Assessment and Bioaccumulation of Heavy Metals in Water, Fish (wild and Farmed) and Associated Human Health Risk.

Worldwide, anthropogenic activities are causing the natural environment and aquaculture systems to become heavily contaminated with heavy metals, which can lead to consumer's health problems. In the current study, wild and farmed fish (n = 30) and water samples (n = 6) have been collected from the Chashma barrage and fish farm to assess the heavy metals concentration, i.e., Cu, Cd, Pb, Zn and Cr, in the water and some important organs (gills, liver, muscle, brain and bones) of wild and farmed fish (Labeo rohita) using Graphite furnace Atomic absorption spectrometry. Bioaccumulation factor and human health risk assessment were calculated to measure the health status of both fish and humans. Results show that in wild and farm fish's gills, muscles and bones, the trend of the heavy metals was Zn > Pb > Cu > Cd > Cr. On the other hand, the brain and liver show Zn > Cu > Pb > Cd > Cr trend. Comparatively, the heavy metals concentration was mostly higher (P < 0.05) in wild fish. Further; in both fish habitats (water) the heavy metals (Cd and Pb) concentration was higher than the WHO standard level, while in the body, Cd was higher (P < 0.05) in all studied organs except the muscle, Cr was only lower (P > 0.05) in muscle and brain. Pb was higher (P < 0.05) in all studied organs of both fish. Bioaccumulation of heavy metals was mostly higher (P < 0.05) in wild fish than in farmed. EDI and THQ were higher in wild fish, but the HI value was lower than 1 for both fish. Moreover, the PCA analysis suggests a positive correlation between heavy metals concentration in fish organs and the water of both fish (wild and farmed). Results indicated that farmed fish showed less potential risk to humans than wild fish.

Human Health Risk Assessment due to the Incidence of Heavy Metals in Different Commercial Feeds Used for the Culturing of Biofloc Fish (Nile tilapia: Oreochromis niloticus).

Fish meat is a major and rich source of white protein; its quality is determined by the fish feed. However, the low-quality feed may contribute to a source of contamination if it does not fulfill the standard protocol. Biofloc is considered one of the most efficient, successful aquacultures, but this system is still under investigation for its efficiency and safety. Thus, current study focused on the heavy metal contamination in biofloc fish fed on different commercial feeds and human health risk analysis. Samples of extensively used three feeds (Supreme™, Hitech™, and MH-Aqua™), tanks water, and biofloc fish (gills, liver muscle) were collected for heavy metals (Cu, Cd, Pb, and Cr) analysis using atomic absorption spectrometry. An experiment was designed by dividing the fish into three groups: group 1 (Supreme™), group 2 (Hitech™), and group 3 (MH-Aqua™). A bioaccumulation factor and human health risk assessment have been calculated to measure fish and human health. Results revealed that most of the heavy metal concentration was higher (P < 0.05) in MH-Aqua™ feed compared to others. Similarly, heavy metal concentration was higher (P < 0.05) in the water of group 3, where fish was cultured on MH-Aqua™ feed. However, in the fish gills, liver, and muscle, the heavy metal concentration was significantly greater in group 3 fed on MH-Aqua™ feed, followed by group 1. Heavy metals in all feeds were positively correlated to the heavy metal concentration of the fish muscles. The bioaccumulation factor for Cu and Pb was higher in the fish liver, Cd and Cr in the case of fish gills, and least in the fish muscle. EDI and THQ values vary in all the groups, while the HI value was found lower than 1 in group 1 and group 2 but higher in group 3 fed on MH-Aqua™ feed. Strict checks and balances in formulating a diet will be helpful to progressively lower the amount of dangerous heavy metals.

Frequency and association of Epstein-Barr Virus genotype in rheumatoid arthritis patients of Khyber Pakhtunkhwa, Pakistan.

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.

Ranuncoside's attenuation of scopolamine-induced memory impairment in mice via Nrf2 and NF-ĸB signaling.

