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This article investigates the intricate associations between apolipoprotein E (APOE) gene alleles variation, insulin resistance (IR) and amyloid-β aggregation in cardiovascular disease (CVD) patients. A cohort of 250 patients exhibiting the symptoms of CVD and 50 control subjects participated in this study. After applying the stringent inclusion and exclusion criteria, the diseased group was further stratified into three categories: CVD+ (Alzheimer's disease) AD, CVD + (diabetes mellitus) DM and CVD + DM + AD. Blood samples were collected from all recruited participants for the biochemical analyses of lipid profile, glycaemic status, liver function enzymes, inflammatory and oxidative stress biomarkers. Tetra amplification-refractory mutation system-polymerase chain reaction (ARMS-PCR) was employed for APOE gene analysis. Biochemical evaluations revealed the significant elevations in the serum levels of glucose, liver enzymes, interleukin-6 (IL-6), cholesterol, low-density lipoproteins (LDL), triglycerides (TG) and malondialdehyde (MDA) in CVD + DM + AD group. Conversely, the serum levels of insulin, HDL and hexokinase decreased in CVD + DM + AD group compared to the controls and other CVD groups. Tetra ARMS-PCR results indicated a higher percentage of the risk allele in CVD + DM + AD group when compared with the other groups. Our study elucidates the multifaceted cardiovascular risk factors contributing to IR and AD in CVD patients. Age-related risk factors, prevalence of APOE risk alleles and the impact of statin use on AD incidences were identified. These findings underscore the need for tailored preventive measures, particularly in APOEε4 and ε3/ε4 carriers with CVD. Further studies should delve into the knowledge-based protocols to comprehend the underlying mechanisms. Focusing on the therapeutic targets to prevent or delay DM and AD progression in CVDs, especially in APOEε4 carriers, is essential., (© 2024 John Wiley & Sons Australia, Ltd.)

One of the venereal tumors that threaten human life is cervical cancer. A2ML1 is detected in advanced-stage cancer patients and is found to be strongly associated with cervical cancer. A2ML1 was shown to be substantially expressed in cervical cancer in this study, which used data from the TCGA database. Those with high A2ML1 expression had a lower chance of survival than patients with low A2ML1 expression. Both univariate and multivariate Cox regression analyses were utilized to investigate the relationship between clinical variables and overall survival rates. An investigation into the link between A2ML1 and immune infiltration was subsequently conducted. Utilizing the immune cell database, research was conducted to investigate the dispersion of 24 immune cells and their correlation to A2ML1 expression. In addition to this, the favorable correlation between immune cells and A2ML1 was validated using all three immune cell methodologies. The Genomics of Drug Sensitivity in Cancer database was used to confirm the idea that there is a link between A2ML1 expression and the efficacy of chemotherapy or immunotherapy. The findings demonstrated that A2ML1 is a potential biomarker for cervical cancer diagnostics. This biomarker may be used to chaperone immunotherapy, as well as to explain the elucidates of cervical cancer caused by the immunological microenvironment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

As newly recognized environmental pollutants, microplastics (MPs, ≤5 mm in length) have been reported in various human tissues and fluids, including the spleen, liver, heart, blood and blood clots, raising global concerns about their impact on human health. This study investigated the characteristics of MPs in intravenous infusion and the removal of MPs from infusion products by infusion sets fitted with different filters using micro-Fourier Transform Infrared Spectroscopy. MPs were detected in infusion products, with an average abundance of 1.24 ± 1.44 items/unit (2.91 ± 3.91 items/L). The primary types of MPs identified were fragmented particles of polyethene and polypropylene, ranging in size from 15-100 µm. Internal filters in infusion sets played a crucial role in removing MPs, particularly fibrous ones, resulting in a reduction in both abundance and particle size of MPs in the human body. Moreover, this study conducted a general assessment of intravenous microplastic exposure among hospital patients and estimated the global per-person input of MPs via intravenous administration. It is an opportunity for us to gain a deeper understanding of MPs in intravenous infusion and provides guides selecting infusion devices, increasing awareness of associated health risks., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Marine microplastics (MPs) are recognized as a growing severe environmental concern. The vertical distribution pattern of MPs in the ocean is still elusive. Meanwhile, different sampling methods have been deployed in previous studies, resulting in difficulties in compiling data. In this study, for the first time, we explored ocean interior MP pollution in the Western Pacific Warm Pool simultaneously using both a CTD (Conductivity-temperature-depth) sampler and a large-volume in-situ filtration system. At the same sampling station, the average abundance of microplastics in the water column obtained by the two sampling methods was 0.37 ± 0.44 n/m 3 (in-situ filtration) and 115.12 ± 64.13 n/m 3 (CTD), respectively, which showed significant differences. Both methods found that the main chemical composition and shape of MPs were PET and fiber. Ocean current was identified as the dominant factor that impacted the horizontal distribution of MPs in the study area. The abundance of MPs in the surface layer was 5.4-703.8 times higher than that of the water column. The similar physical and chemical properties of MPs in the surface water and water column indicated that MPs in the water column originate from the sustained release from the surface layer., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ultrasound imaging is frequently employed to aid with fetal development. It benefits from being real-time, inexpensive, non-intrusive, and simple. Artificial intelligence is becoming increasingly significant in medical imaging and can assist in resolving many problems related to the classification of fetal organs. Processing fetal ultrasound (US) images increasingly uses deep learning (DL) techniques. This paper aims to assess the development of existing DL classification systems for use in a real maternal-fetal healthcare setting. This experimental process has employed two publicly available datasets, such as FPSU23 Dataset and Fetal Imaging. Two novel deep learning architectures have been designed in the proposed architecture based on 3-residual and 4-residual blocks with different convolutional filter sizes. The hyperparameters of the proposed architectures were initialized through Bayesian Optimization. Following the training process, deep features were extracted from the average pooling layers of both models. In a subsequent step, the features from both models were optimized using an improved version of the Generalized Normal Distribution Optimizer (GNDO). Finally, neural networks are used to classify the fused optimized features of both models, which were first combined using a new fusion technique. The best classification scores, 98.5 and 88.6% accuracy, were obtained after multiple steps of analysis. Additionally, a comparison with existing state-of-the-art methods revealed a notable improvement in the suggested architecture's accuracy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Rauf, Attique Khan, Albarakati, Jabeen, Alsenan, Hamza, Teng and Nam.)

Background: Linezolid-resistant Enterococcus faecium (LRE) is a global priority pathogen. Thirteen LRE were reported from clinical specimens between November 2021 and April 2023 at two laboratories in Karachi, Pakistan. We aimed to investigate the strain types and genes associated with linezolid resistance among these isolates. Whole genome sequencing (WGS) was performed and analyzed by multilocus sequence typing (MLST). The presence of linezolid resistance genes was identified using ResFinder v4.1.11 and the LRE-finder tool., Results: Twelve isolates belonged to clonal complex 17 (CC17); ST80 (n = 10), ST612 (n = 1) and ST1380 (n = 1). Six isolates showed the presence of optrA gene and G2576T mutations in the 23S rRNA gene, while six showed poxtA and cfr(D) genes. One isolate showed the combination of optrA, cfr(D) and poxtA genes., Conclusion: Our findings show the circulation of CC17 sequence types with a known outbreak potential and we identified molecular mechanisms of resistance that were not previously reported from Pakistan., (© 2024. The Author(s).)

Background: Oxidative stress mediated by reactive oxygen species (ROS) is a common denominator in arsenic toxicity. Arsenic stress in soil affects the water absorption, decrease stomatal conductance, reduction in osmotic, and leaf water potential, which restrict water uptake and osmotic stress in plants. Arsenic-induced osmotic stress triggers the overproduction of ROS, which causes a number of germination, physiological, biochemical, and antioxidant alterations. Antioxidants with potential to reduce ROS levels ameliorate the arsenic-induced lesions. Plant growth promoting rhizobacteria (PGPR) increase the total soluble sugars and proline, which scavenging OH radicals thereby prevent the oxidative damages cause by ROS. The main objective of this study was to evaluate the potential role of Arsenic resistant PGPR in growth of maize by mitigating arsenic stress., Methodology: Arsenic tolerant PGPR strain MD3 (Pseudochrobactrum asaccharolyticum) was used to dismiss the 'As' induced oxidative stress in maize grown at concentrations of 50 and 100 mg/kg. Previously isolated arsenic tolerant bacterial strain MD3 "Pseudochrobactrum asaccharolyticum was used for this experiment. Further, growth promoting potential of MD3 was done by germination and physio-biochemical analysis of maize seeds. Experimental units were arranged in Completely Randomized Design (CRD). A total of 6 sets of treatments viz., control, arsenic treated (50 & 100 mg/kg), bacterial inoculated (MD3), and arsenic stress plus bacterial inoculated with three replicates were used for Petri plates and pot experiments. After treating with this MD3 strain, seeds of corn were grown in pots filled with or without 50 mg/kg and 100 mg/kg sodium arsenate., Results: The plants under arsenic stress (100 mg/kg) decreased the osmotic potential (0.8 MPa) as compared to control indicated the osmotic stress, which caused the reduction in growth, physiological parameters, proline accumulation, alteration in antioxidant enzymes (Superoxide dismutase-SOD, catalase-CAT, peroxidase-POD), increased MDA content, and H 2 O 2 in maize plants. As-tolerant Pseudochrobactrum asaccharolyticum improved the plant growth by reducing the oxidation stress and antioxidant enzymes by proline accumulation. PCA analysis revealed that all six treatments scattered differently across the PC1 and PC2, having 85.51% and 9.72% data variance, respectively. This indicating the efficiency of As-tolerant strains. The heatmap supported the As-tolerant strains were positively correlated with growth parameters and physiological activities of the maize plants., Conclusion: This study concluded that Pseudochrobactrum asaccharolyticum reduced the 'As' toxicity in maize plant through the augmentation of the antioxidant defense system. Thus, MD3 (Pseudochrobactrum asaccharolyticum) strain can be considered as bio-fertilizer., (© 2024. The Author(s).)

