Human Immunodeficiency Virus Type-1 Genetic Diversity and Drugs Resistance Mutations among People Living with HIV in Karachi, Pakistan.

The human immunodeficiency virus type-1 epidemic in Pakistan has significantly increased over the last two decades. In Karachi, Pakistan, there is a lack of updated information on the complexity of HIV-1 genetic diversity and the burden of drug resistance mutations (DRMs) that can contribute to ART failure and poor treatment outcomes. This study aimed to determine HIV-1 genetic diversity and identify drug-resistance mutations among people living with HIV in Karachi. A total of 364 HIV-positive individuals, with a median age of 36 years, were enrolled in the study. The HIV-1 partial pol gene was successfully sequenced from 268 individuals. The sequences were used to generate phylogenetic trees to determine clade diversity and also to assess the burden of DRMs. Based on the partial pol sequences, 13 distinct HIV-1 subtypes and recombinant forms were identified. Subtype A1 was the most common clade (40%), followed by CRF02_AG (33.2%). Acquired DRMs were found in 30.6% of the ART-experienced patients, of whom 70.7%, 20.7%, and 8.5% were associated with resistance to NNRTIs, NRTIs, and PIs, respectively. Transmitted DRMs were found in 5.6% of the ART-naïve patients, of whom 93% were associated with resistance against NNRTIs and 7% to PIs. The high prevalence of DRMs in ART-experienced patients poses significant challenges to the long-term benefits and sustainability of the ART program. This study emphasizes the importance of continuous HIV molecular epidemiology and drug resistance surveillance to support evidence-based HIV prevention, precise ART, and targeted AIDS care.

Evaluation of the Training Program to Train HIV Treatment Center Staff in Pakistan.

Introduction In Pakistan, HIV training programs, especially for health professionals working in HIV treatment centers, are limited. Consequently, there is little data about HIV awareness among physicians and allied health workers and how it may affect their care for people living with HIV (PLWH). Recently, the Global Fund to Fight AIDS, Tuberculosis, and Malaria (GFATM) grant Principal Recipient UNDP engaged an NGO experienced in HIV/AIDS training, on a competitive basis, to develop a training manual and conduct training of all categories of HIV treatment centers staff. The goal of this study was to assess the training program's influence on trainees' (both physicians and allied health staff) knowledge and abilities and describe its major lessons. Methodology This was a one-group pre-post test study, carried out between January 17 and February 22, 2023. The study was carried out in three phases. In the first phase, a team of experts developed an antiretroviral treatment (ART) training manual. In the second phase, 9- and three-day training workshops were conducted in six different cities of Pakistan, which were attended by physicians and allied health staff working in different HIV treatment centers across Pakistan. The workshops had plenary lectures, discussions, role plays, video cases, and case studies. In the third phase, a quiz, comprising multiple/best choice questions (MCQs/BCQs) and true and false questions, was administered before (pre) and after the workshop (post) to assess the impact of these training sessions in enhancing the level of HIV knowledge, especially related to ART. The workshop was attended by a total of 256 health workers from different cities in Pakistan. The participants had backgrounds in medical science, psychology, laboratory science, nursing, and computer science. Pre-and post-test responses were statistically analyzed to determine the impact of the training program on participant's knowledge. For this, the Shapiro-Wilk test was applied to test data normality, followed by the application of paired t-test or Wilcoxon Signed Rank Test for normally and non-normally distributed data, respectively. Finally, a chi-square test was applied to examine the significant (p<0.05) association between training workshops and improvement in the participant's level of understanding of HIV. In all statistical tests, p<0.05 was considered significant.  Results The results from our study showed that before the training session, both physicians and allied staff possessed limited knowledge about HIV-related domains. After the workshops, participants from all cities demonstrated a uniform enhancement of knowledge related to different HIV-related domains, evident from the improvement in post-test scores compared to pre-test scores (p<0.0001). The chi-square test showed a significant association between training workshops and improvement in the participant's level of understanding about HIV (p-values for BCQ, MCQ, and true and false: 0.001, 0.0047, and 0.0024, respectively). Conclusions Pre- and post-test evaluation provides an objective, data-driven method for measuring the impact of educational interventions in improving healthcare workers' awareness about HIV. The results emphasize the role of continuous workshops and training programs in enhancing the knowledge and understanding of healthcare and allied health workers regarding HIV.

