Range extension and species confirmation of Rhyneptesicus nasutus (Sind Serotine Bat) (Mammalia:Chiroptera) from Bajaur Agency, FATA, Pakistan.

BACKGROUND: The lack of morphological differentiation among chiropteran species and cryptic speciation impedes species identification. DNA-based approaches help species identification and contribute to the discovery of additional species. Rhyneptesicus nasutus (Sind Serotine Bat) is a rare and poorly studied species in Pakistan. METHODS: This study explores the range extension of Sind Bat within the territorial limits of Pakistan from Sind and Baluchistan to Federally Administered Areas of Pakistan. No molecular record exists for the species in Pakistan. In the present study, we for the first time confirm species identification of Rhyneptesicus nasutus from Pakistan using a genetic marker (cytochrome b) along with morphometric analysis. A neighbor-joining tree based on Kimura-2 parameters was created to infer phylogenetic relationships. We sequenced the cytochrome b gene segment and conducted a phylogenetic analysis with previously published data from other countries. RESULTS: Sequences from Pakistan formed a clade with Iranian Rhyneptesicus nasutus specimens suggesting a common ancestry. Various morphometric parameters (mean values) were measured, including Head and Body length (44.3 mm), Tail length (43.4 mm), Hindfoot length (8.3 mm), Forearm length (35.7 mm), and Ear length 36 mm while 5th Metacarpal Length, 4th Metacarpal Length, and 3rd Metacarpal Lengths were 33.2 mm, 34.7 mm, and 35.3 mm. Approaches based on DNA barcoding reveal a high diversity of bat species in the study area. CONCLUSION: The data will enable researchers to build an improved evolutionary framework of the Serotine Bats from this region and subsequently reconstruct a detailed evolutionary history of the genus. Further research is required to test other molecular markers to support the findings of the current study in Pakistan.

Effect of oxidative stress and calcium deregulation on FAM26F (CALHM6) expression during hepatitis B virus infection.

BACKGROUND: Family with sequence similarity 26, member F (FAM26F) is an important innate immunity modulator playing a significant role in diverse immune responses, however, the association of FAM26F expression with HBV infection is not yet known. Thus, the current study aims to explore the differential expression of FAM26F in vitro in HepAD38 and HepG2 cell lines upon HBV infection, and in vivo in HBV infected individuals. The effects of antioxidant and calcium inhibitors on the regulation of FAM26F expression were also evaluated. The expression of FAM26F was simultaneously determined with well-established HBV infection markers: IRF3, and IFN-ß. METHODS: The expression of FAM26F and marker genes was analyzed through Real-time qPCR and western blot. RESULTS: Our results indicate that the differential expression of FAM26F followed the same trend as that of IRF3 and IFN-ß. The in vitro study revealed that, in both HBV infected cell lines, FAM26F expression was significantly down-regulated as compared to uninfected control cells. Treatment of cells with N-acetyl-L-cysteine (NAC), EGTA-AM, BAPTA-AM, and Ru360 significantly upregulated the expression of FAM26F in both the cell lines. Moreover, in in vivo study, FAM26F expression was significantly downregulated in all HBV infected groups as compared to controls (p = 0.0007). The expression was higher in the HBV recovered cases, probably due to the decrease in infection and increase in the immunity of these individuals. CONCLUSION: Our study is the first to show the association of FAM26F with HBV infection. It is proposed that FAM26F expression could be an early predictive marker for HBV infection, and thus is worthy of further investigation.

Correction to: Novel mutation G324C in WNT1 mapped in a large Pakistani family with severe recessively inherited Osteogenesis Imperfecta.

In the original publication of this article [1], there are two errors in the article which the cDNA position of the pathogenic variant WNT1 p.Gly324Cys should be c.970G>T instead of c.1168G>T.

Novel mutation G324C in WNT1 mapped in a large Pakistani family with severe recessively inherited Osteogenesis Imperfecta.

INTRODUCTION: Osteogenesis imperfecta (OI) is a clinically and genetically heterogeneous disease with skeletal fragility and variable extra-skeletal manifestations. To date several point mutations in 18 different genes causing different types of OI have been identified. Mutations in WNT1 compromise activity of the osteoblasts leading to disturbed bone mass accrual, fragility fractures and progressive skeletal abnormalities. The present study was conducted to determine the underlying genetic cause of an autosomal recessive skeletal dysplasia in a large consanguineous family from Chinute, Pakistan. MATERIALS AND METHODS: Blood was collected from 24 individuals of affected family along with clinical data. Homozygosity mapping was performed to confirm consanguinity. SNPs were identified, followed by whole exome and Sanger sequencing. In silico characterization of WNT1 mutation was performed using multiple platforms. RESULTS: Nine affected family members exhibited severe bone deformities, recurrent fractures, short stature and low bone mineral density. SNP array data revealed homozygous segments > 1 Mb in length accounting for 2.1-12.7% of the genome in affected individuals and their siblings and a single 6,344,821 bp homozygous region in all affected individuals on chromosome 12q12-q13. This region includes two potential OI candidate genes WNT1 and VDR. We did whole-exome sequencing for both genes in two patients and identified a novel damaging missense mutation in exon 4 of WNT1: c.1168G > T (NM_005430) resulting in p.G324C. Sanger sequencing confirmed segregation of mutation with the disease in family. CONCLUSION: We report a novel mutation responsible for OI and our investigation expands the spectrum of disease-causing WNT1 mutations and the resulting OI phenotypes.

