up-regulation of IL-10 in the lung may fuel persistent local inflammation thereby promoting lung pathology in TB-DM patients with poorly controlled DM.

Circulating strains of Mycobacterium tuberculosis: 24 loci MIRU-VNTR analysis in Bangladesh.

Bangladesh is among the high burden countries for tuberculosis (TB) and multidrug resistant TB (MDR-TB). As the genetic diversity and distinct phylogeographic distribution of Mycobacterium tuberculosis are responsible for regional differences in drug resistance, this cross sectional study was conducted to identify the circulating M. tuberculosis strains belonging to different lineages among pulmonary tuberculosis and, to investigate the contribution of distinct M. tuberculosis lineages to rifampicin resistant (RR) and rifampicin sensitive (RS) TB. A total of 40 RR and 20 RS isolates were enrolled in this study, all of which confirmed as M. tuberculosis by MPT 64 antigen detection. Furthermore, all isolates were genotyped by 24 loci Mycobacterial Interspersed Repetitive Units-Variable Number of Tandem Repeats (MIRU-VNTR), thus comprising the first study to employ this approach in Bangladesh. Beijing was the predominant lineage (26.8%) followed by EAI (23.2%), Delhi/CAS (16.1%), H37Rv (8.9%), Haarlem (7.1%), LAM (5.4%), Cameroon (3.6%) and a NEW-1 (1.8%). Four (7.1%) isolates remained as unidentified. Beijing strains were the significantly predominant (36.8%; p = 0.0135) among the RR isolates in comparison with other strains whereas EAI was the predominant (38.8%) lineage among RS isolates. Also, approximately 13% RR isolates showed genotypic resistance against fluoroquinolones by LPA and, hence, classed as pre-XDR TB albeit no specific lineage was found associated with these latter strains. A low transmission rate (10.5%) and high genetic diversity was detected in this setting with all the clustered strains herein identified belonging to the Beijing lineage. This study highlights 24 loci MIRU-VNTR analysis as a powerful tool for genotyping of Mycobacterium tuberculosis in this setting as it shows a high discriminatory index (0.81).

Characterization of Genomic Variants Associated with Resistance to Bedaquiline and Delamanid in Naive Mycobacterium tuberculosis Clinical Strains.

The role of mutations in genes associated with phenotypic resistance to bedaquiline (BDQ) and delamanid (DLM) in Mycobacterium tuberculosis complex (MTBc) strains is poorly characterized. A clear understanding of the genetic variants' role is crucial to guide the development of molecular-based drug susceptibility testing (DST). In this work, we analyzed all mutations in candidate genomic regions associated with BDQ- and DLM-resistant phenotypes using a whole-genome sequencing (WGS) data set from a collection of 4,795 MTBc clinical isolates from six countries with a high burden of tuberculosis (TB). From WGS analysis, we identified 61 and 163 unique mutations in genomic regions potentially involved in BDQ- and DLM-resistant phenotypes, respectively. Importantly, all strains were isolated from patients who likely have never been exposed to these medicines. To characterize the role of mutations, we calculated the free energy variation upon mutations in the available protein structures of Ddn (DLM), Fgd1 (DLM), and Rv0678 (BDQ) and performed MIC assays on a subset of MTBc strains carrying mutations to assess their phenotypic effect. The combination of structural and phenotypic data allowed for cataloguing the mutations clearly associated with resistance to BDQ (n = 4) and DLM (n = 35), only two of which were previously described, as well as about a hundred genetic variants without any correlation with resistance. Significantly, these results show that both BDQ and DLM resistance-related mutations are diverse and distributed across the entire region of each gene target, which is of critical importance for the development of comprehensive molecular diagnostic tools.

Prevalence and genetic profiles of isoniazid resistance in tuberculosis patients: A multicountry analysis of cross-sectional data.