Candidate protein biomarkers in chronic kidney disease: a proteomics study.

Proteinuria poses a substantial risk for the progression of chronic kidney disease (CKD) and its related complications. Kidneys excrete hundreds of individual proteins, some with a potential impact on CKD progression or as a marker of the disease. However, the available data on specific urinary proteins and their relationship with CKD severity remain limited. Therefore, we aimed to investigate the urinary proteome and its association with kidney function in CKD patients and healthy controls. The proteomic analysis of urine samples showed CKD stage-specific differences in the number of detected proteins and the exponentially modified protein abundance index for total protein (p = 0.007). Notably, specific urinary proteins such as B2MG, FETUA, VTDB, and AMBP exhibited robust negative associations with kidney function in CKD patients compared to controls. Also, A1AG2, CD44, CD59, CERU, KNG1, LV39, OSTP, RNAS1, SH3L3, and UROM proteins showed positive associations with kidney function in the entire cohort, while LV39, A1BG, and CERU consistently displayed positive associations in patients compared to controls. This study suggests that specific urinary proteins, which were found to be negatively or positively associated with the kidney function of CKD patients, can serve as markers of dysfunctional or functional kidneys, respectively.

Factors Contributing to Rapid Early Infarct Expansion in Acute Ischemic Stroke Patients With Large Vessel Occlusion.

Background Acute ischemic stroke, particularly in cases involving large vessel occlusion (LVO), poses a significant challenge due to the potential for rapid infarct expansion in the early phase. Such expansion, if not managed promptly, can lead to severe neurological deficits and poor clinical outcomes. Understanding the contributing factors that accelerate early infarct expansion is crucial for optimizing treatment strategies and improving patient prognosis. The main aim of the study is to determine the factors contributing to rapid early infarct expansion in acute ischemic stroke patients with LVO. Methodology The retrospective study was conducted at Liaquat National Hospital in Karachi from August 2023 to December 2023. Data were collected from 685 patients with anterior circulation LVO-related acute stroke with witnessed stroke onset and baseline perfusion imaging. Extracted clinical data included age, gender, medical history (hypertension, diabetes, etc.), and baseline National Institutes of Health Stroke Scale (NIHSS) scores. Results The mean age of the included patients was 67.4 years, with a relatively balanced gender distribution, i.e., 48.5% male (n = 332) and 51.5% female (n = 353). The mean baseline NIHSS score was 14.2, reflecting initial neurological severity. Imaging parameters revealed that 294 (42.6%) patients exhibited infarct expansion, with an average penumbra size of 23.5 mL. Hypoperfusion intensity ratio (HIR) quartiles demonstrated a notable association with progression rates, escalating from 27 (4%) patients in the first quartile to approximately 527 (77%) patients in the fourth quartile, highlighting a significant correlation between HIR and infarct expansion (p < 0.001). Conclusions HIR emerged as a pivotal factor strongly associated with rapid infarct expansion, underscoring its significance in predicting the trajectory of ischemic injury.

Identification of a novel first-generation HIV-1 circulating recombinant form (CRF152_DG) among people living with HIV in Karachi, Pakistan.

