(<em>b</em>)SETTING:</<em>b</em>) Tertiary level specialised tu<em>b</em>erculosis (TB) hospital.(<em>b</em>)OBJECTIVE:</<em>b</em>) To determine the prevalence of antimicro<em>b</em>ial resistance in new extra-pulmonary tu<em>b</em>erculosis (EPTB) cases.(<em>b</em>)DESIGN:</<em>b</em>) Prospective cross-sectional study. Presumptive EPTB patients with enlarged lymph nodes or pleural effusion having no history of TB treatment were enrolled. Specimens were tested for smear, Xpert^® MTB/RIF and culture. Indirect drug suscepti<em>b</em>ility testing (DST) was performed using MGIT 960 and line-pro<em>b</em>e assays (LPA).(<em>b</em>)RESULTS:</<em>b</em>) Among 671 cases, 255 were <em>b</em>acteriologically confirmed and 185 DSTs were performed. Multidrug resistance (MDR-TB) was reported in 2.2% (95%CI 0.6-5.4), any resistance to rifampicin (RMP) in 2.7% (95%CI 0.9-6.2), isoniazid (<em>INH</em>) in 7.6% (95%CI 4.1-12.4), etham<em>b</em>utol in 1.1% (95%CI 0.1-3.9), pyrazinamide in 2.2% (95%CI 0.9-5.5) and fluoroquinolones (FQ) in 6.0% (95%CI 3.0-10.4). The sensitivity and specificity of LPA-DST was 100% and >98.8% respectively for RMP, <em>INH</em> and FQ. Among 82 cases with RMP of the results availa<em>b</em>le for all three methods used, five were reported to <em>b</em>e resistant on Xpert <em>b</em>ut all five were suscepti<em>b</em>le on MGIT 960 and four on MTBDR<i>plus</i>.(<em>b</em>)CONCLUSION:</<em>b</em>) Prevalence of RMP resistance in new EPTB cases is 2.7% (95%CI 0.9-6.2). Caution is warranted for RMP resistance detected using Xpert in EPTB samples with a very low <em>b</em>acterial load.

Introduction: Despite the moderate incidence of tuberculosis (TB) in many parts of Iran, Golestan province had a permanently higher TB incidence rate than the national average. Moreover, Golestan province receives immigrants, mainly from TB-endemic areas of Iran and neighbor countries. Here, we aimed to characterize the circulating Mycobacterium tuberculosis complex (MTBC) isolates in terms of the spoligotype and drug resistance patterns, across Golestan province.

Materials and methods: A set of 166 MTBC isolates was collected during July 2014 to July 2015 and subjected to drug susceptibility testing for first- and second-line anti-TB drugs and spoligotyping.

Results: Of 166 MTBC isolates, 139 (83.7%) isolates were assigned to 28 spoligotype international types (SITs). The most frequent SITs were SIT127/Ural-2 (n=25, 15.1%), followed by SIT1/Beijing (n=21, 12.7%) and SIT3427/Ural-2 (n=18, 10.8%). The set of 18 isolates (10.8%) showed resistance to at least one drug, which mainly belonged to SIT1/Beijing (n=7, 38.9%), orphan patterns (n=4, 22.2%) and SIT357/CAS1-Delhi (n=3, 16.7%). In addition, four isolates (2.4%) were resistant to pyrazinamide. The analysis of mutation corresponded to resistance to rifampin and isoniazid showed that two isolates had Ser531Leu substitution in rpoB, four isolates had Ser315Thr substitution in katG and one isolate had [C(-15)T] in inhA locus.

Conclusion: High diversity in spoligotypes of the MTBC isolates and lack of dominant genotype might be due to residence of immigrants in this region and consequent reactivation of latent infection. In addition, due to the presence of extensively drug-resistant (XDR) isolates in Golestan province, it is important to conduct future studies to determine transmission pattern of drug-resistant isolates in this region.

Keywords: Mycobacterium tuberculosis; genotyping; spoligotyping; transmission; tuberculosis.

Few medically-approved excipients are available for formulation strategies to endow microcarriers with improved performance in lung drug targeting. Konjac glucomannan (KGM) is a novel, biocompatible material, comprising mannose units potentially inducing macrophage uptake for the treatment of macrophage-mediated diseases. This work investigated spray-dried KGM microparticles as inhalable carriers of model antitubercular drugs, isoniazid (INH) and rifabutin (RFB). The polymer was characterised and different polymer/drug ratios tested in the production of microparticles for which respirability was assessed in vitro. The swelling of KGM microparticles and release of drugs in simulated lung fluid were characterised and the biodegradability in presence of β-mannosidase, a lung hydrolase, determined. KGM microparticles were drug loaded with 66% - 91% association efficiency and had aerodynamic diameter around 3 µm, which enables deep lung penetration. The microparticles swelled upon liquid contact by 40% - 50% but underwent size reduction (> 62% in 90 min) in presence of β-mannosidase, indicating biodegradability. Finally, drug release was tested showing slower release of RFB compared with INH but complete release of both within 24 h. This work identifies KGM as a biodegradable polymer of natural origin that can be engineered to encapsulate and release drugs in respirable microparticles with physical and chemical macrophage-targeting properties.

Keywords: inhalation; konjac glucomannan; microparticles; pulmonary drug delivery.

