Genomic characterization of the first mpox and varicella-zoster co-infection in Pakistan (2023) through next-generation sequencing.

The global mpox outbreak spanning 2022-2023 has affected numerous countries worldwide. In this study, we present the first report on the detection, whole-genome sequence, and coinfection of the mpox virus and varicella zoster virus (VZV) from Pakistan. During April-May 2023, samples from 20 suspected cases of mpox were tested at the National Institutes of Health, Islamabad among which 4 tested positive. All four cases had a travel history of Saudi Arabia. All the suspected samples were processed by using a Zymo research kit for DNA extraction, followed by qRT-PCR amplification by using a DaAn Gene detection kit for the mpox virus. Further, two of the positive samples with a low Ct value (<20) were subjected to whole-genome sequencing using a metagenomic approach on the iSeq (Illumina) platform. The sequencing results revealed Clade IIb and genotype A.2.1 of MPXV, which clustered with viruses from Slovenia and the UK in July and June 2022, respectively. Our analysis identified two novel nonsynonymous substitutions in mpox virus, namely V98I in OPG046 and P600S in OPG109. Furthermore, we successfully retrieved the complete genome of VZV from the same sample, belonging to Clade 5. This study represents the first positive case of MPXV in Pakistan and the coinfection of mpox and VZV by using a metagenome approach providing insights into their complete genomes. Our results highlight the importance of surveillance at the point of entries, strengthening lab capacities including next-generation sequencing, and using differential diagnosis for timely and accurate detection of mpox cases.

Clinicopathological profile of peritoneal tuberculosis and a new scoring model for predicting mortality: an international ID-IRI study.

Existing literature about peritoneal tuberculosis (TBP) is relatively insufficient. The majority of reports are from a single center and do not assess predictive factors for mortality. In this international study, we investigated the clinicopathological characteristics of a large series of patients with TBP and determined the key features associated with mortality. TBP patients detected between 2010 and 2022 in 38 medical centers in 13 countries were included in this retrospective cohort. Participating physicians filled out an online questionnaire to report study data. In this study, 208 patients with TBP were included. Mean age of TBP cases was 41.4 ± 17.5 years. One hundred six patients (50.9%) were females. Nineteen patients (9.1%) had HIV infection, 45 (21.6%) had diabetes mellitus, 30 (14.4%) had chronic renal failure, 12 (5.7%) had cirrhosis, 7 (3.3%) had malignancy, and 21 (10.1%) had a history of immunosuppressive medication use. A total of 34 (16.3%) patients died and death was attributable to TBP in all cases. A pioneer mortality predicting model was established and HIV positivity, cirrhosis, abdominal pain, weakness, nausea and vomiting, ascites, isolation of Mycobacterium tuberculosis in peritoneal biopsy samples, TB relapse, advanced age, high serum creatinine and ALT levels, and decreased duration of isoniazid use were significantly related with mortality (p < 0.05). This is the first international study on TBP and is the largest case series to date. We suggest that using the mortality predicting model will allow early identification of high-risk patients likely to die of TBP.

Profiles of multidrug-resistant organisms among patients with bacteremia in intensive care units: an international ID-IRI survey.

Evaluating trends in antibiotic resistance is a requisite. The study aimed to analyze the profile of multidrug-resistant organisms (MDROs) among hospitalized patients with bacteremia in intensive care units (ICUs) in a large geographical area. This is a 1-month cross-sectional survey for blood-borne pathogens in 57 ICUs from 24 countries with different income levels: lower-middle-income (LMI), upper-middle-income (UMI), and high-income (HI) countries. Multidrug-resistant (MDR), extensively drug-resistant (XDR), or pan-drug-resistant isolates were searched. Logistic regression analysis determined resistance predictors among MDROs. Community-acquired infections were comparable to hospital-acquired infections particularly in LMI (94/202; 46.5% vs 108/202; 53.5%). Although MDR (65.1%; 502/771) and XDR (4.9%; 38/771) were common, no pan-drug-resistant isolate was recovered. In total, 32.1% of MDR were Klebsiella pneumoniae, and 55.3% of XDR were Acinetobacter baumannii. The highest MDR and XDR rates were in UMI and LMI, respectively, with no XDR revealed from HI. Predictors of MDR acquisition were male gender (OR, 12.11; 95% CI, 3.025-15.585) and the hospital-acquired origin of bacteremia (OR, 2.643; 95%CI, 1.462-3.894), and XDR acquisition was due to bacteremia in UMI (OR, 3.344; 95%CI, 1.189-5.626) and admission to medical-surgical ICUs (OR, 1.481; 95% CI, 1.076-2.037). We confirm the urgent need to expand stewardship activities to community settings especially in LMI, with more paid attention to the drugs with a higher potential for resistance. Empowering microbiology laboratories and reports to direct prescribing decisions should be prioritized. Supporting stewardship in ICUs, the mixed medical-surgical ones in particular, is warranted.