Background: Chromoblastomycosis (CBM) is a chronic infection of skin and subcutaneous tissue. CBM cases have been reported in local literature from Pakistan with heterogenous demographic, diagnostic and therapeutic information. The objective of this study is to share the experience of CBM from a large tertiary care hospital laboratory in Pakistan., Method: This was a retrospective observational study. Histopathology and microbiology data of suspected CBM between 2016 and 2022 was retrieved. Patients' demographics, site of involvement, histopathological findings and positive microbiology cultures were assessed. Literature search on Google Scholar, PubMed and PakMediNet was done between 1990 and 2023 with multiple terms., Result: A total of 16 CBM cases were identified; 14 were histopathology positive and two were both histopathology and culture positive. The median age was 21 years, and 11 patients were male. The predominant site was lower extremities followed by the face. Severe acanthosis, hyperkeratosis and granuloma with sclerotic bodies were observed in all histopathology slides. Alternaria spp. and Phialophora spp. were isolated from two culture-positive cases. A total of nine cases of CBM were reported from Pakistan in PubMed non-indexed journal., Conclusion: CBM is not a commonly thought of disease when evaluating skin lesions in Pakistan. A high index of suspicion when assessing patients who may have a history of trauma, exposure to soil and suggestive lesions is reasonable. An integrated approach between clinicians, histopathologist and microbiologist is required to do early identification and therapeutic interventions., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Candida species are amongst the commensals of the mucosal surfaces of the human body which include the oral cavity, vagina, and intestinal mucosa. Fungal infections are on the rise worldwide. The overall burden of infections due to fungi is difficult to estimate because the majority of them remain undiagnosed. The present study aims to determine the burden of antifungal resistance in low socioeconomic country, Pakistan and the frequency of ERG11 and MDR1 genes involved. A total of 636 Candida isolates were obtained from various tertiary care institutions in Lahore in the form of culture on various culture plates. Sabouraud agar culture plates were used to culture the Candida spp. Antifungal resistance was determined against Fluconazole, Itraconazole, Ketoconazole, and Nystatin via disk diffusion technique. Most resistance was observed against Fluconazole followed by Itraconazole, Ketoconazole, and Nystatin. The Candida isolates recovering from CVP tip and tissue have a high resistance profile. Candida species resistant to at least two antifungals were chosen for further ERG11 and MDR1 detection through real-time PCR. Among 255 Candida isolates, 240 contained ERG11 gene while MDR1 gene is present in 149 Candida isolates. The isolates carrying both genes were tested by the broth microdilution technique for the susceptibility against cycloheximide, all of them were able to grow in cycloheximide. The genetic determinants of antifungal resistance such as ERG11 and MDR1 are as important in the multidrug resistance against a variety of compounds and antifungal drugs., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Background: Pneumocystis jirovecii is the most emerging life-threating health problem that causes acute and fatal pneumonia infection. It is rare and more contagious for patients with leukemia and immune-deficiency disorders. Until now there is no treatment available for this infection therefore, it is needed to develop any treatment against this pathogen., Methods: In this work, we used comparative proteomics, robust immune-informatics, and reverse vaccinology to create an mRNA vaccine against Pneumocystis jirovecii by targeting outer and transmembrane proteins. Using a comparative subtractive proteomic analysis of two Pneumocystis jirovecii proteomes, a distinct non-redundant Pneumocystis jirovecii (strain SE8) proteome was chosen. Seven Pneumocystis jirovecii transmembrane proteins were chosen from this proteome based on hydrophilicity, essentiality, virulence, antigenicity, pathway interaction, protein-protein network analysis, and allergenicity., Objective: The reverse vaccinology approach was used to predict the immunogenic and antigenic epitopes of major histocompatibility complex (MHC) I, II and B-cells from the selected proteins on the basis of their antigenicity, toxicity and allergenicity. These immunogenic epitopes were linked together to construct the mRNA-based vaccine. To enhance the immunogenicity, suitable adjuvant, linkers (GPGPG, KK, and CYY), and PRDRE sequences were used., Results: Through predictive modeling and confirmation via the Ramachandran plot, we assessed secondary and 3D structures. The adjuvant RpfE was incorporated to enhance the vaccine construct's immunogenicity (GRAVY index: -0.271, instability index: 39.53, antigenicity: 1.0428). The physiochemical profiling of vaccine construct was predicted it an antigenic, efficient, and potential vaccine. Notably, strong interactions were observed between the vaccine construct and TLR-3/TLR-4 (-1301.7 kcal/mol-1 and -1374.7 kcal/mol-1)., Conclusions: The results predicted that mRNA-based vaccines trigger a cellular and humoral immune response, making the vaccine potential candidate against Pneumocystis jirovecii and it is more suitable for in-vitro analysis and validation to prove its effectiveness., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Introduction: Salmonella typhi , a gram-negative bacterium responsible for typhoid fever, can infect the inner lining or valves of the heart and cause endocarditis. This systematic review aimed to report cases of S. typhi -associated endocarditis and its clinical features., Methods: This systematic review was reported as per the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) checklist. Only case reports and case series of endocarditis caused by S. typhi , irrespective of age, gender, and demographics, were considered eligible for inclusion. To identify relevant studies, a literature search was conducted using relevant keywords on PubMed, Google Scholar, and the Cochrane Library from inception to 31 December 2023. After selecting the studies, the relevant data were extracted and pooled in terms of frequencies and percentages. A quality assessment was performed using the Joanna Briggs Institute Critical Appraisal Checklist for Case Reports., Results: This review included seven case reports, comprising 22.2% female and 77.8% male patients. The mean age of patients was 27.9 + 12.0 years. Regarding past medical history, 33.3% (3/9) of patients had a previous cardiac pathology. Fever remained the most common complaint, occurring in 88.9% of cases. Transthoracic and transesophageal echocardiography were used to diagnose all cases, with 33.3% identifying vegetation on the mitral, aortic, and tricuspid valves. Ceftriaxone, with or without gentamycin, remained the choice of antibiotic for 88.9% of cases, and all patients responded to the offered treatment., Conclusion: S. typhi -associated endocarditis, though rare, presents unique challenges and requires timely diagnosis. This systematic review of seven cases highlights a predominantly male population affected, with a mean age in the third decade, suggesting a higher invasiveness than other causes. The findings from this study underscore the importance of early recognition and appropriate management, primarily with antibiotic therapy. Further research with larger cohorts is crucial to refine understanding and guide policymaking for this rare but life-threatening condition., Competing Interests: MA was employed by AdventHealth. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Jabeen, Bint Ali, Tufail, Mustafa, Chaudhry, Tahir, Asghar and Ahmed.)

This study aimed to investigate alterations in the expression of four key cytokines (IL-7, IL-11, IL-15, and IL-27) and assess differential FAM26F expression in response to Hepatitis C virus (HCV) infection. Additionally, it sought to analyze changes in these cytokines after treatment in 244 chronic HCV patients and 28 controls undergoing various treatments, including standard interferon, pegylated interferon, and Direct Acting Antivirals (DAAs). The objective was to compare immune system regulation between treatment groups. The expression levels of FAM26F and the cytokines (IL-7, IL-11, IL-15, and IL-27) were evaluated using Real-time qPCR in PBMCs of treatment groups. Results revealed significant downregulation of IL-7 and IL-27 in infected individuals compared to healthy controls, persisting even after treatment. This suggests the crucial roles of these immune modulators in facilitating the necessary T-cell response for viral clearance. IL-11 expression also remained suppressed post-treatment, supporting viral clearance by restoring the Th1 response. The decrease in IL-11 levels during treatment indicates the restoration of the Th1 response, vital for viral clearance. IL-15, the key cytokine regulating cytotoxic cells (NKT and NK cells), displayed consistent expression across all sample groups, indicating maintained IL-15-induced cytotoxicity in both control and infected individuals. Additionally, FAM26F expression was reduced in the HCV-infected group compared to controls, but higher in HCV-recovered cases, potentially due to reduced infection and enhanced immunity. In conclusion, this research unveils the relationship between FAM26F and HCV infection, highlighting the virus's tendency to suppress cytokine and FAM26F expression. An effective treatment strategy for establishing an ideal host immune response may involve restoring FAM26F and cytokine expression to their normal levels., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Jabeen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Listeria monocytogenes , a Gram-positive bacterium, is a prominent foodborne pathogen that causes listeriosis and poses substantial health hazards worldwide. The continuing risk of listeriosis outbreaks underlies the importance of designing an effective prevention strategy and developing a robust immune response by reverse vaccinology approaches. This study aimed to provide a critical approach for developing a potent multiepitope vaccine against this foodborne disease., Methods: A chimeric peptide construct containing 5 B-cell epitopes, 16 major histocompatibility complex I (MHC-I) epitopes, and 18 MHC-II epitopes were used to create a subunit vaccination against L. monocytogenes . The vaccine safety was evaluated by several online methods, and molecular docking was performed using ClusPro to determine the binding affinity. Immune simulation was performed using the C-ImmSimm server to demonstrate the immune response., Results: The results validated the antigenicity, non-allergenicity, and nontoxicity of the chimeric peptide construct, confirming its suitability as a subunit vaccine. Molecular docking showed a good score of 1276.5 and molecular dynamics simulations confirmed the construct's efficacy, demonstrating its promise as a good candidate for listeriosis prophylaxis. The population coverage was as high as 91.04% with a good immune response, indicating good antigen presentation with dendritic cells and production of memory cells., Conclusions: The findings of this study highlight the potential of the designed chimeric peptide construct as an effective subunit vaccine against Listeria , paving the way for future advances in preventive methods and vaccine design., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Introduction and Objective: Pulmonary mucormycosis is a rare but rapidly progressive fatal disease. Limited data exist on the outcomes and factors associated with poor prognosis of pulmonary mucormycosis. The objective of this study was to evaluate clinical characteristics, factors associated with mortality, and outcomes of pulmonary mucormycosis at a tertiary care hospital in Pakistan., Methods: This was a retrospective observational study conducted at a tertiary care hospital in Karachi, Pakistan. Medical records of hospitalized patients diagnosed with proven or probable pulmonary mucormycosis between January 2018 and December 2022 were reviewed. Univariate and regression analyses were performed to identify factors associated with mortality., Results: Fifty-three pulmonary mucormycosis patients (69.8% male) were included, with mean age of 51.19 ± 21.65 years. Diabetes mellitus was the most common comorbidity [ n  = 26 (49.1%)]. Chronic lung diseases were present in [ n  = 5 (9.4%)], and [ n  = 16 (30.2%)] had concurrent coronavirus disease 2019 (COVID-19) pneumonia. The predominant isolated Mucorales were Rhizopus [ n  = 32 (60.3%)] and Mucor species [ n  = 9 (17%)]. Main radiological findings included consolidation [ n  = 39 (73.6%)] and nodules [ n  = 14 (26.4%)]. Amphotericin B deoxycholate was prescribed in [ n  = 38 (71.7%)], and [ n  = 14 (26.4%)] of patients received combined medical and surgical treatment. The median [interquartile range (IQR)] hospital stay was 15.0 (10.0-21.5) days. Intensive care unit (ICU) care was required in [ n  = 30 (56.6%)] patients, with 26 (49.1%) needing mechanical ventilation. Overall mortality was seen in 29 (54.7%) patients. Significantly higher mortality was found among patients requiring mechanical ventilation 20/29 (69%, p  = 0.002). Immunosuppression ( p  = 0.042), thrombocytopenia ( p  = 0.004), and mechanical ventilation ( p  = 0.018) were identified as risk factors for mortality on multivariable analysis., Conclusion: This study provides essential insights into the clinical characteristics, outcomes, and mortality factors associated with pulmonary mucormycosis. The mortality rate was high (54.7%), particularly in patients with immunosuppression, thrombocytopenia, and those who required mechanical ventilation., Competing Interests: The authors declare that there is no conflict of interest., (© The Author(s), 2024.)