The objective of this study was to characterize a novel circulating recombinant form of human immunodeficiency virus type 1 (HIV-1) among people living with HIV in Karachi, Pakistan. We conducted near-full-length genome (NFLG) sequencing on eight samples exhibiting D/G recombination signals in the pol gene region. We successfully obtained NFLG sequences (790-9,614; with reference to the HXB2 genome) from four of the eight samples and then conducted phylogenetic and recombination analyses on them. The four NFLG sequences from our study and one DG unique recombinant form previously identified in the United Kingdom (GenBank accession: MF109700) formed a distinct monophyletic cluster with an Shimodaira-Hasegawa approximate likelihood ratio test node support value of 100%. Bootscan analyses of the five NFLG sequences of DG recombinants showed that all five NFLGs shared the same unique mosaic pattern of recombination breakpoints between D and G clades, with two D fragments in the pol and vif regions inserted into a G backbone. Subregion phylogenetic analyses confirmed these sequences to be a novel circulating recombinant form (CRF) composed of subtypes D and G. The DG recombinant sequences were eventually designated as CRF152_DG by the Los Alamos HIV Sequence Database staff. IMPORTANCE: In Pakistan, the genetic diversity of human immunodeficiency virus type 1 (HIV-1) is becoming increasingly complex, compared to the early years of the epidemic that started after the detection of the first cases of HIV-1 in 1987 in Karachi. Based on the available molecular studies, two dominant HIV-1 clades, sub-subtype A1 and CRF02_AG, have been found to co-circulate with other clades, namely B, C, D, G, CRF01_AE, CRF35_A1D, and CRF56_cpx, in various urban areas of Pakistan. Several novel recombinant forms have also been detected. This first report of CRF152_DG highlights the complex nature of the HIV epidemic in Pakistan and emphasizes the importance of continual molecular surveillance (ideally based on whole-genome sequences) of HIV.

Differential analysis of histopathological and genetic markers of cancer aggressiveness, and survival difference in EBV-positive and EBV-negative prostate carcinoma.

Several studies have shown an association between prostate carcinoma (PCa) and Epstein-Barr virus (EBV); however, none of the studies so far have identified the histopathological and genetic markers of cancer aggressiveness associated with EBV in PCa tissues. In this study, we used previously characterized EBV-PCR-positive (n = 39) and EBV-negative (n = 60) PCa tissues to perform an IHC-based assessment of key histopathological and molecular markers of PCa aggressiveness (EMT markers, AR expression, perineural invasion, and lymphocytic infiltration characterization). Additionally, we investigated the differential expression of key oncogenes, EMT-associated genes, and PCa-specific oncomiRs, in EBV-positive and -negative tissues, using the qPCR array. Finally, survival benefit analysis was also performed in EBV-positive and EBV-negative PCa patients. The EBV-positive PCa exhibited a higher percentage (80%) of perineural invasion (PNI) compared to EBV-negative PCa (67.3%) samples. Similarly, a higher lymphocytic infiltration was observed in EBV-LMP1-positive PCa samples. The subset characterization of T and B cell lymphocytic infiltration showed a trend of higher intratumoral and tumor stromal lymphocytic infiltration in EBV-negative tissues compared with EBV-positive tissues. The logistic regression analysis showed that EBV-positive status was associated with decreased odds (OR = 0.07; p-value < 0.019) of CD3 intratumoral lymphocytic infiltration in PCa tissues. The analysis of IHC-based expression patterns of EMT markers showed comparable expression of all EMT markers, except vimentin, which showed higher expression in EBV-positive PCa tissues compared to EBV-negative PCa tissues. Furthermore, gene expression analysis showed a statistically significant difference (p < 0.05) in the expression of CDH1, AR, CHEK-2, CDKN-1B, and CDC-20 and oncomiRs miR-126, miR-152-3p, miR-452, miR-145-3p, miR-196a, miR-183-3p, and miR-146b in EBV-positive PCa tissues compared to EBV-negative PCa tissues. Overall, the survival proportion was comparable in both groups. The presence of EBV in the PCa tissues results in an increased expression of certain oncogenes, oncomiRs, and EMT marker (vimentin) and a decrease in CD3 ITL, which may be associated with the aggressive forms of PCa.

Microorganism-mediated biodegradation for effective management and/or removal of micro-plastics from the environment: a comprehensive review.