Tu<em>b</em>erculosis remains a leading cause of mortality among infectious diseases worldwide, prompting the need to discover new drugs to fight this disease. We report herein, the design, synthesis and anti-myco<em>b</em>acterial activity of isatin-mono/<em>b</em>is-isoniazid hy<em>b</em>rids. Most of the compounds exhi<em>b</em>ited very high activity against <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> with minimal inhi<em>b</em>itory concentrations in the range of 0.195-0.39 μg/mL and exerted a more potent <em>b</em>actericidal effect than the standard anti-tu<em>b</em>ercular drug isoniazid (<em>INH</em>). Importantly, these compounds were found to <em>b</em>e well tolerated at high doses (>200 μg/mL) on Vero kidney cells, leading to high selectivity indices. Two of the most promising hy<em>b</em>rids were evaluated for activity in THP-1 macrophages infected with <i>M. tu<em>b</em>erculosis</i>, among which (<em>b</em>)11e</<em>b</em>) was found to <em>b</em>e slightly more effective than <em>INH</em>. Overexpression of InhA along with cross-resistance determination of the most potent compounds, selection of resistant mutants and <em>b</em>iochemical analysis allowed us to decipher their mode of action. These compounds effectively inhi<em>b</em>ited mycolic acid <em>b</em>iosynthesis and required KatG to exert their <em>b</em>iological effects. Collectively, this suggests that the synthesized isatin-<em>INH</em> hy<em>b</em>rids are promising anti-tu<em>b</em>ercular molecules for further evaluation in pre-clinical settings.

Acute metabolic acidosis associated with accumulation of lactate has been previously reported in isoniazid (INH) intoxication. To our knowledge, association of INH toxicity with beta-hydroxybutyric acidosis has not been demonstrated previously. The present report documents the occurrence of beta-hydroxybutyric acidosis in patients with INH intoxication. The reason for the lack of previous reports of this association is not clear, although failure to measure plasma beta-hydroxybutyrate levels in previous studies is a likely possibility. Our patients received intravenous sodium bicarbonate, anticonvulsants and dialysis which resulted in complete reversal of metabolic acidosis and other manifestations of INH toxicity.

Herbal tea of Lagerstroemia speciosa Pers., commonly known as banaba, has been traditionally used to treat various ailments including diabetes and obesity due to its antioxidant and anti-inflammatory efficacies. Drug-induced liver injury is a common cause of acute liver failure. Isoniazid (INH) is used as the first-line treatment for tuberculosis; clinical and experimental studies have reported an abnormal liver function after INH therapy. Dapsone (DDS) is used for leprosy and other infections. This study investigates the hepatoprotective effect of ethanolic banaba leaves extract (EBLE) against simultaneously administered INH- and DDS-induced hepatotoxicity in rats. DDS (30 mg/kg, i.p.) and INH (50 mg/kg. p.o.) were administered simultaneously for 30 days. In separate groups, rats were posttreated orally with EBLE (500 mg/kg) and silymarin (100 mg/kg) for 30 days after INH + DDS administration. The marker enzymes of hepatotoxicity, oxidative stress markers, inflammatory markers, and histopathology were done. Simultaneous administration of INH- and DDS-induced significant elevation of marker enzymes of hepatotoxicity in the serum. This treatment also increased lipid peroxidation and pro-inflammatory markers (tumor necrosis factor alpha, transforming growth factor beta, and nuclear factor kappa B) expressions and decreased intracellular antioxidants such as superoxide dismutase, catalase, and glutathione in the liver tissue. All these abnormalities were significantly mitigated after EBLE and SIL posttreatments. The results of this study suggest that EBLE and silymarin can be protective against INH + DDS-induced hepatotoxicity. PRACTICAL APPLICATIONS: Herbal tea contain Lagerstroemia speciosa leaves are used in several Southeast Asian countries due to its rich antioxidant and inflammatory properties. This study showed the hepatoprotective efficacy of L. speciosa ethanolic extract against simultaneously administered dapsone- and isoniazid-induced hepatotoxicity in rats. L. speciosa administration was found to decrease dapsone- and isoniazid-induced oxidative stress and hepatic inflammation. L. speciosa herbal tea can reduce drug-induced hepatic complications as it contains phytochemicals such as corosolic acid, gallic acid, ellagic acid and berberine and are implicated for its hepatoprotective effect. Therefore, L. speciosa extract can be used for drug-induced liver injury.

Keywords: Isoniazid; banaba; corosolic acid; dapsone; drug-induced liver injury.

Orthodontic tooth movement (OTM) is a process depending on the remodeling of periodontal tissues surrounding the roots. Orthodontic forces trigger the conversion of mechanical stimuli into intercellular chemical signals within periodontal ligament (PDL) cells, activating alveolar bone remodeling, and thereby, initiating OTM. Recently, the mechanosensitive ion channel Piezo1 has been found to play pivotal roles in the different types of human cells by transforming external physical stimuli into intercellular chemical signals. However, the function of Piezo1 during the mechanotransduction process of PDL cells has rarely been reported. Herein, we established a rat OTM model to study the potential role of Piezo1 during the mechanotransduction process of PDL cells and investigate its effects on the tension side of alveolar bone remodeling. A total of 60 male Sprague-Dawley rats were randomly assigned into three groups: the OTM + inhibitor (INH) group, the OTM group, and the control (CON) group. Nickel-titanium orthodontic springs were applied to trigger tooth movement. Mice were sacrificed on days 0, 3, 7, and 14 after orthodontic movement for the radiographic, histological, immunohistochemical, and molecular biological analyses. Our results revealed that the Piezo1 channel was activated by orthodontic force and mainly expressed in the PDL cells during the whole tooth movement period. The activation of the Piezo1 channel was essential for maintaining the rate of orthodontic tooth movement and facilitation of new alveolar bone formation on the tension side. Reduced osteogenesis-associated transcription factors such as Runt-related transcription factor 2 (RUNX2), Osterix (OSX), and receptor activator of nuclear factor-kappa B ligand (RANKL)/osteoprotegerin (OPG) ratio were examined when the function of Piezo1 was inhibited. In summary, Piezo1 plays a critical role in mediating both the osteogenesis and osteoclastic activities on the tension side during OTM.

Keywords: Piezo1; alveolar bone; bone remodeling; mechanotransduction; orthodontic tooth movement.

