Stigma is a barrier to care for people affected by TB and HIV in Uganda, where these conditions remain endemic. While scales have been adapted and validated to measure stigma among TB-affected households in Uganda, there is a need for scales that measure the experiences of persons with TB (PWTB).We adapted the Van Rie 12-item individual perspectives TB scale and 10-item individual perspectives HIV scale for use in Uganda through cross-cultural discussions with a multidisciplinary research team and four cognitive interviews with community health workers and PWTB. We then conducted a cross-sectional study administering each scale to 125 PWTB. We performed exploratory factor analysis, evaluated internal validity, and assessed convergent validity with perceived social support.Exploratory factor analysis yielded a one-factor solution for both scales, with marginal model fit (standardised root mean square residual = 0.09 for TB, = 0.07 for HIV). There was evidence of convergent validity through a positive correlation of the TB (r = 0.22, p = 0.01) and HIV stigma (r = 0.22, p = 0.01) scales with perceived social support. Both scales had good internal validity (Cronbach's α = 0.86 for TB, = 0.87 for HIV).Adapted scales to measure perceived HIV and TB stigma among PWTB in Uganda demonstrated promising psychometric properties by removing one and two items, respectively..

BACKGROUND:Neonates admitted to the neonatal intensive care unit (NICU) are at risk for healthcare-associated infections, including central line-associated bloodstream infections. We aimed to characterize the epidemiology of bloodstream infections among neonates with central venous catheters admitted to three Indian NICUs. METHODS:We conducted a prospective cohort study in three tertiary NICUs, from May 1, 2017 until July 31, 2019. All neonates admitted to the NICU were enrolled and followed until discharge, transfer, or death. Cases were defined as positive blood cultures in neonates with a central venous catheter in place for greater than 2 days or within 2 days of catheter removal. RESULTS:During the study period, 140 bloodstream infections were identified in 131 neonates with a central venous catheter. The bloodstream infection rate was 11.9 per 1000 central line-days. Gram-negative organisms predominated, with 38.6% of cases caused by Klebsiella spp. and 14.9% by Acinetobacter spp. Antimicrobial resistance was prevalent among Gram-negative isolates, with 86.9% resistant to third- or fourth-generation cephalosporins, 63.1% to aminoglycosides, 61.9% to fluoroquinolones, and 42.0% to carbapenems. Mortality and length of stay were greater in neonates with bloodstream infection than in neonates without bloodstream infection (unadjusted analysis, p < 0.001). CONCLUSIONS:We report a high bloodstream infection rate among neonates with central venous catheters admitted to three tertiary care NICUs in India. Action to improve infection prevention and control practices in the NICU is needed to reduce the morbidity and mortality associated with BSI in this high-risk population.

These results highlight distinct differences in the cell type composition and cancer epithelial pathways in vaginal vs. cervical SCC. Among upregulated pathways in vaginal SCC, ER and mTORC1 pathway activation may represent targets for therapeutic intervention worthy of further investigation.

Background: P7 viroporin in HCV is a cation-selective ion channel-forming protein, functional in the oligomeric form. It is considered to be a potential target for anti-HCV compounds due to its crucial role in viral entry, assembly, and release. Method: Conserved crucial residues present in HCV p7 protein were delineated from the available literature with a specific focus on the genotypes 3a and 1b prevalent in India. Using the Flex-X docking tool, a library of FDA-approved drugs was docked on the receptor sites prepared around crucial residues. In the present study, we proposed drug repurposing to target viroporin p7, which may help in the rapid development of effective anti-HCV therapies. Results: With our approach of poly-pharmacology, a variety of drugs currently identified as antibiotics, antiparasitic, antiemetic, anti-retroviral, and anti-neoplastic were found to dock successfully on the p7 viroporin. Noteworthy among these are general-purpose cephalosporin antibiotics, leucal, phthalylsulfathiazole, and granisetron, which may be useful in acute HCV infection, and anti-neoplastic sorafenib and nilotinib, which may be valuable in advanced HCV-HCC cases. Conclusion: This study could pave the way for quick repurposing of these compounds as anti-HCV therapeutics.

