Search

Background: Rheumatoid arthritis (RA) is an immune-mediated, polyarthritis linked with various genetic and environmental causative agents. Among environmental triggers, Epstein-Barr Virus (EBV) is considered the most potent etiological agent. Objective: This study aimed to investigate the prevalence of EBV and its genotypes in RA patients and to investigate their association with clinical and laboratory parameters of RA. Methodology: This study included blood samples of RA and control healthy individuals (100 each). Blood samples along with clinical and laboratory parameters were collected from patients after consent in the Department of Rheumatology, at Lady Reading Hospital, in Peshawar Pakistan. Blood samples were processed for DNA extraction followed by PCR amplification for EBV detection and genotype discrimination. Results: RA patients were 85 females and 15 males with a mean age of 40.13±14.05 years. EBV Type-1 was detected in 45% of RA and 9% of control cases. The mean disease duration of RA patients was 6.61±6.23 years. Out of 100 diseased patients, 43% were seropositive rheumatoid arthritis (SPRA) and showed a significant correlation with a family history of RA in EBV-positive individuals (P = 0.017). The demographic, clinical, and laboratory parameters of RA patients showed a non-significant association with EBV. Moreover, only a family history and Serum creatinine of RA patients showed a significant association with EBV (P = 0.0001 and P = 0.022 respectively). Conclusion: It is concluded that EBV-1 is prevalent and associated with RA. Further investigation is required for detailed genetic analysis of EBV to determine its possible role in modulating the immune system in RA. [ABSTRACT FROM AUTHOR]

Tick infestation is a major public and animal health concern causing significant financial losses, especially in tropical and subtropical regions of the world. This study aimed at investigating the epidemiologic profile of ticks infesting cattle and molecular identification of R. microplus in the centrally ignored part of Khyber Pakhtunkhwa, Pakistan. A total of 600 cattle from 20 farms were examined for the tick infestation, among them 358 (59.7%) cattle were infested with ticks. A total of 2118 nymph, larvae and adult tick stages were collected and morphologically identified followed by molecular confirmation of Rhipicephalus microplus. Host-based demographic and ecological parameter analysis revealed significantly higher tick infestation in adult, female, exotic, freely grazing, and with irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, gender, breed, acaricides use, and feeding method were significantly (P < 0.05) associated, whereas multivariate analysis revealed only host breed and feeding method were potential risk factors (P < 0.05) for tick infestation. Microscopy-based examination identified four different species of ticks including R. microplus (44.5%), Hyalomma anatolicum (38.5%), and Hyalomma marginatum (10.5%) and Hyalomma excavatum (6.5%). Tick infestation pattern showed that 55.9% of cattle was found co-infested with R. microplus and H. anatolicum followed by R. microplus and H. anatolicum and H. marginatum (29.3%) then R. microplus, H. anatolicum, H. marginatum, and H. excavatum (11.2%). Sequencing of the second internal transcribed spacer (ITS2-) and 16S rRNA gene fragments also confirmed the molecular identification of Rhipicephalus microplus. Phylogenetic analysis of ITS-2 revealed all sequences clustered in single clade of the R. microplus while the 16S rRNA nucleotide sequences showed that R. microplus in this study was clustered together in clade A along with other isolates from Pakistan, China, and India. The high tick infestation suggests the need for designing strategic and integrated control measures for ticks in order to ensure good health of domestic animals in this region of Pakistan. [ABSTRACT FROM AUTHOR]