(<em>b</em>)Background:</<em>b</em>) Tu<em>b</em>erculosis, mainly caused <em>b</em>y Myco<em>b</em>acterium tu<em>b</em>erculosis (Mt<em>b</em>), is an ancient human disease that gravely affects millions of people annually. We wanted to explore the genetic diversity and lineage-specific association of Mt<em>b</em> with drug resistance among pulmonary tu<em>b</em>erculosis patients. (<em>b</em>)Methods:</<em>b</em>) Sputum samples were collected from pulmonary tu<em>b</em>erculosis patients at six different healthcare institutions in Tigray, Ethiopia, <em>b</em>etween July 2018 and August 2019. DNA was extracted from 74 Mt<em>b</em> complex isolates for whole-genome sequencing (WGS). All genomes were typed and screened for mutations with known associations with antimicro<em>b</em>ial resistance using <i>in silico</i> methods, and results were cross-verified with wet la<em>b</em> methods. (<em>b</em>)Results:</<em>b</em>) Lineage (L) 4 (55.8%) was predominant, followed <em>b</em>y L3 (41.2%); L1 (1.5%) and L2 (1.5%) occurred rarely. The most frequently detected su<em>b</em>lineage was CAS (38.2%), followed <em>b</em>y Ural (29.4%), and Haarlem (11.8%). The recent transmission index (RTI) was relatively low. L4 and Ural strains were more resistant than the other strains to any anti-TB drug (<i>P</i> < 0.05). The most frequent mutations to RIF, <em>INH</em>, EMB, SM, PZA, ETH, FLQs, and 2nd-line injecta<em>b</em>le drugs occurred at <i>rpoB</i> S450L, <i>katG</i> S315T, <i>em<em>b</em>B</i> M306I/V, <i>rps</i>L K43R, <i>pncA</i> V139A, <i>ethA</i> M1R, <i>gyr</i>A D94G, and <i>rrs</i> A1401G, respectively. Disputed <i>rpoB</i> mutations were also shown in four (16%) of RIF-resistant isolates. (<em>b</em>)Conclusion:</<em>b</em>) Our WGS analysis revealed the presence of diverse Mt<em>b</em> genotypes. The presence of a significant proportion of disputed <i>rpoB</i> mutations highlighted the need to esta<em>b</em>lish a WGS facility at the regional level to monitor drug-resistant mutations. This will help control the transmission of DR-TB and ultimately contri<em>b</em>ute to the attainment of 100% DST coverage for TB patients as per the End TB strategy.

Keywords: Ethiopia; Mycobacterium tuberculosis; Tigray; disputed rpoB mutations; drug resistance; whole-genome sequencing.

Objective: To investigate the effect of Danhong Tongjing Prescription (DTP) on sperm quality in patients with bilateral varicocele (VC) after microsurgical varicocelectomy.

Methods: We randomly assigned 68 patients with bilateral VC to receive microsurgical varicocelectomy (the control group, n = 34) or microsurgical varicocelectomy followed by oral administration of DTP for a course of 90 days (the DTP group, n = 34). Before and after treatment, we obtained the sperm concentration, total sperm count, total sperm motility, the percentage of progressively motile sperm (PMS), sperm acrosomal enzyme activity, inhibin B (Inh-B) level, and sperm DNA fragmentation index (DFI) from the patients and compared the parameters between the two groups.

Results: There were no statistically significant differences in sperm concentration, PMS, acrosomal enzyme activity or sperm DFI among the patients with different degrees of VC preoperatively. After 3 months of medication, sperm concentration, total sperm count, total sperm motility, PMS and acrosomal enzyme activity were all increased while DFI decreased in both the control and DTP groups, even more significantly in the DTP group than in the control, and the Inh-B level was also markedly elevated in the DTP group in comparison with the baseline.

Conclusions: The severity of bilateral VC is not correlated with the reduction of semen quality. DTP can improve sperm quality by improving total sperm count, PMS and acrosomal enzyme activity and reducing DFI in VC patients after varicocelectomy. The underlying mechanisms of the prescription may be related to its anti-oxidative stress action and abilities of improving reproductive hypoxia, spermatogenic environment and the function of Sertoli cells, but the specific signaling pathway involved is not yet clear.

Keywords: bilateral varicocele; sperm quality; Danhong Tongjing Prescription.

Conducting clinical trials to understand the exposure-risk/<em>b</em>enefit relationship of canna<em>b</em>is use is not always feasi<em>b</em>le. Alternatively, physiologically-<em>b</em>ased pharmacokinetic (PBPK) models can <em>b</em>e used to predict exposure of the psychoactive canna<em>b</em>inoid, THC, and its active meta<em>b</em>olite 11-OH-THC. Here, we first extrapolated in vitro mechanistic PK information previously quantified to <em>b</em>uild a linked THC/11-OH-THC PBPK model, and verified the model with o<em>b</em>served data after intravenous and <em>inh</em>alation administration of THC in a healthy, non-pregnant population. The in vitro to in vivo extrapolation (IVIVE) of <em>b</em>oth THC and 11-OH-THC disposition was successful. The <em>b</em>ioavaila<em>b</em>ility (F<su<em>b</em>)<em>inh</em></su<em>b</em>)) of THC after <em>inh</em>alation was higher in chronic versus casual canna<em>b</em>is users (F<su<em>b</em>)<em>inh</em></su<em>b</em>) = 0.35 and 0.19, respectively). Sensitivity analysis demonstrated that 11-OH-THC <em>b</em>ut not THC exposure was sensitive to alterations in hepatic intrinsic clearance of the respective compound. Next, we extrapolated the linked THC/11-OH-THC PBPK model to pregnant women. Simulations showed that THC plasma AUC does not change during pregnancy, <em>b</em>ut 11-OH-THC plasma AUC decreases <em>b</em>y up to 41%. Using a maternal-fetal PBPK model, maternal and fetal THC serum concentrations were simulated and compared to the o<em>b</em>served THC serum concentrations in pregnant women at term. In order to recapitulate the o<em>b</em>served THC fetal serum concentrations, active placental efflux of THC needed to <em>b</em>e invoked. In conclusion, we <em>b</em>uilt and verified a linked THC/11-OH-THC PBPK model in healthy non-pregnant population and demonstrated how this mechanistic physiological and pharmacokinetic platform can <em>b</em>e extrapolated to a special population, such as pregnant women. (<em>b</em>)Significance Statement</<em>b</em>) While the pharmacokinetics of canna<em>b</em>inoids has <em>b</em>een extensively studied clinically, there are limited mechanistic PK models. Here we present the development and verification of a physiologically-<em>b</em>ased pharmacokinetic (PBPK) model for (-)-Δ⁹-tetrahydrocanna<em>b</em>inol (THC) and its active meta<em>b</em>olite, 11-OH-THC. The PBPK model was verified in healthy non-pregnant population after intravenous and <em>inh</em>alation administration of THC, and then extrapolated to pregnant women. The THC/11-OH-THC PBPK model can <em>b</em>e used to predict exposure in special populations, predict drug-drug interactions, or impact of genetic polymorphism.

