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Hepatitis C virus (HCV) is the main cause of severe liver disease, including hepatocellular carcinoma, cirrhosis and end stage liver disease. In Pakistan most of HCV positive patients have history of facial/armpit shaving from barbers. 79% of barbers are rubbing Potash Alum stone on facial shaving cuts. Dark blood spots are analyzed on Potash Alum stones being used at different barber shops. The aim of the study was to check the viability of hepatitis C virus on potash alum stone being used at barber shops. Blood samples from HCV positive patients were taken and treated with 0.1, 0.2, 0.3, 0.4 and 0.5 molar concentrations of Potash Alum for different periods of time. Blood was centrifuged to isolate the serum; HCV RNA was extracted from serum and subjected to first strand synthesis and PCR. PCR fragments were confirmed by sequencing. PCR amplification was observed in all the samples, treated with different concentrations of Potash Alum, indicated that the virus remains alive on Potash Alum stone for a long period of time. Potash Alum being used by barbers on facial shaving cuts has definite role in HCV transmission in Pakistani population. Therefore use of Potash Alum stone should be banned on facial shaving cuts at barber shops. [ABSTRACT FROM AUTHOR]

Introduction: Enteric viruses, including Hepatitis E virus (HEV), are able to persist under environmental conditions and may cause public health problems by contaminating natural and drinking water resources. Routine procedures for monitoring viruses in water samples have not been established for the water microbiology screening panel. Methodology: Eighty-six raw sewerage samples were collected from the different regions of Islamabad and Rawalpindi, the twin cities of Pakistan. Samples were concentrated for HEV, using a polyethylene glycol-based method followed by viral RNA extraction using a commercial kit-based method. Reverse transcription polymerase chain reaction (RT-PCR) with HEV specific primers was used for the detection of HEV. Results: The present investigation focused on 86 raw sewerage water samples taken from different locations of drainage outlets of Islamabad and Rawalpindi. After careful experimentation, 35 samples were found to be RT-PCR positive. Nineteen (44.7%) out of 47 samples from Rawalpindi city were HEV positive while 16 (41.02%) out of 39 samples from Islamabad were HEV positive. All positive samples were found in the highly congested areas. Conclusions: The high detection rate of HEV in this study shows that HEV circulates at a relatively high frequency in the sewerage waters in Pakistan. This study is the first report on detection of HEV from sewerage waste water from Pakistan and suggests that HEV might be a potent indicator of viral pollution in environmental specimens. [ABSTRACT FROM AUTHOR]

In Pakistan and other developing countries, the available data on the association of T2DM and HCV is limited. We therefore made an attempt to report the association of HCV and diabetes in Pakistani population through this meta-analysis. HCV and diabetes related studies were identified using various key words, from a number of databases including CINAHL, PubMed, Web of Science and Embase. Using RevMan5, the main outcome was regarded as type 2 diabetes associations with hepatitis c virus infection in Pakistan. Independent analyses were made for "HCV in diabetic patients" and "diabetes cases in hepatitis C virus patients". Using random effect model, odds ratios were calculated with 95% CIs (dichotomous data). I2 statistics were used to calculate heterogeneity. From a total of 53 studies, we finally selected 6 studies for the meta-analysis. Using random effects model, hepatitis c virus patients in 3 studies (n = 1,902) demonstrated that HCV is a risk factor in developing diabetes, contrary to patients with no HCV infection (OR 0.01, 95% CI: 0.00-0.06, I2 = 0%; RR 0.01, 95% CI: 0.00-0.07, I2 = 0%). The remaining 3 studies (n = 13,710) had reported HCV infections in type 2 diabetic patients and patients with no diabetes. Similarly, our meta-analysis revealed higher prevalence of HCV infections in patients with type 2 diabetes (OR 0.7, 95% CI: 0.17-0.42, I2 = 32%; RR 0.30, 95% CI: 0.20-0.46, I2 = 32%) as compared to patients with no type 2 diabetes mellitus. Our meta-analysis demonstrates a significant link between HCV and T2DM. Further studies are recommended with adequate sample sizes [ABSTRACT FROM AUTHOR]

