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This study aimed to evaluate the effects of an organic acid blend and essential oils individually and in combination on growth performance, blood biochemical profile, gut health, and nutrient digestibility of broilers fed a higher level of an animal protein concentrate. Five hundred day-old Ross-308 male broiler chicks (average body weight, 39 ± 1.2 g) were randomly assigned to five replicated (5 replicates/treatment; 20 birds/replica) dietary treatments (100 birds/treatment). Birds in these group were given five different experimental diets that were prepared and designated as (i) basal diet (negative control, NC); (ii) basal diet plus Enramycin (positive control, PC), 50 mg/kg feed; (iii) basal diet with the addition of organic acid (OA) at 200 mg/kg feed; (iv) basal diet plus essential oils (EO) at 150 mg/kg feed; (v) basal diet plus combination of OA and EOs 200 and150 mg/kg feed (OA + EO). Experimental diets were prepared and fed in two phases i.e. starter (1–21 d) and finisher (22–42 d) phases of rearing. The findings of the present study revealed that feed intake did not vary significantly among the treatment groups, however, better (p

This study examined the effects of dried sprouted wheat grains (SWGs) on the growth performance, serum biochemical profile and liver histopathology of broiler chickens at starter to finisher phases of growth. A total of 320, day-old Ross-308 as hatched broiler chicks (43.23 ± 2 g) were randomly assigned to four different replicated (n = 4; 20 birds/replicate) dietary treatments. All birds were reared in cages (5l × 5w × 3h feet) that served as an experimental unit and fitted with drinker, feeder, bulb and saw dust as bedding material. Dietary treatments were designated as TRT1 that served as control (CON) and birds in this group were fed a standard diet only, TRT2 (CON + 5 g/kg SWG), TRT3 (CON + 10 g/kg SWG) and TRT4 (CON + 15 g/kg SWG). It was noticed that birds supplemented on higher level of SWG (15 g/kg of diet) had significantly improved body weight gain (BWG) (1806 g) and feed efficiency (1.67) compared to birds in other groups. Similarly, birds in TRT4 had significantly higher high-density lipoprotein both at starter (99.4 mg/dL) and finisher (110 mg/dL), and lower (p

Animal industry seeks cost-effective solutions to enhance performance and health of domestic animals. This study investigated the effects of supplementing Bacillus spp. probiotics and xylanase on 2000 one-day-old Japanese quails, randomly assigned to four treatment groups (10 replicates). The control group received no supplementation, while the others were supplemented with a Bacillus-based probiotic at 7.5 × 107 cfu/kg of feed, xylanase enzyme (2,000 U/kg) alone or in combination. Quails receiving both probiotic and enzyme exhibited significantly (p

This study aimed to evaluate the effect of varying levels of sorghum-based diets as an alternative to maize in broiler nutrition. A total of 320 one-day-old male Ross 708 broiler chickens were randomly allocated to four treatment groups (5 pens per treatment and 16 birds per pen), comprising a control group with a basal diet and groups receiving sorghum-based diets with 20%, 40%, and 100% maize replacement. The overall weight gain was significantly (p

The study aimed to evaluate the effects of multi-strain probiotic (MSP) on broiler growth, health, and nutrient utilization under high animal protein concentrate conditions. Male Ross-308 broilers were assigned to four groups: a control (C), MSP at 50 mg/kg (MSP50), MSP at 100 mg/kg (MSP100), and Zinc bacitracin (ZB) as a positive control. MSP supplementation significantly improved feed intake, weight gain, and feed conversion efficiency, with MSP100 demonstrating the best feed conversion ratio during both starter and finisher phases. MSP also enhanced dressing percentages and positively influenced blood lipid profiles. The supplementation reduced harmful bacterial growth (E. coli, Salmonella, Clostridium perfringens) while promoting beneficial bacteria (Lactobacillus). Moreover, MSP positively impacted gut histomorphology, showing higher villus height, width, and crypt depth in the MSP100 group. Digestibility of dry matter and crude protein was enhanced in birds fed the MSP100 diet. Overall, MSP supplementation exhibited positive effects on broiler performance, health, and nutrient digestibility, indicating its potential as an antibiotic alternative for improving practices in the poultry feed industry.