Keywords: cannabinoids; in vitro-in vivo prediction (IVIVE); metabolite disposition; modeling and simulation; physiologically-based pharmacokinetic modeling/PBPK.

Complement, contact activation, coagulation, and fi<em>b</em>rinolysis are serum protein cascades that need strict regulation to maintain human health. Serum glycoprotein, a C1 inhi<em>b</em>itor (C1-<em>INH</em>), is a key regulator (inhi<em>b</em>itor) of serine proteases of all the a<em>b</em>ove-mentioned pathways. Recently, an autotransporter protein, virulence-associated gene 8 (Vag8), produced <em>b</em>y the whooping cough pathogen, <i>Bordetella pertussis</i>, was shown to <em>b</em>ind to C1-<em>INH</em> and interfere with its function. Here, we present the structure of the Vag8-C1-<em>INH</em> complex determined using cryo-electron microscopy at a 3.6-Å resolution. The structure shows a unique mechanism of C1-<em>INH</em> inhi<em>b</em>ition not employed <em>b</em>y other pathogens, where Vag8 sequesters the reactive center loop of C1-<em>INH</em>, preventing its interaction with the target proteases.(<em>b</em>)IMPORTANCE</<em>b</em>) The structure of a 10-kDa protein complex is one of the smallest to <em>b</em>e determined using cryo-electron microscopy at high resolution. The structure reveals that C1-<em>INH</em> is sequestered in an inactivated state <em>b</em>y <em>b</em>urial of the reactive center loop in Vag8. By so doing, the <em>b</em>acterium is a<em>b</em>le to simultaneously pertur<em>b</em> the many pathways regulated <em>b</em>y C1-<em>INH</em>. Virulence mechanisms such as the one descri<em>b</em>ed here assume more importance given the emerging evidence a<em>b</em>out dysregulation of contact activation, coagulation, and fi<em>b</em>rinolysis leading to COVID-19 pneumonia.

Keywords: Bordetella pertussis; C1-INH; autotransporters; bacterial pathogenicity; complement system; immune evasion; serpin; single-particle cryo-EM; three-dimensional structure.

(<em>b</em>)<em>B</em>ACKGROUND:</<em>b</em>) T<em>B</em> is the leading cause of death from a single infectious disease, particularly among people living with HIV (PLHIV). Molecular epidemiology provides information on prevalent genotypes of <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> and disease transmission dynamics, which aid in T<em>B</em> control. Identification of mutations that confer drug resistance is essential for the rapid diagnosis of drug-resistant T<em>B</em>, especially in high T<em>B</em> <em>b</em>urden settings, like the Philippines.(<em>b</em>)METHODS:</<em>b</em>) This study aimed to determine mutations in <i>M. tu<em>b</em>erculosis</i> drug resistance-conferring genes and circulating genotypes in PLHIV. MIRU-VNTR (myco<em>b</em>acterial interspersed repetitive unit-varia<em>b</em>le num<em>b</em>er of tandem repeats) typing using a set of 24-loci and sequencing of drug resistance-conferring genes were performed in 22 <i>M. tu<em>b</em>erculosis</i> isolates from T<em>B</em>-HIV co-infected patients.(<em>b</em>)RESULTS:</<em>b</em>) The prevalence of resistance to any drug was 31.8%, 18.2% for isoniazid monoresistance, 4.5% for streptomycin monoresistance and 9.1% for multidrug resistance. The identified mutations in the <i>kat</i>G<i>, rpo</i><em>B</em><i>, pnc</i>A<i>, rps</i>L and <i>gyr</i>A genes have <em>b</em>een reported in the literature; none was found in the <i><em>inh</em></i>A and <i>em<em>b</em></i><em>B</em> genes. All isolates <em>b</em>elonged to the EAI2-Manila family and were grouped into four clusters <em>b</em>ased on their phenotypic drug resistance and mutation profiles.(<em>b</em>)CONCLUSION:</<em>b</em>) The use of 24-loci set may <em>b</em>e used as a more discriminatory MIRU-VNTR typing in settings where the East African-Indian lineage is predominant, like the Philippines.

(<em>b</em>)Background and study aims</<em>b</em>) The risk of aerosolization of <em>b</em>ody fluids during endoscopic procedures should <em>b</em>e evaluated during the COVID-19 era, as this may contri<em>b</em>ute to serious disease transmission. Here, we aimed to investigate if use of endoscopic tools during flexi<em>b</em>le endoscopy may permit gas leakage from the scope or tools. (<em>b</em>)Material and methods</<em>b</em>) Using a fresh 35-cm porcine rectal segment, a colonoscope tip, and manometer were placed intraluminally at opposite ends of the segment. The colonoscope handle, including the <em>b</em>iopsy valve, was su<em>b</em>merged in a water <em>b</em>ath. Sequentially, various endoscopic devices (forceps, clips, snares, endoscopic su<em>b</em>mucosal dissection (ESD) knives) were inserted into the <em>b</em>iopsy valve, simultaneously su<em>b</em>merging the device handle in a water <em>b</em>ath. The <em>b</em>owel was slowly inflated up to 74.7 mmHg (40 <em>inH</em> <su<em>b</em>)2</su<em>b</em>) O) and presence of gas leakage, leak pressure, and gas leakage volume were measured. (<em>b</em>)Results</<em>b</em>) Gas leakage was o<em>b</em>served from the <em>b</em>iopsy valve upon insertion and removal of all endoscopic device tips with jaws, even at 0 mmHg (60/60 trials). The insertion angle of the tool affected extent of gas leakage. In addition, gas leakage was o<em>b</em>served from the device handles (8 of 10 devices) with continuous gas leakage at low pressures, especially two snares at 0 mmHg, and an injecta<em>b</em>le ESD knife at 0.7 ± 0.8 mmHg). (<em>b</em>)Conclusions</<em>b</em>) Gas leakage from the <em>b</em>iopsy valve and device handles commonly occur during endoscopic procedures. We recommend protective measures <em>b</em>e considered during use of any tools during endoscopy.

