A multiplex PCR assay to detect mislabelling in fish products.

Fish substitution in fish products is an important issue in fish markets, as it is a widespread practice. An authentication protocol for Rohu, Thaila and Tilapia was developed by multiplex PCR. Three species-specific and one degenerate common forward primer were designed using the Cytb gene of the mitochondrial genome. These primers for Labeo rohita, Labeo catla and Oreochromis niloticus showed the fragment size of 235 bp, 186 bp and 506 bp on the agarose gel, respectively. The primers for L. rohita and L. catla were sensitive to 0.1 ng of DNA template, while for O. niloticus this value was 1 ng of DNA template. A total of 230 commercial samples (160 fried and 70 processed fish products) were screened, where 60% mislabeling in fried and 30% mislabeling in processed fish were found. This multiplex PCR protocol could give useful insights for food inspection and enforcement of regulatory food control.

Delving the vitamin D receptor variation and expression profiles in the context of type 2 diabetes among families.

BACKGROUND: Vitamin D is essential for insulin secretion and sensitivity. Consequently, its inadequacy is linked to higher insulin resistance and Type 2 Diabetes (T2D). The Vitamin D receptor (VDR) gene is one potential candidate for T2D, and multiple polymorphisms in VDR have been examined in various populations, but no conclusive answers have been provided. OBJECTIVES: This study was designed to evaluate the susceptibility of VDR gene polymorphism and its expression in diabetic families in Pakistan. METHODOLOGY: In this family-based study, twenty diabetic families with a positive family history of T2D and at least three T2D patients were recruited from outpatient clinics and public hospitals. The current study comprised 143 individuals with 55 affected and 88 unaffected individuals. Blood samples of the selected families were collected. DNA was extracted from the collected samples and the PCR-RFLP method was followed to identify the genotyping and RT-qPCR for expression. Phenotypic and genotypic pedigrees of the families were developed by the progeny online tool. The association values of SNPs were determined by TDT and DFAM analysis implemented on Plink software. RESULTS: The results explained a significant familial aggregation among phenotypic characters including Age, Gender, BMI (body mass index), age of disease diagnosis, disease duration, and blood pressure in the probands, affected FDRs (First Degree Relatives) and affected SDRs (Second Degree Relatives). A significant association of rs731236 C/T (OR = 1.522), rs2228570 C/T (OR = 1.327) with p < 0.05. Whereas, for rs1544410 G/A (OR = 0.9706) and rs7975232 T/G (OR = 0.7368) no considerable association evidence was seen (p > 0.05) in families. The mRNA expression of VDR increased threefold (p = 0.0204) in patients compared to controls. Variation-based expression analysis exhibited that the rs2228570 genotype influences the expression. CONCLUSION: A linkage was found among the FDRs with probands. Variation in the gene VDR at loci rs731236 and rs2228570 was associated with familial T2D. However further research is required to explore more genetic factors that could influence T2D risks in families.

Exosomes in Cancer: Diagnostic and Therapeutic Applications.

Small extracellular vesicles called exosomes are produced by cells and contain a range of biomolecules, including proteins, lipids, and nucleic acids. Exosomes have been implicated in the development and spread of cancer, and recent studies have shown that their contents may be exploited as biomarkers for early detection and ongoing surveillance of the disease. In this review article, we summarize the current knowledge on exosomes as biomarkers of cancer. We discuss the various methods used for exosome isolation and characterization, as well as the different types of biomolecules found within exosomes that are relevant for cancer diagnosis and prognosis. We also highlight recent studies that have demonstrated the utility of exosomal biomarkers in different types of cancer, such as lung cancer, breast cancer, and pancreatic cancer. Overall, exosomes show great promise as noninvasive biomarkers for cancer detection and monitoring. Exosomes have the ability to transform cancer diagnostic and therapeutic paradigms, providing promise for more efficient and individualized. This review seeks to serve as an inspiration for new ideas and research in the never-ending fight against cancer. Moreover, further studies are needed to validate their clinical utility and establish standardized protocols for their isolation and analysis. With continued research and development, exosomal biomarkers have the potential to revolutionize cancer diagnosis and treatment.

Geographic distribution and genetic diversity of Newcastle disease virus in pigeons from Pakistan.