This study assessed the potential of a commercial probiotic mixture on performance, gut health and nutrient utilisation of broilers-fed poultry by-product meal (PBM)-based diet. Three-hundred-and-twenty Ross-308-day-old broiler chicks were obtained from a commercial poultry hatchery. The chicks were randomly assigned to four different treatment groups, each consisting of four replicates (n = 10) in a randomised complete design. The positive control group (CON) was fed commercial poultry ration, and the negative control group (N-CON) was fed PBM partially replacing soybean meal at a 65% level. The MSP1 and MSP2 groups contained the same diet as the N-CON group but supplemented with multi-strain probiotics (MSP) at 50 mg/kg and 100 mg/kg. Overall N-CON group performed poorly, but the addition of probiotics in group MSP1 and MSP2 diets significantly improved performance. Feed efficiency and bird weight gain were improved (p

This study investigated the impact of ginger-derived phyto-protease enzyme on growth performance, immunity, gut health and nutrient digestibility in broilers. A total of 360 day-old broiler chickens were randomly assigned to four treatments with six replicates each: a control group (T1) and groups supplemented with 50 g/ton (T2), 100 g/ton (T3) and 150 g/ton (T4) of the enzyme. Over a 35-day period, birds were provided with free access to feed and water under controlled conditions. The highest feed intake was observed in T3 during the second week, while T4 showed the lowest intake overall. Body weight gain was significantly highest in T4, demonstrating enhanced growth performance. Feed conversion ratio (FCR) was lowest in T4, indicating improved feed efficiency. Carcase quality analysis revealed higher dressing percentages and organ weights (bursa, spleen and thymus) in T4 compared to the control. Gut pH levels were significantly lower in T3 and T4 across different segments of the gastrointestinal tract, enhancing nutrient absorption. Antibody titres against Newcastle disease virus were significantly higher in T4 on days 21 and 35, indicating improved immunity. Histomorphological analysis of the intestines showed increased villus height and width in T4, with reduced crypt depth, further supporting enhanced nutrient absorption. These findings suggest that the highest inclusion rate of ginger-derived phyto-protease enzyme (150 g/ton) optimally enhances growth performance, immunity, gut health and nutrient digestibility in broilers. This study highlights the potential benefits of incorporating ginger-derived phyto-protease in poultry diets to improve overall health and performance.

We hypothesized that both performance and bone mineralization would show improvements when cholecalciferol supplementation was increased above the recommended NRC (1994) [Nutrient requirements of poultry, 9th rev. ed. Washington, DC: National Academy Press] level. In this study, male Ross 308 chickens (n = 540) were supplemented with diets containing either a low level of 200 IU/kg or a higher dose of 5,000 IU/kg of cholecalciferol (VD). The results showed that during the starter phase, weight gain was significantly higher in VD5000 and the control compared to VD0, with significantly lower feed intake in VD0. Consequently, FCR was significantly better in VD200 and VD5000 compared to VD0. In the finisher phase, weight gain, feed intake, and FCR were significantly higher in VD200 and VD5000 compared to VD0. Tibia ash, Ca, and P were significantly higher in VD5000 compared to VD200. Carcass weight was significantly higher in VD200 and VD5000 compared to VD0. Total tract N retention and GE were significantly higher in VD5000 compared to VD0. The study concludes that cholecalciferol supplementation at 5000 IU/kg enhances growth performance, tibia mineralization, and nutrient digestibility in broilers fed with 30% reduced Ca and P in their diets.

