Assessment of ubiquitin specific Peptidase-18 gene in peripheral blood of chronic hepatitis C patients treated with direct-acting antiviral drugs.

Hepatitis C virus (HCV) infection remains one of the leading causes of liver complications globally. Ubiquitin Specific Peptidase-18 (USP18) is a ubiquitin-specific protease that cleaves interferon-stimulated gene 15 (ISG15) from ISGylated protein complexes and is involved in regulating interferon responsiveness. To study the effect of direct-acting antivirals (DAAs) on the USP18 gene using qPCR, 132 participants were recruited and classified into different groups based on treatment duration. USP18 expression was raised compared to rapid virologic response (RVR) and early virologic response (EVR) groups with P = 0.0026 and P = 0.0016, respectively. USP18 was found to be 7.36 folds higher in naïve patients than those with RVR and sustained viral response (SVR). In RVR and SVR groups where patients had cleared HCV RNA after treatment with direct-acting antiviral agents (DAA) therapy, the expression of USP18 was found to be low, with a fold change of 1.3 and 1.4 folds, respectively. Expression of USP18 was significantly higher in the non-RVR group than in the RVR group. In the No EVR group, gene expression was significantly higher than in the EVR group. It is concluded that targeting HCV proteins using DAAs can cause USP18 expression to be normalized more effectively. Moreover, USP18 is a vital marker indicating treatment resistance and distinguishing responders from non-responders during DAA therapy.

Large cell calcifying Sertoli cell tumour: molecular and immunohistochemical assessment of a series comprising non-metastasising and metastasising neoplasms.

Large cell calcifying Sertoli cell tumour (LCCSCT) is a type of testicular sex cord-stromal tumour that may occur sporadically or in the context of Carney complex and other genetic syndromes. A subset is clinically malignant, and the molecular mechanisms that drive such aggressive behaviour remain unknown. METHODS AND RESULTS: We analysed 21 samples from 20 patients with LCCSCT (12 non-metastasising and eight metastasising) using PRKAR1A immunohistochemistry (IHC) and next-generation sequencing. All tumours except two (cases 17 and 20, both metastasising) demonstrated loss of PRKAR1A expression. Among 11 cases with interpretable sequencing results, all harboured pathogenic single nucleotide variants of PRKAR1A. Evidence of loss of heterozygosity (LOH) of PRKAR1A was present in all tumours with interpretable zygosity data, but the mechanisms of LOH were different for non-metastasising and metastasising tumours. Non-metastasising tumours demonstrated only copy-neutral LOH, while metastasising tumours demonstrated a spectrum of mechanisms of LOH, including copy-loss LOH, two concurrent mutations or copy-neutral LOH. Relevant molecular findings in non-metastasising LCCSCT were limited to PRKAR1A variants. In contrast, all metastasising LCCSCTs with interpretable data harboured additional pathogenic variants, including (but not restricted to) BRCA2 mutations with evidence of LOH and bi-allelic CDKN2A/B deletions. Three patients harboured PRKAR1A variants of inferred germline origin, including one with Carney complex and two without known syndromic features. CONCLUSIONS: This study further confirms that PRKAR1A IHC is a useful diagnostic tool for both non-metastasising and metastasising tumours and suggests that molecular analyses can be helpful to identify non-metastasising tumours with malignant potential in selected patients. Importantly, these results highlight that germline assessment could be beneficial for all patients presenting with LCCSCT.

Assessment of Attitudes and Intentions towards COVID-19 Vaccines and Associated Factors among General Populations of Pakistan: A Cross-Sectional Study.

OBJECTIVE: The goal of public health in combatting COVID-19 is to increase herd immunity. However, vaccine reluctance makes attaining herd immunity a worldwide challenge. This investigation aimed to identify negative and positive attitudes and intentions about COVID-19 vaccinations. METHODS: A cross-sectional online survey was conducted once free COVID-19 vaccines became available in Pakistan in 2021. 4392 Pakistanis aged 18 and older were surveyed from seven administrative units between 1 July and 30 August 2021. Online structured questionnaires were utilized to collect data using a simple sampling procedure. The questionnaires were divided into three major sections: sociodemographic, health factors, and attitudes toward COVID-19. RESULTS: The survey link was shared with approximately 4500 participants. 97.6%(4392) completed the survey once begun. Frequency, percentage and Chi-square tests were used to analyze statistical data. Most of the participants in the research were men (2703 (61.54%)), 3277 (74.61%) were aged 18-29 years, and 1824 (41.53%) were residents of the Khyber Pakhtunkhwa province. (18.69%) Respondents expressed COVID-19 vaccine hesitancy, whereas 36.66% of participants liked getting the Sinopharm and Sinovac vaccines and (35.84%) of participants preferred the Pfizer vaccine. A significant number of participants (38.05%) were concerned about the vaccine's unexpected side effects Thus, it is essential to realize that many participants were concerned about the vaccine's unexpected side effects. CONCLUSIONS: The overall high level of concern about the unforeseen side effects of COVID-19 vaccines, as well as widespread vaccine hesitancy among Pakistani populations and its predictors, should be taken into account if public health intervention campaigns in Pakistan are changing negative attitudes and improving compliance with regard to COVID-19 vaccines.