RESEARCH HIGHLIGHTS: Virulent NDV genotypes were repeatedly isolated from pigeons.Evidence of epidemiological links among viruses isolated from various locations.Distinct phylogenetic branches suggest separate, simultaneous evolution of NDVs.Study information could be helpful in the development of an effective vaccine.

Phylogenetic and antigenic analysis of infectious bronchitis virus isolated from commercial and backyard chickens in Pakistan, 2015-2018.

Infectious bronchitis virus (IBV) is a rapidly evolving virus affecting both vaccinated and unvaccinated poultry flocks and is responsible for significant economic losses globally; hence, it is imperative to obtain a deeper understanding of this pathogen. In this study, seven IBV strains were isolated from commercial and backyard poultry flocks during 2015-2018. We obtained full-length IBV genomes of two viruses using the Illumina sequencing method, while five additional viruses were genetically characterized through full-length spike (S1) gene sequencing. Phylogenetic and distance analysis based on complete S1 gene and full-length genome sequences revealed that one IBV isolate belonged to genotype GI-1 and six viruses were clustered within genotype GI-13. Deduced amino acid sequences of GI-13 strains exhibited 31.8-37.2 % divergence with the commonly used classic vaccine strains (M41) and 2.7-12.6 % with variant vaccine strains (4/91) in Pakistan. High evolutionary distances suggest that the IBV viruses circulating in Pakistan are under continuous evolutionary pressure. Moreover, ch/IBV/Pak/AW-2/2017 was found to have originated from an intra-genotypic recombination event between the variant group (GI-23 lineage as a major parent) and variant vaccine strain (4/91-like as a minor parent) and is the first example of recombination within genotype GI-13 in Pakistan. Together, these findings provide genetic and evolutionary insights into the currently circulating IBV genotypes in Pakistan, which could help to better understand the origin, spread and evolution of IBVs, and to ascertain the importance of disease monitoring as well as re-evaluation forof currently used vaccines and vaccination programmes.

Hepatitis C virus/Hepatitis B virus coinfection: Current prospectives.

In endemic areas, hepatitis C virus (HCV)/hepatitis B virus (HBV) coinfection is common, and patients with coinfection have a higher risk of developing liver disease such as hepatocellular carcinoma, liver fibrosis and cirrhosis. In such cases, HCV predominates, and HBV replication is suppressed by HCV. HCV core proteins and interferons that are activated by HCV are responsible for the suppression of HBV. Immunosuppression is also seen in patients with HCV and HBV coinfections. A decrease in HCV-neutralizing antibody response and circulation of Th1-like Tfh cells is observed in patients with HCV and HBV coinfection. Both viruses interacted in the liver, and treatment of HCV/HBV coinfection is genotype-based and complex due to the interaction of both viruses. In HCV-dominant cases, direct-acting antiviral drugs and peg interferon plus ribavirin are used for the treatment, with continuous monitoring of AST and ALT. HBV-dominant cases are less common and are treated with peg interferon and nucleoside nucleotide analogues with monitoring of AST and ALT. The SVR rate in HCV-HBV coinfection is higher than that in monoinfection when treated with direct-acting antiviral drugs. But there is a risk of reactivation of HBV during and after therapy. The rate of reactivation is lower in patients treated with direct-acting antiviral drugs as compared to those treated with peg interferon plus ribavirin. Biomarkers of HBV such as HBcrAg, HBV DNA and HBVpg RNA are not effective in the prediction of HBV reactivation; only the hepatitis B surface antigen titre can be used as a biomarker for HBV reactivation. HCV can also be reactive, but this is found in very rare cases in which HBV is present and is treated first.

Parasitological, molecular, and epidemiological investigation of Trypanosoma evansi infection among dromedary camels in Balochistan province.

Surra is a zoonotic disease caused by Trypanosoma evansi (T. evansi), which affects a wide variety of animals worldwide. The disease has a severe impact on the productivity, health, and working capacity of camels and causes mortality and extensive economic losses if not diagnosed early. This is the first comprehensive report on the prevalence of T. evansi infection in dromedaries in Balochistan province. In the present study, 393 blood samples (indigenous, n = 240; imported, n=153) were collected from one-humped camels (Camelus dromedarius) and were tested by molecular methods to estimate the prevalence of T. evansi in three districts (Pishin, Nushki, and Lasbella) of Balochistan province. The overall prevalence of T. evansi among examined camel samples was 28.24% (95% confidence interval (CI): 24.02-32.89%). The risk of T. evansi infection in adult camels (> 10 years) is higher than that in young ones (odd-ration (OR) = 2.7; 95% CI: 1.3357-5.3164%). Moreover, male camels were six times more likely to get an infection than female camels. The detection of T. evansi infection in camels sampled in summer and spring was 3.12- and 5.10-fold higher, respectively, than in camels sampled in winter. In conclusion, our findings showed a high rate of T. evansi infection in camels from the three districts. Our study emphasizes the need for a strict surveillance program and risk assessment studies as prerequisites for control measures.