Background: Hepatitis B Virus (HBV) is one of the most common human infectious agents, and the mutations in its genome may cause chronic hepatitis (CH), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). This study was designed to characterize the enhancer-II (Enh-II) region of X gene in HBV positive patients to assess the association of such mutations with CH, LC, and HCC. Methods: HBV positive samples (N = 200) with patients' demographic and clinical data were collected from different regions of Khyber Pakhtunkhwa (KP), Pakistan. The Enh-II region of the HBx gene was sequenced and zanalyzed for polymorphism associated with advanced liver disease. Univariate and logistic regression analyses were performed to evaluate potent mutations associated with a risk for LC and HCC. Results: HBV Enh-II region sequences analysis revealed 25 different mutations. The highest frequency of mutations S101F (62.2%), A102V/R/G/I (56.25%), M103L/A (68.75%)were found in HCC, followed in LC and CH patients as 57.1%, 42.8%, 28.52% 16%, 15.2% and 18.4% respectively. H94 deletion in the α-box of the Enh-II region, associated with a high risk of HCC was found in half of the HCC patients. This deletion was present in 28.5% of LC and 6.5% of CH patients. Importantly, the high frequency of some notable mutations such as E109A/Y, A110S/K, Y111D/E, and F112L was first time reported in the entire study population. The frequencies of these mutations were high in HCC (43.75%, 37.5%, 50% and 43.75% respectively) as compared to LC (14.28%, 14.28%, 28.2% and 42.8%) and CH patients (12.8%, 15.2%, 16.8% and 16% respectively). Conclusion: Mutations associated with LC and HCC are prevalent in the Enh-II region in Pakistani HBV isolates. The mutations found are alarming in CH patients as these may progress to LC and HCC in a large number of patients. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is an important parasite that belongs to the phylum Apicomplexa, distributed globally, causing major health issues for a wide range of hosts, including humans, native and wild animals. Methods: In the present study, we detected IgG and IgM antibodies through an ELISA kit and DNA of T. gondii through PCR in 197 pets and stray cats in Peshawar, Charsadda, Mardan, and Kohat districts of Khyber Pakhtunkhwa (Pakistan) to estimate the existence of feline toxoplasmosis. Results: The current study revealed that stray cats have a significant infection rate of T. gondii (74.6%) as compared to pet cats (25.4%). In all the four districts, the prevalence of T. gondii was pointedly higher in district Kohat (95.5%) in the feline population. In comparison to the female (75.18%) and male (both pets and stray) cats have a maximum infection of (81.66%) non-significantly. The prevalence of T. gondii was observed to be significantly higher (91.66%) in the older and greater than 4 year old population of cats as compared to the younger ones. In poor health condition, the cat populations has a higher risk of infection of 92.3% as compared to healthy and poor body condition (73.91%) and (82.6%) respectively. The chronic and reactivated chronic conditions of toxoplasmosis were higher (58.37%) as compared to the acute condition. Conclusion: It has been concluded that toxoplasmosis is widely spread in the studied population.The outcomes of the present study show that T. gondii infection has a significant impact on the type of cat, age, and area, which implies a serious threat to human beings. Therefore, genotyping of T. gondii strains from different hosts is needed to forecast the current approach for prevention and control of this zoonotic parasite. [ABSTRACT FROM AUTHOR]

Background: Understanding the genetic diversity of Plasmodium species through polymorphic studies can assist in designing more effective control strategies of malaria like new drug formulation and development of a vaccine. Pakistan is moderate endemic for Plasmodium falciparum , but little is known about the genetic diversity of this parasite. This study aimed to investigate the molecular diversity of P. falciparum based on msp-1 and msp-2 genes in the malaria-endemic regions of Khyber Pakhtunkhwa, Pakistan. Methods: A total of 199/723 blood samples, tested positive by microscopy for falciparum malaria, were collected from four districts (Dera Ismail Khan, Karak, Mardan, and Peshawar) of Khyber Pakhtunkhwa. Nested PCR amplification technique was employed to target block 2 of msp-1 and the central domain of msp-2 genes, including their respective allelic families K1, MAD20, RO33, FC27, and 3D7/IC, and to detect the extent of genetic diversity of P. falciparum clinical isolates. Results: Among the 199 microscopy-positive P. falciparum samples, a total of 192 were confirmed using PCR. Ninety-seven amplicons were observed for msp - 1 and 95 for msp-2. A total of 33 genotypes, 17 for msp-1 (eight K1, six MAD20, and three RO33) and 16 for msp-2 (nine FC27 and seven 3D7/IC), were identified. The specific allelic frequency of the K1 family was higher (44.3%) than that of MAD20 (33.0%) and RO33 (23.0%) for msp-1 , while the FC27 allelic family was dominant (60.0%) compared with 3D7/IC (40.0%) for msp-2. No polyclonal infection was observed in msp-1 and msp-2. The expected heterozygosity was 0.98 and 0.97 for msp-1 and msp-2 , respectively. Conclusion: It was concluded that the P. falciparum populations are highly polymorphic, and diverse allelic variants of msp-1 and msp-2 are present in Khyber Pakhtunkhwa, Pakistan. [ABSTRACT FROM AUTHOR]