Stimuli-responsive transmembrane anion transport by AIE-active fluorescent probes.

Anticancer drug resistance implicates multifunctional mechanisms, and hypoxia is one of the key factors in therapeutic resistance. Hypoxia-specific therapy is considered an extremely effective strategy to fight against cancer. The development of small molecule-based synthetic anion transporters has also recently drawn attention for their potential therapeutic applications against several ion-transport-associated diseases, such as cancer and others. Herein, we describe the development of a hypoxia-responsive proanionophore to trigger controlled transport of anions across membranes under pathogenic conditions. Herein, we report the development of tetraphenylethene (TPE)-based anion transporters. The sulfonium-linked p-nitrobenzyl containing TPE-based proanionophore could be converted into a lipophilic fluorescent Cl- ion carrier in a hypoxic or reductive environment. Stimuli such as nitroreductase (NTR) and glutathione (GSH) mediated regeneration of the TPE-based active Cl- ion transporter also showed aggregation-induced emission (AIE) properties. We hypothesize that such hypoxia and reductive stimuli activatable proanionophores have tremendous potential to fight against channelopathies, including cancer.

Translation and cross-cultural adaptation of WHOQOL-HIV Bref among people living with HIV/AIDS in Pakistan.

BACKGROUND: Reliable Health-Related Quality of Life (HRQoL) assessment will be useful in identifying health issues and in identifying health care actions. Due to the lack of a psychometrically valid tool in Urdu, we aim to translate and examine the psychometric and cross-cultural adaptation of WHOQOL HIV Bref among people living with HIV/AIDS (PLWHA) in Pakistan. METHODS: The standard forward-backwards translation technique was used to convert English version of the WHOQOL HIV Bref into Urdu. After cognitive debriefing, final Urdu version of instrument was developed. Based on the principle of at least 5 subjects for each item, a sample of 182 patients was used using a universal random sampling technique from the Pakistan Institute of Medical Sciences, Islamabad. The Cronbach's alpha and intra-class correlation coefficients (ICC) were estimated to assess internal validity and reliability of the translated version. Exploratory factor analysis was carried out to determine the factor structure and independent associations between the instrument domains and CD-4T-cell count were assessed using multivariable linear regression RESULTS: High Cronbach alpha 0.93 was found for all WHOQOL HIV Bref facets. The test-retest reliability demonstrated a statistically significant ICC ranged from 0.88 to 0.98 (p < 0.001). In known group validity, lower CD-4 lymphocytes count was significantly related to poor scores for all six domains (p < 0.001). Similarly, symptomatic subjects had significantly lower scores compared to asymptomatic subjects on the physical, psychological, social relationship and independence domains (p < 0.05). Statistically significant positive correlation of all six domains of instrument with CD4 cells count (p < 0.001), exhibiting patients with higher CD-4 cells will have higher mean scores of all domains. Factor analysis revealed 5 domains, including physical health, psychological health, social relationship, environmental, and spiritual health. Multivariable linear regression analysis reported; only physical, psychological health and environment health domains were found significantly associated with higher CD-4 lymphocytes count (Beta = 0.121, p < 0.001, Beta = 0.103, p = 0.002, and Beta = 0.032, p = 0.032). CONCLUSION: Findings suggested that the Urdu version of WHOQOL HIV Bref is a psychometrically valid and culturally well-adapted HRQoL measurement tool for PLWHA in Pakistan.