Micro- plastics (MPs) pose significant global threats, requiring an environment-friendly mode of decomposition. Microbial-mediated biodegradation and biodeterioration of micro-plastics (MPs) have been widely known for their cost-effectiveness, and environment-friendly techniques for removing MPs. MPs resistance to various biocidal microbes has also been reported by various studies. The biocidal resistance degree of biodegradability and/or microbiological susceptibility of MPs can be determined by defacement, structural deformation, erosion, degree of plasticizer degradation, metabolization, and/or solubilization of MPs. The degradation of microplastics involves microbial organisms like bacteria, mold, yeast, algae, and associated enzymes. Analytical and microbiological techniques monitor microplastic biodegradation, but no microbial organism can eliminate microplastics. MPs can pose environmental risks to aquatic and human life. Micro-plastic biodegradation involves fragmentation, assimilation, and mineralization, influenced by abiotic and biotic factors. Environmental factors and pre-treatment agents can naturally degrade large polymers or induce bio-fragmentation, which may impact their efficiency. A clear understanding of MPs pollution and the microbial degradation process is crucial for mitigating its effects. The study aimed to identify deteriogenic microorganism species that contribute to the biodegradation of micro-plastics (MPs). This knowledge is crucial for designing novel biodeterioration and biodegradation formulations, both lab-scale and industrial, that exhibit MPs-cidal actions, potentially predicting MPs-free aquatic and atmospheric environments. The study emphasizes the urgent need for global cooperation, research advancements, and public involvement to reduce micro-plastic contamination through policy proposals and improved waste management practices.

Diosgenin loaded cellulose nanoonion impedes different stages of protein aggregation induced cell death via alleviating mitochondrial dysfunction and upregulation of autophagy.

Protein aggregation is a multifaceted phenomenon prevalent in the progression of neurodegenerative diseases, yielding aggregates of diverse sizes. Recently, increased attention has been directed towards early protein aggregates due to their pronounced toxicity, largely stemming from inflammation mediated by reactive oxygen species (ROS). This study advocates for a therapeutic approach focusing on inflammation control rather than mere ROS inhibition in the context of neurodegenerative disorders. Here, we introduced Camellia sinensis cellulose nanoonion (CS-CNO) as an innovative, biocompatible nanocarrier for encapsulating the phytosteroid diosgenin (DGN@CS-CNO). The resulting nano-assembly, manifesting as spherical entities with dimensions averaging ~180-220 nm, exhibits a remarkable capacity for the gradual and sustained release of approximately 39-44 % of DGN over a 60-hour time frame. DGN@CS-CNO displays a striking ability to inhibit or disassemble various phases of hen egg white lysozyme (HEWL) protein aggregates, including the early (HEWLEA) and late (HEWLLA) stages. In vitro experiments employing HEK293 cells underscore the potential of DGN@CS-CNO in mitigating cell death provoked by protein aggregation. This effect is achieved by ameliorating ROS-mediated inflammation and countering mitochondrial dysfunction, as evidenced by alterations in TNFα, TLR4, and MT-CO1 protein expression. Western blot analyses reveal that the gradual and sustained release of DGN from DGN@CS-CNO induces autophagy, a pivotal process in dismantling intracellular amyloid deposits. In summary, this study not only illuminates a path forward but also presents a compelling case for the utilization of phytosteroid as a formidable strategy against neuroinflammation incited by protein aggregation.

Digital Therapeutics for COPD Patient Self-Management: Needs Analysis and Design Study.

Timely management of Chronic Obstructive Pulmonary Disease (COPD) exacerbations can improve recovery and reduce the risk of hospitalization. Digital therapeutics are digital interventions, based on best evidence, designed to provide home-based, patient-centered and pervasive self-management support to patients. Digital therapeutics can be effectively used to offer personalized and explainable self-management and behaviour modification resources to patients to reduce the burden of COPD, especially the prevention of acute COPD exacerbations. The functionalities of COPD specific digital therapeutics for self-management need to be grounded in clinical evidence and behavioral theories, in keeping with the self-management needs of COPD patients and their care providers. In this paper, we report the functionalities of a COPD digital therapeutic mobile application based on a needs analysis qualitative study involving both COPD patients and physicians, and, based on the study's finding, we present a knowledge-driven digital therapeutic for COPD self-management.

Predicting Urgent Dialysis at Ambulance Transport to the Emergency Department Using Machine Learning Methods.

Hemodialysis patients frequently require ambulance transport to the hospital for dialysis. Some patients require urgent dialysis (UD) within 24 hours of transport to hospital to avoid morbidity and mortality. UD is not available in all hospitals; therefore, predicting patients who need UD prior to hospital transport can help paramedics with destination planning. In this paper, we developed machine learning models for paramedics to predict whether a patient needs UD based on patient characteristics available at the time of ambulance transport. This paper presented a study based on ambulance data collected in Halifax, Canada. Given that relatively few patients need UD, a class imbalance problem is addressed by up-sampling methods and prediction models are developed using multiple machine learning methods. The achieved prediction scores are F1-score=0.76, sensitivity=0.76, and specificity=0.97, confirming that models can predict UD with limited patient characteristics.