Trajectories of maternal depressive symptoms from pregnancy to 11 years postpartum: Findings from Avon Longitudinal Study of Parents and Children (ALSPAC) cohort.

BACKGROUND: Maternal depressive symptoms are common in pregnancy and may extend to the perinatal period and beyond for some women. To date, few longitudinal studies have investigated maternal depressive symptoms from pregnancy to eleven years postpartum. Drawing data from a large population-based study cohort the aims of this study were to 1) identify distinct groups of mothers defined by their trajectories of depressive symptoms spanning from pregnancy to eleven years following the birth of the child, and 2) to identify psychosocial risk factors during pregnancy and in the first few postnatal years that are associated with these trajectories. METHODS: Data were analyzed from 14,170 mothers who participated in Avon Longitudinal Study of Parents and Children (ALSPAC). The Edinburgh Postnatal Depression Scale (EPDS) was used to capture maternal depressive symptoms across 10 time points including two prenatal (18 and 32 weeks), and eight postnatal (2, 8, 21, 33, 61, 73, 97 and 134 months) time points. The latent growth model was created to describe the course of maternal depressive symptoms across the preceding time points followed by a latent growth mixture modelling (LGMM) to identify distinct trajectories of depressive symptoms over time within the overall sample. The predictors of maternal depressive symptoms trajectories were categorized into sociodemographic, child, and psychosocial factors. The multinomial regression analyses were conducted to explore associations between the risk factors and depressive symptoms trajectories. RESULTS: LGMM identified four distinct trajectories of maternal depressive symptoms over time: minimal symptoms, increasing symptoms, persistent symptoms, and decreasing symptoms. Predictors of all patterns of depression - persistent, increasing and decreasing symptoms include smoking during pregnancy, and partner conflict. The strongest predictors of the persistent symptom trajectory included maternal history of depression and inadequate social support. LIMITATIONS: The use of self-reported maternal mental health symptoms and under representation of ethnic minorities are our study's limitations. CONCLUSIONS: The study findings highlight the importance of early identification and treatment for mothers experiencing depressive symptoms from pregnancy to the perinatal period and beyond.

Genomic and comparative clinico-pathological assessment of two Pakistani pigeon-derived newcastle disease virus sub-genotypes XXI.1.1 and XXI.1.2 isolated in 2017.

Pigeon paramyxovirus type-1 (PPMV-1) is an antigenic-variant of Newcastle disease virus (NDV) which is associated with infection in Columbidae family. In this study, we isolated two pigeon-derived strains pi/Pak/Lhr/SA_1/17 (designed as SA_1) and pi/Pak/Lhr/SA_2/17 (designed as SA_2) from diseased pigeons collected in Punjab province in 2017. We performed the whole genome, phylogenetic analysis and comparative clinico-pathological evaluation of two viruses in pigeons. Phylogenetic analysis based on fusion (F) gene and complete genome sequences showed that SA_1 belonged to sub-genotype XXI.1.1 and SA_2 clustered in sub-genotype XXI.1.2. SA_1 and SA_2 viruses contributed to morbidity and mortality in pigeons. Remarkably, although the two viruses resulted in comparatively similar pattern of pathogenesis and replication ability in various tissues of infected pigeons, SA_2 could cause more severe histopathological lesions and had comparatively high replication ability in pigeons than SA_1. Moreover, pigeons infected with SA_2 had higher shedding efficiency than that of pigeons infected with SA_1. Moreover, several aa substitutions in the major functional domains of the F and HN proteins might be contributed to the pathogenic differences between the two isolates in pigeons. Overall, these findings provide us with important insight into the epidemiology and evolution of PPMV-1 in Pakistan and laid the foundation for the further elucidation of the mechanism underlying the pathogenic difference of PPMV-1 in pigeons.