Scopolamine is a well-known pharmacological agent responsible for causing memory impairment in animals, as well as oxidative stress and neuroinflammation inducer which lead to the development of Alzheimer disease. Although a cure for Alzheimer's disease is unavailable. Ranuncoside, a metabolite obtained from a medicinal plant has demonstrated antioxidant and anti-inflammatory properties in vitro, making it a promising treatment with potential anti-Alzheimer disease properties. However, as ranuncoside has not been evaluated for its antioxidant and anti-neuroinflammatory properties in any in vivo model, our study aimed to evaluate its neurotherapeutic efficacy against scopolamine-induced memory impairment in adult male albino mice. Mice were randomly divided into four experimental groups. Mice of group I was injected with saline, group II was injected with scopolamine (1 mg/kg/day) for 3 weeks. After receiving a daily injection of scopolamine for 1 week, the mice of group III were injected with ranuncoside (10 mg/kg) every other day for 2 weeks along with scopolamine daily and group IV were injected with ranuncoside on 5th alternate days. Behavioral tests (i.e., Morris water maze and Y-maze) were performed to determine the memory-enhancing effect of ranuncoside against scopolamine's memory deleterious effect. Western blot analysis was also performed to further elucidate the anti-neuroinflammatory and antioxidant effects of ranuncoside against scopolamine-induced neuroinflammation and oxidative stress. Our results showed memory-enhancing, anti-neuroinflammatory effect, and antioxidant effects of ranuncoside against scopolamine by increasing the expression of the endogenous antioxidant system (i.e., Nrf2 and HO-1), followed by blocking neuroinflammatory markers such as NF-κB, COX-2, and TNF-α. The results also revealed that ranuncoside possesses hypoglycemic and hypolipidemic effects against scopolamine-induced hyperglycemia and hyperlipidemia in mice as well as scopolamine's hyperglycemic effect. In conclusion, our findings suggest that ranuncoside could be a potential agent for the management of Alzheimer's disease, hyperglycemia, and hyperlipidemia.

Investigation of the incidence of heavy metals contamination in commonly used fertilizers applied to vegetables, fish ponds, and human health risk assessments.

Overuse of fertilizers on agricultural lands and fish ponds may result in serious pollution problems, such as heavy metals that can enter the food chain and pose serious health problems. Due to this, the present study investigates the incidence of heavy metals in commonly used fertilizers and its association with heavy metals in vegetables, soil, fish species, and pond water. Samples were collected from different sites (fields and ponds) in district Kohat, where the application of fertilizers was common and control groups (no fertilizers used). Heavy metal analysis was carried out through a spectrophotometer. Results showed higher Cd and Cr concentrations in triple superphosphate (TSP), Cu and Pb in nitrogen, phosphorus, and potassium (NPK), while lower concentrations were found in gypsum. In vegetables (onion, tomato, brinjal, and potato) and associated soil, most of the heavy metals concentrations were significantly higher (P < 0.05) in fertilizer-applied sites than in the control. Also, the Cd concentration in potatoes and Pb level in all vegetables obtained from sites were greater than the WHO/FAO standard limit. In the case of fish species (Hypophthalmichthys molitrix and Cyprinus carpio) muscles and their habitat (water), all the understudy heavy metals were notably higher (P < 0.05) in fertilizer-applied sites (ponds) than the control group. Collectively, in all vegetables and muscles of fish species, the bioaccumulation factor was higher in sites compared to the control. The estimated daily intake (EDI) and target hazard quotient (THQ) values were also higher in fertilizer-applied sites (fields and ponds) than control. The health index (HI) value was > 1 in vegetables (onion, tomato, and potato) and fish muscles collected from different sites compared to the control. Thus, there is the possibility of severe health risks. The use of fertilizers must be carefully monitored in order to ensure that humans and animals are safe from exposure to heavy metals.

Folicitin abrogates scopolamine induced oxidative stress, hyperlipidemia mediated neuronal synapse and memory dysfunction in mice.