Characterizing Cluster-Based Frailty Phenotypes in a Multicenter Prospective Cohort of Kidney Transplant Candidates.

Frailty is associated with a higher risk of death among kidney transplant candidates. Currently available frailty indices are often based on clinical impression, physical exam or an accumulation of deficits across domains of health. In this paper we investigate a clustering based approach that partitions the data based on similarities between individuals to generate phenotypes of kidney transplant candidates. We analyzed a multicenter cohort that included several features typically used to determine an individual's level of frailty. We present a clustering based phenotyping approach, where we investigated two clustering approaches-i.e. neural network based Self-Organizing Maps (SOM) with hierarchical clustering, and KAMILA (KAy-means for MIxed LArge data sets). Our clustering results partition the individuals across 3 distinct clusters. Clusters were used to generate and study feature-level phenotypes of each group.

Ensemble Clustering to Generate Phenotypes of Kidney Transplant Donors and Recipients.

In this paper we investigate the generation of phenotypes for kidney transplant donors and recipients to assist with decision making around organ allocation. We present an ensemble clustering approach for multi-type data (numerical and categorical) using two different clustering approaches-i.e., model based and vector quantization based clustering. These clustering approaches were applied to a large, US national deceased donor kidney transplant recipient database to characterize members of each cluster (in an unsupervised fashion) and to determine whether the subsequent risk of graft failure differed for each cluster. We generated three distinct clusters of recipients, which were subsequently used to generate phenotypes. Each cluster phenotype had recipients with varying clinical features, and the risk of kidney transplant graft failure and mortality differed across clusters. Importantly, the clustering results by both approaches demonstrated a significant overlap. Utilization of two distinct clustering approaches may be a novel way to validate unsupervised clustering techniques and clustering can be used for organ allocation decision making on the basis of differential outcomes.

Unveiling the silent threat of new onset atrial fibrillation in covid-19 hospitalized patients: A retrospective cohort study.

BACKGROUND: COVID-19, a highly infectious respiratory disease, has been associated with a range of cardiovascular complications. One of the most commonly reported cardiovascular issues in COVID-19 patients is the development of arrhythmias. Among all types of arrhythmias, atrial fibrillation is the most frequently observed. Atrial fibrillation is characterized by an irregular and often rapid heartbeat, and it can be a serious and potentially life-threatening condition. OBJECTIVE: To investigate the incidence and association of new onset atrial fibrillation in COVID-19 hospitalized patients and its impact on survival. METHOD: A retrospective cross-sectional study that encompassed all patients, both positive and negative for COVID-19, who were consecutively admitted to the Aga Khan University Hospital in Karachi, a tertiary care facility, from June 2021 to December 2021. RESULTS: A total of 1,313 patients who met the inclusion criteria of our study were enrolled as participants. These patients were then stratified into two groups based on COVID-19 status: the study group (COVID-19 positive) comprised 626 (47.7%) patients and the control group (COVID-19 negative) consisted of 687 (52.3%) patients. The incidence of new-onset atrial fibrillation was 85 (13.6%) in COVID-19 positive compared to 43 (5.2%) in COVID-19 negative group. The study found a strong association between COVID-19 and new-onset atrial fibrillation in both univariate (unadjusted odd ratio 2.35 [95% CI, 1.60-3.45], p-value < 0.01) and a multiple-adjusted regression analysis (adjusted odd ratio 3.86 [95% CI, 2.31-6.44], p-value < 0.01). CONCLUSION: These findings highlight the importance of vigilant monitoring of cardiovascular complications in COVID-19 patients, especially those with pre-existing conditions that predispose them to the development of atrial fibrillation. The study underscores the need for prompt recognition and management of new onset atrial fibrillation in COVID-19 patients, as this may mitigate the risk of adverse outcomes and improve overall prognosis.