Gestational Antibodies to C. pneumoniae, H. pylori and CMV in Women with Preeclampsia and in Matched Controls.

OBJECTIVES: Some research has suggested a possible role for past infection in the development of preeclampsia. The objective of this study was to explore the role of Helicobacter pylori, cytomegalovirus, and Chlamydophila pneumoniae in the development of preeclampsia in a prospective pregnancy sample. METHODS: We conducted a nested case-control study in The Archive for Child Health (ARCH), a pregnancy cohort of 867 unselected women enrolled at the first prenatal visit with archived blood and urine in pregnancy. We matched 21 cases of preeclampsia to 52 unaffected controls on maternal age (±4 years), race, parity, and gestational age at blood draw. Using conditional logistic regression, we examined the association between preeclampsia status and immunoglobulins G (IgG) tested by indirect ELISA to each of the three microorganisms, adjusting for potential confounders. RESULTS: No significant difference was found between cases and controls. The unadjusted odds ratio was 1.5 (95%CI: 0.2-9.1), 0.6 (95%CI: 0.2-1.9), and 1.9 (95%CI: 0.6-5.6) for H. pylori, cytomegalovirus and C. pneumoniae respectively. After controlling for confounders analysis found increased odds of H. pylori IgG (AOR: 1.9; 95% CI: 0.2-15.3) and C. pneumoniae IgG (AOR: 2.3; 95% CI: 0.6-9.2) for preeclampsia, albeit being not significant. Conversely, cytomegalovirus IgG had lower odds for preeclampsia (AOR: 0.4; 95% CI: 0.1-1.7). CONCLUSIONS: Past infection with H. pylori, and C. pneumoniae in early pregnancy showed a higher risk of preeclampsia, but the findings failed to achieve statistical significance. Cytomegalovirus was not associated with preeclampsia in these data. These preliminary findings encourage future research in populations with high prevalence of these infections.

Surveillance and Assessment of Risk Factors for Newcastle Disease Virus from Live Bird Retail Stalls in Lahore District of Pakistan.

Live bird markets (LBMs) in Asian countries are considered hubs for the spread of several poultry viruses. In Pakistan, there is a lack of uniformity in practices used in LBMs, which leads to the spread of poultry diseases. A cross-sectional survey was conducted in June-October 2017 to determine the circulation of Newcastle disease virus (NDV) in chickens being sold in live bird retail stalls (LBRSs) and to identify potential risk factors associated with estimated prevalence. A total of 189 stalls (n = 1134 birds) distributed in eight administrative towns of Lahore were visited. A pool of six oropharyngeal swabs was collected from each stall and tested by real-time reverse transcriptase PCR for the presence of NDV. Forty-two out of 189 swabs were found positive with an overall prevalence of 22.22% (95% confidence interval [Cl]: 16.88%-28.67%). Data for 11 potential risk factors acquired through questionnaires were analyzed by survey-weighted logistic regression and prevalence odds ratios (ORs) for associated risk factors were calculated. A final multivariable model identified three risk factors for NDV prevalence in LBRSs, including trading other poultry breeds alongside broilers (OR = 2.41; 95% confidence interval [CI] = 1.5-6.1), purchasing birds from mixed sources (OR = 3.12; 95% CI = 1.4-11.9), and number of birds sold per day (OR = 6.32; 95% CI = 1.9-23.5). Additionally, 24 selected samples were sequenced and phylogenetic analysis of the complete fusion gene (1662 bp) revealed that all isolates belonged to Subgenotype VII.2. This study provides important information on the epidemiology of NDV in Pakistan and highlights the importance of implementing surveillance and biosecurity practices in LBRSs.