BACKGROUND: The surveillance of drug resistance among tuberculosis (TB) patients is central to combatting the global TB epidemic and preventing the spread of antimicrobial resistance. Isoniazid and rifampicin are two of the most powerful first-line anti-TB medicines, and resistance to either of them increases the risk of treatment failure, relapse, or acquisition of resistance to other drugs. The global prevalence of rifampicin resistance is well documented, occurring in 3.4% (95% CI 2.5%-4.4%) of new TB patients and 18% (95% CI 7.6%-31%) of previously treated TB patients in 2018, whereas the prevalence of isoniazid resistance at global and regional levels is less understood. In 2018, the World Health Organization (WHO) recommended a modified 6-month treatment regimen for people with isoniazid-resistant, rifampicin-susceptible TB (Hr-TB), which includes rifampicin, pyrazinamide, ethambutol, and levofloxacin. We estimated the global prevalence of Hr-TB among TB patients and investigated associated phenotypic and genotypic drug resistance patterns. METHODS AND FINDINGS: Aggregated drug resistance data reported to WHO from either routine continuous surveillance or nationally representative periodic surveys of TB patients for the period 2003-2017 were reviewed. Isoniazid data were available from 156 countries or territories for 211,753 patients. Among these, the global prevalence of Hr-TB was 7.4% (95% CI 6.5%-8.4%) among new TB patients and 11.4% (95% CI 9.4%-13.4%) among previously treated TB patients. Additional data on pyrazinamide and levofloxacin resistance were available from 6 countries (Azerbaijan, Bangladesh, Belarus, Pakistan, the Philippines, and South Africa). There were no cases of resistance to both pyrazinamide and levofloxacin among Hr-TB patients, except for the Philippines (1.8%, 95% CI 0.2-6.4) and Belarus (5.3%, 95% CI 0.1-26.0). Sequencing data for all genomic regions involved in isoniazid resistance were available for 4,563 patients. Among the 1,174 isolates that were resistant by either phenotypic testing or sequencing, 78.6% (95% CI 76.1%-80.9%) had resistance-conferring mutations in the katG gene and 14.6% (95% CI 12.7%-16.8%) in both katG and the inhA promoter region. For 6.8% (95% CI 5.4%-8.4%) of patients, mutations occurred in the inhA promoter alone, for whom an increased dose of isoniazid may be considered. The main limitations of this study are that most analyses were performed at the national rather than individual patient level and that the quality of laboratory testing may vary between countries. CONCLUSIONS: In this study, the prevalence of Hr-TB among TB patients was higher than the prevalence of rifampicin resistance globally. Many patients with Hr-TB would be missed by current diagnostic algorithms driven by rifampicin testing, highlighting the need for new rapid molecular technologies to ensure access to appropriate treatment and care. The low prevalence of resistance to pyrazinamide and fluoroquinolones among patients with Hr-TB provides further justification for the recommended modified treatment regimen.

Sero-prevalence of toxoplasmosis among pregnant women attending an ante-natal clinic at a teaching hospital in Al Kharj, Saudi Arabia.

Toxoplasma gondii (T. gondii) is a zoonotic infection that may be transmitted to human beings either by consumption of raw or uncooked meat or by ingesting oocysts. Toxoplasma organisms can cross blood placenta barrier and may result in congenital toxoplasmosis. About 80% of immunocompetent individuals do not show any clinical manifestations and are silent carriers of this disease. Pregnant women especially in highly prevalent areas are recommended to be screened for this disease in order to prevent the potential vertical transmission. To our knowledge no such study has been conducted in this region of Saudi Arabia. This study attempted to carry out two objectives: first, to find out the seroprevalence of T. gondii infection in pregnant women attending prenatal care services in our hospital; second, to find out risk factors associated with T. gondii seroprevalence in our patients. It was carried out in Teaching Hospital in Al-Kharj over a period of one year. All 306 pregnant women attending antenatal clinic were involved in the study. A pretested selfexplanatory questionnaire was filled out by the patients and their sera were collected to be tested for IgG and/or IgM against T. gondii. The results were then statistically analyzed using SPSS software and p-value was calculated using Pearson Chi Square test. Out of the 306 blood samples tested, 99 (32.4%) were seropositive for specific anti T. gondii IgG antibodies and 3(1%) were seropositive for IgM. This show that seroprevalence of T. gondii antibodies was high among pregnant women and the prevalence showed a significant association with age. The study recommends conducting educational programs to raise awareness among women about risk factors and precautions to be taken.

Sero-prevalence of toxoplasmosis among pregnant women attending an ante-natal clinic at a teaching hospital in Al Kharj, Saudi Arabia