In this study, we compared the predisposing factors, key demographic and clinical characteristics, clinical outcomes, and factors associated with poor prognosis in pneumocystis pneumonia (PCP) infection among the human immunodeficiency virus (HIV)-positive and non-HIV patient populations. This retrospective analysis was conducted at the Aga Khan University Hospital, Karachi, via the collection and analysis of patient records with a diagnosis of "pneumocystosis" between January 2015 and October 2020. Additionally, the laboratory database was evaluated, and patients with a laboratory-confirmed diagnosis of PCP were included. During the study period, 52 laboratory-confirmed hospitalized PCP patients were identified. Of these, 23 and 29 patients were diagnosed using microscopy and polymerase chain reaction, respectively. 34.6% of our patients were HIV positive, with a median CD4 count of 20.5 cells/mm3 (range: 10.7-50.5). Other conditions identified were corticosteroid use, autoimmune diseases, malignancy, radiation, and chemotherapy. On chest imaging, consolidation was found in 30%, ground-glass opacities in 24%, and nodular infiltrates in 20% of the cases. HIV-positive patients had a lower hemoglobin level and a higher level of β-D-glucan at the time of admission, whereas non-HIV patients were found to have more co-morbid conditions than HIV patients. We observed no difference in clinical outcomes between the two populations. Factors associated with a poor prognosis among our patients included concomitant infections at the time of diagnosis, the need for invasive mechanical ventilation, and a longer duration of stay in the hospital as well as the intensive care unit.

River systems are important recipients of environmental plastic pollution and have become key pathways for the transfer of mismanaged waste from the land to the ocean. Understanding the sources and fate of plastic debris, including plastic litter (>5 mm) and microplastics (MPs) (<5 mm), entering different riverine systems is essential to mitigate the ongoing environmental plastic pollution crisis. We comprehensively investigated the plastic pollution in the catchments of two rivers in the Yangtze River basin: an urban river, the Suzhou section of the Beijing-Hangzhou Grand Canal (SZ); and a pristine rural river, the Jingmen section of the Hanjiang River (JM). The abundance of plastic pollutants in SZ was significantly higher than in JM: 0.430 ± 0.450 items/m 3 and 0.003 ± 0.003 items/m 3 of plastic litter in the water; 23.47 ± 25.53 n/m 3 and 2.78 ± 1.55 n/m 3 MPs in the water; and 218.82 ± 77.40 items/kg and 5.30 ± 1.99 items/kg of MPs in the sediment, respectively. Plastic litter and MPs were closely correlated in abundance and polymer composition. Overall, the polymer type, shape and color of MPs were dominant by polypropylene (42.5%), fragment (60.4%) and transparent (40.0%), respectively. Source tracing analysis revealed that packaging, shipping, and wastewater were the primary sources of plastic pollutants. The mantel analysis indicated that socio-economic and geospatial factors play crucial roles in driving the hotspot formation of plastic pollution in river networks. The composition of the MP communities differed significantly between the sediments and the overlying water. The urban riverbed sediments had a more pronounced pollutant 'sink' effect compared with the pristine rivers. These findings suggested that the modification of natural streams during urbanization may influence the transport and fate of plastic pollutants in them. Our results offer pivotal insights into effective preventive measures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Contamination of the environment due to speedup of anthropogenic activities has become a serious threat to modern humanity. Among the contaminants, the new emerging concern is the heavy metal (HM) contamination in the environment. Because the persistence and harmfulness of heavy metals affect the ecosystem and the health of plants, animals, and humans, they are the most toxic substances in the environment. Among them, Arsenic (As) emerged as major environmental constraint leading to enormous negative effects on the plant, animal, and human health. Even in minute quantity, As is known to cause various critical diseases in humans and toxicity in plants. Research was performed to observe the capability of plant growth-promoting strains of bacteria in enhancing Zea mays (L.) growth in arsenic polluted soil. Total 30 bacterial strains were isolated from the polluted soils, screened for plant growth promotion potential and arsenic tolerance. Eighteen isolates showed resistance to different levels of sodium arsenate (ranging from 0 to 50 mM) in agar plate using LB media. Of 18 isolates, 83.3% produced IAA, methyl red, and hydrogen cyanide; 55.5% exhibited catalase activity; 61.1% showed siderophore production; 88.8% showed phosphate solubilization; and 44.4% showed oxidase, Voges proskauer activity, and KOH solubility. The most efficient isolates SR3, SD5, and MD3 with significant arsenic tolerance and plant growth-promoting (PGP) activity were examined via sequencing of amplified 16S rRNA gene. Isolates of bacteria, i.e., SR3, SD5, and MD3, showing multiple PGP-traits were identified as Bacillus pumilus (NCBI accession number: OR459628), Paenibacillus faecalis (NCBI accession number: OR461560), and Pseudochrobactrum asaccharolyticum (NCBI accession number: OR458922), respectively. Maize seeds treated with these PGPR strains were grown in pots contaminated with 50 ppm and 100 ppm sodium arsenate. Compared to untreated arsenic stressed plants, bacterial inoculation P. asaccharolyticum (MD3) resulted 20.54%, 18.55%, 33.45%, 45.08%, and 48.55% improvement of photosynthetic pigments (carotenoid content, chlorophyll content, stomatal conductance (gs), substomatal CO 2 , and photosynthetic rate), respectively. Principal component analysis explained that first two components were more than 96% of the variability for each tested parameter. The results indicate that in comparison to other isolates, P. asaccharolyticum isolate can be used as efficient agent for improving maize growth under arsenic polluted soil., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

With over 2.1 million new cases of breast cancer diagnosed annually, the incidence and mortality rate of this disease pose severe global health issues for women. Identifying the disease's influence is the only practical way to lessen it immediately. Numerous research works have developed automated methods using different medical imaging to identify BC. Still, the precision of each strategy differs based on the available resources, the issue's nature, and the dataset being used. We proposed a novel deep bottleneck convolutional neural network with a quantum optimization algorithm for breast cancer classification and diagnosis from mammogram images. Two novel deep architectures named three-residual blocks bottleneck and four-residual blocks bottle have been proposed with parallel and single paths. Bayesian Optimization (BO) has been employed to initialize hyperparameter values and train the architectures on the selected dataset. Deep features are extracted from the global average pool layer of both models. After that, a kernel-based canonical correlation analysis and entropy technique is proposed for the extracted deep features fusion. The fused feature set is further refined using an optimization technique named quantum generalized normal distribution optimization. The selected features are finally classified using several neural network classifiers, such as bi-layered and wide-neural networks. The experimental process was conducted on a publicly available mammogram imaging dataset named INbreast, and a maximum accuracy of 96.5% was obtained. Moreover, for the proposed method, the sensitivity rate is 96.45, the precision rate is 96.5, the F1 score value is 96.64, the MCC value is 92.97%, and the Kappa value is 92.97%, respectively. The proposed architectures are further utilized for the diagnosis process of infected regions. In addition, a detailed comparison has been conducted with a few recent techniques showing the proposed framework's higher accuracy and precision rate., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Jabeen, Khan, Hameed, Alqahtani, Alouane and Masood.)

