Background Artificial intelligence (AI) algorithms can be trained to recognise tuberculosis-related abnormalities on chest radiographs. Various AI algorithms are available commercially, yet there is little impartial evidence on how their performance compares with each other and with radiologists. We aimed to evaluate five commercial AI algorithms for triaging tuberculosis using a large dataset that had not previously been used to train any AI algorithms.Methods Individuals aged 15 years or older presenting or referred to three tuberculosis screening centres in Dhaka, Bangladesh, between May 15, 2014, and Oct 4, 2016, were recruited consecutively. Every participant was verbally screened for symptoms and received a digital posterior-anterior chest x-ray and an Xpert MTB/RIF (Xpert) test. All chest x-rays were read independently by a group of three registered radiologists and five commercial AI algorithms: CAD4TB (version 7), InferRead DR (version 2), Lunit INSIGHT CXR (version 4.9.0), JF CXR-1 (version 2), and qXR (version 3). We compared the performance of the AI algorithms with each other, with the radiologists, and with the WHO's Target Product Profile (TPP) of triage tests (≥90% sensitivity and ≥70% specificity). We used a new evaluation framework that simultaneously evaluates sensitivity, proportion of Xpert tests avoided, and number needed to test to inform implementers' choice of software and selection of threshold abnormality scores. Findings Chest x-rays from 23 954 individuals were included in the analysis. All five AI algorithms significantly outperformed the radiologists. The areas under the receiver operating characteristic curve were 90•81% (95% CI 90•33-91•29) for qXR, 90•34% (89•81-90•87) for CAD4TB, 88•61% (88•03-89•20) for Lunit INSIGHT CXR, 84•90% (84•27-85•54) for InferRead DR, and 84•89% (84•26-85•53) for JF CXR-1. Only qXR (74•3% specificity [95% CI 73•3-74•9]) and CAD4TB (72•9% specificity [72•3-73•5]) met the TPP at 90% sensitivity. All five AI algorithms reduced the number of Xpert tests required by 50% while maintaining a sensitivity above 90%. All AI algorithms performed worse among older age groups (>60 years) and people with a history of tuberculosis.Interpretation AI algorithms can be highly accurate and useful triage tools for tuberculosis detection in high-burden regions, and outperform human readers.Funding Government of Canada.

BackgroundGeneXpert MTB/RIF (Xpert) and Genotype MTBDRplus (DRplus) are two World Health Organization (WHO) endorsed probe based molecular drug susceptibility testing (DST) methods for rapid diagnosis of drug resistant tuberculosis. Both methods target the same 81 bp Rifampicin Resistance Determining Region (RRDR) of bacterial RNA polymerase β subunit (rpoB) for detection of Rifampicin (RIF) resistance associated mutations using DNA probes. So there is a correspondence of the probes of each other and expected similarity of probe binding.MethodsWe analyzed 92 sputum specimens by Xpert, DRplus and LJ proportion method (LJ-DST). We compared molecular DSTs with gold standard LJ-DST. We wanted to see the agreement level of two molecular methods for detection of RIF resistance associated mutations. The 81bp RRDR region of rpoB gene of discrepant cases between the two molecular methods was sequenced by Sanger sequencing.ResultsThe agreement of Xpert and DRplus with LJ-DST for detection of RIF susceptibility was found to be 93.5% and 92.4%, respectively. We also found 92.4% overall agreement of two molecular methods for the detection of RIF susceptibility. A total of 84 out of 92 samples (91.3%) had agreement on the molecular locus of RRDR mutation by DRplus and Xpert. Sanger sequencing of 81bp RRDR revealed that Xpert probes detected seven of eight discrepant cases correctly and DRplus was erroneous in all the eight cases.ConclusionAlthough the overall concordance with LJ-DST was similar for both Xpert and DRplus assay, Xpert demonstrated more accuracy in the detection of RIF susceptibility for discrepant isolates compared with DRplus. This observation would be helpful for the improvement of probe based detection of drug resistance associated mutations especially rpoB mutation in M. tuberculosis.