Tracking the Variations in Trace Elements, Some Nutrients, Phenolics, and Anthocyanins in Grewia asiatica L. (Phalsa) at Different Fruit Development Stages.

Grewia asiatica L. (phalsa) is a very prevalent berry in Pakistan and is consumed extensively as raw or in the form of juice. Here, for the first time, we assessed phalsa from Pakistan in terms of variations in macro and micro minerals, nutrients, and bio-active phyto-constituents including total phenolic and anthocyanin contents at different fruit developmental stages. It was found that the sugars in phalsa increased from D1 (small at the initial fruit setting stage) to D6 development stage (fully ripened fruit) where sugars at D5 (near to fully ripe) and D6 stages were many times greater than at D1, D2 (unripe close to full-size completion), D3 (close to semi ripe), and D4 stage (semi ripened and full-size attainment). Total acidity of was declined in all developmental stages, where the D1 stage displayed maximum and D6 with the lowest acidity. Ascorbic acid was decreased from D1 to D2 and then increased gradually from D3 to D5 stages. At the D6 stage, again a steep decline in ascorbic acid was observed. The total phenolics (mg gallic acid equivalents/100g) at stage D6 were higher (136.02 ± 1.17), whereas D1 being the lowermost in total phenolic content (79.89 ± 1.72). For anthocyanins (mg/100g), an increasing pattern of changes was observed in all stages of phalsa fruit where the D1 stage showed lower (13.97 ± 4.84) anthocyanin contents which then increased gradually at stage D2 (67.79 ± 6.73), but increased sharply at D3 (199.66 ± 4.90), D4 (211.02 ± 18.85), D5 (328.41 ±14.96) and D6 (532.30 ± 8.51) stages. A total of four anthocyanins such as cyanidin, delphidine-3-glucoside, pelargonidin, and malvidin in phalsa were identified using HPLC procedures, and a significant > 90 % DPPH inhibition in phalsa was observed at the D5 and D6 development stages. The macro and micro minerals including Ni, Zn, Fe, Ca, Cu, Mg, Na, P, and K contents were decreased from initial (D1) stage to the final (D6) development stage, while only Fe displayed an increasing trend from the initial to final fruit development stages (D1-D6). Conclusively, these findings could be of great interest for patients who are intended to consume phalsa as adjuvant therapy against diabetes and metabolic syndromes and other diseases involving reactive oxygen species with minimum metal toxicity.

Spatial Hotspots and Sociodemographic Profiles Associated With Traumatic Brain Injury in Nova Scotia.

Traumatic brain injury (TBI) is a leading cause of death and disability, primarily caused by falls and motor vehicle collisions (MVCs). Although many TBIs are preventable, there is a notable lack of studies exploring the association of geographically defined TBI hotspots with social deprivation. Geographic information systems (GIS) can be used to identify at-risk neighborhoods (hotspots) for targeted interventions. This study aims to determine the spatial distribution of TBI by major causes and to explore the sociodemographic and economic characteristics of TBI hotspots and cold spots in Nova Scotia. Patient data for TBIs from 2003 to 2019 were obtained from the Nova Scotia Trauma Registry. Residential postal codes were geocoded and assigned to dissemination areas (DA). Area-based risk factors and deprivation status (residential instability [RI], economic dependency [ED], ethnocultural composition [EC], and situational vulnerability [SV]) from the national census data were linked to DAs. Spatial autocorrelation was assessed using Moran's I, and hotspot analysis was performed using Getis-Ord Gi* statistic. Differences in risk factors between hot and cold spots were evaluated using the Mann-Whitney U test for numerical variables and the χ2 test or Fisher's exact test for categorical variables. A total of 5394 TBI patients were eligible for inclusion in the study. The distribution of hotspots for falls exhibited no significant difference between urban and rural areas (p = 0.71). Conversely, hotspots related to violence were predominantly urban (p = 0.001), whereas hotspots for MVCs were mostly rural (p < 0.001). Distinct dimensions of deprivation were associated with falls, MVCs, and violent hotspots. Fall hotspots were significantly associated with areas characterized by higher RI (p < 0.001) and greater ethnocultural diversity (p < 0.001). Conversely, the same domains exhibited an inverse relationship with MVC hotspots; areas with low RI and ethnic homogeneity displayed a higher proportion of MVC hotspots. ED and SV exhibited a strong gradient with MVC hotspots; the most deprived quintiles displayed the highest proportion of MVC hotspots compared with cold spots (ED; p = 0.002, SV; p < 0.001). Areas with the highest levels of ethnocultural diversity were found to have a significantly higher proportion of violence-related hotspots than cold spots (p = 0.005). This study offers two significant contributions to spatial epidemiology. First, it demonstrates the distribution of TBI hotspots by major injury causes using the smallest available geographical unit. Second, we disentangle the various pathways through which deprivation impacts the risk of main mechanisms of TBI. These findings provide valuable insights for public health officials to design targeted injury prevention strategies in high-risk areas.