No effective drug treatment is available for Alzheimer disease, thus the need arise to develop efficient drugs for its treatment. Natural products have pronounced capability in treating Alzheimer disease therefore current study aimed to evaluate the neuro-protective capability of folicitin against scopolamine-induced Alzheimer disease neuropathology in mice. Experimental mice were divided into four groups i.e. control (single dose of 250 µL saline), scopolamine-administered group (1 mg/kg administered for three weeks), scopolamine plus folicitin-administered group (scopolamine 1 mg/kg administration for three weeks followed by folicitin administration for last two weeks) and folicitin-administered group (20 mg/kg administered for 5 alternate days). Results of behavioral tests and Western blot indicated that folicitin has the capability of recovering the memory against scopolamine-induced memory impairment by reducing the oxidative stress through up-regulating the endogenous antioxidant system like nuclear factor erythroid 2-related factor and Heme oxygenase-1 while prohibiting phosphorylated c-Jun N-terminal kinase. Similarly, folicitin also improved the synaptic dysfunction by up-regulating SYP and PSD95. Scopolamine-induced hyperglycemia and hyperlipidemia were abolished by folicitin as evidenced through random blood glucose test, glucose tolerance test and lipid profile test. All these results revealed that folicitin being a potent anti-oxidant is capable of improving synaptic dysfunction and reducing oxidative stress through Nrf-2/HO-1 pathway, thus plays a key role in treating Alzheimer disease as well as possess hyperglycemic and hyperlipidemic effect. Furthermore, a detailed study is suggested.

Genetic diversity in enhancer II region of HBV genotype D and its association with advanced liver diseases.

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.

Peripheral blood T cells response in human parainfluenza virus-associated lower respiratory tract infection in children.

Human Parainfluenza virus (HPIV) causes lower respiratory tract infections (LRTI) mostly in young children. Respiratory viral infections may decline T cells in circulation and display enhanced pathogenicity. This study is aimed to analyze T cells alterations due to HPIV in children with LRTIs. Children (N = 152) with bronchitis or pneumonia, admitted in tertiary care hospitals were included in the study. Respiratory samples (throat or nasopharyngeal swabs) were taken and HPIV genotypes (1-4) were analyzed through RT-PCR. Peripheral blood T cells, CD3+, CD4+, CD8+, and CD19+, were analyzed in confirmed HPIV positive and healthy control group children through flow cytometry. The positivity rate of HPIV was 24.34% and the most prevalent genotype was HPIV-3 (20.40%). HPIV-1 and HPIV-2 were detected in 0.66% and 02% children respectively. The T lymphocyte counts were observed significantly reduced in children infected with HPIV-3. CD4+ cell (1580 ± 97.87) counts did not change significantly but the lowest CD8+ T cell counts (518.5 ± 74.00) were recorded. Similarly, CD3+ and CD19 cell ratios were also reduced. The CD4/CD8 ratio was significantly higher (3.12 ± 0.59) in the study population as compared to the control group (2.18 ± 0.654). Changes in the count of CD8+ T cells were more pronounced in patients with bronchiolitis and pneumonia. It is concluded that CD8+ T cells show a reduced response to HPIV-3 in children with severe LRTIs suggesting a strong association of these cells with disease severity.

Molecular identification of Toxoplasma gondii in domesticated and broiler chickens (Gallus domesticus) that possibly augment the pool of human toxoplasmosis.

Toxoplasma gondii (T. gondii) is a protozoan parasite that infects all warm-blooded animals including domesticated birds and humans. Birds normally get infected by ground feeding and human beings contract the disease by consumption of undercooked chicken meat. This study aimed to analyze seroprevalence and DNA of T. gondii in chickens (domesticated and broiler) and to assess possible transfer to humans by review of available literature from Pakistan. Blood from and tissues from domesticated and broilers chickens were analyzed for Toxo-IgM/IgG and Toxoplasma DNA through ELISA and PCR respectively. Furthermore, research articles published during 1990-2019 on the prevalence of T. gondii in humans from Pakistan, were analyzed to assess the possible infection burden in the area in connection to transmission from chickens. The overall prevalence of IgM and IgG for T. gondii was 17.83% and 8.8% respectively in the study areas. Significant seroprevalence was found in domesticated chickens than broilers. In domesticated chickens, the prevalence was high in age ≥ 2 years. Toxoplasma DNA was detected in tissues with an overall prevalence of 10.84%. Higher prevalence was observed in liver (10.50%) than heart (9.5%) and muscles (7.11%). Only 4.78% broiler and 2.38% domesticated chickens were positive for both IgM and DNA, 1.2% domesticated and 1.30% broilers were positive for IgG and DNA, while 2.98% domesticated and 2.17% broilers were positive for IgM, IgG, and DNA. Available literature showed that 25.8% of human beings were infected with T. gondii in Pakistan. The prevalence was 20.64% in male and 26.82%in the female. The rate of infections increases with age and high (37.36%) was found in humans of age range 40 to 60 years. A high prevalence of T. gondii is found in both domesticated and broiler chickens in the study area. Moreover, the literature survey indicates that a high seroprevalence of T. gondii is present in human beings of Pakistan. It is concluded that the high prevalence of T. gondii in humans may be associated with the parasite transmission through infected chicken's meat in Pakistan.