Hybrid nanofluids (HNFs) of metallic oxide-based nanoparticles (NPs) have been prepared in different basefluids (BFs) employing the thermal plasma technique. NPs of ZnO-MgO were directly dispersed into pristine coolant, engine oil, distilled water (DW), and coconut oil. Plasma was generated between two identical electrodes applying 8.0 kV at the ambient conditions and proved economically viable in preparing stable HNFs. X-ray Diffractometry (XRD) showed ZnO and MgO NPs possessed hexagonal and cubic crystal structures, respectively. The band gap is calculated through UV-visible spectroscopy. The thermal conductivity (TC) of the HNFs has been measured using a thermal conductivity analyzer based on the transient hot wire method. The band gaps of pristine coolant and its HNFs were obtained to be 3.35 eV and 3.33 eV, respectively. In engine oil and its HNFs, band gaps of 3.16 eV and 3.02 eV have been extracted. There appears to be a slight reduction in band gap for coolant and engine oil-based HNFs. The band gap value of coconut oil-based HNFs was 4.05 eV, which showed a higher value than the pristine coconut oil-based HNFs (3.95 eV). The band gap calculated in the case of DW-based HNFs was 3.79 eV. TC of HNFs with volume concentration of 0.019 % for DW, 0.020 % for coolant, 0.016 % for engine oil, and 0.017 % for coconut oil were tested between 20 and 60 °C. An increase in TC was observed with the rise in temperature of the HNFs. Maximum increment in TC was observed at 60 °C for coolant-based HNFs, which was 19 %, followed by DW (18%), coconut oil (18%), and engine oil (16%), respectively. DW-based HNFs can be used as a coolant and optical filter for optoelectronics devices like photovoltaic cells for better performance. The study underscores precise control of NPs size as pivotal for band gap influence. HNFs hold promise as the next-gen heat transfer fluids (HTFs), revolutionizing thermal conductivity across industries. This research lays a firm foundation for plasma-synthesized HNFs' application in enhanced heat transfer and optoelectronic devices. Coolant-based HNFs excel in thermal conductivity, addressing heat transfer challenges., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

More than 95% of patients fall victim to primary amoebic meningoencephalitis (PAM), a fatal disease attacking the central nervous system. Naegleria fowleri, a brain-eating microorganism, is PAM's most well-known pathogenic ameboflagellate. Despite the use of antibiotics, the fatality rate continues to rise as no clinical trials have been conducted against this disease. To address this, we mined the UniProt database for pathogenic proteins and selected assumed epitopes to create an mRNA-based vaccine. We identified thirty B-cell and T-cell epitopes for the vaccine candidate. These epitopes, secretion boosters, subcellular trafficking structures, and linkers were used to construct the vaccine candidate. Through predictive modeling and confirmation via the Ramachandran plot (with a quality factor of 92.22), we assessed secondary and 3D structures. The adjuvant RpfE was incorporated to enhance the vaccine construct's immunogenicity (GRAVY index: 0.394, instability index: 38.99, antigenicity: 0.8). The theoretical model of immunological simulations indicated favorable responses from both innate and adaptive immune cells, with memory cells expected to remain active for up to 350 days post-vaccination, while the antigen was eliminated from the body within 24 h. Notably, strong interactions were observed between the vaccine construct and TLR-4 (- 11.9 kcal/mol) and TLR-3 (- 18.2 kcal/mol)., (© 2024. The Author(s).)

Aim: To determine the efficacy of manuka honey against multidrug-resistant (MDR) and extensively drug-resistant (XDR) clinical strains of Salmonella Typhi. Materials & methods: Clinical isolates were processed using the Bactec blood culture system, identification and antibiogram by Vitek 2 and antibiotic resistance genes through polymerase chain reaction (PCR). Microbroth dilution assays evaluated the antibacterial activity of manuka honey. Results: MDR and XDR- S. Typhi was susceptible to azithromycin. These strains carried the H58, gyrA , gyrB , bla CTX-M-15  , and bla TEM-1 genes. At 100% honey, the zone of inhibition for MDR (15-23 mm) and XDR (15-24 mm) strains. 18/50 MDR and 14/50 XDR strains inhibited at 3.125 v/v% killed at 6.25 v/v% concentration respectively. Conclusion: Manuka honey could be an alternative option for treating S. Typhi infections.

Background: Recent reports of the emergence of fluconazole resistance in Candida parapsilosis species complex poses a challenge, more specifically in settings where echinocandin-based treatment regime is not feasible., Objective: This study reported emergence of fluconazole resistance in C. parapsilosis species complex strains isolated from blood cultures., Materials and Methods: This retrospective observational study was conducted from 2018 to 2020 at a tertiary care laboratory from Pakistan. Fluconazole-resistant C. parapsilosis species complex fungemia cases were identified from laboratory database and clinical details were collected. Identification of C. parapsilosis species complex was done using API 20C AUX and Cornmeal Tween80 agar morphology. Minimum inhibitory concentrations (MICs) were determined using Sensititre YeastONE and interpretation was done with CLSI M60 ED1:2017. ERG11 gene region was amplified and sequenced by Sanger sequencing and analysed by MEGA 11 Software., Results: A total of 13 (8.5%) fluconazole-resistant isolates were identified from 152 C. parapsilosis species complex candidemia cases. Fluconazole MICs of resistant isolates ranged between 8 and 256 μg/mL. Analysis of ERG11 gene revealed nonsynonymous mutations at position Y132F in 86% of the fluconazole-resistant isolates. Diabetes and hospitalization were important risk factors for candidemia with fluconazole-resistant C. parapsilosis complex., Conclusion: This is the first report of the emergence and molecular mechanisms of fluconazole resistance in C. parapsilosis species complex from Pakistan. Y132F mutation in the ERG11 gene was the most common mutation in fluconazole-resistant strains. These findings are concerning and necessitate better diagnostics, newer antifungals, ongoing surveillance and further insights on resistance mechanisms in the country., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

Background and Objective: Fungal empyema is a rare entity which is associated with high mortality. It is mostly seen in immune-compromised hosts. However, there is limited data available on fungal empyema from developing countries regarding risk factors, treatment, and outcome. This study was conducted to determine the risk factors, clinical features, treatment, and outcome of fungal empyema., Methods: A retrospective observational study was performed on proven fungal empyema cases, admitted at Aga Khan University Hospital, Karachi, Pakistan during January 2018 to May 2021. We excluded all those patients with polymicrobial bacterial and fungal empyema or with negative pleural fluid cultures. A preformed questionnaire was filled out for each case., Results: A total of 26 patients were diagnosed with fungal empyema with a mean age of 43.6 ± 20.3 years. Of these, 16 (61.5%) patients were male. Diabetes mellitus was the most frequent comorbidity ( n  = 11, 42.3%), followed by hypertension ( n  = 9, 34.6%), malignancy ( n  = 6, 23.1%), and asthma ( n  = 1, 3.8%). Ten (38.5%) patients had multiple comorbidities. Candida spp. was isolated in 21 (80.8%) patients and Aspergillus spp. in 7 (26.9%) patients. Fusarium spp. was isolated from one (3.9%) patient. Video-assisted thoracoscopy surgery was done in 14 (53.8%) patients and 12 (46.1%) patients were managed with tube thoracostomy. Twenty-one (80.8%) patients received antifungal agents. Overall, in-hospital mortality was 38.5% ( n  = 10) and all patients developed respiratory failure. Clinical improvement was seen in 16 (61.5%) patients., Conclusion: Our data suggest that fungal empyema has a poor outcome as almost one-third of our patients died. Early diagnosis and intervention can improve outcome., Competing Interests: The authors declare that there is no conflict of interest., (© The Author(s), 2024.)

Klebsiella pneumonia is a Gram negative facultative anaerobic bacterium involved in various community-acquired pneumonia, nosocomial and lungs associated infections. Frequent usage of several antibiotics and acquired resistance mechanisms has made this bacterium multi-drug resistance (MDR), complicating the treatment of patients. To avoid the spread of this bacterium, there is an urgent need to develop a vaccine based on immuno-informatics approaches that is more efficient than conventional method of vaccine prediction or development. Initially, the complete proteomic sequence of K. pneumonia was picked over for specific and prospective vaccine targets. From the annotation of the whole proteome, eight immunogenic proteins were selected, and these shortlisted proteins were interpreted for CTL, B-cells, and HTL epitopes prediction, to construct mRNA and multi-epitope vaccines. The Antigenicity, allergenicity and toxicity analysis validate the vaccine's design, and its molecular docking was done with immuno-receptor the TLR-3. The docking interaction showed a stronger binding affinity with a minimum energy of -1153.2 kcal/mol and established 23 hydrogen bonds, 3 salt bridges, 1 disulfide bond, and 340 non-binding contacts. Further validation was done using In-silico cloning which shows the highest CAI score of 0.98 with higher GC contents of 72.25% which represents a vaccine construct with a high value of expression in E. coli. Immune Simulation shows that the antibodies (IgM, IgG1, and IgG2) production exceeded 650,000 in 2 to 3 days but the response was completely neutralized in the 5 th day. In conclusion, the study provides the effective, safe and stable vaccine construct against Klebsiella pneumonia, which further needs in vitro and in vivo validations.Communicated by Ramaswamy H. Sarma.