Humoral and T cell responses to SARS-CoV-2 reveal insights into immunity during the early pandemic period in Pakistan.

BACKGROUND: Protection against SARS-CoV-2 is mediated by humoral and T cell responses. Pakistan faced relatively low morbidity and mortality from COVID-19 through the pandemic. To examine the role of prior immunity in the population, we studied IgG antibody response levels, virus neutralizing activity and T cell reactivity to Spike protein in a healthy control group (HG) as compared with COVID-19 cases and individuals from the pre-pandemic period (PP). METHODS: HG and COVID-19 participants were recruited between October 2020 and May 2021. Pre-pandemic sera was collected before 2018. IgG antibodies against Spike and its Receptor Binding Domain (RBD) were determined by ELISA. Virus neutralization activity was determined using a PCR-based micro-neutralization assay. T cell - IFN-γ activation was assessed by ELISpot. RESULTS: Overall, the magnitude of anti-Spike IgG antibody levels as well as seropositivity was greatest in COVID-19 cases (90%) as compared with HG (39.8%) and PP (12.2%). During the study period, Pakistan experienced three COVID-19 waves. We observed that IgG seropositivity to Spike in HG increased from 10.3 to 83.5% during the study, whilst seropositivity to RBD increased from 7.5 to 33.3%. IgG antibodies to Spike and RBD were correlated positively in all three study groups. Virus neutralizing activity was identified in sera of COVID-19, HG and PP. Spike reactive T cells were present in COVID-19, HG and PP groups. Individuals with reactive T cells included those with and without IgG antibodies to Spike. CONCLUSIONS: Antibody and T cell responses to Spike protein in individuals from the pre-pandemic period suggest prior immunity against SARS-CoV-2, most likely from cross-reactive responses. The rising seroprevalence observed in healthy individuals through the pandemic without known COVID-19 may be due to the activation of adaptive immunity from cross-reactive memory B and T cells. This may explain the more favourable COVID-19 outcomes observed in this population.

Characterization of HIV-1 CRF02_AG/A3/G unique recombinant forms identified among children in Larkana, Pakistan.

Co-circulation of different human immunodeficiency virus type 1 HIV-1 subtypes among infected populations can lead to the generation of new recombinants. In Pakistan, subtype A1 and CRF02_AG are the dominant strains circulating among key populations. The high prevalence of new HIV infections among the key populations highlights the possibility of recombination between the dominant strains, which can lead to the generation of new recombinants. Here, we identified a recombinant cluster composed of CRF02_AG, sub-subtype A3, and subtype G among HIV-infected children in Larkana. For the study, 10 retrospectively collected samples, with recombination signals in the pol gene, were used to perform a near full-length genome NFLG sequencing. Of the 10 samples, NFLG was successfully sequenced from seven samples. Phylogenetic analysis of the seven NFLGs showed that all recombinants formed a distinct monophyletic cluster and were distinct from known HIV-1 circulating recombinant forms CRFs. Recombination analyses showed that all seven NFLGs shared a similar recombinant structure consisting of CRF02_AG, sub-subtype A3, and subtype G, with a sub-subtype A3 fragment inserted into pol and vif regions spanning from (HXB2: 4218-5518), and a subtype G fragment inserted into vpu, rev, tat and env regions spanning from (HXB2: 5957-8250) of the CRF02_AG backbone. The identification of unique recombinant forms may indicate the presence and transmission of several co-circulating lineages in Larkana, giving rise to newer CRFs. This study also highlights the importance of continuous molecular surveillance to fully understand HIV-1 genetic diversity in Pakistan, particularly in Larkana, which is the epicenter of HIV outbreaks.