Vigilancia y evaluación de factores de riesgo para el virus de la enfermedad de Newcastle en puestos de venta al menudeo de aves vivas en el distrito de Lahore en Pakistán. Los mercados de aves vivas (LBM, por sus siglas en inglés) en los países asiáticos se consideran centros de propagación de varios virus aviares. En Pakistán, existe una falta de uniformidad en las prácticas utilizadas en los mercados de aves vivas, lo que conduce a la propagación de enfermedades avícolas. Se realizó una encuesta transversal de junio a octubre del 2017 para determinar la circulación del virus de la enfermedad de Newcastle (NDV) en pollos que se venden en puestos minoristas de aves vivas y para identificar posibles factores de riesgo asociados con la prevalencia estimada. Se visitó un total de 189 puestos (n = 1134 aves) distribuidos en ocho ciudades administrativas de Lahore. Se recolectó un grupo de seis hisopos orofaríngeos de cada puesto y se analizó mediante transcripción reversa y PCR en tiempo real para detectar la presencia del virus de Newcastle. Cuarenta y dos de los 189 hisopos resultaron positivos con una prevalencia general del 22.22 % (intervalo de confianza [IC] del 95 % = 16.88­28.67). Los datos para 11 factores de riesgo potenciales adquiridos a través de cuestionarios se analizaron mediante regresión logística ponderada por encuesta y se calcularon las razones de probabilidad (OR) de prevalencia para los factores de riesgo asociados. Un modelo multivariable final identificó tres factores de riesgo para la prevalencia del virus de Newcastle en puestos minoristas de aves vivas, incluido el comercio de otras razas de aves de corral junto con pollos de engorde (OR = 2.41; IC del 95 % = 1.5­6.1), la compra de aves de fuentes mixtas (OR = 3.12; IC del 95 % = 1.4 ­11.9), y número de aves vendidas por día (OR = 6.32; IC 95% = 1.9­23.5). Además, se secuenciaron 24 muestras seleccionadas y el análisis filogenético del gene de fusión completo (1662 pb) reveló que todos los aislamientos pertenecían al subgenotipo VII.2. Este estudio brinda información importante sobre la epidemiología del virus de Newcastle en Pakistán y destaca la importancia de implementar prácticas de vigilancia y bioseguridad en los en puestos minoristas de aves vivas.

Performance Analysis of Wearable Dual-Band Patch Antenna Based on EBG and SRR Surfaces.

This paper presents the performance comparison of a dual-band conventional antenna with a split-ring resonator (SRR)- and electromagnetic bandgap (EBG)-based dual-band design operating at 2.4 GHz and 5.4 GHz. The compactness and dual-frequency operation in the legacy Wi-Fi range of this design make it highly favorable for wearable sensor network-based Internet of Things (IoT) applications. Considering the current need for wearable antennas, wash cotton (with a relative permittivity of 1.51) is used as a substrate material for both conventional and metamaterial-based antennas. The radiation characteristics of the conventional antenna are compared with the EBG and SRR ground planes-based antennas in terms of return loss, gain, and efficiency. It is found that the SRR-based antenna is more efficient in terms of gain and surface wave suppression as well as more compact in comparison with its two counterparts. The compared results are found to be based on two distinct frequency ranges, namely, 2.4 GHz and 5.4 GHz. The suggested SRR-based antenna exhibits improved performance at 5.4 GHz, with gains of 7.39 dbi, bandwidths of 374 MHz, total efficiencies of 64.7%, and HPBWs of 43.2 degrees. The measurements made in bent condition are 6.22 db, 313 MHz, 52.45%, and 22.3 degrees, respectively. The three considered antennas (conventional, EBG-based, and SRR-based) are designed with a compact size to be well-suited for biomedical sensors, and specific absorption rate (SAR) analysis is performed to ensure user safety. In addition, the performance of the proposed antenna under bending conditions is also considered to present a realistic approach for a practical antenna design.