Can IFNL3 polymorphisms predict response to interferon/ribavirin treatment in hepatitis C patients with genotype 3?

Favourable genotypes of IFNL3 polymorphism CC for rs12979860 and TT for rs8099917 are strongly associated with the interferon/ribavirin treatment outcome in hepatitis C virus (HCV) patients with genotypes 1 and 4. Contrarily, conflicting results have been reported for patients with HCV genotypes 2 and 3. Therefore, we sought to investigate the association between IFNL3 with sustained virological response (SVR) after treatment to ascertain the predictive value of IFNL3 single-nucleotide polymorphisms (SNPs) in HCV patients with genotype 3. For this purpose, we genotyped five IFNL3 SNPs, rs12980275, rs12979860, rs9109886, rs8099917 and rs7248668, in HCV patients with genotype 3 and assessed its association with SVR, individually and in haplotype. Interestingly, we report that the IFNL3 SNPs we genotyped have shown no association with SVR following treatment, either individually or in haplotype, indicating that genotyping IFNL3 SNPs have limited predictive value in HCV patients with genotype 3. Therefore, we propose that IFNL3 genotyping can be excluded from a patient's pre-treatment workup for subsequent treatment choice. This will greatly reduce the economic burden for HCV patients with genotype 3 in resource-limited regions, especially South Asia where genotype 3 is predominant.

Evaluation of the thiazole Schiff bases as ß-glucuronidase inhibitors and their in silico studies.

Twenty eight (28) derivatives 2-29 were synthesized and four analogs were found to exhibit single-digit IC(50) values as ß-glucuronidase inhibitors. Molecular modeling indicates that three factors: substituent R, lone pair on the nitrogen of azomethine part, and the interactions made by the main skeleton of the molecule, determined the enzyme inhibitory potential of these compounds. The planar conformation of the molecules allows them to fit deep inside the pocket while blocking the entry of other physiological substrates seems to play an important role in their activity.

Host immune players and their response to Hepatitis C therapies.

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.

Expression of IFN-Gamma is significantly reduced during severity of covid-19 infection in hospitalized patients.

Cytokines play an important role in SARS-CoV-2 infection progression and severity. A number of inflammatory cytokines have been directly associated with disease severity including IL-6 (interleukin-6), IL-10, TNF-α (tumor necrosis factor alpha), IFN-γ (interferon-gamma). Here, in this study, the aim was to better understand the interplay between host immune response mediated by cytokines and severity of SARS-CoV-2 infection by assessing cytokine expression. Therefore, we measured expression levels of a total of 12 genes (IFNA-1, IFN-γ, IL-1α, IL-1ß, IL-4, IL-6, IL-7, IL-10, IL-11, IL-13, IL-15, and IL-27) encoding inflammatory, anti-inflammatory and regulatory cytokines using QRT-PCR in hospitalized patients with severe infection compared to mildly infected. IFN-γ was identified as a potent marker of disease severity as indicated previously. Moreover, levels of IL-7 were also found to be partially reduced in patients compared to the healthy controls and linked negatively to disease severity. Identification of these cytokines may be helpful in not only understanding disease pathogenesis but also in better management of the patients after covid infection.

Efficient treatment of tannery wastewater through aeration, coagulation, and algal pond.

BACKGROUND: Tannery wastewater effluents contain many toxic and carcinogenic heavy metals and physiochemical parameters that need to be removed before these effluents enter in the main water bodies or rivers. In this study, the effluents from the tannery industry are treated through aeration, coagulation, and Chlorella vulgaris pond treatment processes for the removal of physiochemical: parameters only. METHODS: The effect of removal efficiencies (%) was studied on the physicochemical parameters, including salinity, electrical conductivity (EC), total dissolved solids (TDS), turbidity, total suspended solids (TSS), biochemical oxygen demand (BOD), and chemical oxygen demand (COD). RESULTS: The key results showed that the removal of EC, TDS, turbidity, TSS, BOD, and COD was 80.2%, 67%, 81%, 80.8%, 68.6%, and 100%, respectively, in raw wastewater treatment having 25, 50, and 70 g of algae C. vulgaris doses. The removal efficiencies (%) of salinity, EC, TDS, turbidity, TSS, BOD, and COD were 83%, 87.1%, 77.1%, 80%, 40%, 97%, and 98%, respectively, during coagulated wastewater treatment with three doses of algae. The observed improvement in treated wastewater indicated that the removal efficiencies (%) of salinity, EC, TDS, turbidity, TSS, BOD, and COD were 85.7%, 39.3%, 81.3%, 67.8%, 50.3%, 97%, and 98%, with C. vulgaris. CONCLUSION: This study confirmed that the treatment of tannery wastewater by these processes increased the pollutant removal efficiencies as all the physiochemical parameters were exceeding the permissible limits. RESULTS CONTRIBUTION IN FUTURE: This research will be helpful to treat the industrial wastewaters or effluents before it further mixes up in the main water streams. In this way, water quality will be better, aquatic life will be saved, and further researchers can analyze more ways for efficient treatments as they have a baseline data through this study findings. PRACTITIONER POINTS: One of the most pollutant sources in terms of both physical and chemical parameters is the produced wastewater from tannery industries. The effluents from tannery industry are treated through aeration, coagulation, and algae ponds treatment processes. These treatment made the tannery wastewater as environmental friendly.