The presence of plastic and microplastic pollution in freshwater systems receives extensive concerns for its accumulative trend and potential ecological impacts. This is the first annual study that investigated the monthly profile of plastic pollution in the mouth of the Soan River. Plastic pollutants comprising microplastic content up to 91.7 % were abundantly found during different seasons around the year, ranging from 132.7 items/m 3 to 641.3 items/m 3 . The average abundance of plastics was significantly higher in August (641.3 ± 23.7 items/m 3 ) than in other months. Overall, fibers, large microplastics (L-MPs), and transparent items were dominant by shape (57.7 %), size (61.9 %), and color (24.6 %), respectively. The highest average number of fibers (374.3/m 3 ) and L-MPs (396 items/m 3 ) were recorded during May and remained higher in the surface water from December to May. Fragments (432.3/m 3 ) and S-MPs were observed higher (362.3 items/m 3 ) during the peak rainy month of the summer monsoon season (August). Variations in the abundance and morphotypes were seemingly not only influenced by the seasonal change but also might be due to hydromorphological characteristics of the river, especially riverbed morphology, and the flow of the water. Only 5.2 % of the total items found were identified using μ-FTIR (micro-Fourier Transformed Infrared Spectroscope) which consisted of 70.7 % plastic items. Spectroscopy revealed that polyethylene terephthalate was an abundantly found polymer that largely prevailed in the form of fibers, followed by polypropylene and polyethylene. Most of the fragments, foams, and films were composed of polypropylene, polystyrene, and rayon respectively. Being an urban river, the polymeric profile demonstrated that anthropogenic activities had a significant impact on polluting the river. These findings are a very important source to understand the profile of plastic pollution in the Soan River and also a significant reference for policy-making in controlling plastic pollution among the riverine networks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Despite a worldwide decline in maternal mortality over the past two decades, a significant gap persists between low- and high-income countries, with 94% of maternal mortality concentrated in low and middle-income nations. Ultrasound serves as a prevalent diagnostic tool in prenatal care for monitoring fetal growth and development. Nevertheless, acquiring standard fetal ultrasound planes with accurate anatomical structures proves challenging and time-intensive, even for skilled sonographers. Therefore, for determining common maternal fetuses from ultrasound images, an automated computer-aided diagnostic (CAD) system is required. A new residual bottleneck mechanism-based deep learning architecture has been proposed that includes 82 layers deep. The proposed architecture has added three residual blocks, each including two highway paths and one skip connection. In addition, a convolutional layer has been added of size 3 × 3 before each residual block. In the training process, several hyper parameters have been initialized using Bayesian optimization (BO) rather than manual initialization. Deep features are extracted from the average pooling layer and performed the classification. In the classification process, an increase occurred in the computational time; therefore, we proposed an improved search-based moth flame optimization algorithm for optimal feature selection. The data is then classified using neural network classifiers based on the selected features. The experimental phase involved the analysis of ultrasound images, specifically focusing on fetal brain and common maternal fetal images. The proposed method achieved 78.5% and 79.4% accuracy for brain fetal planes and common maternal fetal planes. Comparison with several pre-trained neural nets and state-of-the-art (SOTA) optimization algorithms shows improved accuracy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Rauf, Khan, Bashir, Jabeen, Hamza, Alzahrani, Alalwan and Masood.)

There is limited data available about allergic bronchopulmonary aspergillosis (ABPA) in Pakistan. The aim of the study was to describe the radiological and microbiological profile of ABPA patients presenting to the outpatient pulmonary clinic of a tertiary care hospital in Karachi, Pakistan. A retrospective study was conducted on ABPA patients who presented to the pulmonary outpatient clinic at Aga Khan University Hospital, Karachi, Pakistan, from January 2017 to December 2019. Data was collected on microbiology and radiology features on predesigned proforma. A total of 7759 asthmatic patients presented at the outpatient pulmonology clinic during the study period. Of the 245 patients labeled as ABPA, 167 fulfilled the inclusion criteria, and 91 (54.5%) were female (mean age 41.9±13.0 years). A high resolution computed tomography scan of the chest was available for 126 patients. Of these, 104 (82.5%) patients had bronchiectasis. Central bronchiectasis was noted in 98 (94.2%), mucus plugging in 71 (56.3%), and hyperinflation was seen in 30 (23.4%) patients. Microbiological testing was available in 103/167 (61.7%) patients. The most common bacterial pathogen was Pseudomonas aeruginosa 32 (31.1%), followed by Hemophilus influenzae 16 (15.5%), and Moraxella catarrhalis 7 (9.7%). Aspergillus fumigatus 17 (23.6%) was the most common mold, followed by Aspergillus flavus 16 (22.2%) and Aspergillus niger 11 (15.3%). Co-infection (bacterial and fungal) was found in 18 (17.45%) patients. Bronchiectasis was frequently observed in our cohort of patients with ABPA. Pseudomonas aeruginosa was found to be common among bacterial pathogens. Isolation of fungus is not uncommon in these patients.

Neurodegenerative complications, like Alzheimer's disease (AD) exert adverse effects i.e. psychological and physiological in the central nervous system. The synthetic drugs used for these complications have negative effects on body health and therefore natural remedies are a good and targeted approach to counter such complications. Alternatively, fruits and a variety of biochemicals which are an important source of diet, can be used for remedial purposes. Due to the antioxidant properties of polyphenolic compounds, several companies utilize this property to advertise polyphenol-rich beverages. Pomegranate ( Punica granatum L.), is one such fruit that is well known for its medical usage due to its antioxidant properties. In the cuurent study a literature search survey was performed on traditional uses, phytochemicals on pomegranate and their medical applications especaily in neurodegenerative deasese using electronic data bases like PubMed, Google Scholar, Scopus, Science Direct Wikipedia and Springer Nature. Based on previous preclinical and clinical studies, pomegranate juice, extracts, and its bioactive constituents have shown many mitigating properties, including suppression of inflammatory cell signaling, reduction in expression of genes associated with oxidative stress as well as pro-inflammatory cytokines in neurons, decreased production of inflammatory and oxidative stress biomarkers and increased expression of endothelial nitric oxide synthase. It also decreases the expression of soluble amyloid protein procurer β (sAPPβ), β-secretase and carboxyl terminal fragment β (CTFβ). Similarly, during an in-vivo study on APP/PS1 mice, pomegranate supplementation has been shown to impart cognitive aid by the protection of neurons and triggering neurogenesis through anti-inflammatory signaling pathway. In conclusion, pomegranate supplementation can be a promising source of protection against Alzheimer's disease., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Importance: Candida glabrata is a major fungal pathogen, which is able to lose mitochondria and form small and slow-growing colonies, called "petite." This attenuated growth rate has created controversies and questioned the clinical importance of petiteness. Herein, we have employed multiple omics technologies and in vivo mouse models to critically assess the clinical importance of petite phenotype. Our WGS identifies multiple genes potentially underpinning petite phenotype. Interestingly, petite C. glabrata cells engulfed by macrophages are dormant and, therefore, are not killed by the frontline antifungal drugs. Interestingly, macrophages infected with petite cells mount distinct transcriptomic responses. Consistent with our ex vivo observations, mitochondrial-proficient parental strains outcompete petites during systemic and gut colonization. Retrospective examination of C. glabrata isolates identified petite prevalence a rare entity, which can significantly vary from country to country. Collectively, our study overcomes the existing controversies and provides novel insights regarding the clinical relevance of petite C. glabrata isolates., Competing Interests: The authors declare no conflict of interest.

Aims: Salinity adversely affects okra [Abelmoschus esculentus (L.) Moench] plants by inducing osmotic and oxidative stresses. This study was designed to enhance salinity-induced osmotic and oxidative stress tolerance in okra plants by applying organic amendments., Methods: The effects of different organic amendments (municipal solid waste compost, farmyard manure (FYM) and press mud) on osmotic potential, water use efficiency, activities of antioxidant enzymes, total soluble sugar, total soluble proline, total soluble protein and malondialdehyde (MDA) contents of okra plants grown under saline conditions (50 mM sodium chloride) were evaluated in a pot experiment. The organic amendments were applied each at the rate of 5% and 10% per pot or in various combinations (compost + FYM, FYM + press mud and compost + press mud each at the rate of 2.5% and 5% per pot)., Results: As compared to control, high total soluble sugar (60.41), total soluble proline (33.88%) and MDA (51%) contents and increased activities of antioxidant enzymes [superoxide dismutase (83.54%), catalase (78.61%), peroxidase (53.57%] in salinity-stressed okra plants, were indicative of oxidative stress. Salinity significantly reduced the osmotic potential (41.78%) and water use efficiency (4.75%) of okra plants compared to control. Under saline conditions, 5% (farmyard manure + press mud) was the most effective treatment, which significantly improved osmotic potential (27.05%), total soluble sugar (4.20%), total soluble protein (73.62%) and total soluble proline (23.20%) contents and superoxide dismutase activity (32.41%), compared to saline soil. Application of 2.5% (FYM + press mud), 5% press mud, and 10% compost significantly reduced MDA content (27%) and improved activities of catalase (38.64%) and peroxidase (48.29%), respectively, compared to saline soil, thus facilitated to alleviate oxidative stress in okra plants., Conclusions: Using organic amendments (municipal solid waste compost, farmyard manure and press mud) was a cost-effective approach to improve salinity-induced osmotic and oxidative stress tolerance in okra plants., (© 2023. The Author(s).)

Candida auris is a multidrug-resistant pathogen, that is a well-known cause of nosocomial infections. This pathogen is being identified using advanced diagnostic approaches and epidemiological typing procedures. In underdeveloped nations, several researchers developed and validated a low-cost approach for reliably identifying Candida auris. The goal of this study was to assess the burden of Candida auris in different teaching hospitals of Lahore and to limit its spread to minimize hospital-related illnesses. Candida isolates were obtained from various tertiary care institutions in Lahore in the form of culture on various culture plates. Sabouraud agar culture plates were used to culture the Candida spp. Fluconazole-resistant Candida species were chosen for further identification using VITEK 2 Compact ID and molecular identification using species-specific PCR assay. The current study obtained 636 Candida samples from several tertiary care institutions in Lahore. Fluconazole resistance was found in 248 (38.9%) of 636 Candida samples. No isolate was identified as Candida auris by VITEK 2 Compact ID and real-time PCR-based molecular identification. Thus with limited resources, these two methods may serve as useful screens for Candida auris. However, it should be screened all over the country to limit its spread to break the chain of nosocomial infections., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Huma et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The global burden of dengue infections has increased dramatically. Early diagnosis of dengue infection is critical to proper medical management to avoid further complications in patients. This study was geared to assess the severity of dengue infections based on clinical and hematological examinations. A cross-sectional study was conducted among febrile patients with dengue infection in a teaching hospital in Pakistan. Blood samples were investigated for dengue-specific antibodies (IgM and IgG) and the nonstructural 1 antigen. The clinical findings of each subject were noted to assess the severity of the infection. Tests for hematological parameters were performed. Of 130 patients with confirmed dengue infection, 23 had severe and 107 had nonsevere dengue. Patients with severe dengue experienced mucosal bleeding (71.4%), fluid accumulation (57.1%), shock (35.7%), and gastrointestinal bleeding (28.6%). The most significant hematological findings among severe and nonsevere patients with dengue infection were thrombocytopenia, leukopenia, and a raised hematocrit level (P < 0.001). Patients with severe dengue infection showed marked thrombocytopenia, with a mean platelet count of 49.96 × 109 platelets/L. The clinical presentation of patients with dengue infection along with hematological markers are the most important clues for the diagnosis of, prognosis of, and therapy for dengue infection. Thrombocytopenia, leukopenia, and raised hematocrit levels were the most significant hematological parameters when assessing the severity of dengue infection.