Delayed admissions and efficacy of steroid use in patients with critical and severe COVID-19: an apprehensive approach.

BACKGROUND: Inadvertent delays in access to appropriate therapeutic interventions in high-risk group coronavirus disease 2019 (COVID-19) patients contribute to mortality in patients with severe/critical disease presentation. OBJECTIVES: The aim of this study was to determine the effect of timely admission to the hospital on mortality of patients with severe/critical COVID-19. Another secondary aspect of this study was to observe the efficacy of time-dependent use of corticosteroids on mortality of critical/severe COVID-19 patients. METHODS: Clinical data of 659 patients with severe/critical COVID-19, admitted to four major tertiary care hospitals from the Islamabad-Rawalpindi region of Pakistan was retrospectively collected from a period February-August 2020. Multivariate logistic regression analysis was carried out to determine the predictors of mortality in severe/critical COVID-19 patients. RESULTS: Out of a total of 659 patients, 469 (71.2%) patients died. Age > 60 years, presence of hypertension, heart disease and kidney disease along with late admission (>5 days) were significant predictors of mortality in patients with severe/critical COVID-19. CONCLUSIONS: The study highlights the importance of well-timed provision of appropriate medical interventions control COVID-19-associated mortality.

A cross sectional study to assess nasal carriage of methicillin resistant Staphylococcus aureus in healthcare professionals in a tertiary care hospital.

OBJECTIVE: To determine the nasal carriage of staphylococcus aureus and methicillin-resistant staphylococcus aureus among healthcare workers in a tertiary care setting. METHODS: The cross-sectional study was conducted at the Pakistan Institute of Medical Sciences, Islamabad, Pakistan, from April to July 2018, and comprised healthcare workers at the institution. Nasal swabs were collected and cultured on Mannitol salt agar. Mannitol fermenting colonies which were gram-positive cocci, catalase-positive and coagulase-positive were identified as staphylococcus aureus. Antibiotic susceptibility test was performed by modified Kirby-Bauer disc diffusion method. Methicillin resistance was detected using cefoxitin disc diffusion method. Data was analysed using SPSS 23. RESULTS: Of the 210 nasal swabs, 52(24.76%) had a staphylococcus aureus growth, and of them, 15(7.1%) were methicillin-resistant. No association could be established with either any single category of healthcare worker or an inter-department variation (p>0.05). Likewise, there was no association with age, gender, duration of service, smoking, co-morbidities, use of antibiotics in the preceding six months, treating a patient with methicillin-resistant staphylococcus aureus in the preceding six months and hospitalisation in the preceding year (p>0.05). CONCLUSIONS: The frequency of nasal carriage of methicillin resistant staphylococcus aureus amongst healthcare workers was regardless of the nature of their professional engagement.

Multi-stimuli controlled release of a transmembrane chloride ion carrier from a sulfonium-linked procarrier.

In recent times, anion transporters have received substantial consideration due to their ability to disrupt the ionic equilibrium across membrane bilayers. While numerous Cl- ion transporters were developed for channelopathies, unfortunately, poor aqueous solubility precluded their bioapplicability. Herein, we demonstrate the development of a multi-stimuli activatable anion transport approach to induce regulated transport of Cl- ions across membranes under specific conditions. The sulfonium-based procarrier was initially inactive, but the transmembrane transport of Cl- ions was activated in the presence of stimuli such as glutathione (GSH), reactive oxygen species (ROS) and light. The release of the hydrophobic anionophore from the aqueous-soluble procarrier under specific conditions leads to the successful transport of Cl- ions. Under physiological conditions, these anion carriers follow an antiport exchange mechanism to transport Cl- ions across lipid bilayers. Such multi-stimuli activatable procarriers have great potential to combat various types of channelopathies, including cancer, cystic fibrosis, kidney stones, myotonia, and others.

Primary metabolites from overproducing microbial system using sustainable substrates.