Income inequality, financial development, and ecological footprint: fresh evidence from an asymmetric analysis.

The emerging environmental concerns are entrenched in social issues, largely stem from income differences and power disparity. Income distribution and environmental disruption are increasingly pointed as obstacles in securing sustainable development goals and environmental preservation. The existing empirical studies have explored the environmental pollution impact of income inequality. However, the results are conflicting, and little attention has been paid to explore the short and long-run environmental impacts from a national viewpoint. Similarly, the role of aggregate income and financial sector for environmental quality has attracted considerable attention and many studies have provided conflicting empirical evidence. The literature generally ignores the importance of relative income in explaining environmental outcomes and also assumes symmetric association, ignoring asymmetric shocks. The present study explores the role of nonlinear associations in forming the links between income distribution and environmental quality using linear and nonlinear autoregressive distributed lag models from 1972 to 2018. The study follows the extended environmental Kuznets curve (EKC) approach. The results suggest that inequality promotes environmental pollution. Further financial development also escalates carbon emissions. The nonlinear analysis confirms the asymmetric effect of inequality on ecological footprint. The EKC, however, is not validated for Pakistan. The results suggest important policy implications.

New Parameterized Inequalities for η-Quasiconvex Functions via (p, q)-Calculus.

In this work, first, we consider novel parameterized identities for the left and right part of the (p,q)-analogue of Hermite-Hadamard inequality. Second, using these new parameterized identities, we give new parameterized (p,q)-trapezoid and parameterized (p,q)-midpoint type integral inequalities via η-quasiconvex function. By changing values of parameter µ∈[0,1], some new special cases from the main results are obtained and some known results are recaptured as well. Finally, at the end, an application to special means is given as well. This new research has the potential to establish new boundaries in comparative literature and some well-known implications. From an application perspective, the proposed research on the η-quasiconvex function has interesting results that illustrate the applicability and superiority of the results obtained.

Health risk assessment of heavy metals via consumption of dietary vegetables using wastewater for irrigation in Swabi, Khyber Pakhtunkhwa, Pakistan.

Health assumptions to the population due to the utilization of contaminated vegetables have been a great concern all over the world. In this study, an investigation has been conducted to ascertain metal concentrations in the wastewater, soil and commonly consumed vegetables from the vicinity of Gadoon Industrial Estate Swabi, Khyber Pakhtunkhwa Pakistan. Physicochemical parameters such as pH, electrical conductivity (EC), total dissolved solids (TDS), total suspended solids (TSS) and total solids (TS) and heavy metals such as Pb, Cr, Cd, Ni, Zn, Cu, Fe, Mn were determined using Atomic Absorption Spectrophotometer (AAS). Moreover, possible health risks due to the consumption of vegetables have also been estimated. pH and TSS in wastewater were found to be higher than the permissible limit set by WHO (1996). These results revealed that Cr concentration in the wastewater was above the permissible limits of United States Environmental Protection Agency (USEPA) which may lead to a detrimental effect on soil quality deterioration, ultimately leading to food contamination. ANOVA analysis demonstrated a significant difference in soil samples for Pb, Cr, Cd, Ni, Zn and Cu at p ≤ 0.001, for Mn at p ≤ 0.05 while no significant difference was observed for Fe respectively. ANOVA analysis also exhibited the highest mean value for Pb, Cr, Cd and Zn in vegetables. A substantial positive correlation was found among the soil and vegetable contamination. The transfer factor for Cr, Pb, Zn, Mn, Ni, Cd and Cu was greater than 0.5 due to contamination caused by domestic discharges and industrial effluents. Health assessment via consumption of dietary vegetables revealed a higher level than the permissible limit (HRI > 1) for Pb and Cd in children and adults. Enrichment factor (EF) due to consumption of vegetables was found higher for Pb and Cr respectively. Based on the findings of this study, there would be a significant risk to the consumers associated with consumptions of vegetables being cultivated in Gadoon Industrial Estate area of district Swabi. Therefore, strict regulatory control measures are highly recommended for the safety of vegetables originated from the study area.