Background/aims: Hepatitis B virus induces mitochondrial damage via the production of reactive oxygen species and concomitant with deregulation of calcium homeostasis. The current study evaluates the potential of antioxidant and calcium modulators for inhibition of hepatitis B virus-induced mitochondrial damage using in vitro cell culture models., Materials and Methods: Hepatitis B virus-induced mitochondrial fragmentation was observed by immunofluorescence confocal micros- copy in hepatitis B virus-infected cell lines (HepG2 and HepAD38). Differential protein expression of mitochondrial fragmentation mark- ers, dynamin-related protein 1 and phospho-dynamin-related protein 1, were evaluated both pre- and posttreatment with antioxidant N-acetyl-l-cysteine and calcium modulators like 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakisacetoxymethyl ester, ethylene-bis (oxyethylenenitrilo) tetraacetic acid glycol ether diamine tetraacetic acid-acetoxymethyl ester, and ruthenium amine complex by western blot analysis., Results: A slight reduction in mitochondrial fragmentation in both cell lines was observed post-antioxidant treatment with a partial prevention observed with calcium modulators. The expression of phospho-dynamin-related protein 1 was significantly upregulated (P = .0007, P = .003) in both hepatitis B virus-infected cell lines compared to uninfected cells. In line with these observations, the expres- sion of dynamin-related protein 1 and phospho-dynamin-related protein 1 was found to be significantly downregulated with N-acetyl- l-cysteine treatment in both cell lines (P = .003, P = .002), respectively. A nonsignificant trend was observed in the case of calcium modulators treatment., Conclusions: Current study indicates that the mitochondrial fragmentation induced by hepatitis B virus infection can be reduced after antioxidant treatment pointing toward exploring better drug targets for the prevention of hepatitis B virus-induced mitochondrial frag- mentation and associated liver damage.

Avena sativa L. a cereal crop that is badly affected by several abiotic and biotic stresses. In the current study, silicon nanoparticles are used to mitigate the harmful effects of root rot disease caused by Rhizoctonia solani Kuhn on the growth of A. sativa. In vitro (Petri plates) and in vivo (pots experiment) were performed to measure the various physiological and biochemical parameters i.e. osmotic potential, chlorophyll, proline content, growth parameters, sugar, fresh and dry weight, and disease index. Results revealed that physiological and biochemical parameters were reduced under fungal stress with silicon nanoparticles treatment as compared to the control group. Si nanoparticles helped to alleviate the negative effects caused by fungus i.e. germination percentage upto 80%, germination rate 4 n/d, radical and plumule length was 4.02 and 5.46, dry weight 0.08 g, and relative water content was (50.3%) increased. Fungus + Si treatment showed the maximum protein content, i.e. 1.2 µg/g as compared to Fungus (0.3 µg/g) treated group. The DI was maximum (78.82%) when the fungus directly attacked the target plant and DI reduced (44.2%) when the fungus was treated with Si nanoparticles. Thus, silicon nanoparticles were potentially effective against the stress of R. solani and also used to analyze the plant resistance against fungal diseases. These particles can use as silicon fertilizers, but further studies on their efficacy under field conditions and improvement in their synthesis are still needed., (© 2023. Springer Nature Limited.)

Cardiovascular diseases (CVDs) and neurodegenerative disorders, such as diabetes mellitus and Alzheimer's disease, share a common pathophysiological link involving insulin resistance (IR), inflammation, and hypertension. Aluminium chloride (AlCl 3 ), a known neurotoxicant, has been associated with neurodegeneration, cognitive impairment, and various organ dysfunctions due to the production of reactive oxygen species (ROS) and oxidative stress. In this study, we aimed to investigate the potential protective effects of metformin and vitamin E against AlCl 3 -induced neuroinflammation and cardiometabolic disturbances in rat models. Rats were divided into five groups: a normal control group, an AlCl 3 -treated diseased group without any treatment, and three groups exposed to AlCl 3 and subsequently administered with metformin (100 mg/kg/day) alone, vitamin E (150 mg/kg/day) orally alone, or a combination of metformin (100 mg/kg/day) and vitamin E (150 mg/kg/day) for 45 days. We analyzed serum biomarkers and histopathological changes in brain, heart, and pancreatic tissues using H&E and Masson's trichrome staining and immunohistochemistry (IHC). Electrocardiogram (ECG) patterns were observed for all groups. The AlCl 3 -treated group showed elevated levels of inflammatory biomarkers, MDA, and disturbances in glycemic and lipid profiles, along with reduced insulin levels. However, treatment with the combination of metformin and vitamin E resulted in significantly reduced glucose, cholesterol, LDL, and TG levels, accompanied by increased insulin and HDL levels compared to the individual treatment groups. Histopathological analyses revealed that combination therapy preserved neuronal structures, muscle cell nuclei, and normal morphology in the brain, heart, and pancreatic tissues. IHC demonstrated reduced amyloid plaques and neurofibrillary tangles in the combination-treated group compared to the AlCl 3 -treated group. Moreover, the combination group showed a normal ECG pattern, contrasting the altered pattern observed in the AlCl 3 -treated group. Overall, our findings suggest that metformin and vitamin E, in combination, possess neuroprotective and cardiometabolic effects, alleviating AlCl 3 -induced neuroinflammation and metabolic disturbances.

Small colony variants (SCVs) are relatively common among some bacterial species and are associated with poor prognosis and recalcitrant infections. Similarly, Candida glabrata - a major intracellular fungal pathogen - produces small and slow-growing respiratory-deficient colonies, termed "petite." Despite reports of clinical petite C . glabrata strains, our understanding of petite behavior in the host remains obscure. Moreover, controversies exist regarding in-host petite fitness and its clinical relevance. Herein, we employed whole-genome sequencing (WGS), dual-RNAseq, and extensive ex vivo and in vivo studies to fill this knowledge gap. WGS identified multiple petite-specific mutations in nuclear and mitochondrially-encoded genes. Consistent with dual-RNAseq data, petite C . glabrata cells did not replicate inside host macrophages and were outcompeted by their non-petite parents in macrophages and in gut colonization and systemic infection mouse models. The intracellular petites showed hallmarks of drug tolerance and were relatively insensitive to the fungicidal activity of echinocandin drugs. Petite-infected macrophages exhibited a pro-inflammatory and type I IFN-skewed transcriptional program. Interrogation of international C . glabrata blood isolates ( n =1000) showed that petite prevalence varies by country, albeit at an overall low prevalence (0-3.5%). Collectively, our study sheds new light on the genetic basis, drug susceptibility, clinical prevalence, and host-pathogen responses of a clinically overlooked phenotype in a major fungal pathogen., Importance: Candida glabrata is a major fungal pathogen, which is able to lose mitochondria and form small and slow-growing colonies, called "petite". This attenuated growth rate has created controversies and questioned the clinical importance of petiteness. Herein, we have employed multiple omicstechnologies and in vivo mouse models to critically assess the clinical importance of petite phenotype. Our WGS identifies multiple genes potentially underpinning petite phenotype. Interestingly, petite C. glabrata cells engulfed by macrophages are dormant and therefore are not killed by the frontline antifungal drugs. Interestingly, macrophages infected with petite cells mount distinct transcriptomic responses. Consistent with our ex-vivo observations, mitochondrial-proficient parental strains outcompete petites during systemic and gut colonization. Retrospective examination of C. glabrata isolates identified petite prevalence a rare entity, can significantly vary from country to country. Collectively, our study overcomes the existing controversies and provides novel insights regarding the clinical relevance of petite C. glabrata isolates.

Background: The emergence of extensively drug-resistant (XDR) typhoid in Pakistan has endangered the treatment options available to manage this infection. Third generation cephalosporin were the empiric choice to treat typhoid fever in Pakistan, but acquisition of ESBLs have knocked them out of the arsenal. The current empiric choice is azithromycin which is vulnerable to resistance too. This study aimed to assess the burden of XDR typhoid and the frequency of resistance determinants in blood culture samples collected from different hospitals in Lahore, Pakistan., Methods: A total of 835 blood cultures were collected from different tertiary care hospitals in Lahore during January 2019 to December 2021. Among 835 blood cultures, 389 Salmonella Typhi were identified, and 150 were XDR S . Typhi (resistant to all recommended antibiotics). Antibiotics resistance genes of the first-line drugs ( bla TEM-1 , cat A1, sul 1, and dhfR7) and second line drugs ( gyrB, gyrA, qnrS, ParC and ParE ) were investigated among XDR S . Typhi. There were different CTX-M genes isolated using the specific primers, bla CTX-M-U , bla CTX-M-1, bla CTX-M-15, bla CTX-M-2, bla CTX-M-8 and bla CTX-M-9 ., Results: Antibiotic resistant genes of the first-line drugs were isolated with different frequency, bla TEM-1 (72.6%), catA1 (86.6%), sul 1 (70%), and dhfR7 (56%). Antibiotics resistance genes of second-line drugs were isolated as: gyrB (60%), gyrA (49.3%), qnrS (32.6%), parC (44%) and parE (28%). Among CTX-M genes, bla CTX-M-U (63.3%) was the most frequent followed by bla CTX-M-15 (39.3%) and bla CTX-M-1 (26%)., Conclusion: Our study concluded that XDR isolates circulating in Pakistan have acquired first-line and second-line antibiotic resistant genes quite successfully along with CTX-M genes (ESBLs) rendering them resistant to the third generation cephalosporins as well. Emergence of azithromycin resistance in XDR S . Typhi which is currently used as an empiric treatment option is worrisome and needs to be monitored carefully in endemic countries like Pakistan., Competing Interests: The authors report no conflicts of interest in this work., (© 2023 Jabeen et al.)