Sociodemographic and genetic determinants of nonalcoholic fatty liver disease in type 2 diabetes mellitus patients.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) showed significant association with PNPLA3 rs738409 polymorphism in unrelated individuals. However, it is still unknown whether the relationship of NAFLD with PNPLA3 variant exists or not among subjects with type 2 diabetes mellitus (T2DM). Therefore, the study aimed to evaluate sociodemographic and genetic determinants of NAFLD in type 2 diabetics. METHODS: The cross-sectional analytical study was conducted at the Department of Molecular Biology, Virtual University of Pakistan, Lahore, Pakistan, during 2019-2020. A total of 153 known cases of T2DM were enrolled using convenience sampling. After excluding patients (n = 24) with HCV, alcoholism, or missing information, data from 129 eligible diabetics with and without NAFLD were analyzed using SPSS. Odds ratios using crosstabs and adjusted odds ratios using binary and multinomial logistic regression were calculated to measure the risk of NAFLD. RESULTS: Adults 18-35 years were 7.0%, 36-55 years were 64.3%, ≥ 56 years were 28.7%, and females were 66.7%. A total of 41.1% of patients had obesity, 52.7% had NAFLD, and 29.05% carried mutant G allele of rs738409 polymorphism. Among overall diabetics, NAFLD showed association with female (OR = 2.998, p = 0.007), illiterate (OR = 3.067, p = 0.005), and obese (OR = 2.211, p = 0.046) but not with PNPLA3 genotype under any model (all p = > 0.05). Among obese diabetics, NAFLD showed association with female (AOR = 4.010, p = 0.029), illiterate (AOR = 3.506, p = 0.037), GG + CG/CC (AOR = 3.303, p = 0.033), and GG/CG + CC (AOR = 4.547, p = 0.034) using binary regression and with female (AOR = 3.411, p = 0.051), illiterate (AOR = 3.323, p = 0.048), GG + CG/CC (AOR = 3.270, p = 0.029), and GG/CG + CC (AOR = 4.534, p = 0.024) using multinomial regression. CONCLUSIONS: NAFLD and obesity were the most common comorbid diseases of T2DM. Gender female, being illiterate, and PNPLA3 rs738409 polymorphism were significant risk factors of NAFLD among obese diabetic patients.

Patterns and predictors of depressive and anxiety symptoms in mothers affected by previous prenatal loss in the ALSPAC birth cohort.

BACKGROUND: Studies investigating the patterns or predictors of psychological distress in expecting and postpartum mothers affected by previous prenatal loss, are limited. The study objectives were to explore longitudinal trajectory patterns of depressive and anxiety symptoms in mothers affected by previous prenatal loss from early in a subsequent pregnancy up to pre-adolescence, and to identify early factors predictive of elevated symptom trajectory patterns. METHODS: A total of 2854 mothers from the Avon Longitudinal Study of Parents and Children self-identified as having experienced a previous prenatal loss. A latent class analysis identified trajectory patterns of symptoms across 10 timepoints from 18-weeks' gestation up to 134-months postpartum, multivariate regression analysis identified predictors of elevated symptom trajectories, and hierarchical regression analysis determined predictive accuracy between predictors and elevated trajectory patterns. RESULTS: Three distinct longitudinal trajectory patterns of depressive and anxiety symptoms reflected low (54%), sub-clinical (34%), and clinical symptoms (12%). Key factors that predicted elevated symptom trajectory patterns better than increased symptom scores early in subsequent pregnancy include history of severe depression or other psychiatric problem, experiencing three or more stressful events from mid-pregnancy, inadequate social support, history of induced abortion, and history of abuse. Predictive accuracy of elevated trajectories was 0.542 (depression) and 0.432 (anxiety). LIMITATIONS: Generalizability may be compromised by attrition, under-reporting, and recall bias. CONCLUSION: Including factors predictive of long-term sub-clinical or clinical depressive and anxiety symptoms in early assessments will improve clinician's ability to identify mothers who may benefit from immediate and/or ongoing monitoring, and psychotherapeutic intervention after prenatal loss.

Isolation and phylogenetic analysis of Avian orthoavulavirus 1 sub-genotypes VII.2 and XXI.1.2 from caged birds in the Lahore district, Pakistan - Short communication.

In this study, the prevalence of Avian orthoavulavirus-1 (AOAV-1) (also commonly known as Newcastle disease virus) was investigated in caged birds kept in bird markets in the Lahore district of Pakistan. A total of 354 swab samples were obtained from 14 different species of clinically healthy birds. The overall virus prevalence was 12.7% in 9 out of the 14 species. Phylogenetic analysis of the complete fusion protein (F) gene showed that 23 isolates from different avian species belonged to sub-genotype VII.2 while three isolates of pigeon origin clustered with sub-genotype XXI.1.2. The VII.2 viruses isolated had a high nucleotide identity to viruses repeatedly isolated from poultry in Pakistan from 2011 to 2018. To date, sub-genotype XXI.1.2 viruses have only been identified in Pakistan. These findings suggest that the Newcastle disease (ND) outbreaks occurring in Pakistan involve multiple hosts and environments. The study emphasises the importance of continuing to monitor multiple avian species for the presence of AOAV-1s and implementing effective ND control strategies.

Identification of Cleavage Sites Proteolytically Processed by NS2B-NS3 Protease in Polyprotein of Japanese Encephalitis Virus.