Low-cost green recyclable biomaterial for energy-dependent electrical switching and intact biofilm with antibacterial properties.

A highly cost-effective recycled biomaterial extracted from lime peel has been made biocompatible and has been coated on a commercial fluorine-doped tin oxide (FTO) substrate of glass using the spin coating method. Structural, morphologic, electronic, and antibacterial measurements were thoroughly characterized as a green biomaterial thin film using X-rays (XRD), PL, FTIR, Raman, SEM, HRTEM, AFM, I-V, and antibacterial diffusion techniques. The comprehensive analysis of structures of recyclable waste in the form of lime peel extract (LPE) as thin films showed the crystalline cellulose structure that corresponds to the lattice fringe (0.342 nm) exposed by HRTEM. The K+1 interstitial active sites or vacancies in LPE/FTO thin films are confirmed by the PL spectra that show important evidence about conduction mechanisms, and hence facilitates Ag+1 ion migration from the top to the bottom electrode. The AFM investigations revealed the minor surface roughness (169.61 nm) of the LPE/FTO film, which controls the current leakage that is associated with surface defects. The designed memory cell (Ag/LPE/FTO) exhibits stable, reproducible electrical switching under low operational voltage and is equipped with excellent retention over 5 × 103 s. Furthermore, owing to presence of flavonoids and their superior antioxidant nature, lime peel extract powder shows tremendous antimicrobial activity against gram-positive and Gram-negative bacterial strains.

Heavy Metals Level, Health Risk Assessment Associated with Contamination of Black Tea; A Case Study from Khyber Pakhtunkhwa (KPK), Pakistan.

In the present study, 15 different commercial tea brands sold in Khyber Pakhtunkhwa were collected from the markets. The samples were analyzed for the concentrations of ten selected heavy metals. The metal concentration showed a random distribution in all samples. The mean concentration of Cd, Cu, Mn, Pb, Zn, and Fe was found in the range of 0.029-0.094 mg kg-1, 7.11-12.30 mg kg-1, 20.73-24.17 mg kg-1, 0.159-0.824 mg kg-1, 1.136-2.938 mg kg-1, and 0.670-118.30 mg kg-1 respectively. Co, Cr, Ni, and Sb were found below the detection limit of the instrument. Cu and Mn were found to be the abundant metals with a high concentration in the collected samples. The estimated daily intake (EDI), target hazard quotients (THQs), and hazard index (HI) were used for the assessment of health risks associated with the intake of metals. The metal transfer rates to tea infusion were reported from previous studies. Except for Cu, the EDI values of all the elements were found to be lower than the RfD values. The corresponding HI values of metals, in the different tea brands, were found to be below 1 suggesting that the consumption of mature tea infusions in the studied area could cause no carcinogenic risk. The principle cluster analysis (PCA) was used to reduce the number of variables to a new set which extracted three factors. For the assessment of health risks associated with dietary metal exposure, constant determination of heavy metals in all food is necessary. The present study provides valuable information to the general public about the consumption of tea infusions.

A computational framework and sensitivity analysis for the hormonal treatment of bone.

BACKGROUND: Osteoporosis is a bone disease identified by disordering of bone formation and resorption cells. It increases the risk of bone fragility and fracture. Autocrine and paracrine signalling of osteoclasts and osteoblasts plays an important role in the regulation of bone remodelling. Calcitonin is an approved pharmacologic agent for the treatment of osteoporosis. METHODS: A novel mathematical model comprising the interaction among osteoclasts and osteoblasts cells with intermittent administration of calcitonin has been presented to study the dynamics of osteoporotic bone. The stability of model and sensitivity of parameters are also discussed. FINDINGS: The population of osteoclastic and osteoblastic cells has been predicted via numerical simulation. The results of Monte Carlo sensitivity analysis are shown via tornado diagram. INTERPRETATION: It is concluded that intermittent administration of calcitonin is an effective therapy for the treatment of osteoporosis.

Environmental risk assessment of chronic arsenic in drinking water and prevalence of type-2 diabetes mellitus in Pakistan.