Esophageal lung is a type of Group-II communicating bronchopulmonary foregut malformations (CBPFM) usually diagnosed beyond neonatal period during investigation for recurrent respiratory symptoms and persistent radiographic features suggesting pneumonia or bronchiectasis. In our case, we noticed bronchiectasis and disproportionately severe volume loss in an infant with associated multisystem anomalies in the absence of “significant” lower respiratory tract symptoms. A detailed evaluation with repeat imaging confirmed a Group-II CBPFM, a congenital pathology instead of an infective cause. Pneumonectomy is a more prudent option instead of undertaking major airway reconstruction for the dysplastic “dysfunctional” tissue. Amongst the various associated anomalies reported till now, the associated rib and renal anomalies noted by us have not been described earlier to the best of our knowledge.

This study aimed to estimate the ICU admission rate and related cost in patients taking Molnupiravir versus Nirmatrelvir+Ritonavir, the oral antiviral therapies to receive Emergency Use Authorization from the US FDA in December 2021 for the treatment of mild to moderate COVID-19 patients. In this retrospective claim data study, Optum's de-identified Clinformatics® Data Mart Database (Mainly Commercial and Medicare Advantage) is used to identify the patients who have been prescribed Molnupiravir or Nirmatrelvir+Ritonavir with NDC codes. The patient cohort is divided into two groups: one has been prescribed Molnupiravir and the other one has been prescribed Nirmatrelvir+Ritonavir. The patients in this study cohort have both medical and pharmacy claims. The study period is considered from the approval of these drugs, which is December 22nd and 23rd, 2021, to May 31st, 2022. The first fill date of the prescription for Molnupiravir and Nirmatrelvir+Ritonavir is considered the index date. All patients were followed for 30 days post index to look for any ICU admission and its related costs. The data was analyzed, and two tailed t-test and chi square test were applied. The percentage of patients who have been prescribed Molnupiravir is 25% (N= 4,851), while the patient cohort prescribed with Nirmatrelvir+Ritonavir is 75% (N= 14,479). Patients who have been prescribed Molnupiravir have a significantly higher ICU admission rate (1.2%) than patients who have been prescribed Nirmatrelvir+Ritonavir (0.2%) (P<0.001). The average cost related to ICU admission in patients who have taken Molnupiravir (mean-$15,439) does not show a difference from the cost of ICU admission in patients who have taken Nirmatrelvir+Ritonavir (mean-$13,383) (P = 0.72). This study shows that the ICU admission rate is noticeably different in the cohort of patients taking Molnupiravir compared to the Nirmatrelvir+Ritonavir combination, where ICU-related costs are almost similar in both the cohorts.

The objective of this study is to analyze the impact of COVID-19 vaccination in reducing standard hospitalization costs among COVID-19 patients in the US who required admission to the ICU or usage of ventilators. In this retrospective study, Optum's de-identified Clinformatics® Data Mart database was used to identify all the patients above 18 years who underwent treatment for COVID-19 in the US between 1 Jan 2021 to 30 Sep 2021. The occurrence of the ICD-10 code for COVID-19 infection in the claims database was defined as the index event. These patients were then checked for admission to the ICU or usage of ventilators. The average length of stay and standard costs were compared between unvaccinated patients, and fully vaccinated patients (who had COVID-19 after 28 days of the second dose of the COVID-19 vaccine). Wilcoxon Whitney U test was applied to analyze the level of significance. Out of 76,320 COVID-19 patients who required admission to the ICU or usage of ventilators, 75,839 (99.4%) were unvaccinated while 481 (0.4%) were fully vaccinated patients. The average length of stay among the unvaccinated (11.2 days) was significantly higher (p<.0001) as compared to fully vaccinated patients (8.8 days). Also, the mean standard costs among the unvaccinated ($30,025 for 49,724 patients) was significantly higher (p = .03) as compared to fully vaccinated patients ($23,170 for 205 patients). COVID-19 vaccines significantly reduced the average length of stay and standard hospitalization cost among the COVID-19 patients who required admission to the ICU or usage of ventilators.