bGiven the increases in drug-resistant tuberculosis, laboratory capacities for drug susceptibility testing are being scaled up worldwide. A laboratory must decide among several endorsed methodologies. We evaluated 87 Mycobacterium tuberculosis isolates for concordance of susceptibility results across six methods: the L-J proportion method, MGIT 960 SIRE AST, Gene/Xpert MTB/ RIF, GenoType MTBDRplus line probe assay, MycoTB MIC plate, and a laboratory-developed mycobacteriophage quantitative PCR (qPCR)-based method. Most (80%) isolates were multidrug resistant. Of the culture-based methods, the mycobacteriophage qPCR method was fastest, the L-J proportion method was the slowest, and the MGIT method required the most repeat testing (P < 0.05). For isoniazid (INH), 82% of isolates were susceptible by all methods or resistant by all methods, whereas for rifampin (RIF), ethambutol (EMB), and streptomycin (STR), such complete concordance was observed in 77%, 50%, and 51% of isolates, respectively (P < 0. G iven the increasing rates of multidrug-resistant tuberculosis (MDR-TB) isolates worldwide and the emergence of extensively drug-resistant TB, the development of rapid and accurate methods for drug susceptibility testing (DST) of Mycobacterium tuberculosis isolates is a global priority. The culture-based proportion method that employs Löwenstein-Jensen medium is a World Health Organization (WHO)-recommended method that has been in wide use for over 50 years (1, 2). Such solid medium-based DST methods are slow, requiring readings at 4 to 6 weeks, which delays the detection of drug resistance and risks inappropriate treatment and spread of drug-resistant strains. This deficiency has led to the development of newer DST methodologies, including liquid culture systems and molecular line probe assays, which have also received recommendations from WHO (3, 4).As a consequence, laboratories have seen an accumulation of methods from which they must choose, and a given specimen or isolate may be tested across a variety of methods. A natural consequence is that discrepancies between methods may be encountered. Such discrepancies may be of little consequence in certain scenarios, such as streptomycin (STR) resistance in settings that use primarily isoniazid (INH), rifampin (RIF), pyrazinamide (PZA), and ethambutol (EMB), but they may have critical implications for treating MDR-TB in areas where the arsenal of drugs is limited in number and potency. Discordance is becoming a vexing aspect for TB clinicians and will likely increase in frequency as new methodologies are adopted, yet its extent has received little attention. Most diagnostic evaluations examine one new method against one gold standard reference method, not several methods against each other. Additionally, most diagnostic evaluations are performed on predominantly drug-susceptible isolates, often highly susceptible clinical isolates or reference strains, and thus discrepancies between methods would be expected to be rare.For this reason, we prospectively examined 87 mos...

Coronavirus disease 2019 (COVID-19), an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been declared a global pandemic by the World Health Organization, and the situation worsens daily, associated with acute increases in case fatality rates. The main protease (Mpro) enzyme produced by SARS-CoV-2 was recently demonstrated to be responsible for not only viral reproduction but also impeding host immune responses. The element selenium (Se) plays a vital role in immune functions, both directly and indirectly. Thus, we hypothesised that Se-containing heterocyclic compounds might curb the activity of SARS-CoV-2 Mpro. We performed a molecular docking analysis and found that several of the selected selenocompounds showed potential binding affinities for SARS-CoV-2 Mpro, especially ethaselen (49), which exhibited a docking score of −6.7 kcal/mol compared with the −6.5 kcal/mol score for GC376 (positive control). Drug-likeness calculations suggested that these compounds are biologically active and possess the characteristics of ideal drug candidates. Based on the binding affinity and drug-likeness results, we selected the 16 most effective selenocompounds as potential anti-COVID-19 drug candidates. We also validated the structural integrity and stability of the drug candidate through molecular dynamics simulation. Using further in vitro and in vivo experiments, we believe that the targeted compound identified in this study (ethaselen) could pave the way for the development of prospective drugs to combat SARS-CoV-2 infections and trigger specific host immune responses.

This research describes an investigation of the antipyretic and hepatoprotectiveproperties of both a crude organic extract and various subfractions of the ethnomedicinal plant Tinospora crispa, using appropriate animal models. In an attempt to identify potential lead hepatoprotective compounds, in silico experiments were utilized.Antipyretic activity was assessed via the Brewer's yeast-induced pyrexia method, while hepatoprotective effects were evaluated in a carbon tetrachloride (CCl 4 )induced animal model. A computer-aided prediction of activity spectra for substances (PASS) model was applied to a selection of documented phytoconstituents, with the aim of identifying those compounds with most promising hepatoprotective effects.Results were analyzed using Molinspiration software. Our results showed that both the methanol extract (METC) and various subfractions (pet ether, PEFTC; n-hexane, NHFTC; and chloroform, CFTC) significantly (p < .05) reduced pyrexia in a dose-dependent manner. In CCl 4 -induced hepatotoxicity studies, METC ameliorated elevated hepatic markers including serum alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP), and total bilirubin. Malondialdehyde (MDA) levels were significantly reduced, while superoxide dismutase (SOD) levels were significantly increased. Among a selection of metabolites of T. crispa, genkwanin was found to be the most potent hepatoprotective constituent using PASS predictive models. These results demonstrate that both the methanolic extract of T. crispa and those fractions containing genkwanin may offer promise in reducing pyrexia and as a source of potential hepatoprotective agents. K E Y W O R D S antipyretic, CCl 4 -induced hepatotoxicity, hepatoprotective, in silico studies, PASS prediction, Tinospora crispa, yeast-induced pyrexia How to cite this article: Rakib A, Ahmed S, Islam MA, et al. Antipyretic and hepatoprotective potential of Tinospora crispa and investigation of possible lead compounds through in silico approaches. Food Sci Nutr. 2020;8:547-556. https ://doi.