Understanding the proteolytic processing of polyprotein mediated by NS2B-NS3 protease contributes to the exploration of the mechanisms underlying infection of Japanese encephalitis virus (JEV), a zoonotic flavivirus. In this study, eukaryotic and prokaryotic cell models were employed to identify the cleavage sites mediated by viral NS2B-NS3 protease in JEV polyprotein. Artificial green fluorescent protein (GFP) substrates that contained the predicted cleavage site sequences of JEV polyprotein were expressed in swine testicle (ST) cells in the presence and absence of JEV infection, or co-expressed in E. coli with the recombinant NS2B-NS3 protease that was generated by fusing the N-terminal protease domain of NS3 to the central hydrophilic domain of NS2B. The cleavage of GFP substrates was examined by western blot. Among twelve artificial GFP substrates containing the cleavage site sequences predictively processed by host cell and/or NS2B-NS3 proteases, all sites were found to be cleaved by host cell proteases with different efficiencies. The sites at internal C, NS2A/NS2B, NS2B/NS3 and NS3/NS4A junctions, but not the sites at internal NS3, internal NS4A and NS4B/NS5 junctions were identified to be cleaved by JEV NS2B-NS3 protease. These data provide insight into the proteolytic processing of polyprotein, which is useful for understanding JEV replication and pathogenesis.

Genomic and biological characteristics of Avian Orthoavulavirus-1 strains isolated from multiple wild birds and backyard chickens in Pakistan.

Circulation of the dominant sub-genotype VII.2 of Avian Orthoavulavirus-1 (AOAV-1) is affecting multiple poultry and non-poultry avian species and causing significant economic losses to the poultry industry worldwide. In countries where ND is endemic, continuous monitoring and characterization of field strains are necessary. In this study, genetic characteristics of eleven AOAV-1 strains were analyzed isolated from wild birds including parakeets (n = 3), lovebird parrot (n = 1), pheasant (n = 1), peacock (n = 1), and backyard chickens (n = 5) during 2015-2016. Genetic characterization (genome size [15,192 nucleotides], the presence of typical cleavage site [112-RRQKRF-117]) and biological assessment (HA log 27 to 29 and intracerebral pathogenicity index [ICPI] value ranging from 1.50 to 1.86) showed virulent AOAV-1. Phylogenetic analysis showed that the studied isolates belonged to sub-genotype VII.2 and genetically very closely related (> 98.9%) to viruses repeatedly isolated (2011-2018) from commercial poultry. These findings provide evidence for the existence of epidemiological links between poultry and wild bird species in the region where the disease is prevalent. The deduced amino acid analysis revealed several substitutions in critical domains of fusion and hemagglutinin-neuraminidase genes. The pathogenesis and transmission potential of wild bird-origin AOAV-1 strain (AW-Pht/2015) was evaluated in 21-day-old chickens that showed the strain was highly virulent causing clinical signs and killed all chickens. High viral loads were detected in different organs of the infected chickens correlating with the severity of lesions developed. The continuous monitoring of AOAV-1 isolates in different species of birds will improve our knowledge of the evolution of these viruses, thereby preventing possible panzootic.

Investigation of oxidative stress enzymes and histological alterations in tilapia exposed to chlorpyrifos.

Chlorpyrifos (ChF) is an organophosphate pesticide that is widely used in agricultural fields and indoor for controlling pests. Aquatic ecosystems are the recipients of various pesticide residues due to leaching spray drift and agricultural runoff and pose toxicity for aquatic organisms. Therefore, the current study was designed to investigate the oxidative stress enzymes and histological alterations in the vital organs of tilapia due to ChF exposure. LC50 (24 h) was calculated as 52.78 µg/l by exposing tilapia with different acute concentrations of ChF. For assessment of sub-lethal toxicity of ChF, the fish were divided into four groups (ChF1, ChF2, ChF3, and control group). ChF1 group was treated with 1/15th of LC50, whereas ChF2 and ChF3 groups were treated with 1/10th and 1/5th of LC50, respectively for 14 days. After that, ChF induced changes in oxidative stress enzymes and histological alterations were evaluated. It was found that the level of glutathione S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD) increased significantly in the liver of ChF-treated tilapia. Histological study of liver tissues showed an increased number of Kupffer cells, hydropic degeneration, necrosis, and hemorrhage. In the spleen of treated fish, increased melanomacrophage centers, necrosis, and congestion were detected. Disorganized muscle fibers, cardiac muscle fiber degeneration, and coagulative necrosis were observed in the heart of ChF-treated fish. It is concluded that sub-lethal concentrations of ChF can induce oxidative stress and histological alterations in the tissues of tilapia.