@#Toxoplasma gondii (T. gondii) is a zoonotic infection that may be transmitted to human beings either by consumption of raw or uncooked meat or by ingesting oocysts. Toxoplasma organisms can cross blood placenta barrier and may result in congenital toxoplasmosis. About 80% of immunocompetent individuals do not show any clinical manifestations and are silent carriers of this disease. Pregnant women especially in highly prevalent areas are recommended to be screened for this disease in order to prevent the potential vertical transmission. To our knowledge no such study has been conducted in this region of Saudi Arabia. This study attempted to carry out two objectives: first, to find out the seroprevalence of T. gondii infection in pregnant women attending prenatal care services in our hospital; second, to find out risk factors associated with T. gondii seroprevalence in our patients. It was carried out in Teaching Hospital in Al-Kharj over a period of one year. All 306 pregnant women attending antenatal clinic were involved in the study. A pretested selfexplanatory questionnaire was filled out by the patients and their sera were collected to be tested for IgG and/or IgM against T. gondii. The results were then statistically analyzed using SPSS software and p-value was calculated using Pearson Chi Square test. Out of the 306 blood samples tested, 99 (32.4%) were seropositive for specific anti T. gondii IgG antibodies and 3(1%) were seropositive for IgM. This show that seroprevalence of T. gondii antibodies was high among pregnant women and the prevalence showed a significant association with age. The study recommends conducting educational programs to raise awareness among women about risk factors and precautions to be taken.

Fatal Infection Caused by Chromobacterium violaceum: A Case Report from a Tertiary Care Hospital in Bangladesh.

Chromobacterium violaceum is a Gram-negative bacterium, found in tropical and subtropical regions. C. violaceum infection rarely occurs, but once occurs, it is associated with significant mortality due to severe systemic infection. Since the first human case from Malaysia in 1927, >150 cases of C. violaceum infection have been reported worldwide. We have described here a fatal case of C. violaceum infection in a tertiary care hospital in Dhaka, Bangladesh. To the best of our knowledge, this is the first case of C. violaceum infection in Bangladesh.

Immune responses in the treatment of drug-sensitive pulmonary tuberculosis with phenylbutyrate and vitamin D3 as host directed therapy.

BACKGROUND: We have previously shown that 8 weeks' treatment with phenylbutyrate (PBA) (500mgx2/day) with or without vitamin D3 (vitD3) (5000 IU/day) as host-directed therapy (HDT) accelerated clinical recovery, sputum culture conversion and increased expression of cathelicidin LL-37 by immune cells in a randomized, placebo-controlled trial in adults with pulmonary tuberculosis (TB). In this study we further aimed to examine whether HDT with PBA and vitD3 promoted clinically beneficial immunomodulation to improve treatment outcomes in TB patients. METHODS: Cytokine concentration was measured in supernatants of peripheral blood mononuclear cells (PBMC) from patients (n = 31/group). Endoplasmic reticulum stress-related genes (GADD34 and XBP1spl) and human beta-defensin-1 (HBD1) gene expression were studied in monocyte-derived-macrophages (MDM) (n = 18/group) from PBMC of patients. Autophagy in MDM (n = 6/group) was evaluated using LC3 expression by confocal microscopy. RESULTS: A significant decline in the concentration of cytokines/chemokines was noted from week 0 to 8 in the PBA-group [TNF-α (ß = - 0.34, 95% CI = - 0.68, - 0.003; p = 0.04), CCL11 (ß = - 0.19, 95% CI = - 0.36, - 0.03; p = 0.02) and CCL5 (ß = - 0.08, 95% CI = - 0.16, 0.002; p = 0.05)] and vitD3-group [(CCL11 (ß = - 0.17, 95% CI = - 0.34, - 0.001; p = 0.04), CXCL10 (ß = - 0.38, 95% CI = - 0.77, 0.003; p = 0.05) and PDGF-ß (ß = - 0.16, 95% CI = - 0.31, 0.002; p = 0.05)] compared to placebo. Both PBA- and vitD3-groups showed a decline in XBP1spl mRNA on week 8 (p < 0.03). All treatment groups demonstrated increased LC3 expression in MDM compared to placebo over time (p < 0.037). CONCLUSION: The use of PBA and vitD3 as adjunct therapy to standard TB treatment promoted favorable immunomodulation to improve treatment outcomes. TRIALS REGISTRATION: This trial was retrospectively registered in clinicaltrials.gov, under identifier NCT01580007 .

Comparison of nebulised dexmedetomidine, ketamine, or midazolam for premedication in preschool children undergoing bone marrow biopsy.