Despite their limitations, unfractionated heparin (UFH) and bivalirudin remain standard-of-care parenteral anticoagulants for percutaneous coronary intervention (PCI). We discovered novel direct thrombin inhibitors (DTIs) from tick salivary transcriptomes and optimised their pharmacologic activity. The most potent, ultravariegin, inhibits thrombin with a Ki of 4.0 pM, 445-fold better than bivalirudin. Unexpectedly, despite their greater antithrombotic effect, variegin/ultravariegin demonstrated less bleeding, achieving a 3-to-7-fold wider therapeutic index in rodent thrombosis and bleeding models. When used in combination with aspirin and ticagrelor in a porcine model, variegin/ultravariegin reduced stent thrombosis compared with antiplatelet therapy alone but achieved a 5-to-7-fold lower bleeding time than UFH/bivalirudin. Moreover, two antibodies screened from a naïve human antibody library effectively reversed the anticoagulant activity of ultravariegin, demonstrating proof-of-principle for antidote reversal. Variegin and ultravariegin are promising translational candidates for next-generation DTIs that may reduce peri-PCI bleeding in the presence of antiplatelet therapy.

Background The purpose of this study was to evaluate the outcome of stereotactic body radiation therapy (SBRT) in patients of unresectable hepatocellular carcinoma (HCC ) complicated with portal vein tumor thrombosis (PVTT) who are also unsuitable for other locoregional therapies. Materials and methods Between May 2018 and January 2020, twenty-nine patients with advanced unresectable HCC s, treated with SBRT, were enrolled in this retrospective audit. Patients of Child status A5–B7 and with healthy liver volume, ≥ 700 ccs were treated. Local control (LC), overall survival (OS), progression-free survival (PFS), PVTT opening rate, and effect of prognostic factors were analyzed. Results The median tumor diameter was 8.6 cm (5–14), and the median tumor volume was 275 cc (151–1196). The median SBRT dose prescription was 48 Gy in 6 fractions (32–50 Gy in 5–6 fractions). The median follow up was eight months (1–20), 1-year local control, progression-free survival, and overall survival were 95%, 53.4%, and 60%, respectively. Overall rate of grade III toxicity was less than 5%, and the most common toxicity was lymphocytopenia. Tumors of more than 350cc had worse OS and PFS when compared to tumors < 350 cc (median OS and PFS of tumors > 350 cc was 4 months and two months, p = .01 and .003, respectively). A total of fifteen patients progressed with the disease and the median time to progression was two months [1–4]. Conclusion SBRT is safe and provides excellent local control in advanced HCC complicated with PVTT. The out of field failure pattern and time to failure in these patients highlights the need for adjuvant systemic therapy after completion of local treatment. Our data warrant the need for multimodality trials in this patient cohort.

Emulsion PCR (ePCR) is an important technique that permits amplification of DNA molecules in physically separated picoliter volume water-in-oil droplets, and thus avoids formation of unproductive chimera and other artifacts between similar DNA sequences.However, the recovery of ePCR products involves repeated extraction with hazardous organic solvents followed by purification using silica-based columns, making the overall process cumbersome. In this report, we have described a ‘Quick ePCR extraction protocol’ for the purification of ePCR products, which directly employs silica-based DNA purification columns and the products purified using this method have been found to be compatible with gene cloning and NGS applications. The method described here makes ePCR easy, safe and within the reach of every laboratory.

Efficient spin injection from epitaxial ferrimagnetic NiFe_2O_4 thin films into a Pd layer is demonstrated via spin Seebeck effect measurements in the longitudinal geometry. The NiFe_2O_4 films (60 nm to 1 μm) are grown by pulsed laser deposition on isostructural spinel MgAl_2O_4, MgGa_2O_4, and CoGa_2O_4 substrates with lattice mismatch varying between 3.2 the spin Seebeck voltage is observed with decreasing lattice mismatch, which correlates well with a decrease in the Gilbert damping parameter as determined from ferromagnetic resonance measurements. High resolution transmission electron microscopy studies indicate substantial decrease of antiphase boundary and interface defects that cause strain-relaxation, i.e., misfit dislocations, in the films with decreasing lattice mismatch. This highlights the importance of reducing structural defects in spinel ferrites for efficient spin injection. It is further shown that angle-dependent spin Seebeck effect measurements provide a qualitative method to probe for in-plane magnetic anisotropies present in the films.