Acquired angioedema due to C1 inhibitor deficiency (C1-INH-AAE) is a rare disease characterized by adult-onset recurrent non-urticarial angioedema with low levels of C1-INH. It is associated with lymphoproliferative disorders, and treatments are off-label with variable success. We conducted a systematic literature review to include patients with C1-INH-AAE identified in PubMed and Embase databases between January 2006 and February 2021. Clinical features of these patients were summarized, and factors associated with disease remission were explored. A total of 121 patients were included in the current study with a median age at diagnosis of 64 years and 45.5% being male. An associated disease was recorded in 94 patients (77.7%), and lymphoproliferative disorder was the most reported (59/94, 62.8%). Anti-C1-INH autoantibodies were present in 45 of 71 patients (63.4%). Factors impacting disease remissions included age (odds ratio [OR] 0.951, 95% confidence interval [CI] 0.909-0.994, p = 0.027), male (OR 0.327, 95% CI 0.124-0.866, p = 0.025), presence of monoclonal gammopathy (OR 0.133, 95% CI 0.041-0.429, p = 0.001), requirement of specific on-demand treatment (OR 0.216, 95% CI 0.066-0.709, p = 0.012) and rituximab use (OR 2.865, 95% CI 1.038-7.911, p = 0.042). A total of nine patients (7.4%) died at last follow up and most were unrelated to C1-INH-AAE. Our results imply that C1-INH-AAE is primarily associated with underlying B or plasma cell abnormalities, and clone-directed therapies could be promising options for its long-term management.

Keywords: Acquired angioedema; C1 inhibitor; Lymphoproliferative disorder; Rituximab.

Novel chemotherapeutic agents against multidrug resistant-tuberculosis (MDR-TB) are urgently needed at this juncture to save the life of TB-infected patients. In this work, we have synthesized and characterized novel isatin hydrazones 4(a-o) and their thiomorpholine tethered analogues 5(a-o). All the synthesized compounds were initially screened for their anti-mycobacterial activity against the H₃₇Rv strain of Mycobacterium tuberculosis (MTB) under level-I testing. Remarkably, five compounds 4f, 4h, 4n, 5f and 5m (IC₅₀ = 1.9 µM to 9.8 µM) were found to be most active, with 4f (IC₅₀ = 1.9 µM) indicating highest inhibition of H₃₇Rv. These compounds were further evaluated at level-II testing against the five drug-resistant strains such as isoniazid-resistant strains (INH-R1 and INH-R2), rifampicin-resistant strains (RIF-R1 and RIF-R2) and fluoroquinolone-resistant strain (FQ-R1) of MTB. Interestingly, 4f and 5f emerged as the most potent compounds with IC₅₀ of 3.6 µM and 1.9 µM against RIF-R1 MTB strain, followed by INH-R1 MTB strain with IC₅₀ of 3.5 µM and 3.4 µM, respectively. Against FQ-R1 MTB strain, the lead compounds 4f and 5f displayed excellent inhibition at IC₅₀ 5.9 µM and 4.9 µM, respectively indicating broad-spectrum of activity. Further, molecular docking, ADME pharmacokinetic and molecular dynamics simulations of the compounds were performed against the DNA gyrase B and obtained encouraging results.

Keywords: Docking; Isatin; MD simulations; Multidrug-resistant tuberculosis; Synthesis.

<strong class="su<em>b</em>-title"> Background: </strong> Tu<em>b</em>erculosis (<em>b</em>)(</<em>b</em>)TB) and drug-resistant TB (DR-TB) continue to persist as a serious pu<em>b</em>lic health challenges in Ghana. Although several research has evaluated the drug resistance of <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> complex (MTBc) strains across the country, there is a paucity of data on its magnitude as well as the various lineages circulating in the Eastern region of Ghana.

Objective: This study therefore evaluated the distribution of the various lineages of MTBc in the Eastern region of the country and the associated drug resistance.

<strong class="su<em>b</em>-title"> Materials and methods: </strong> One hundred and forty-three (143) patients with pulmonary TB attending the Eastern Regional Hospital, Koforidua/Ghana were included in the study. The BACTEC MGIT 960 tu<em>b</em>e media was used for <em>b</em>oth sputum culture and drug suscepti<em>b</em>ility of streptomycin (STR), isoniazid (INH), rifampicin (RIF) and Etham<em>b</em>utol (ETH). Isolates were initially typed using IS<i>6110</i>, followed <em>b</em>y large sequence polymorphisms analysis and spoligotyping.

Results: The majority [108 (75.5%)] of the 143 patients were male gender and the 45-54 years [46 (32.2%)] age range had the highest frequency. Forty-one (28.7%) of the 143 isolates were IS6110 negative. Of the 102 spoligotyped isolates, the main sub-lineages included 45 (44.1%) Cameroon and 23 (22.5%) Ghana. SITs 61 and 53 represented the major cluster with 22/102 (21.6%) and 13/102 (12.7%) isolates respectively, while 59/65 (90.8%) isolates belonged to Lineage 4 with 27/65 (41.5%) LAM10_CAM. MDR-TB occurred in 26/79 (32.9%) isolates, and was not associated with neither gender [20/58 (34.5%) male vs 6/21 (28.6%) female, OR = 1.31; 95%CI, 0.44-3.92; p = 0.624)] nor age. No association was found between MDR-TB and the major sub-lineages [8/25 (32%) Cameroon (OR = 0.94; 95%CI, 0.34-2.59; p = 0.920) and 5/11 (45.5%) Ghana (OR = 1.87; 95%CI, 0.51-6.80; p = 0.489)], or previously treated [8/23 (34.8%), OR = 0.89; 95%CI, 0.32-2.48; p = 0.823)] patients.