The trial investigated the effects of anise seed powder and protease, alone and combined, in Japanese quail diets. 540 seven-day-old quails were divided into four groups: Control (basal diet), Anise Seed Group (1000 mg/kg anise seed powder), Protease Enzyme Group (30,000 IU/kg Bacillus subtilis-derived protease enzyme), and a combination of anise powder and protease. The combination significantly increased weight gain and dressing percentage compared to the control. Quails receiving protease, anise powder, or both showed higher antibody titers against infectious diseases. The population of beneficial Lactobacillus increased in groups receiving protease and the combination. Additionally, the presence of harmful bacteria like Escherichia coli and Salmonella decreased significantly in quails supplemented with both protease enzyme and anise powder. In summary, combining protease and anise powder improved growth performance, immune responses, increased Lactobacillus population, and decreased pathogenic bacteria in quails.

Objective: To determine the frequency and etiology of Early Return Visits within 3 days of discharge to the Emergency Department. Study Design: Prospective longitudinal study. Place and Duration of Study: Emergency Department of Combined Military Hospital, Rawalpindi Pakistan, from Jan to Jun 2022. Methodology: All patients visiting the Emergency Department within the defined 6-month period were included in this study, which came to 101’113. To determine overall incidence of Early Return Visits, total number of patients revisiting within 72 hours of Emergency Department discharge was used. To find out the possible factors responsible for ERVs, a sample size of 80 patients revisiting the Emergency Department was calculated through World Health Organization calculator and selected through random sampling technique. Factors responsible for unscheduled Early Return Visits were divided into three broad categories: Doctor-associated, Disease-associated, and Patient-associated. Results: The total number of patients revisiting Emergency Department within 72 hours was 2,880 (2.9%). Out of these 52.5% patients reported back to the Emergency Department due to disease-associated reasons, 27.5% due to doctor-related reasons, and 20% revisited for patient-related reasons. Conclusion: Two-point nine percent patients made ERVs to the Emergency Department within 72 hours. The most common causes of Early Return Visits in this study are disease-related issues followed by doctor-related issues.

Objective: To compare post-operative analgesic requirements in patients undergoing hemorrhoidectomy under caudal epidural anaesthesia versus saddle anaesthesia. Study Design: Quasi Experimental Study. Place and Duration of Study: Combined Military Hospital, Rawalpindi Pakistan, Feb to Nov 2021. Methodology: The study was conducted on 200 patients who underwent hemorrhoidectomy during the study period. Patients were randomized into two groups. Group-A received caudal epidural anaesthesia, while Group-B received saddle anaesthesia. The pain at the surgical site was recorded on a visual analogue scale 04 hours and 08 hours after the surgical procedure. The difference in significant post-operative pain and the requirement of opioid analgesia was compared. Results: Out of 200 patients randomized into two groups, 96(48%) were categorized into Group-A, and 104(52%) were categorized into Group-B. The mean age of patients who were operated for hemorrhoids in our study was 41.56±6.98 years. Patients who received caudal epidural anaesthesia had more chances of not having significant post-operative pain at 04 and 08 hours after the surgery as compared to those who received saddle anaesthesia (p-value

ABSTRACT: Citrus citrus peeling has a wide range of vitamins and trace minerals that have antioxidant and antimicrobial properties. It is hypothesized that the addition of citrus peeling to broiler quail diets can improve their production performance, humoral immunity, nutrients, and energy utilization. A trial was performed to study the impact of a methanolic extract of citrus peeling (Citrus sinensis) on production performance, humoral immunity, nutrients, and energy utilization of broiler quails. A healthy day-old 300 quails were randomly assigned with 5 replicates each replicate had 15 birds and used CRD for the trial. Different dietary supplementations were presented to different groups. The control group was not supplemented with any supplementation in their feed. While, the 3 other groups were supplemented by 0.5 mL/kg, 1 mL/kg, and 1.5 mL/kg of methanolic extract of dried Citrus sinensis peel (DCSP) in the basal diet (DCSP0.5, DCSP1, and DCSP1.5 groups, respectively). All the birds were allowed ad libitum feeding and water. The feed intake and FCR were significantly higher in the control group, followed by DCSP0.5, and then DCSP1. The significantly lowest feed intake and FCR were observed in the DCSP1.5 group. The weight gain and dressing % were significantly improved with the increasing level of methanolic extract of Citrus sinensis. The significantly highest weights of thymus, spleen, and bursa were recorded in the DCSP1.5 group, followed by the DCSP1. The antibody titers against infectious bursal disease, New Castle disease, and infectious bronchitis disease were significantly higher in the DCSP1.5, DCSP1, and DCSP0.5 groups than in the control group. It was concluded from the study that supplementation of quails with methanolic extract of citrus at a dose rate of 0.5 to 1.5 mL/kg of feed can improve feed intake, weight gain, FCR, dressing percentage, relative weight of lymphoid organs, and digestibility coefficient. Supplementation of Citrus sinensis has also concluded positive impacts on antibody titers against various viral diseases. The best improvement in the evaluated parameters was observed at a dose of extract of citrus was 1.5 mL/kg of feed.