Pre-existing resistance associated polymorphisms to NS3 protease inhibitors in treatment naïve HCV positive Pakistani patients.

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.

PCR/RFLP-based analysis of genetically distinct Plasmodium vivax population of Pvmsp-3α and Pvmsp-3ß genes in Pakistan.

BACKGROUND: Plasmodium vivax is one of the widespread human malarial parasites accounting for 75% of malaria epidemics. However, there is no baseline information about the status and nature of genetic variation of Plasmodium species circulating in various parts of Pakistan. The present study was aimed at observing the molecular epidemiology and genetic variation of Plasmodium vivax by analysing its merozoite surface protein-3α (msp-3α) and merozoite surface protein-3ß (msp-3ß) genes, by using suballele, species-specific, combined nested PCR/RFLP detection techniques. METHODS: A total of 230 blood samples from suspected subjects tested slide positive for vivax malaria were collected from Punjab, Sindh, Khyber Pakhtunkhwa, and Balochistan during the period May 2012 to December 2013. Combined nested PCR/RFLP technique was conducted using Pvmsp-3α and Pvmsp-3ß genetic markers to detect extent of genetic variation in clinical isolates of P. vivax in the studied areas of Pakistan. RESULTS: By PCR, P. vivax, 202/230 (87.82%), was found to be widely distributed in the studied areas. PCR/RFLP analysis showed a high range of allelic variations for both msp-3α and msp-3ß genetic markers of P. vivax, i.e., 21 alleles for msp-3α and 19 for msp-3ß. Statistically a significant difference (p ≤ 0.05) was observed in the genetic diversity of the suballelic variants of msp-3α and msp-3ß genes of P. vivax. CONCLUSION: It is concluded that P. vivax populations are highly polymorphic and diverse allelic variants of Pvmsp-3α and Pvmsp-3ß are present in Pakistan.

Aurora kinase-C-T191D is constitutively active mutant.

BACKGROUND: Aurora kinases (Aurora-A, B and C) belong to a family of conserved serine/threonine kinases which are key regulators of cell cycle progression. Aurora-A and Aurora-B are expressed in somatic cells and involved in cell cycle regulation while aurora-C is meiotic chromosome passenger protein. As Aurora kinase C is rarely expressed in normal somatic cells and has been found over expressed in many cancer lines. It is suggested that Aurora-C-T191D is not hyperactive mutant. RESULT: Aurora-C-T191D variant form was investigated and compared with wild type. The overexpression of Aurora-C-T191D was observed that it behaves like Aurora-C wild type (aurC-WT). Both Aurora-C-T191D and aurC-WT induce abnormal cell division resulting in centrosome amplification and multinucleation in transiently transfected cells as well as in stable cell lines. Similarly, Aurora-C-T191D and aurC-WT formed foci of colonies when grown on soft agar, indicating that a gain of Aurora-C activity is sufficient to transform cells. Furthermore, we reported that NIH-3 T3 stable cell lines overexpressing Aurora-C-T191D and its wild type partner induced tumour formation when injected into nude mice, demonstrating the oncogenic activity of enzymatically active Aurora kinase C. Interestingly enough tumour aggressiveness was positively correlated with the rate of kinase activity, making Aurora-C a potential anti-cancer therapeutic target. CONCLUSION: These findings proved that Aurora C-T191D is not hyperactive but is constitutively active mutant.