Conclusion: Despite the serious threat posed by MDR in the study area, no sub-lineage was shown to be associated with drug resistance. Nonetheless, a sustained surveillance of drug resistance pattern is advocated. A lower proportion of M. africanum was observed in the Eastern region of Ghana and will require further evaluation.

Keywords: Cameroon sub-lineage; Eastern region; Ghana; MDR; Tuberculosis.

High-throughput centralized testing for tu<em>b</em>erculosis (TB) and drug resistance is important, <em>b</em>ut comparative data are limited. In this retrospective cross-sectional study, participants were recruited from Johannes<em>b</em>urg, South Africa, and T<em>b</em>ilisi, Georgia. The index tests, A<em>b</em><em>b</em>ott RealTi<i>m</i>e MTB (RT-MTB) and RealTi<i>m</i>e MTB RIF/<em>INH</em> (RT-MTB RIF/<em>INH</em>), were performed on specimens stored frozen for an extended period of time (<em>b</em>eyond manufacturer-validated specifications) and compared to paired Xpert MTB/RIF Ultra (Xpert Ultra) and Xpert MTB/RIF (Xpert) results o<em>b</em>tained with fresh specimens. The detection reference standard was the Myco<em>b</em>acterium tu<em>b</em>erculosis complex culture, and for resistance detection, it was phenotypic drug suscepti<em>b</em>ility testing. The median age of 474 participants was 39 (interquartile range [IQR], 31 to 51) years. On decontaminated sputum, Xpert Ultra had a sensitivity of 91%, compared to 77% for RT-MTB, with a difference of +14% (95% confidence interval [CI], +9.2 to +21%; 18/127). On raw sputum, Xpert Ultra exhi<em>b</em>ited a sensitivity of 89% and Xpert one of 88%, compared to 80% for RT-MTB, exhi<em>b</em>iting differences of +10% (95% CI, +3.3 to +18%; 9/93) and +8.6% (95% CI, +2.4 to +17%; 8/93), respectively. Specificity was ≥98% for all tests. All three tests showed high sensitivity and specificity for detection of rifampin resistance. A<em>b</em><em>b</em>ott assays may have lower sensitivity than Xpert and Xpert Ultra for TB detection <em>b</em>ut similar performance for detection of resistance. The differences in TB detection may <em>b</em>e attri<em>b</em>uta<em>b</em>le to differences in testing of frozen (A<em>b</em><em>b</em>ott) versus fresh (Xpert) samples. Studies in compliance with manufacturer's instructions are required to compare performance. (<em>b</em>)IMPORTANCE</<em>b</em>) In 2019, 10 million people fell ill with tu<em>b</em>erculosis (TB), of whom 1.4 million died. There are few comparative studies of diagnostic assays, particularly those aiming to <em>b</em>e used in high-throughput la<em>b</em>oratories. One such assay is the A<em>b</em><em>b</em>ott RealTi<i>m</i>e MTB (RT-MTB) and RealTi<i>m</i>e MTB RIF/<em>INH</em> (RT-MTB RIF/<em>INH</em>), which uses the m2000 platform already in use in many settings for HIV load testing and allows the diagnosis of TB and resistance to two first-line drugs, rifampin and isoniazid. Our study compared the RT-MTB and RT-MTB RIF/<em>INH</em> to the WHO-recommended Xpert MTB/RIF Ultra and Xpert MTB/RIF. The study is the largest comparative study to date and was performed independent of the manufacturer. The study results suggest that the A<em>b</em><em>b</em>ott RealTi<i>m</i>e MTB may have a lower sensitivity, <em>b</em>ut the study may have placed the A<em>b</em><em>b</em>ott test at a disadvantage <em>b</em>y using frozen samples and comparing the results to those for fresh samples for the Xpert.

Keywords: diagnostics; pulmonary; pulmonary infection; tuberculosis.

Oriental herbal medicine with the two bioactive constituents, β-eudesmol (BE) and atractylodin (AT), has been used as a remedy for gastrointestinal disorders. There was no scientific evidence reporting their antidiarrheal effect and underpinning mechanisms. Therefore, we aimed to investigate the anti-secretory activity of these two compounds in vitro. The inhibitory effect of BE and AT on cAMP-induced Cl^- secretion was evaluated by Ussing chamber in human intestinal epithelial (T84) cells. Short-circuit current (I_SC) and apical Cl^- current (I_Cl-) were measured after adding indirect and direct cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activator. MTT assay was used to determine cellular cytotoxicity. Protein-ligand interaction was investigated by in silico molecular docking analysis. BE, but not AT concentration-dependently (IC₅₀ of ~1.05 µM) reduced cAMP-mediated, CFTR_inh-172 inhibitable Cl^- secretion as determined by transepithelial I_SC across a monolayer of T84 cells. Potency of CFTR-mediated I_Cl- inhibition by BE did not change with the use of different CFTR activators suggesting a direct blockage of the channel active site(s). Pretreatment with BE completely prevented cAMP-induced I_Cl-. Furthermore, BE at concentrations up to 200 µM (24 h) had no effect on T84 cell viability. In silico studies indicated that BE could best dock onto dephosphorylated structure of CFTR at ATP-binding pockets in nucleotide-binding domain (NBD) 2 region. These findings provide the first evidence for the anti-secretory effect of BE involving inhibition of CFTR function. BE represents a promising candidate for the therapeutic or prophylactic intervention of diarrhea resulted from intestinal hypersecretion of Cl.

Keywords: CFTR; Diarrhea; β-eudesmol.