Objective: To evaluate probiotics' effects in reducing hyperbilirubinemia in late preterm and term neonates. Study Design: Quasi-experimental study Place and Duration of Study: Department of Neonatology, Pakistan Institute of Medical Sciences (PIMS), Islamabad Pakistan, from Oct 2018 to Mar 2019. Methodology: The quasi-experimental study was conducted on 60 neonates with hyperbilirubinemia. They were randomly divided into two Groups. Group-A (n=30) received phototherapy. Group-B (n=30) received probiotics containing Lactobacillus reuteri at a dose of 5 drops daily for three days, in addition to phototherapy. Total serum bilirubin was calculated at 0, 24 and 72 hours for both Groups. Results: In Group-A, the mean total bilirubin in mg/dl at 0, 24 and 72 hours were 18.14±2.35, 15.28±2.93 and 12.31±2.29, respectively. Similarly, in Group-B, it was 19.86±5.85, 16.23±4.01 and 10.75±2.23 at 0, 24 and 72 hours, respectively. There was an insignificant statistical difference in mean total bilirubin between Groups A and B at 0 and 24 hours (p<0.001). However, when comparison was made at 72 hours, a significant statistical difference was observed in Group-B (Intervention Group) (p=0.005) Conclusion: Probiotics significantly lowered total serum bilirubin in neonates with hyperbilirubinemia. Further studies are required to evaluate its role in physiological and pathological jaundice. [ABSTRACT FROM AUTHOR]

Abstract: The effective standard therapeutic regimen for patients with chronic hepatitis C is pegylated interferon plus ribavirin. The efficacy of treatment in chronic hepatitis C is defined as absence of detectable virus at six months after treatment. Analysis of patient dependent and virus related factors that enable us to predict the response to antiviral treatment is very important. We prospectively studied 403 patients who received PEG-IFN alpha-2b 1.5μg/kg/body weight plus ribavirin. Treatment was administrated for 24 weeks and 48 weeks for hepatitis C virus (HCV) genotypes 3 and 1, respectively. Out of 403 treated patients, 301 patients (74.7%) showed a sustained virologic response (SVR). Seven variables (age, sex, ethnic group, pretreatment viral load, HCV genotyping and pretreatment ALT) were chosen as possible predictors of SVR and were analysed by means of univariable and multivariable logistic regression analysis. Five variables were statistically significant (p <0.005) on univariable analysis: age, ethnic group, pretreatment viral load, response rate at week 4, and HCV genotype. In multivariable analysis independent factors associated with SVR were low pretreatment viral load (1.97; 95%CI, 1.06–3.66; p =0.03) and attainment of rapid virological response (RVR) (7.19; 95%CI, 4.15–12.45; p <0.001). Our findings support the association between viral load and SVR to PEG-IFN-alpha-2b plus ribavirin therapy. No achievement of RVR is an unfavorable marker for SVR. These findings suggest that all patients considered for treatment should have quantification of serum HCV RNA levels. The result can be used to counsel patients on the likelihood of achieving SVR and may influence the patient''s decision on treatment. Future studies should confirm and explore this observation in other ethnic groups and in relation to HCV genotypes 1 and 3. [Copyright &y& Elsevier]

Background: Hepatitis C virus (HCV) is the cause of high morbidity and mortality worldwide, inflicting around one million people in Pakistan alone. The HCV genomic RNA harbors conserved structural elements that are indispensable for its replication. The 3’ untranslated region (UTR) contains several of these elements essentially involved in regulating the major steps of the viral life cycle. Objectives: Differences in regulatory elements of HCV may contribute towards differential infectivity of local isolates. The present study explicates sequence analysis and secondary structure prediction of HCV 3'UTR region of subtype 3a from Pakistan to characterize this particular region. Patients and Methods: HCV 3'UTR region was amplified, cloned and sequenced from five different patients. Sequence and structural analysis was performed and phylogenetic analysis was carried out using the 3'UTR sequence reported in NCBI nucleotide data base (http://www.ncbi. nlm.nih.gov/nuccore) by other studies. Results: Sequence analysis of the amplified fragment from five patients indicated that the 3'UTR is composed of 214-235 nts. Its sequence contains a type-specific variable region followed by a poly U/UC region and a highly conserved X-tail of 98 nts. The variable region reported here has 26 nts and one stem loop at the secondary structure that differentiate it from HCV genotype 1a ( GT1a) 3'UTR which contains additional 14 nts and two stem loops. The poly U/UC region varied in length (100-79 nts) and nucleotide sequence within the Pakistani isolates, and among different genotypes. Some substitutions found in the X-tail do not affect secondary structure of this element suggesting that this region might play an important role in replication, stabilization and packaging of HCV genome. Additionally, U residues are not present at the end of the X-tail in Pakistani 3a isolates as otherwise reported for the variants of genotype 1b. Conclusions: Sequence and structural diversity of the 3'UTR variable region and Poly U/UC region found in the local isolates indicate specificity in the regulating elements of 3'UTR that might be associated with differential replication efficacy of the HCV Pakistani isolates. The study necessitates functional characterization of these regulating elements to elucidate variable viral efficiency and pathogenicity associated with inter-geographical isolates. [ABSTRACT FROM AUTHOR]