The comparative genomic analysis of Lactiplantibacillus plantarum YW11 ( L. plantarum YW11) isolated from Tibetan kefir involves comparison of the complete genome sequences of the isolated strain with other closely related L. plantarum strains. This type of analysis can be used to identify the genetic diversity among strains and to explore the genetic characteristics of the YW11 strain. The genome of L. plantarum YW11 was found to be composed of a circular single chromosome of 4,597,470 bp with a G + C content of 43.2%. A total of 4,278 open reading frames (ORFs) were identified in the genome and the coding density was found to be 87.8%. A comparative genomic analysis was conducted using two other L. plantarum strains, L. plantarum C11 and L. plantarum LMG21703. Genomic comparison revealed that L. plantarum YW11 shared 72.7 and 75.2% of gene content with L. plantarum C11 and L. plantarum LMG21703, respectively. Most of the genes shared between the three L. plantarum strains were involved in carbohydrate metabolism, energy production and conversion, amino acid metabolism, and transcription. In this analysis, 10 previously sequenced entire genomes of the species were compared using an in-silico technique to discover genomic divergence in genes linked with carbohydrate intake and their potential adaptations to distinct human intestinal environments. The subspecies pan-genome was open, which correlated with its extraordinary capacity to colonize several environments. Phylogenetic analysis revealed that the novel genomes were homogenously grouped among subspecies of l Lactiplantibacillus. L. plantarum was resistant to cefoxitin, erythromycin, and metronidazole, inhibited pathogens including Listeria monocytogenes, Clostridium difficile, Vibrio cholera, and others, and had excellent aerotolerance, which is useful for industrial operations. The comparative genomic analysis of L. plantarum YW11 isolated from Tibetan kefir can provide insights into the genetic characteristics of the strain, which can be used to further understand its role in the production of kefir., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Aziz, Naveed, Jabeen, Shabbir, Sarwar, Zhennai, Alharbi, Alshammari and Alasmari.)

One of the most frequent cancers in women is breast cancer, and in the year 2022, approximately 287,850 new cases have been diagnosed. From them, 43,250 women died from this cancer. An early diagnosis of this cancer can help to overcome the mortality rate. However, the manual diagnosis of this cancer using mammogram images is not an easy process and always requires an expert person. Several AI-based techniques have been suggested in the literature. However, still, they are facing several challenges, such as similarities between cancer and non-cancer regions, irrelevant feature extraction, and weak training models. In this work, we proposed a new automated computerized framework for breast cancer classification. The proposed framework improves the contrast using a novel enhancement technique called haze-reduced local-global. The enhanced images are later employed for the dataset augmentation. This step aimed at increasing the diversity of the dataset and improving the training capability of the selected deep learning model. After that, a pre-trained model named EfficientNet-b0 was employed and fine-tuned to add a few new layers. The fine-tuned model was trained separately on original and enhanced images using deep transfer learning concepts with static hyperparameters' initialization. Deep features were extracted from the average pooling layer in the next step and fused using a new serial-based approach. The fused features were later optimized using a feature selection algorithm known as Equilibrium-Jaya controlled Regula Falsi. The Regula Falsi was employed as a termination function in this algorithm. The selected features were finally classified using several machine learning classifiers. The experimental process was conducted on two publicly available datasets-CBIS-DDSM and INbreast. For these datasets, the achieved average accuracy is 95.4% and 99.7%. A comparison with state-of-the-art (SOTA) technology shows that the obtained proposed framework improved the accuracy. Moreover, the confidence interval-based analysis shows consistent results of the proposed framework.

This study aimed to evaluate the minimum inhibitory concentration (MIC) of azithromycin (AZM) in clinical isolates of extensively drug-resistant (XDR) Salmonella Typhi (i.e., resistant to chloramphenicol, ampicillin, trimethoprim-sulfamethoxazole, fluoroquinolones, and third-generation cephalosporin) using the E-test versus the broth microdilution method (BMD). From January to June 2021, a retrospective cross-sectional study was carried out in Lahore, Pakistan. Antimicrobial susceptibility was performed initially by the Kirby-Bauer disk diffusion method for 150 XDR Salmonella enterica serovar Typhi isolates, and MICs of all the recommended antibiotics were determined by the VITEK 2 (BioMérieux) fully automated system using Clinical Laboratory Standard Institute (CLSI) 2021 guidelines. The E-test method was used to determine AZM MICs. These MICs were compared with the BMD, which is the method recommended by the CLSI but not adopted in routine laboratory reporting. Of 150 isolates, 10 (6.6%) were resistant by disk diffusion. Eight (5.3%) of these had high MICs against AZM by the E-test. Only three isolates (2%) were resistant by E-test, having an MIC of 32 μg/mL. All eight isolates had a high MIC by BMD with different MIC distributions, but only one was resistant, having an MIC of 32 μg/mL by BMD. The sensitivity, specificity, negative predictive value, positive predictive value, and diagnostic accuracy of the E-test method versus BMD were 98.65%,100%, 99.3%, 33.3%, and 98.6%, respectively. Similarly, the concordance rate was 98.6%, negative percent agreement was 100%, and positive percent agreement was 33%. The BMD is the most reliable approach for reporting AZM sensitivity in XDR S. Typhi compared with the E-test and disk diffusion methods. Potentially, AZM resistance in XDR S. Typhi is around the corner. Sensitivity patterns should be reported with MIC values, and if possible, higher values should be screened for the presence of any potential resistance genes. Antibiotic stewardship should be strictly implemented.

Introduction: Invasive fungal infections (IFIs) in Asia/Pacific are a particular threat to patients with malignancies, uncontrolled diabetes mellitus or undiagnosed/untreated human immunodeficiency virus infection and acquired immunodeficiency syndrome (HIV/AIDS). Adequate and early access to diagnostic tools and antifungals is essential for IFI clinical management and patient survival., Methods: Details on institution profile, self-perception on IFI, and access to microscopy, culture, serology, antigen detection, molecular testing, and therapeutic drug monitoring for IFI were collected in a survey., Results: As of June 2022, 235 centres from 40 countries/territories in Asia/Pacific answered the questionnaire. More than half the centres were from six countries: India (25%), China (17%), Thailand (5%), Indonesia, Iran, and Japan (4% each). Candida spp. (93%) and Aspergillus spp. (75%) were considered the most relevant pathogens. Most institutions had access to microscopy (98%) or culture-based approaches (97%). Furthermore, 79% of centres had access to antigen detection, 66% to molecular assays, and 63% to antibody tests. Access to antifungals varied between countries/territories. At least one triazole was available in 93% of the reporting sites (voriconazole [89%] was the most common mould-active azole), whereas 80% had at least one amphotericin B formulation, and 72% had at least one echinocandin., Conclusion: According to the replies provided, the resources available for IFI diagnosis and management vary among Asia/Pacific countries/territories. Economical or geographical factors may play a key role in the incidence and clinical handling of this disease burden. Regional cooperation may be a good strategy to overcome shortcomings., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Lead (Pb) contamination continues to contribute to world-wide morbidity in all countries, particularly low- and middle-income countries. Despite its continued widespread adverse effects on global populations, particularly children, accurate prediction of elevated household dust Pb and the potential implications of simple, low-cost household interventions at national and global scales have been lacking. A global dataset (∼40 countries, n = 1951) of community sourced household dust samples were used to predict whether indoor dust was elevated in Pb, expanding on recent work in the United States (U.S.). Binned housing age category alone was a significant (p < 0.01) predictor of elevated dust Pb, but only generated effective predictive accuracy for England and Australia (sensitivity of ∼80%), similar to previous results in the U.S. This likely reflects comparable Pb pollution legacies between these three countries, particularly with residential Pb paint. The heterogeneity associated with Pb pollution at a global scale complicates the predictive accuracy of our model, which is lower for countries outside England, the U.S., and Australia. This is likely due to differing environmental Pb regulations, sources, and the paucity of dust samples available outside of these three countries. In England, the U.S., and Australia, simple, low-cost household intervention strategies such as vacuuming and wet mopping could conservatively save 70 billion USD within a four-year period based on our model. Globally, up to 1.68 trillion USD could be saved with improved predictive modeling and primary intervention to reduce harmful exposure to Pb dust sources., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objectives: Hepatitis B virus (HBV) infection alters the cytokines production to establish persistent infection. A reversion of cytokines back to their normal state can be a promising therapeutic approach to establish an optimal host immune response., Materials and Methods: We investigated the alteration in expression of IL-15 and IL-11 after HBV infection in vitro and in vivo in PBMCs of 63 individuals divided into various HBV-infected patient groups. The mRNA expression was evaluated post-anti-oxidant and calcium modulators treatment by Real-time qPCR., Results: In vitro mRNA expression of both cytokines, post-infection was down-regulated considerably. Interestingly, in line with in vitro results, both cytokines' in vivo expression was intensively down-regulated in chronic HBV-infected individuals rather than healthy controls. Both cytokines' expression was up-regulated in cases of recovery compared with the inactive carriers and chronic HBV-infected individuals. IL-15 mRNA expression was significantly up-regulated in both cell lines post EGTA and Ru360 treatment while a significant increase was observed in the HepAD38 cell line with NAC and BAPTA treatment. IL-11 mRNA expression was significantly up-regulated in the HepG2 cell line after all modulator treatments, whereas in the HepAd38 cell line it was observed after BAPTA treatment. Our results thus indicate that viral infection tends to down-regulate the expression of cytokines and an in vivo up-regulation is an indication of recovery., Conclusion: Treatment of anti-oxidants and calcium modulators has resulted in the successful restoration of these cytokines thus pointing towards the use of calcium modulators to boost natural antiviral cytokine production., Competing Interests: The authors have no conflicts of interest.