BackgroundSputum smear microscopy is fast and inexpensive technique for detecting tuberculosis (TB) in high incidence areas but has low sensitivity. Physical and chemical sputum processing along with centrifugation have been found to show promise in overcoming this limitation. Our objective was to compare the sensitivity of smear microscopy obtained with smears made directly from respiratory specimens to those from concentrated specimens.MethodsBy active screening, 915 TB suspects were identified from Dhaka Central Jail and sputum specimens were aseptically collected. Direct smears were prepared by taking a small portion of the purulent part of the sputum with a sterile loop. The specimens were then processed by a standard N-acetyl-L-cysteine-NaOH digestion-decontamination method to prepare concentrated specimens. Both smears were then air dried, heat fixed, and stained by the Ziehl-Neelsen staining technique. The stained slides were examined under oil immersion and were graded following International Union Against Tuberculosis and Lung Diseases guidelines. All the specimens were inoculated into Lowenstein-Jensen (L-J) media and culture results were considered as gold standard to calculate sensitivity.ResultsOf 915 specimens, 73 (8%) specimens were positive both on direct and concentrated methods, one sample was positive on direct microscopy but was negative on concentrated method. An extra 14 (1.5%) samples were positive on concentrated method which were negative on direct smear. In L-J media 105 specimens were found positive for TB bacilli and of them, 74 (70.5%) and 87 (82.9%) were positive in direct and concentrated smear, respectively. The sensitivity of direct and concentrated smear microscopy was different when using positive culture as the gold standard (71% vs. 83%).ConclusionsThe results showed that concentrated technique increases the sensitivity of microscopy up to 12%. Therefore, the national programs in high TB burden countries may consider incorporating the technique into their guidelines at least in the district and higher level laboratories to improve case finding strategy.

With an increasing fatality rate, severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) has emerged as a promising threat to human health worldwide. Recently, the World Health Organization (WHO) has announced the infectious disease caused by SARS-CoV-2, which is known as coronavirus disease-2019 (COVID-2019), as a global pandemic. Additionally, the positive cases are still following an upward trend worldwide and as a corollary, there is a need for a potential vaccine to impede the progression of the disease. Lately, it has been documented that the nucleocapsid (N) protein of SARS-CoV-2 is responsible for viral replication and interferes with host immune responses. We comparatively analyzed the sequences of N protein of SARS-CoV-2 for the identification of core attributes and analyzed the ancestry through phylogenetic analysis. Subsequently, we predicted the most immunogenic epitope for the T-cell and B-cell. Importantly, our investigation mainly focused on major histocompatibility complex (MHC) class I potential peptides and NTASWFTAL interacted with most human leukocyte antigen (HLA) that are encoded by MHC class I molecules. Further, molecular docking analysis unveiled that NTASWFTAL possessed a greater affinity towards HLA and also available in a greater range of the population. Our study provides a consolidated base for vaccine design and we hope that this computational analysis will pave the way for designing novel vaccine candidates.

Background: Rifampicin resistance (RR) is a key indicator of multidrug-resistant tuberculosis (MDR-TB) and 95% of the RR is associated with the mutation in the 81-bp rifampicin resistance determining region (RRDR) of the rpoB gene of Mycobacterium tuberculosis complex (MTBC). The Xpert MTB/RIF (Xpert) assay uses five overlapping molecular beacon probes (A-E) complementary to RRDR region that detect MTBC and mutations associated with RR. The objective of the study was to investigate the distribution and frequency of mutations detected by Xpert assay among Beijing and non-Beijing RR-TB isolates. Methods: A total of 205 randomly selected RR-TB specimens detected by Xpert assay were included in this study. A portion of specimens was further subjected to culture, MTBDRplus test and the positive culture isolates were genotyped by spoligotyping. Results: We found that the most frequent mutation occurred at probe E (S531L) binding region in both Beijing and non-Beijing isolates (61.9% and 66.9%, respectively). The Beijing family had higher mutation rates than non-Beijing (19.0% vs 12.4%) at probe B (D516V) while the non-Beijing family had higher mutations at probe D (H526D or H526Y) than the Beijing (13.2% vs 10.7%) family. Mutations at probes Aand C were less common in both Beijing and non-Beijing isolates. There was no significant difference (P=0.36) in the occurrence of mutations at different probes between Beijing and non-Beijing isolates. Conclusions: The study results revealed that the most frequent mutation occurs in the region of probe E and the least common mutations at probe A and C among both Beijing and non-Beijing RR-TB cases. This first insight into the probe mutation variation and frequencies among the RR-TB cases in Bangladesh forms the baseline information for further investigation.