Abstract: Background: Rapid diagnosis of HIV/AIDS enables the development of prevention and treatment programmes but accurate, reliable and cost effective testing strategies should be used for testing of HIV/AIDS from a large population. Objective: To evaluate the performance and effectiveness of three assays for the diagnosis of HIV in comparison with Western blot and to formulate an alternative cost-effective confirmatory approach for HIV diagnosis. Study design: 472 specimens (serum) from a Pakistani population were evaluated. Two rapid HIV testing kits (Capillus, SD Bioline) and one ELISA (Vironostika Ag/Ab) kit were used to detect HIV. Results were compared with Western blot against which sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of all HIV assays were assessed. Results: 280/472 (59.3%) of the samples were positive for antibodies against purified HIV-1 viral proteins. The sensitivity of SD Bioline and Vironostika ELISA was 100% (95% CI; 98–100) while that of anti-HIV Capillus™ kit was 94.6% (95% CI; 91–96.8). The specificity of the Vironostika ELISA and anti-HIV Capillus™ kit was 100% (95% CI; 97–100) while specificity of SD Bioline was 98.4% (95% CI; 95–99). PPV was 100% (95% CI; 98–100%) for the anti-HIV Capillus™ and Vironostika ELISA and 98.9% (95% CI; 96–99%) for SD Bioline. NPV for SD Bioline and Vironostika ELISA was 100% (95% CI; 98–100%) and 92.7% for anti-HIV Capillus™ (95% CI; 88–96%). Conclusion: The sensitivity and specificity of all three kits were satisfactory compared to Western blot and could be used for effective diagnosis of HIV/AIDS in Pakistani population. [Copyright &y& Elsevier]

Summary: Background: Hepatitis C virus (HCV) is a causative agent of chronic liver disease, cirrhosis, and hepatocellular carcinoma. In Pakistan, more than 10 million people are living with HCV. Very little is known about the genotype distribution in Punjab Province, the largest province of Pakistan. Pretreatment genotype identification is very important, as different HCV genotypes respond differently to interferon therapy. Methods: In this study we performed HCV genotyping for 1537 HCV-infected patients from different districts of Punjab Province, Pakistan. Sequencing of partial HCV NS5B sequences from 14 samples belonging to genotypes 3 and 1 was also done. A sequence comparison was made of our reported sequences with those reported in the National Center for Biotechnology Information (NCBI), and a phylogenetic tree was constructed. Results: Our study showed that the most frequent HCV genotype was 3a (in 88.1% of infected individuals), followed by 1a (3.5%), 3b (3.0%), 1b (0.8%), and 2a (1.0%). A mixed genotype infection was found in 3.6% of infected individuals, with 0.3% living with 1a+1b co-infection, 3.1% with 3a+3b, and 0.2% suffering from 3a+1b co-infection. The sequence comparison showed that HCV NS5B motif B residues G283, T287, and N291 were highly conserved in both genotype 1 and genotype 3 sequences, while the motif B residue T286 was mutated to proline in all the genotype 3 sequences. The GDD motif, which forms the catalytic pocket and binding site for the divalent cations, was highly conserved in all the reported sequences. The phylogenetic tree suggests clustering of genotype 1 sequences with sequences from the USA, UK, Germany, and France, while genotype 3 sequences are clustered with sequences from Japan and the UK. Conclusions: The major prevalent genotype in Punjab Province of Pakistan was genotype 3a, followed by genotype 1a, and only 3.6% of infected individuals had a mixed genotype infection. Sequencing of the HCV NS5B gene suggested that the active site residues were highly conserved in all the reported sequences. Our sequences, which are clustered with sequences from the USA, UK, France, and Japan, show the diversity in origin of the different genotypes prevalent in Pakistan. [Copyright &y& Elsevier]