Background . Concurrent coronavirus disease 2019 (COVID-19) and Pneumocystis jirovecii pneumonia (PJP) has been described in various reports, with a recent study describing a 9.3 % P . jirovecii detection rate in critically ill COVID-19 patients. Methods . Patients with PCR-confirmed PJP following COVID-19 infection who were admitted to Aga Khan University Hospital, Karachi, Pakistan from March 2020-June 2021 were identified through a laboratory database. Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus was performed by RT-PCR Cobas SARS-CoV-2 qualitative assay. P. jirovecii PCR was performed using the RealStar Pneumocystis jirovecii PCR kit. Clinical, radiological and laboratory data for PJP patients were recorded. Results . During the study period, 3707 patients were admitted with COVID-19 at our hospital. P. jirovecii PCR was requested for 90 patients and was positive in 10 (11 %). Five out of 10 patients were discharged from the hospital and later developed cough and dyspnoea. Five patients remained hospitalized with severe COVID-19 and developed PJP. Eight patients in our study received systemic steroids. The trends of lymphocyte counts of all patients showed a lymphocyte count of <1000 mm -3 (<1.0×10 6  cells µl -1 ) in the week of PJP diagnosis. Four patients did not survive; one of these patients did not receive co-trimoxazole due to late diagnosis, one patient had concomitant nosocomial pneumonia and bacteraemia with multidrug-resistant Acinetobacter species, and two patients had concomitant aspergillosis. Conclusion . In summary, invasive fungal infections such as PJP should be considered as a complication in COVID-19 patients, with prompt evaluation and management., Competing Interests: The authors declare that there are no conflicts of interest., (© 2023 The Authors.)

Background: Early identification of COVID-19-associated pulmonary aspergillosis (CAPA) is particularly challenging in low- middle-income countries where diagnostic capabilities are limited, and risk factors for CAPA have not been identified. It is also essential to recognise CAPA patients who are likely to have a poorer outcome to decide on aggressive management approaches. Therefore, this study aimed to identify risk factors and outcomes for CAPA among admitted moderate to critical COVID-19 patients at our centre in Pakistan., Methods: An unmatched case-control study with ratio of 1:2 was conducted on hospitalised adult patients with COVID-19 from March 2020-July 2021. Cases were defined according to European Confederation of Medical Mycology and the International Society for Human and Animal Mycology consensus criteria. Controls were defined as patients hospitalised with moderate, severe or critical COVID-19 without CAPA., Results: A total of 100 CAPA cases (27 probable CAPA; 73 possible CAPA) were compared with 237 controls. Critical disease at presentation (aOR 5.04; 95% CI 2.18-11.63), age ≥ 60 years (aOR 2.00; 95% CI 1.20-3.35) and underlying co-morbid of chronic kidney disease (CKD) (aOR 3.78; 95% CI 1.57-9.08) were identified as risk factors for CAPA. Patients with CAPA had a significantly greater proportion of complications and longer length of hospital stay (p-value < .001). Mortality was higher in patients with CAPA (48%) as compared to those without CAPA (13.5%) [OR = 6.36(95% CI 3.6-11)]., Conclusions: CAPA was significantly associated with advanced age, CKD and critical illness at presentation, along with a greater frequency of complications and higher mortality., (© 2022 Wiley-VCH GmbH.)

Objectives: To assess the diagnostic accuracy of different cut-off values of pleural fluid adenosine deaminase levels as a diagnostic method for tuberculous pleural effusion., Methods: The prospective study was conducted from 2014 to 2016 at the Aga Khan University Hospital, Karachi, and comprised pleural fluid samples of adult patients with and without tuberculosis which were tested for adenosine deaminase levels, and divided into tuberculosis group A and non-tuberculosis group B. Sensitivity, specificity, negative predictive value and positive predictive value were calculated using different cut-offs. Data was analysed using IBM SPSS (Statistical Package for Social Sciences) version 21.0 (IBM Corp., Armonk, NY)., Results: Of 155 patients, 46(29.7%) had tuberculosis; 30(65.2%) males and 16(34.8%) females. Those who did not have tuberculosis were 109(70.3%); 69(63.3%) males and 40(36.7%) females. The adenosine deaminase levels were elevated in group A compared to group B (p<0.001). The cut-off of 30U/L showed the highest sensitivity (71.7%) and negative predictive value (87.4%), and a specificity of 82.6%. The cut-off of 50U/L showed the highest specificity (89.9%) with sensitivity 52.2%, and the cut-off of 40U/L showed the highest positive predictive value of 68.9% with sensitivity 67.4% and specificity 87.2%., Conclusions: Pleural fluid adenosine deaminase testing for diagnosing tuberculosis pleuritis revealed highest sensitivity and moderate specificity for cut-off value of 30U/L.

Introduction: The gradual increase in caspofungin usage in Pakistan raises a concern of emergence of echinocandin resistance in local Candida glabrata strains. We sequenced and determined mutations in fks1 and fks2 genes in invasive Candida glabrata strains from Pakistan., Material and Methods: Thirty-six invasive C. glabrata strains were selected with median (min-max) minimum inhibitory concentrations (MICs) of 0.06 (0.015-0.25) mg/L for caspofungin, 0.015 (0.008-0.06) mg/L for micafungin and 0.06 (0.015-0.12) mg/L for anidulafungin. fks1 and fks2 gene fragments were sequenced using Sanger methodology. Sequences were analysed with MEGA-6 software to identify specific single-nucleotide polymorphisms (SNP) against wild-type sequences of C. glabrata., Results: In fks1 gene, non-synonymous mutation D632H was observed in one isolate with caspofungin MIC of 0.25 mg/L. Synonymous mutation at position A742 was observed in 26/36 (72%) of the isolates. 34/36 (94.5%) isolates analysed for fks2 gene were observed as wild type. A novel non-synonymous mutation at I661T was observed in fks2 gene in one isolate with caspofungin MIC of 0.12 mg/L and anidulafungin and micafungin MIC of 0.06 and 0.015 mg/L, respectively. Novel fks2 synonymous mutations at position T647, K652 and I706 were observed in 16/36 (44%), 25/36 (69%) and 23/36 (63%) isolates, respectively., Conclusion: Low frequencies of both non-synonymous and synonymous polymorphisms were observed in invasive C. glabrata strains. Since S663P in fks2 gene is associated with caspofungin resistance, a novel mutation at 661 codon identified in our study needs correlation with treatment outcome data and mandates periodic genomic surveillance., (© 2022 Wiley-VCH GmbH.)

Comorbidities in human beings signify the numerous risk factors that increase the incidences of neuro- and cardio-metabolic disorders. Experimental models depicting comorbidities are important to explore the molecular pathophysiology that can help suggest appropriate treatment strategies. Tissue-accumulating potential and pathological effects of aluminium chloride (AlCl 3 )and sodium azide (NaN 3 ) are well recognized. Hence, in the current work, we have for the first time aimed to investigate the unexplored potential of graded dose effects of AlCl 3 and NaN 3 in inducing early inflammation and cardiometabolic toxicity via comparative biochemical analysis of AlCl 3 and/or NaN 3 . Rats were allocated into seven groups ( n = 6). Group 1 was normal control. Remaining groups were given graded doses of AlCl 3 and/or NaN 3 , as LD-AlCl 3 (AlCl 3 40 mg), MD-AlCl 3 (AlCl 3 45 mg), and HD-AlCl 3 (AlCl 3 50 mg) representing low dose, medium dose, and high dose of AlCl 3 , respectively, and the remaining as LD-NaN 3 (NaN 3 13 mg), MD-NaN 3 (NaN 3 15 mg), and HD-NaN 3 (NaN 3 17 mg) representing low dose, medium dose, and high dose of NaN 3 , respectively. Serum levels of glucose, insulin, lipid profile, inflammatory mediators like IL-6 and oxidative stress marker, and malondialdehyde (MDA) were analyzed. Likewise, subacute toxicity parameters were analyzed. Immunohistochemistry (IHC) and histopathology (H&E/Masson's trichrome staining) of brain, heart, and pancreatic tissues were done. ECG pattern of all groups was observed. HD-AlCl 3 was associated with elevated levels of inflammatory biomarkers, MDA, and glycemic and lipid profiles, whereas it decreased the insulin levels. HD-NaN 3 also showed the similar effects of aggravated inflammatory biomarkers, impaired glycemic and lipid profiles, but depicted the maximum mortality rate as compared to HD-AlCl 3 . IHC showed prominent amyloid plaques and neurofibrillary tangle formation with MD-AlCl 3 and HD-AlCl 3 as compared to NaN 3 -treated groups. Likewise, in brain tissues, vacuolation of white matter, vascular congestion, and hemorrhage were seen in HD-AlCl 3 treated group, while HD-NaN 3 induced death in animals. AlCl 3 exposure resulted in an inverted QRS complex, while exposure to NaN 3 showed ST depression but with increased mortality. AlCl 3 has better controlled results as compared to NaN 3 for induction of comorbid experimental animal model depicting early neuroinflammation and cardiometabolic disruption. These determined efforts facilitate the researchers for the development of clinically effective treatment strategies using such experimental models., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)