Emulsion PCR (ePCR) is an important technique that permits amplification of DNA molecules in physically separated picoliter-volume water-in-oil droplets, and thus avoids formation of unproductive chimeras and other artifacts between similar DNA sequences. However, the recovery of ePCR products involves repeated extraction with hazardous organic solvents followed by purification using silica-based columns, making the overall process cumbersome. In this benchmark, we have described a quick ePCR extraction protocol for the purification of ePCR products, which directly employs silica-based DNA purification columns; products purified using this method have been found to be compatible with gene cloning and next-generation sequencing applications. The method described here makes ePCR easy, safe and within the reach of every laboratory.

Macrolide antibiotics, such as erythromycin and josamycin, are natural polyketide products harboring 14- to 16-membered macrocyclic lactone rings to which various sugars are attached. These antibiotics are used extensively in the clinic because of their ability to inhibit bacterial protein synthesis. More recently, some macrolides have been shown to also possess anti-inflammatory and other therapeutic activities in mammalian cells. To better understand the targets and effects of this drug class in mammalian cells, we used a genome-wide shRNA screen in K562 cancer cells to identify genes that modulate cellular sensitivity to josamycin. Among the most sensitizing hits were proteins involved in mitochondrial translation and the mitochondrial unfolded protein response, glycolysis, and the mitogen-activated protein kinase signaling cascade. Further analysis revealed that cells treated with josamycin or other antibacterial agents exhibited impaired oxidative phosphorylation and metabolic shifts to glycolysis. Interestingly, we observed that knockdown of the mitogen-activated protein kinase kinase kinase 4 (MAP3K4) gene, which contributes to p38 mitogen-activated protein kinase signaling, sensitized cells only to josamycin but not to other antibacterial agents. There is a growing interest in better characterizing the therapeutic effects and toxicities of antibiotics in mammalian cells to guide new applications in both cellular and clinical studies. To our knowledge, this is the first report of an unbiased genome-wide screen to investigate the effects of a clinically used antibiotic on human cells.

Genome-wide analysis of the mode of action of GSK983, a potent antiviral agent, led to the identification of dihydroorotate dehydrogenase as its target along with the discovery that genetic knockdown of pyrimidine salvage sensitized cells to GSK983. Because GSK983 is an ineffective antiviral in the presence of physiological uridine concentrations, we explored combining GSK983 with pyrimidine salvage inhibitors. We synthesized and evaluated analogs of cyclopentenyl uracil (CPU), an inhibitor of uridine salvage. We found that CPU was converted into its triphosphate in cells. When combined with GSK983, CPU resulted in large drops in cellular UTP and CTP pools. Consequently, CPU-GSK983 suppressed dengue virus replication in the presence of physiological concentrations of uridine. In addition, the CPU-GSK983 combination markedly enhanced the effect of RNA-dependent RNA polymerase (RdRp) inhibition on viral infection. Our findings highlight a new host-targeting strategy for potentiating the antiviral activity of RdRp inhibitors.

Chikungunya virus (CHIKV) a re-emerging mosquito-borne alpha virus causes significant distress which is further accentuated in the lack of specific therapeutics or a preventive vaccine, mandating accelerated research for anti-CHIKV therapeutics. In recent years, drug repositioning has gained recognition for the curative interventions for its cost and time efficacy. CHIKV envelope proteins are considered to be the promising targets for drug discovery because of their essential role in viral attachment and entry in the host cells. In the current study, we propose structure-based virtual screening of drug molecule on the crystal structure of mature Chikungunya envelope protein (PDB 3N41) using a library of FDA approved drug molecules. Several cephalosporin drugs docked successfully within two binding sites prepared at E1-E2 interface of CHIKV envelop protein complex with significantly low binding energies. Cefmenoxime, ceforanide, cefotetan, cefonicid sodium and cefpiramide were identified as top leads with a cumulative score of -67.67, -64.90, -63.78, -61.99, and - 61.77, forming electrostatic, hydrogen and hydrophobic bonds within both the binding sites. These shortlisted leads could be potential inhibitors of E1-E2 hetero dimer in CHIKV, hence might disrupt the integrity of envelope glycoprotein leading to loss of its ability to form mature viral particles and gain entry into the host.