(<em>b</em>)<i>Background:</i></<em>b</em>) The severity of airway o<em>b</em>struction may affect patient's a<em>b</em>ility to perform an effective drug <em>inh</em>alation from a dry powder <em>inh</em>aler (DPI). Also, an incorrect <em>inh</em>alation technique may negatively affect the efficacy of asthma treatment. The aims of the study were (1) to analyze and compare <em>inh</em>alation profiles recorded with the use of different <em>inh</em>alation techniques, and thus, (2) to esta<em>b</em>lish model <em>inh</em>alation profiles representative for healthy su<em>b</em>jects and su<em>b</em>jects with mild and moderate-to-severe asthma. (<em>b</em>)<i>Methods:</i></<em>b</em>) This study was performed in healthy volunteers, patients with mild and moderate-to-severe asthma. A modified flow-volume test to define two different expiratory levels (to residual volume and half-way to residual volume) was performed. Inspiratory flow parameters were extracted: peak inspiratory flow rate (PIF<su<em>b</em>)<em>inh</em></su<em>b</em>)), time at which peak inspiratory flow rate occurs (tPIF<su<em>b</em>)<em>inh</em></su<em>b</em>)), total <em>inh</em>alation time (<i>T</i>), and <em>inh</em>aled volume (<i>V</i>). Test of frequency for tPIF<su<em>b</em>)<em>inh</em></su<em>b</em>)100% and tPIF<su<em>b</em>)<em>inh</em></su<em>b</em>)50% <em>b</em>y asthma severity was performed, to provide information a<em>b</em>out initial flow accelerations. The impact of two different expiratory levels preceding <em>inh</em>alation (with severity of asthma as a categorical factor) on inspiratory flow parameters was examined. (<em>b</em>)<i>Results:</i></<em>b</em>) PIF<su<em>b</em>)<em>inh</em></su<em>b</em>) was dependent upon asthma severity (<i>p</i> = 0.046). Type of exhalation <em>b</em>efore <em>inh</em>alation had no effect on PIF<su<em>b</em>)<em>inh</em></su<em>b</em>) values. <i>V</i> value was significantly affected <em>b</em>oth <em>b</em>y asthma severity (<i>p</i> = 0.024) and type of exhalation <em>b</em>efore <em>inh</em>alation (<i>p</i> < 0.0001). Mean <i>T</i> value was influenced <em>b</em>y type of exhalation <em>b</em>efore <em>inh</em>alation (<i>p</i> = 0.0003), <em>b</em>ut not <em>b</em>y asthma severity. Mean tPIF<su<em>b</em>)<em>inh</em></su<em>b</em>) value was affected <em>b</em>y the type of exhalation <em>b</em>efore <em>inh</em>alation only in healthy su<em>b</em>jects (<i>p</i> = 0.01). (<em>b</em>)<i>Conclusions:</i></<em>b</em>) Both asthma severity and type of exhalation <em>b</em>efore <em>inh</em>alation have little impact on the dynamics of <em>inh</em>alation through a DPI. An alternative form of equation descri<em>b</em>ing <em>inh</em>alation profiles demonstrating a relationship <em>b</em>etween lung mechanics and dynamics of inspiratory profile has <em>b</em>een proposed.

Keywords: asthma; dry powder inhaler; in vitro/in vivo correlations; inhalation profile; inhalation technique; inspiratory flow parameters.

Osteoarticular Tuberculosis (TB) is a challenging issue because of its chronicity and recurrence. Many drug delivery systems (DDSs) have been developed for general chemotherapy. Herein, we take advantage of instant hydrogelation to in situ encapsulate drugs onto implants intraoperatively, optimizing the drug release profile against osteoarticular TB. First-line chemodrugs, i.e. rifampicin (RFP) and isoniazid (INH) are firstly loaded on tricalcium phosphate (TCP). Then, the encapsulating hydrogel is fabricated by dipping in chitosan (CS) and β-glycerophosphate (β-GP) solution and heating at 80 °C for 40 min. The hydrogel encapsulation inhibits explosive drug release initially, but maintains long-term drug release (INH, 158 days; RFP, 53 days) in vitro. Therefore, this technique could inhibit bone destruction and inflammation from TB effectively in vivo, better than our previous ex situ prepared DDSs. The encapsulating technology, i.e. instant hydrogelation of drug-loaded implants, shows potential for regulating the type and ratio of drugs, elastic and viscous modulus of the hydrogel according to the state of illness intraoperatively for optimal drug release.

Tuberculosis (TB), caused by the Mycobacterium tuberculosis infection, continues to be a leading cause of morbidity and mortality in developing countries. Resistance to the first-line anti-TB drugs, isoniazid (INH) and rifampicin (RIF), is a major drawback to effective TB treatment. Genetic mutations in the β-subunit of the DNA-directed RNA polymerase (rpoB) are reported to be a major reason of RIF resistance. However, the structural basis and mechanisms of these resistant mutations are insufficiently understood. In the present study, thirty drug-resistant mutants of rpoB were initially modeled and screened against RIF via a comparative molecular docking analysis with the wild-type (WT) model. These analyses prioritized six mutants (Asp441Val, Ser456Trp, Ser456Gln, Arg454Gln, His451Gly, and His451Pro) that showed adverse binding affinities, molecular interactions, and RIF binding hinderance properties, with respect to the WT. These mutant models were subsequently analyzed by molecular dynamics (MD) simulations. One-hundred nanosecond all-atom MD simulations, binding free energy calculations, and a dynamic residue network analysis (DRN) were employed to exhaustively assess the impact of mutations on RIF binding dynamics. Considering the global structural motions and protein-ligand binding affinities, the Asp441Val, Ser456Gln, and His454Pro mutations generally yielded detrimental effects on RIF binding. Locally, we found that the electrostatic contributions to binding, particularly by Arg454 and Glu487, might be adjusted to counteract resistance. The DRN analysis revealed that all mutations mostly distorted the communication values of the critical hubs and may, therefore, confer conformational changes in rpoB to perturb RIF binding. In principle, the approach combined fundamental molecular modeling tools for robust "global" and "local" level analyses of structural dynamics, making it well suited for investigating other similar drug resistance cases.