Objective: To associate the findings of CT scan with the Glasgow coma scale (GCS) of cases presenting with head trauma in the Emergency Department. Study Design: Cross-sectional study. Place and Duration of Study: Emergency Department, Combined Military Hospital, Rawalpindi Pakistan, from Jul to Dec 2021. Methodology: Ninty head injury cases of either gender, aged 12-60 years were included using non-probability consecutive sampling. The severity of the head injury was determined via GCS. CT scan findings were noted. Results: The mean age was 22.21±11.10 years. The highest frequency of patients was noted in the age group of (13-29 years).The most common cause of head injury in men was road traffic accidents n=39/58(67.24%). As per GCS scoring, 63.3% of patients had mild brain injury. A strong association was observed between GCS and CT scan findings in cases with brain oedema (p=0.002), skull fracture (0.018) and no findings on CT scan (p-value≤0.001). No association between findings on CT scan and GCS scoring was noted in cases with epidural, subdural hematoma, cerebral contusions, intracranial and subarachnoid haemorrhage. Conclusions: There is a lack of association between brain injury based on GCS scoring and CT scan findings. Using GCS scoring only for assessing the degree of brain injury is insufficient. Keywords: Computed tomography (CT) scan, Glasgow coma scales (GDS), Head injury.

In the past, taurine was not considered essential in poultry diet; however, heavy reliance on plants derived feed ingredients and environmental stresses have compelled its application inevitable. Six hundred day old male broilers (Hubbard) were supplementation with taurine at the level of 0 (TAN0), 2.5 (TAN2.5), 5 (TAN5) and 7.5 g/kg (TAN7.5) in drinking water during heat stressed conditions for a period of 35 days. On overall basis, feed intake was not significantly (p>.05) different between the treatments. At the end of the experiment, weight gain was significantly (p

Dengue neurological disease is an uncommon yet severe complication of dengue infection. It can manifest as encephalitis, encephalopathy, neuro-ophthalmic complications, or neuromuscular disorders. Severe infection can result in viral shedding across multiple body sites. We describe a case of severe neuro-ophthalmic dengue infection in an otherwise healthy returned traveller, presenting with prolonged multiple-body-site viral detections by PCR. The dengue virus (DENV) dynamics and serological response support a direct DENV neuropathogenicity. A retrospective review of the laboratory data at the Victorian Infectious Diseases Reference Laboratory (VIDRL) suggests that blood is the most frequent sample type with DENV detection (92% of all DENV-positive samples). Genotype variation is seen across different sample types. The similarity of CSF and nasopharyngeal DENV subtypes (genotype 1 and 3) suggests a possible correlation between nasopharyngeal replication and neurological complications. The case presented highlights the direct neuropathogenicity of DENV early in the course of infection, and a potential correlation between nasopharyngeal replication and neurological disease. [ABSTRACT FROM AUTHOR]

This study examined the effects of dietary supplementation with animal protein concentrate (APC) and ginger phyto-protease (GPZ) on growth performance, carcase yield, litter quality and gut microbiota in broiler chicks over a 35–day period. A total of 700-day-old chicks were divided into give groups with five replicates as follow: control (standard diet), diet with 4% APC, diet with 6% APC, diet with 4% APC + GPZ at 0.02% and diet with 6% APC + GPZ at 0.02%. The results showed that GPZ supplementation improved body weight gain (BWG) and feed conversion ratio (FCR) in diets containing APC. Carcase yield analysis revealed that diet with 4% APC + GPZ, and diet with 6% APC + GPZ had the highest yields (p