Primary (or secondary) metabolites are produced by animals, plants, or microbial cell systems either intracellularly or extracellularly. Production capabilities of microbial cell systems for many types of primary metabolites have been exploited at a commercial scale. But the high production cost of metabolites is a big challenge for most of the bioprocess industries and commercial production needs to be achieved. This issue can be solved to some extent by screening and developing the engineered microbial systems via reconstruction of the genome-scale metabolic model. The predicted genetic modification is applied for an increased flux in biosynthesis pathways toward the desired product. Wherein the resulting microbial strain is capable of converting a large amount of carbon substrate to the expected product with minimum by-product formation in the optimal operating conditions. Metabolic engineering efforts have also resulted in significant improvement of metabolite yields, depending on the nature of the products, microbial cell factory modification, and the types of substrate used. The objective of this review is to comprehend the state of art for the production of various primary metabolites by microbial strains system, focusing on the selection of efficient strain and genetic or pathway modifications, applied during strain engineering.

pH-Regulated anion transport activities of bis(iminourea) derivatives across the cell and vesicle membrane.

Recently, synthetic anion transporters have gained considerable attention because of their ability to disrupt cellular anion homeostasis and promote cell death. Herein, we report the development of bis(iminourea) derivatives as a new class of selective Cl- ion carrier. The bis(iminourea) derivatives were synthesized via a one-pot approach under mild reaction conditions. The presence of iminourea moieties suggests that the bis(iminourea) derivatives can be considered as unique guanidine mimics, indicating that the protonated framework could have much stronger anion recognition properties. The cooperative interactions of H+ and Cl- ions with these iminourea moieties results in the efficient transport of HCl across the lipid bilayer in an acidic environment. Under physiological conditions these compounds weakly transport Cl- ions via an antiport exchange mechanism. This pH-dependent gating/switching behavior (9-fold) within a narrow window could be due to the apparent pKa values (6.2-6.7) of the compounds within the lipid bilayer. The disruption of ionic homeostasis by the potent compounds was found to induce cell death.

Risk factors and clinical outcomes for vancomycin-resistant enterococcus bacteraemia in hospitalised cancer patients in Pakistan: A case-control study.

OBJECTIVE: To compare the risk factors and outcomes of vancomycin-resistant enterococcus with vancomycin-sensitive enterococcus bacteraemia among hospitalised cancer patients. METHODS: The retrospective, case-control study was conducted at Shaukat Khanum Memorial Cancer Hospital and Research Centre, Lahore, Pakistan, and comprised data of cancer patients whose blood culture grew either vancomycin-sensitive or vancomycin-resistant enterococcus from January 2012 to December 2014. Multivariable logistic regression analyses were used to determine the factors associated with the development of vancomycin-resistant enterococcus bacteraemia and 12-week mortality. Stata 11 was used for data analysis. RESULTS: Of the 138 cases, 111(80%) were selected, of which 46(41.44%) were of vancomycin-resistant and 65(58.55%) were of vancomycin-sensitive enterococcus. Length of hospital stay prior to bacteraemia (adjusted odds ratio 1.18; 95% confidence interval 1.08-1.28) and use of vancomycin 30 days before the onset of bacteraemia (adjusted odds ratio 9.4; 95% confidence interval 1.70-52.19) were significant risk factors for the development of vancomycin-resistant enterococcus bacteraemia. The overall 12-week mortality rate was 29(63%) for patients with vancomycin-resistant bacteraemia and 28(43.1%) for vancomycin-sensitive enterococcus bacteraemia patients. Risk factors for mortality included the presence of shock at the time of the onset of bacteraemia (adjusted odds ratio 32.91; 95% confidence interval 3.02-358.81). CONCLUSIONS: The length of hospital stay and prior exposure to vancomycin were significant risk factors for the occurrence of vancomycin-resistant enterococcus bacteraemia.

Comparing machine learning screening approaches using clinical data and cytokine profiles for COVID-19 in resource-limited and resource-abundant settings.