Background: Cystic echinococcosis (CE) is one of the principal causes of economic loss to the livestock industry because of its morbidity and mortality of food-producing animals and condemnation of important visceral organs. Pakistan being an agricultural country having an extensive livestock sector, is mostly practiced by poor people, which has a fundamental role in the economy. The present study was aimed to conduct a cross-sectional survey and PCR based confirmation of Echinococcus granulosus in sheep, goats, cows, and buffaloes from southern regions (three districts: Lakki Marwat, Bannu, and Karak) of Khyber Pakhtunkhwa, Pakistan. During the study, a total of 2833 animals were examined randomly including; sheep (n = 529), goats (n = 428), cows (n = 1693), and buffaloes (n = 183). Hydatid cysts were collected and examined for the presence of protoscoleces using microscopy. Detection of DNA was performed by using PCR and two mitochondrial genetic markers namely; NAD-1 and COX-1 were amplified. Results: The overall prevalence of CE was found to be (9%) among the examined animals. The hydatid cyst infection was highly prevalent in buffaloes (12%), followed by sheep (10%), cows (9%), and goats (5.1%). Cystic echinococcosis was more prevalent (10%; 96/992) in district Lakki Marwat followed by district Bannu (9%; 112/1246) and Karak (7%; 39/595). Female animals were more likely to be infected with CE (11.6%) than male animals (5.3%) (p = 0.001). Similarly, the infection was higher in the older group of animals as compared to younger (p = 0.001). Mostly (52.2%; n = 129) of hydatid cysts were found in the liver, while (64.4%; n = 159) cysts of the infected animals were infertile. PCR based identification confirmed the presence of E. granulosus sensu stricto (s.s) in the study area. Conclusion: Cystic echinococcosis was found to be highly prevalent in southern regions of Khyber Pakhtunkhwa and could be a potential threat to human health. Moreover, molecular sequencing and phylogenetic analyses should be carried out in future to identify the prevailing genotype (s) of E. granulosus s.s. [ABSTRACT FROM AUTHOR]

Chronic Hepatitis C Virus (HCV) infection is still a major health issue especially in endemic areas where fewer direct-acting virals (DAAs) are treatment options. Some HCV variants are associated with resistance and it reduces DAAs success where pre-existing variants prevail. In this study, we investigated resistance-associated polymorphisms (RAPs) in the HCV NS3 region from DAAs naïve Pakistani patients. 277 chronic HCV treatment naïve patients infected with genotype 1a, 3a and 3b were selected from various clinical centers in the capital city of Khyber Pakhtunkhwa province Pakistan. All the patients were included in this study after taking informed consent. HCV NS3 region was amplified and Sanger sequencing was performed to analyze RAPs to NS3 protease inhibitors. Of the total 29.24% (81/277) patients had detected with known RAPs viz V36A/G/L, T54S, V55A/D/I, Q80K/R, S122G/T/R, R155K/T/I, V158I, D168T/Q, and I170V. Among HCV-1a subjects overall RAPs found were 26.09% (12/46) and most prevalent substitutions were V36A/G (10.87%, 5/46) and R155K/T/I (8.70%, 4/46). Of the total HCV-3a infected patients, 30.95% were observed with RAPS. Ammon these, the most frequent substitutions were Q80R (13.69%, 23/168) followed by V36L (18.33%, 14/168) and V55I (5.95%, 10/168). Among HCV-3b patients, 26.98% were found with RAPs and S122R and Q80R were the dominant variants detected in 17.46 (11/63) and 12.70% (8/63) patients respectively. All these substitutions were associated with Boceprevir, Simeprevir, Telaprevir, and Paritaprevir. Single substitution in one sequence was found in 18.77% (52/277) and multiple in 10.46% (29/277). More than one RAP was frequent in HCV-3a sequences. Natural RAPs are common in chronic HCV patients infected with genotype 1a, 3a and 3b, the most prevalent subtypes in Pakistan. High prevalence of HCV NS3 RAPs suggested a large scale study of the NS3 gene before the introduction of NS3 protease inhibitors in Pakistan. [ABSTRACT FROM AUTHOR]