Chronic arsenic (As) unprotection in drinking water can lead to Type 2 Diabetes Mellitus (T2DM). The purpose of this study was to investigate the association between chronic As in drinking water and the prevalence of T2DM. A study was conducted in targeted urban areas of Peshawar city of KPK, Pakistan, where drinking water is heavily contaminated with chronic arsenic. Participants protected to arsenic were selected from Kohat city of KPK, Pakistan (where people consumed water that is free from As contamination) and treated as the control group. People with arsenic-related skin lesions were defined as participants unprotected to arsenic. T2DM was diagnosed using a glucometer following the fasting blood glucose ≥6.0 mmol L-1 from the WHO guideline. The common odds ratio for T2DM among participants unprotected to arsenic was <4. The Mantel-Haenszel weighted prevalence ratio with 95% of confidence interval for confounding factors were (age <4 m femininity <4 and body mass index >4). The results revealed that designated association were important. The findings suggested that unprotected chronic arsenic in drinking water may be a risk factor of T2DM.

DOG1 immunohistochemical staining of testicular biopsies is a reliable tool for objective assessment of infertility.

Testicular biopsy may be a component of the work-up of male infertility. However, no reliable diagnostic tools are available for objective quantitative assessment of spermatogenic cells. It is well known that MAGE-A4 is selectively expressed in spermatogonia and our group has previously demonstrated that DOG1 differentially stains germ cells. Therefore, we performed DOG1 and a double stain cocktail (DOG1 and 57b murine monoclonal anti-MAGE-A4) immunohistochemical stains on 40 testicular infertility biopsies (10 each with active spermatogenesis, Sertoli cell-only, hypospermatogenesis, and maturation arrest), 25 benign seminiferous tubules from radical orchiectomies, and 5 spermatocytic tumors (ST). In biopsies/resections with active spermatogenesis, DOG1 stained spermatocytes and spermatids and was absent in spermatogonia, while MAGE-A4 stained spermatogonia and primary spermatocytes (weak). In hypospermatogenesis, DOG1 highlighted decreased spermatocytes/spermatids and MAGE-A4 highlighted decreased spermatogonia. DOG1 staining confirmed decreased to absent spermatocytes in maturation arrest and MAGE-A4 staining established the presence of preserved spermatogonia in all cases. All STs were negative for DOG1 and positive for MAGE-A4, while all Sertoli cell-only cases were negative for DOG1 and the double stain cocktail. In conclusion, we confirmed that DOG1 is expressed in spermatocytes and spermatids and MAGE-A4 highlights primarily spermatogonia. Usage of these stains facilitates confirmation of maturation arrest, assessment of the percentage of testis involvement in hypospermatogenesis and identification of mixed patterns. Finally, this study supports that the differentiation of STs is more closely related to spermatogonia than the more mature spermatocytes.

Detection of Anti-Hepatitis B Virus Drug Resistance Mutations Based on Multicolor Melting Curve Analysis.

Detection of anti-hepatitis B virus (HBV) drug resistance mutations is critical for therapeutic decisions for chronic hepatitis B virus infection. We describe a real-time PCR-based assay using multicolor melting curve analysis (MMCA) that could accurately detect 24 HBV nucleotide mutations at 10 amino acid positions in the reverse transcriptase region of the HBV polymerase gene. The two-reaction assay had a limit of detection of 5 copies per reaction and could detect a minor mutant population (5% of the total population) with the reverse transcriptase M204V amino acid mutation in the presence of the major wild-type population when the overall concentration was 104 copies/µl. The assay could be finished within 3 h, and the cost of materials for each sample was less than $10. Clinical validation studies using three groups of samples from both nucleos(t)ide analog-treated and -untreated patients showed that the results for 99.3% (840/846) of the samples and 99.9% (8,454/8,460) of the amino acids were concordant with those of Sanger sequencing of the PCR amplicon from the HBV reverse transcriptase region (PCR Sanger sequencing). HBV DNA in six samples with mixed infections consisting of minor mutant subpopulations was undetected by the PCR Sanger sequencing method but was detected by MMCA, and the results were confirmed by coamplification at a lower denaturation temperature-PCR Sanger sequencing. Among the treated patients, 48.6% (103/212) harbored viruses that displayed lamivudine monoresistance, adefovir monoresistance, entecavir resistance, or lamivudine and adefovir resistance. Among the untreated patients, the Chinese group had more mutation-containing samples than did the Pakistani group (3.3% versus 0.56%). Because of its accuracy, rapidness, wide-range coverage, and cost-effectiveness, the real-time PCR assay could be a robust tool for the detection if anti-HBV drug resistance mutations in resource-limited countries.