Trend of transfusion transmitted infections frequency in blood donors: provide a road map for its prevention and control.

BACKGROUND: Transfusion transmitted infections create significant burden on health care system. Donor selection is of paramount importance because infected individuals serve as an asymptomatic reservoir and a potential source of transmission. METHODS: A retrospective study was carried out in healthy blood donors in the Lady Reading Hospital Peshawar, Pakistan over a period of three and a half years i.e., from January 2008 to June 2011, to determine the prevalence of HBV, HCV, HIV and syphilis in order to provide information for relevant polices. RESULTS: Out of 1,27,828 sample of blood donors, recorded mean prevalence for HBs Ag, anti-HCV, anti-HIV and syphilis was 2.68%, 2.46%, 0.06% and 0.43%, respectively, with an increasing trend in frequencies of transfusion transmitted infections (TTIs). CONCLUSIONS: This study reflects that blood transfusion is one of the leading risk factor of spread of the TTIs, which showed the need and importance of the mandatory screening of these infectious markers in blood donations.

Rising burden of Hepatitis C Virus in hemodialysis patients.

AIM: High prevalence of Hepatitis C virus (HCV) has been reported among the dialysis patients throughout the world. No serious efforts were taken to investigate HCV in patients undergoing hemodialysis (HD) treatment who are at great increased risk to HCV. HCV genotypes are important in the study of epidemiology, pathogenesis and reaction to antiviral therapy. This study was performed to investigate the prevalence of active HCV infection, HCV genotypes and to assess risk factors associated with HCV genotype infection in HD patients of Khyber Pakhtunkhwa as well as comparing this prevalence data with past studies in Pakistan. METHODS: Polymerase chain reaction was performed for HCV RNA detection and genotyping in 384 HD patients. The data obtained was compared with available past studies from Pakistan. RESULTS: Anti HCV antibodies were observed in 112 (29.2%), of whom 90 (80.4%) were HCV RNA positive. In rest of the anti HCV negative patients, HCV RNA was detected in 16 (5.9%) patients. The dominant HCV genotypes in HCV infected HD patients were found to be 3a (n = 36), 3b (n = 20), 1a (n = 16), 2a (n = 10), 2b (n = 2), 1b (n = 4), 4a (n = 2), untypeable (n = 10) and mixed (n = 12) genotype. CONCLUSION: This study suggesting that i) the prevalence of HCV does not differentiate between past and present infection and continued to be elevated ii) HD patients may be a risk for HCV due to the involvement of multiple routes of infections especially poor blood screening of transfused blood and low standard of dialysis procedures in Pakistan and iii) need to apply infection control practice.

Hepatitis C virus genotypes in Pakistan: a systemic review.

BACKGROUND AND AIM: Phylogenetic analysis has led to the classification of hepatitis C virus (HCV) into 1-6 major genotypes. HCV genotypes have different biological properties, clinical outcome and response to antiviral treatment and provide important clues for studying the epidemiology, transmission and pathogenesis. This article deepens the current molecular information about the geographical distribution of HCV genotypes and subgenotypes in population of four provinces of Pakistan. 34 published papers (1996-2011) related to prevalence of HCV genotypes/serotypes and subgenotypes in Pakistan were searched. RESULT: HCV genotype/s distribution from all 34 studies was observed in 28,400 HCV infected individuals in the following pattern: 1,999 (7.03%) cases of genotype 1; 1,085 (3.81%) cases of genotype 2; 22,429 (78.96%) cases of genotype 3; 453 (1.59%) cases of genotype 4; 29 (0.10%) cases of genotype 5; 37 (0.13%) cases of genotype 6; 1,429 (5.03%) cases of mixed genotypes, and 939 (3.30%) cases of untypeable genotypes. Overall, genotype 3a was the predominant genotype with a rate of 55.10%, followed by genotype 1a, 3b and mixed genotype with a rate of 10.25%, 8.20%, and 5.08%, respectively; and genotypes 4, 5 and 6 were rare. Genotype 3 occurred predominately in all the provinces of Pakistan. Second more frequently genotype was genotype 1 in Punjab province and untypeable genotypes in Sindh, Khyber Pakhtunkhwa and Balochistan provinces.