Selenium (Se) is an essential trace mineral required for growth, development, immune function, and metabolism. Selenium exerts its biological effects as an integral component of selenoproteins (SePs). Deficiency or low Se status leads to marked changes in many biochemical pathways and a range of pathologies and disorders which are associated with SeP function. Animals, and presumably humans, are able to efficiently utilize nutritionally adequate levels of Se in both organic and inorganic forms. It is now clear that the bioavailability of Se varies depending on the source and chemical form of the Se supplement. There are a range of products available for dietary Se supplementation, however, organic sources have been shown to be assimilated more efficiently than inorganic compounds and are considered to be less toxic and more appropriate as a feed supplement. Yeast enriched with Selenohomoalanthionine (SeHLan) has recently become commercially available, and initial research suggests that it may be an efficacious source for the production of Se enriched animal products.

The use of fiber-reinforced polymer (FRP) jackets as external confinement is becoming popular, especially in seismic areas, because of its ability to enhance the strength and ductility of reinforced concrete to perform as a sustainable symmetric structural member. Therefore, various researchers have worked out for the prediction of strength and strain models of FRP-confined concrete. This study presents the improved strain models for the FRP confined cylindrical concrete members. Different previously proposed models of axial strain of FRP-confined concrete were evaluated based on a large database of 678 specimens from previous experiments and an improved model was proposed using the general regression analysis technique. Furthermore, the proposed model was validated using the previous experimental work of FRP-wrapped concrete cylinders and their finite elements analysis (FEA) using the ABAQUS software. The accuracy of the proposed strain model was quite satisfactory in comparison with the previous experimental and FEA results of the present study. Moreover, the proposed empirical strain model was used for the parametric study to investigate the effect of different geometric and material parameters such as the compressive strength of unconfined concrete, diameter of the cylinder, elastic modulus and thickness of the FRP layers, on the axial strain of FRP-wrapped cylinders. A close agreement among the proposed strain models and experimental outputs was observed. This study will help in understanding the behavior of sustainable FRP-confined symmetric concrete members.

Present study explored the potentials of medicinal plants (Aloe barbedensis, Pimpinella anisum, Berberis lycium, Trigonella foenum-graecum, Allium sativum) mixture in broiler chicks at a ratio of 1:3:1:2:1, respectively. For this purpose 240 chicks were randomly assigned into four major groups, namely; A, B, C and D. Each group was further divided into two subgroups with three replicates of 10 chicks each. One of the subgroups was vaccinated against Newcastle disease (ND), Infectious bronchitis (IB) and Infectious bursal disease (IBD) according to locally adopted vaccination schedule keeping the other subgroup as non vaccinated control. Experimental birds in groups A, B, and C were provided with mentioned infusion @ 20, 10 and 5ml per liter of drinking water while group D was maintained as control. Relevant data were recorded throughout the experiment and or at the termination of the experiment and subjected to statistical analysis. Significantly low mean feed intake (3258.3g) and better FCR (1.87) was recorded in group A. Other parameters like weight gain (1739.7g), antibody titer against ND (7.1), IBD (3300.5) and high density lipoprotein (71.6 g/dl) were significantly increased in chicks from group A. Similarly, blood cholesterol (145.6 g/dl), triglyceride (145.8 g/dl) and low density lipoprotein (57.5 g/dl) were significantly reduced in group A. Overall vaccinated group had higher (P