BACKGROUND: The aim of our study was to compare the efficacy of dexmedetomidine, ketamine, and midazolam for sedative premedication administered by nebuliser 30 min before general anaesthesia in preschool children undergoing bone marrow biopsy and aspiration. METHODS: Ninety children aged 3-7 yr were randomly allocated into three equal groups to be premedicated with either nebulised ketamine 2 mg kg-1 (Group K), dexmedetomidine 2 µg kg-1 (Group D), or midazolam 0.2 mg kg-1 (Group M). The primary endpoint was a five-point sedation score on arrival in the operating room 30 min after end of study drug administration. Secondary outcomes included: parental separation anxiety scale; medication and mask acceptance scales; haemodynamic variables; recovery time; postoperative face, legs, activity, cry, and consolability scale; emergence agitation scale; and adverse effects. RESULTS: The median (range) sedation score on arrival in the operating room was 3.5 (1-4), 2.0 (2-3) and 2.0 (1-3) in Groups M, D, and K, respectively (P=0.000). Subjects in Group D showed higher medication (P<0.03) and mask acceptance scores (P<0.015) and more satisfactory parental separation anxiety scale (P<0.044). The median (range) recovery time was significantly shorter in Group D [5.5 (4-8) min] compared with Group K [10.0 (5-15) min, P=0.000] and M [8.0 (6-15) min, P=0.000]. The incidence of emergence agitation was lower in Group D (P<0.008). CONCLUSIONS: Preschool children premedicated with nebulised dexmedetomidine had more satisfactory sedation, shorter recovery time, and less postoperative agitation than those who received nebulised ketamine or midazolam. CLINICAL TRIAL REGISTRATION: NCT02935959.

Genetic sequencing for surveillance of drug resistance in tuberculosis in highly endemic countries: a multi-country population-based surveillance study.

BACKGROUND: In many countries, regular monitoring of the emergence of resistance to anti-tuberculosis drugs is hampered by the limitations of phenotypic testing for drug susceptibility. We therefore evaluated the use of genetic sequencing for surveillance of drug resistance in tuberculosis. METHODS: Population-level surveys were done in hospitals and clinics in seven countries (Azerbaijan, Bangladesh, Belarus, Pakistan, Philippines, South Africa, and Ukraine) to evaluate the use of genetic sequencing to estimate the resistance of Mycobacterium tuberculosis isolates to rifampicin, isoniazid, ofloxacin, moxifloxacin, pyrazinamide, kanamycin, amikacin, and capreomycin. For each drug, we assessed the accuracy of genetic sequencing by a comparison of the adjusted prevalence of resistance, measured by genetic sequencing, with the true prevalence of resistance, determined by phenotypic testing. FINDINGS: Isolates were taken from 7094 patients with tuberculosis who were enrolled in the study between November, 2009, and May, 2014. In all tuberculosis cases, the overall pooled sensitivity values for predicting resistance by genetic sequencing were 91% (95% CI 87-94) for rpoB (rifampicin resistance), 86% (74-93) for katG, inhA, and fabG promoter combined (isoniazid resistance), 54% (39-68) for pncA (pyrazinamide resistance), 85% (77-91) for gyrA and gyrB combined (ofloxacin resistance), and 88% (81-92) for gyrA and gyrB combined (moxifloxacin resistance). For nearly all drugs and in most settings, there was a large overlap in the estimated prevalence of drug resistance by genetic sequencing and the estimated prevalence by phenotypic testing. INTERPRETATION: Genetic sequencing can be a valuable tool for surveillance of drug resistance, providing new opportunities to monitor drug resistance in tuberculosis in resource-poor countries. Before its widespread adoption for surveillance purposes, there is a need to standardise DNA extraction methods, recording and reporting nomenclature, and data interpretation. FUNDING: Bill & Melinda Gates Foundation, United States Agency for International Development, Global Alliance for Tuberculosis Drug Development.

Comparative yielding of BACTEC MGIT 960 and GeneXpert MTB/RIF Assay for Rapid Diagnosis of Drug Resistance Tuberculosis from Sputum Specimen.

Among communicable disease, tuberculosis is the second leading cause of death worldwide, killing nearly two million people each year. Several diagnostic techniques are currently used to detect mycobacteria in respiratory specimens. Therefore an alternative, rapid and most effective method is required for diagnosis of pulmonary tuberculosis which is relatively more sensitive and specific. A total of 107 sputum samples of suspected TB patients were enrolled attending the National Tuberculosis Reference Laboratory (NTRL), Mohakhali, Dhaka, Bangladesh from July 2014 to July 2015. Liquid culture of 107 sputum samples, yielded 76.63% culture positive and 1.86% contamination was observed. Antibiotic susceptibility test was done for 82 culture positive sputum samples among which 28.04% cases were resistant to both Rifampicin and Isoniazid. GeneXpert assay detected 85.04% M. tuberculosis among which 25.23% were found to be Rifampicin Resistance.