Accurate screening of COVID-19 infection status for symptomatic patients is a critical public health task. Although molecular and antigen tests now exist for COVID-19, in resource-limited settings, screening tests are often not available. Furthermore, during the early stages of the pandemic tests were not available in any capacity. We utilized an automated machine learning (ML) approach to train and evaluate thousands of models on a clinical dataset consisting of commonly available clinical and laboratory data, along with cytokine profiles for patients (n = 150). These models were then further tested for generalizability on an out-of-sample secondary dataset (n = 120). We were able to develop a ML model for rapid and reliable screening of patients as COVID-19 positive or negative using three approaches: commonly available clinical and laboratory data, a cytokine profile, and a combination of the common data and cytokine profile. Of the tens of thousands of models automatically tested for the three approaches, all three approaches demonstrated > 92% sensitivity and > 88 specificity while our highest performing model achieved 95.6% sensitivity and 98.1% specificity. These models represent a potential effective deployable solution for COVID-19 status classification for symptomatic patients in resource-limited settings and provide proof-of-concept for rapid development of screening tools for novel emerging infectious diseases.

Stimuli-responsive release of active anionophore from RGD-peptide-linked proanionophore.

Integrin-mediated cellular delivery was attempted to optimize practical applications of hydrophobic ionophores. The potent ionophore preferentially transports H+/Cl- across the lipid bilayers following a symport mechanism. The RGD-peptide-appended tag was stimulated by glutathione to generate the active ionophore, prompting the transport of Cl- under the cellular environment.

Effective hydrolysis for waste plant biomass impacts sustainable fuel and reduced air pollution generation: A comprehensive review.

Among various natural biowastes availability in the environment, agricultural residues showed great impacts. It is due to huge availability and cheap carbon source, creating big challenges for their utility and systematic reduction. Objective of this review is to address the waste biomass availability and huge quantities issues and also put effort to minimize this nutrient load via biotransforming into value-added products. Different wastes (organic/inorganic) generation with their negative issues are due to numbers of developmental and social activities, reported. Currently, various efforts are found for these wastes minimization via generation of different types of value-added products (biogas, bioH2, alcoholic fuel, organic acids and others products) and these wastes in municipal cities are also reported with production of advanced biofuels as promising outcomes. For hydrolysis of complex organic resources including lignocellulosic biomasses, physicochemical, structural or compositional changes are needed that aid in conversion into sugar and organic compounds such as biofuels. So, efficient and effective pretreatment processes selection (physical, biological, chemical or combined one) is critical to achieve these hydrolysis goals and resultant cellulose or hemicellulose components can be accessible by biological catalysis. These can achieve final hydrolysis and fermentative or monomer sugars. And later, synthesis of fuels or value-added products during microbial fermentation or biotransformation processes can be achieved. This review discusses pretreatment techniques for improved hydrolysis for fermentative sugar with emphasis on reduced quantities of toxic compounds (furfural compound) in hydrolyzed biomasses. Minimum deterioration fuel economy also reported with production of different bioproducts including biofuels. Additionally, impacts of toxic products and gasses emission are also discussed with their minimization.

Quinoline Thiourea-Based Zinc Ionophores with Antibacterial Activity.

The increasing resistance of bacteria to commercially available antibiotics threatens patient safety in healthcare settings. Perturbation of ion homeostasis has emerged as a potential therapeutic strategy to fight against antibacterial resistance and other channelopathies. This study reports the development of 8-aminoquinoline (QN) derivatives and their transmembrane Zn2+ transport activities. Our findings showed that a potent QN-based Zn2+ transporter exhibits promising antibacterial properties against Gram-positive bacteria with reduced hemolytic activity and cytotoxicity to mammalian cells. Furthermore, this combination showed excellent in vivo efficacy against Staphylococcus aureus. Interestingly, this combination prevented bacterial resistance and restored susceptibility of gentamicin and methicillin-resistant S. aureus to commercially available ß-lactam and other antibiotics that had lost their activity against the drug-resistant bacterial strain. Our findings suggest that the transmembrane transport of Zn2+ by QN derivatives could be a promising strategy to combat bacterial infections and restore the activity of other antibiotics.

Analysing the role of gender and place of residence in acceptability and satisfaction towards e-learning among university students' during COVID-19 pandemic in India.