The high-affinity interaction between biotin and streptavidin has opened avenues for using recombinant proteins with site-specific biotinylation to achieve efficient and directional immobilization. The site-specific biotinylation of proteins carrying a 15 amino acid long Biotin Acceptor Peptide tag (BAP; also known as AviTag) is effected on a specific lysine either by co-expressing the E. coli BirA enzyme in vivo or by using purified recombinant E. coli BirA enzyme in the presence of ATP and biotin in vitro. In this paper, we have designed a T7 promoter-lac operator-based expression vector for rapid and efficient cloning, and high-level cytosolic expression of proteins carrying a C-terminal BAP tag in E. coli with TEV protease cleavable N-terminal deca-histidine tag, useful for initial purification. Furthermore, a robust three-step purification pipeline integrated with well-optimized protocols for TEV protease-based H10 tag removal, and recombinant BirA enzyme-based site-specific in vitro biotinylation is described to obtain highly pure biotinylated proteins. Most importantly, the paper demonstrates superior sensitivities in indirect ELISA with directional and efficient immobilization of biotin-tagged proteins on streptavidin-coated surfaces in comparison to passive immobilization. The use of biotin-tagged proteins through specific immobilization also allows more efficient selection of binders from a phage-displayed naïve antibody library. In addition, for both these applications, specific immobilization requires much less amount of protein as compared to passive immobilization and can be easily multiplexed. The simplified strategy described here for the production of highly pure biotin-tagged proteins will find use in numerous applications, including those, which may require immobilization of multiple proteins simultaneously on a solid surface.

“This article uses bioethical and cross-cultural lenses to examine the narratives of 11 mental health professionals (psychiatrists, psychologists, and counselors) regarding their perceptions of, and experiences in providing mental health services in Kuwait. Given that there is no legal ethical body governing mental health service delivery in Kuwait, and that there have been recent reports of negative personal experiences with mental health professionals, this study sought to understand the types of narratives and treatment approaches that may contribute to inadequate service delivery. This study drew on interpretive phenomenological analysis (IPA) and critical discourse analysis in its analysis. The analyses indicated that ideology (either patient-centered or disease-centered) can be shaped by educational background and professional experiences, which can, in turn, shape how mental health professionals deliver mental health services to the Kuwaiti community. Findings also indicate that mainstream western medical discourses are actively transforming the landscape of mental health care in Kuwait; while this western transformation is welcomed (and even imposed) by some clinicians, it is critiqued by others who feel that: a) indigenous forms of healing are beginning to wane; and b) local clinicians can be pressured to assimilate to North American standards of mental health care. Research limitations and directions for clinical education and practice are also discussed.”

This report describes a rare case of type IIA2 sagittal urethral duplication. The presentation, investigation, and management of this rare anomaly are briefly discussed. A 3½-year-old boy presented with urinary obstruction and recurrent urinary tract infection due to a stenosed dorsal urethra and segmental stenosis of the dominant ventral urethra. The child also had left-sided vesicoureteric reflux. Staged surgical management consisted of an initial vesicostomy followed by serial dilatation of the ventral urethral stricture, left ureteric reimplantation, and a 2-cm long distal urethrourethrostomy between the dorsal urethra, opening at the tip of the penis, and the ventral urethra, which had a hypospadic opening at the base of the glans. The functional and cosmetic outcomes were satisfactory. The management needs to be individualized as best suited for the patient.Keywords: urethra, urethral duplication, urethral stricture