Impact of Treviamet® & Treviamet XR® on quality of life besides glycemic control in type 2 DM patients.

BACKGROUND: Maintaining the quality of life is the main objective of managing type 2 diabetes (T2DM) (QoL). Since it is a key factor in patient motivation and adherence, treatment-related QoL has always been considered when choosing glucose-lowering medicines. The objective of the study was to evaluate the quality of life besides glycemic control among type 2 diabetes mellitus patients receiving Treviamet® & Treviamet XR® (Sitagliptin with Metformin) in routine care. METHODS: It was a prospective, open-label, non-randomized clinical trial including T2DM patients uncontrolled on Metformin therapy. All patients received Treviamet® & Treviamet XR® for six months. Sequential changes in QoL, fasting plasma glucose, HbA1c, body weight, and blood pressure were monitored from baseline to 3 consecutive follow-up visits. The frequency of adverse events (AEs) was also noted throughout the study. RESULTS: A total of 504 patients were screened; 188 completed all three follow-ups. The mean QoL score significantly declined from 57.09% at baseline to 33.64% at the 3rd follow-up visit (p < 0.01). Moreover, a significant decline in mean HbA1c and FPG levels was observed from baseline to 3rd follow-up visit (p < 0.01). Minor adverse events were observed, including abdominal discomfort, nausea, flatulence, and indigestion. Gender, HbA1c, diarrhea, and abdominal discomfort were significant predictors of a patient's QoL, as revealed by the Linear Regression Model (R2 = 0.265, F(16, 99) = 2.231). CONCLUSION: Treviamet® & Treviamet XR® significantly improved glycemic control (HbA1c levels) and QoL in T2DM patients without serious adverse events. TRIAL REGISTRATION: ClinicalTrials.gov identifier (NCT05167513), Date of registration: December 22, 2021.

Epidemiology Of Male Urethral Strictures In Pakistan.

Urethral stricture disease is relatively common in Pakistan, constituting 4-5% of the urological workload. Despite the high prevalence, little is known about its epidemiology in the country. The current narrative review comprised search on PubMed, Pak MediNet and Google Scholar databases for studies done in Pakistan and published between January 1, 2000, and December 31, 2021. The search yielded 30 local publications on stricture urethra. Demographic data as well as causes and management pattern of male urethral stricture were noted and analysed. There were 5,021 patients, with 3850 (76.6%) being from the province of Sindh. The disease had the greatest impact on younger patients aged up to 40 years (n=1572), while after the age of 60 years, 248 (9%) patients had the disease. The common cause was trauma due to road traffic accidents in both anterior and posterior strictures compared to idiopathic cause reported in the West. Infection 170 (6.9%) and Lichen sclerosis 123(4.5%) as a cause was found to decline in our region. A clinic-based regular urethral dilatation was still in practice at some centres to manage such cases. Vast majority of stricture patients were being treated by endoscopic procedures, and only 1154 (23%) cases underwent urethroplasty.

Acute Porphyria: An Unusual Case Of Quadriparesis, Hypertension, Recurrent Severe Cyclic Abdominal Pain, And Seizures.

Porphyria refers to a rare group of genetically inherited or acquired disorders that arise due to reduced metabolic activity of any of the enzymes in the haem biosynthetic pathway. Defect in any enzyme causes the presentation of symptoms of porphyria. The epidemiology of Acute Intermittent Porphyria (AIP) is complicated because of its rarity and delay in diagnosis. We present the case of a seven-year-old girl who presented with multisystem involvement; her symptoms were quadriparesis, hypertension, recurrent severe cyclic abdominal pain, and seizures. These symptoms together were not explained by the differentials taken into account. She presented before puberty with no family history of such conditions, while being born of consanguineous marriage. Her symptoms along with urinary porphobilinogen positivity test helped to reach the diagnosis of AIP in the absence of cutaneous manifestations. This case highlights the variable presentation of porphyria and emphasises the importance of appropriate and timely diagnosis and management in these patients.