Prevalence of hepatitis B virus genotypes in HBsAg positive individuals of Afghanistan.

BACKGROUND: The structural and functional differences between hepatitis B virus (HBV) genotypes are the mainstay to severity, complications, treatment and possibly vaccination against the virus. This study was conducted to determine the HBV genotypes in HBsAg positive patients of Afghanistan as no such large scale data available previously. METHODS: Two hundred and fourteen HBsAg-positive patients were included in this study. All patients were anti-HCV and anti-HIV negative. All the samples were confirmed for HBV DNA with nested PCR while HBV DNA positive samples were subjected to type specific PCR for HBV genotyping (A-F). RESULTS: Of the total samples, 168 (78.5%) were males and 46 (21.49%) females, aged ranged between 18 to 71 years. This study demonstrated that genotype D (35.67%) is the predominant genotype circulating in Afghani's population. Genotype C was observed in 32.16% followed by genotype A (19.30%), and genotype B (7.02%) while 6.07% of the individuals were not typed. CONCLUSION: This study has shown a heterogeneous distribution of HBV genotypes. Further more, extensive studies are required to investigate genetic and geographical divergence and characteristics of the virus in the country, as no such large sample sized study has been carried out so far in this country.

Prevalence of HBV and HBV vaccination coverage in health care workers of tertiary hospitals of Peshawar, Pakistan.

BACKGROUND: Hepatitis B Virus (HBV) may progress to serious consequences and increase dramatically beyond endemic dimensions that transmits to or from health care workers (HCWs) during routine investigation in their work places. Basic aim of this study was to canvass the safety of HCWs and determine the prevalence of HBV and its possible association with occupational and non-occupational risk factors. Hepatitis B vaccination coverage level and main barriers to vaccination were also taken in account. RESULTS: A total of 824 health care workers were randomly selected from three major hospitals of Peshawar, Khyber Pakhtunkhwa. Blood samples were analyzed in Department of Zoology, Kohat University of Science and Technology Kohat, and relevant information was obtained by means of preset questionnaire. HCWs in the studied hospitals showed 2.18% prevalence of positive HBV. Nurses and technicians were more prone to occupational exposure and to HBV infection. There was significant difference between vaccinated and non-vaccinated HCWs as well as between the doctors and all other categories. Barriers to complete vaccination, in spite of good knowledge of subjects in this regard were work pressure (39.8%), negligence (38.8%) un-affordability (20.9%), and unavailability (0.5%). CONCLUSIONS: Special preventive measures (universal precaution and vaccination), which are fundamental way to protect HCW against HBV infection should be adopted.

Response to combination therapy of HCV 3a infected Pakistani patients and the role of NS5A protein.

BACKGROUND: Hepatitis C virus (HCV) genotype 3a is known to show comparatively better response to combination therapy than genotype 1 and 4. Mutations within NS5A gene of HCV have earlier been implicated with response to interferon (IFN) therapies in chronic HCV patients among various populations. As response to therapy are available in different populations because of the ethnic and viral factors and there was no study available on the phenomenon of resistivity to IFN. RESULTS: Chronic HCV 3a infected Pakistani patients were kept on IFN-α and ribavirin therapy for six months. NS5A gene of HCV was amplified and sequenced in the case of all the patients prior to therapy and the sequences were analysed for mutations. Out of the total 27 patients, 20 (74.07%) were observed with sustained virological response (SVR), 4 (14.81%) patients were non responder (NR) while 3 (11.11%) patients exhibited in end of treatment response (ETR). Three (3/20) (15%) SVR patients and two (2/3) ETR patients had mutations (ranging from I-V amino acids) within the NS5A ISDR regions. While the rest of the SVR patients (85%) and the NR had no mutations at ISDR region when compared with HCV K3a ISDR. CONCLUSIONS: Mutations within the NS5A gene of HCV 3a genotype may not influence the outcome of combination therapy in Pakistani populations.