This paper has two broad objectives; the first is to examine the challenges of e-learning faced by the students keeping in view their place of residence and gender in India, particularly during the second-wave of Covid-19. The second objective is to examine the role of place of residence and gender of students in the acceptance and satisfaction towards e-learning. The data has been obtained through an online survey of the students of University of Jamia Millia Islamia, New Delhi, India, in which a total of 490 students participated. Selection of students has been done through stratified sampling technique. Initially the obtained data was analysed and discussed through simple statistical analysis. Later, a chi-square test of independence was applied to find out the dependency of psychological stress, level of acceptance and level of satisfaction towards e-learning on the place of residence and the gender. The major finding of the paper reveals that the gender and the place of residence of the students is significantly associated with their psychological stress, acceptance and satisfaction towards e-learning. Extra money spent on the purchase of online learning resources was greater in case of rural students.

Self-reported antibiotic stewardship and infection control measures from 57 intensive care units: An international ID-IRI survey.

We explored the self-reported antibiotic stewardship (AS), and infection prevention and control (IPC) activities in intensive care units (ICUs) of different income settings. A cross-sectional study was conducted using an online questionnaire to collect data about IPC and AS measures in participating ICUs. The study participants were Infectious Diseases-International Research Initiative (IDI-IR) members, committed as per their institutional agreement form. We analyzed responses from 57 ICUs in 24 countries (Lower-middle income (LMI), n = 13; Upper-middle income (UMI), n = 33; High-income (HI), n = 11). This represented (~5%) of centers represented in the ID-IRI. Surveillance programs were implemented in (76.9%-90.9%) of ICUs with fewer contact precaution measures in LMI ones (p = 0.02); (LMI:69.2%, UMI:97%, HI:100%). Participation in regional antimicrobial resistance programs was more significantly applied in HI (p = 0.02) (LMI:38.4%,UMI:81.8%,HI:72.2%). AS programs are implemented in 77.2% of institutions with AS champions in 66.7%. Infectious diseases physicians and microbiologists are members of many AS teams (59%&50%) respectively. Unqualified healthcare professionals(42.1%), and deficient incentives(28.1%) are the main barriers to implementing AS. We underscore the existing differences in IPC and AS programs' implementation, team composition, and faced barriers. Continuous collaboration and sharing best practices on APM is needed. The role of regional and international organizations should be encouraged. Global support for capacity building of healthcare practitioners is warranted.

Isolation and characterization of secondary metabolites from Plumeria obtusa.

Chromatographic purification of the methanolic extract of Plumeria obtusa yielded two new iridoid obtusadoids A (1) and B (2), along with eight known compounds plumieridin A (3), plumieridine (4), 1α-plumieride (5), 15-demethylplumieride (6), rel-(3R,3'S,4R,4'S)-3,3',4,4'-tetrahydro-6,6'-dimethoxy[3,3'-bi-2H-benzopyran]-4,4'-diol (7), glochiflavanoside B (8), oleanolic acid (9), and methyl coumarate (10). The structures of all the isolates (1-10) were determined by NMR spectroscopy and mass spectrometry. The data of known compounds (3-10) were further compared with the reported data for these compounds.

Prevalence and risk factors for diabetes mellitus in a selected urban population of a city in Punjab.

OBJECTIVE: To estimate the prevalence of diabetes mellitus (diagnosed and undiagnosed), impaired fasting glucose and possible risk factors for diabetes mellitus among Pakistani population. METHODS: This cross sectional study was performed in Rawalpindi which is one of the cities in Northern Punjab of Pakistan in July 2008. An area was selected in Rawalpindi city, with mixed population representative of almost all provinces with different socioeconomic groups. Three hundred and thirteen houses were selected through systematic random sampling technique and fasting blood glucose was obtained and subjects were labeled to have diabetes according to WHO criteria of diagnosing diabetes mellitus. The statistical analysis was performed by using Stata version 10. RESULTS: There were 1091 respondents who were selected after cleaning the data, among them 293 were males and 798 were females. Of the total 15.41% of the males and 12.31% of females were found to have diabetes mellitus. Thus making a total prevalence of 13.14%. Impaired fasting glucose (IFG) was found in 5.14% males and 5.78% females making a total prevalence of 5.61%. Over all (DM & IFG) was found to be 20.55% in males and 18.09% in females. The main risk factors identified were obesity, family history, hypertension and increasing age. CONCLUSION: There is an increased prevalence of Type 2 diabetes in Pakistan and main risk factors identified were obesity, overweight, family history of diabetes mellitus, and hypertension.