Interpersonal Psychotherapy to Reduce Psychological Distress in Perinatal Women: A Systematic Review.

BACKGROUND: Interpersonal psychotherapy (IPT) is a psychological intervention with established efficacy in the prevention and treatment of depressive disorders. Previous systematic reviews have not evaluated the effectiveness of IPT on symptoms of stress, anxiety, depression, quality of life, relationship satisfaction/quality, social supports, and an improved psychological sense of wellbeing. There is limited information regarding moderating and mediating factors that impact the effectiveness of IPT such as the timing of the intervention or the mode of delivery of IPT intervention. The overall objective of this systematic review was to evaluate the effectiveness of IPT interventions to treat perinatal (from pregnancy up to 12 months postpartum) psychological distress. METHODS: MEDLINE(R) and Epub Ahead of Print, In-Process & Other Non-Indexed Citations and Daily (Ovid), EMBASE (Ovid), PsycINFO (Ovid), Cochrane Central Register of Controlled Trials (OVID), CINAHL with Full Text (Ebsco), Social Work Abstracts (Ebsco), SocINDEX with Full Text (Ebsco), Academic Search Complete (Ebsco), Family & Society Studies Worldwide (Ebsco), Family Studies Abstracts (Ebsco), and Scopus databases were searched from inception until 31 January 2019. Two researchers independently screened articles for eligibility. Of the 685 screened articles, 43 met the inclusion criteria. The search was re-run on 11 May 2020. An additional 204 articles were screened and two met the inclusion criteria, resulting in a total of 45 studies included in this review. There were 25 Randomized Controlled Trials, 10 Quasi-experimental studies, eight Open Trials, and two Single Case Studies. All included studies were critically appraised for quality. RESULTS: In most studies (n = 24, 53%), the IPT intervention was delivered individually; in 17 (38%) studies IPT was delivered in a group setting and two (4%) studies delivered the intervention as a combination of group and individual IPT. Most interventions were initiated during pregnancy (n = 27, 60%), with the remaining 18 (40%) studies initiating interventions during the postpartum period. LIMITATIONS: This review included only English-language articles and peer-reviewed literature. It excluded government reports, dissertations, conference papers, and reviews. This limited the access to grassroots or community-based recruitment and retention strategies that may have been used to target smaller or marginalized groups of perinatal women. CONCLUSIONS: IPT is an effective intervention for the prevention and treatment of psychological distress in women during their pregnancy and postpartum period. As a treatment intervention, IPT is effective in significantly reducing symptoms of depression and anxiety as well as improving social support, relationship quality/satisfaction, and adjustment. Systematic Review Registration: PROSPERO CRD42019114292.

Outbreak investigation of foot and mouth disease in Nangarhar province of war-torn Afghanistan, 2014.

Afghanistan has long history of ongoing conflicts, resulting in massive destruction of the country's infrastructure. Illegal trade of livestock between Afghanistan and Pakistan boosted the spread of Foot & Mouth Disease (FMD). Current study was conducted to investigate outbreaks of FMD occurred between April-August 2014 in Nangarhar, Afghanistan. Descriptive data about suspected FMD cases were collected from the Civil Veterinary Hospital, Nangarhar to analyze spatio-temporal pattern of FMD. Case farms (n = 137) were selected from list of clinically confirmed FMD outbreaks available in the hospital. Control farms (n = 137) were enrolled from neighboring premises of case farms. The epidemic curve showed that the virus is continuously circulating among susceptible population. The mean age of the oldest lesion was 2.8 days. Foot & Mouth Disease was more likely to occur in female animals compared to male animals (p < 0.001). Farmers having no ability to clinically recognize FMD (OR 5.8, 95% CI 1.4-23.8); previously having any FMD case in herd (OR 11.8, 95% CI 3.0-45.8), farms where animals leave shed during day (OR 15.4, 95% CI 5.6-42.0), and farms, where neighboring farmers used to visit the premises (OR 3.5, 95% CI 1.2-9.9) were identified as risk factors. Current findings may be used to create awareness of concerned veterinary health authorities about FMD control.