Revisiting the AIDS Epidemic in Pakistan: Where We Stand and What We Must Aim For.

Human immunodeficiency virus type 1 (HIV-1) is the major cause of acquired immunodeficiency syndrome (AIDS) in humans, where the immune system totally succumbs to the virus. A large proportion of the AIDS infected belong to developing countries and AIDS prevalence is intensified by severe poverty, malnutrition, and famine; fatal illnesses with a scorn shortage of medical amenities complemented with the lack of education and development. Current Pakistani health system setting is in a dire need of improvement. Low literacy rates, high birth rates, and associated maternal mortality plus a lack of clean drinking water and appropriate sanitation system have a serious impact on general living conditions contributing to a relatively short lifespan. HIV is, therefore, becoming a growing health concern in Pakistan with a rapid rise in the reported cases. AIDS is most prevalent among injection drug users (IDUs), male/female sex workers, and unchecked deported migrant workforce. To combat this virus, the Pakistan Government has been working hard over the past few years with local bodies and international organizations in an effort to combat this menace. This review aims to discuss the risk factors for the rise of this epidemic in the country and the recommendations, efforts to be done to address this alarming issue.

Heterogeneous genomic locations within NS3, NS4A and NS4B identified for genotyping and subtyping of Hepatitis C virus: A simple genome analysis approach.

Hepatitis C virus (HCV) displays excessive genetic heterogeneity and exists in several genotypes and subtypes. Characterizing these genotypes and subtypes becomes extremely important for diagnostic and epidemiological reasons. Present study analyzed HCV genome using a simple genome analysis approach. We combined manual sectioning of a reference genome alignment (RA) followed by a comprehensive comparative phylogenetic analysis. The main aim was to identify heterogeneous locations on HCV genome suitable for genotyping/subtyping. HCV reference dataset, comprising of whole genome sequences from all HCV genotypes and subtypes, was aligned into an RA. The RA was manually clipped into overlapping sections of 500 bases, each 50 bases apart. Phylogeny for each section and RA was estimated using neighbor-joining phylogenetic method. Clustering pattern between section phylogenies and RA phylogeny was compared for similarity. Sections (locations on genome) with clustering similar to whole genome were selected since it displays comparable genetic heterogeneity making these sections suitable for genotyping/subtyping. Based on this conception, we identified new genomic locations on NS3, NS4A and NS4B suitable for genotyping and subtyping. Exact genomic positions for known genotyping locations, core and NS5B were also identified. Furthermore, phylogenetic analyses at such small genomic scale provided opportunities to explore evolutionary relationships usually overlooked.

BCR-ABL1 in leukemia: disguise master outplays riding shotgun.

Leukemia is a many-sided molecular disorder that arises because of over expression of oncogenes, suppression of tumor suppressor genes, and chromosomal translocations. These chromosomal rearrangements are nonetheless among the many determinants that underlie transformation of cells from normal to a cancerous phenotype and predispose cells to refractoriness against interventions by reduced drug influx and substantial drug efflux. This review unfolds current understanding of BCR-ABL1 (break point cluster region-c-abl oncogene 1, non-receptor tyrosine kinase) signaling with a focus on apoptotic suppressive mechanisms and alternative approaches to chronic myeloid leukemia therapy.

Formal modeling and analysis of the MAL-associated biological regulatory network: insight into cerebral malaria.

The discrete modeling formalism of René Thomas is a well known approach for the modeling and analysis of Biological Regulatory Networks (BRNs). This formalism uses a set of parameters which reflect the dynamics of the BRN under study. These parameters are initially unknown but may be deduced from the appropriately chosen observed dynamics of a BRN. The discrete model can be further enriched by using the model checking tool HyTech along with delay parameters. This paves the way to accurately analyse a BRN and to make predictions about critical trajectories which lead to a normal or diseased response. In this paper, we apply the formal discrete and hybrid (discrete and continuous) modeling approaches to characterize behavior of the BRN associated with MyD88-adapter-like (MAL)--a key protein involved with innate immune response to infections. In order to demonstrate the practical effectiveness of our current work, different trajectories and corresponding conditions that may lead to the development of cerebral malaria (CM) are identified. Our results suggest that the system converges towards hyperinflammation if Bruton's tyrosine kinase (BTK) remains constitutively active along with pre-existing high cytokine levels which may play an important role in CM pathogenesis.