Keywords: drug resistance; dynamic residue network analysis; molecular dynamics simulations; mutations; rifampicin; rpoB.

Background: Prothrombotic and inflammatory variables decrease after obesity surgery. The contact activation system may be a common denominator of these changes.

Objective: To characterize the contact system before and 6 months after Roux-en-Y gastric bypass (RYGB) and to evaluate associations with changes (post-surgery minus pre-surgery) in metabolic variables.

Methods: Women (n = 42) and men (n = 18) with obesity underwent RYGB, and measures of kallikrein generation, factor XII (FXII), prekallikrein, high molecular weight kininogen (HK), and C1 esterase inhibitor (C1-inh) were determined before and 6 months after surgery. Associations were evaluated using correlation and multivariate regression analyses.

Results: After RYGB, the endogenous kallikrein potential (EKP), peak kallikrein generation, FXII, and prekallikrein were reduced, and kallikrein generation lag time was prolonged (all p < 0.0005). Before and after RYGB, absolute values of EKP, lag time, and peak kallikrein generation correlated consistently with contact system proteins (range of correlation coefficients (r_S): -0.43 to -0.28 and 0.24 to 0.45 (pre-surgery); -0.43 to -0.30 and 0.28 to 0.50 (post-surgery)). RYGB-associated changes in EKP correlated with C1-inh (r_S = -0.29, p = 0.025), but also with triglycerides (r_S = 0.34, p = 0.007) and cholesterol (r_S = 0.28, p = 0.029), and independently associated with changes in C1-inh (β = -0.40) and triglycerides (β = 0.39). Changes in C1-inh associated with reductions in body weight (β = -0.39) and HbA1c (β = 0.38).

Conclusion: The contact system was affected 6 months after RYGB. Absolute values of kallikrein generation before and after RYGB correlated with contact system proteins, whereas changes after RYGB associated with changes in C1-inh and metabolic variables.

Keywords: Bariatric surgery; Hemostasis; Inflammation; Metabolism; Obesity.

Objective: To evaluate the clinical efficacy of bovine serum albumin nanoparticles loaded with isoniazid and rifampicin (INH-RFP-BSA-NPs) in the treatment of spinal tuberculosis in rabbits.

Methods: 35 spinal tuberculosis rabbit models were grouped into three groups, including 14 in group A and group B respectively and 7 in group C.All rabbits in group A were treated by INH-RFP-BSA-NPs's injection and in group B were treated with classic dosage form of INH and RFP, while in group C normal saline was given as the blank control. After intervention, the body weighing and CT scan, as well as concentration's measurement of INH and RFP in blood and tissues, were performed in all rabbits at the time of the 6thweek and 12th week, respectively.

Results: In group A, rabbits' weight increased by 0.44 kg and 0.27 kg within 6 weeks and 12 weeks' treatment respectively. The bactericidal concentrations of 1.64 µg•g^-1 for INH and 21.36 µg•g^-1 for RFP were measured in focus vertebral body 6 weeks post-injection and six weeks later the concentrations of INH and RFP in vertebral body still maintained at the level of 1.96 µg•g^-1 and 22.35 µg•g^-1respectively. After 12 weeks therapy, CT-scanned showed all the necrotic tissue was replaced by normal bone tissue. In group B, all rabbits had no significant increment of body weight and 4 rabbits had paralysis of hind leg. The concentrations of INH and RFP in vertebral body and focus were much lower than group A. CT-scanned showed the focus vertebral body was only partially repaired after 12 weeks' therapy.

Conclusion: The INH-RFP-BSA-NPs has the characteristics of sustained release in vivo and target biodistribution in focus vertebral body. Its therapeutic effect in rabbit spinal tuberculosis is much better than common INH and RFP.

Keywords: Spinal tuberculosis; bovine serum albumin; isoniazid; nanoparticles; rifampin.

Background: Rabies is a disease that still exists in developing countries and leads to more fatalities than other zoonotic diseases. Our study aimed to describe the profile of human exposures to animals over fifteen years and to assess the post-exposure prophylaxis (PEP) practices in the governorate of Kasserine (Tunisia) on pre- and post-revolution (2011).

Methods: We carried out a descriptive study using surveillance data from a region in Central-West Tunisia. All humans exposed to animals, residents in Kasserine Governorate and declared to the regional directorate of primary health care (RDPH) from January 1st, 2004 to December 31st, 2018 were included.

Results: A total of 45,564 cases of human exposures to animals were reported over the fifteen-year period of the study with an annual average of 3089.2 ± 403.1. The standardized incidence rate (SIR) of human exposures to animals was 694 per year per 100,000 inhabitants (inh). The most listed offending animal was the dog (91.3%) and the most reported type of exposure was bites (63.7%). The trend in human exposures to animals increased significantly over time. The number of exposures by vaccinated dogs decreased significantly and by unvaccinated and stray dogs increased steeply. When comparing pre-and post-revolution periods, the yearly average of animal exposures post-2011 was significantly greater than the average prior to 2011 (3200 ± 278.5 vs 2952.8 ± 483) (p < 0.001). The yearly average of animal bites post-2011 was significantly greater than the average prior to 2011 (2260.5 ± 372.1 vs 1609.8 ± 217.9) (p < 0.001). The average number of vaccine doses per animal exposure was 2.4. Concerning PEP protocols, protocol A (2 and 3 doses) was indicated in 79% of animal exposures cases. From 2004 to 2018, a downward trend was noted for protocol A (r = - 0.29, p < 0.001) and an upward trend for protocol B (3 and 5 doses) (r = 0.687, p < 0.001). During our study period, 5 fatal cases of human rabies were declared.

Conclusion: Rabies remains a major public health problem in Tunisia. The political dynamics had an impact on the health care system and rabies control. Preventive measures should be applied adequately to decrease the burden of this disease.

Keywords: Animal bite; Post-exposure prophylaxis; Rabies; Tunisia.