Milk thistle was added in aflatoxin B1 contaminated poultry feed to investigate and compare its hepatoprotective effects with a commercial toxin binder. Two hundred and forty, day-old broilers were randomly allocated into four major groups A, B, C and D. Group A was kept as control, having aflatoxin free feed, while group B was fed aflatoxin contaminated feed, group C was raised on aflatoxin contaminated feed with toxin binder “Mycoad” @ 3g/kg of feed, while group D was provided aflatoxin contaminated feed along with milk thistle @10g/kg of feed. Aflatoxin B1 was present at the level of 80 µg/kg feed during the first week and 520 µg/kg feed in the remaining experimental period. Serum total protein was significantly (P

Objective: To determine the frequency and etiology of Early Return Visits within 3 days of discharge to the Emergency Department. Study Design: Prospective longitudinal study. Place and Duration of Study: Emergency Department of Combined Military Hospital, Rawalpindi Pakistan, from Jan to Jun 2022. Methodology: All patients visiting the Emergency Department within the defined 6-month period were included in this study, which came to 101'113. To determine overall incidence of Early Return Visits, total number of patients revisiting within 72 hours of Emergency Department discharge was used. To find out the possible factors responsible for ERVs, a sample size of 80 patients revisiting the Emergency Department was calculated through World Health Organization calculator and selected through random sampling technique. Factors responsible for unscheduled Early Return Visits were divided into three broad categories: Doctor-associated, Disease-associated, and Patient-associated. Results: The total number of patients revisiting Emergency Department within 72 hours was 2,880 (2.9%). Out of these 52.5% patients reported back to the Emergency Department due to disease-associated reasons, 27.5% due to doctor-related reasons, and 20% revisited for patient-related reasons. Conclusion: Two-point nine percent patients made ERVs to the Emergency Department within 72 hours. The most common causes of Early Return Visits in this study are disease-related issues followed by doctor-related issues. [ABSTRACT FROM AUTHOR]

Objective: To compare post-operative analgesic requirements in patients undergoing hemorrhoidectomy under caudal epidural anaesthesia versus saddle anaesthesia. Study Design: Quasi Experimental Study. Place and Duration of Study: Combined Military Hospital, Rawalpindi Pakistan, Feb to Nov 2021. Methodology: The study was conducted on 200 patients who underwent hemorrhoidectomy during the study period. Patients were randomized into two groups. Group-A received caudal epidural anaesthesia, while Group-B received saddle anaesthesia. The pain at the surgical site was recorded on a visual analogue scale 04 hours and 08 hours after the surgical procedure. The difference in significant post-operative pain and the requirement of opioid analgesia was compared. Results: Out of 200 patients randomized into two groups, 96(48%) were categorized into Group-A, and 104(52%) were categorized into Group-B. The mean age of patients who were operated for hemorrhoids in our study was 41.56±6.98 years. Patients who received caudal epidural anaesthesia had more chances of not having significant post-operative pain at 04 and 08 hours after the surgery as compared to those who received saddle anaesthesia (p-value<0.05). The requirement of opioid analgesia at 02 hours was also statistically significantly less in these patients as well (p-value<0.05). Conclusion: The Use of caudal epidural anaesthesia during the haemorrhoidectomy emerged as better management for early post-operative pain as compared to the use of saddle anaesthesia. The requirement for opioid analgesia was higher in patients after two hours of surgery who received saddle block than those who were administered caudal epidural anaesthesia. [ABSTRACT FROM AUTHOR]

Abstract Background Hepatitis C virus (HCV) is one of the major health concerns globally, with genotype 3a as the most prevalent in Pakistan. Lack of efficient HCV genotype 3a small animal models as well as genomic replicons has hampered the complete understanding of its life cycle, pathogenesis and therapeutic options. In this study we aimed to develop a persistent HCV genotype 3a infectious cell culture model. Methods We inoculated Huh-7 cells with HCV genotype 3a serum. Cells and media supernatant were collected at different time periods up to 40th day post infection. Culture media supernatant was also collected to find out its ability to infect naive Huh-7 cells. Results HCV replication was confirmed at both RNA and protein level through Real Time RCR and western blot using HCV core as marker. In order to validate the persistence of our model for HCV genotype 3a replication we inhibited the HCV replication through core specific siRNAs. The HCV RNA was detected intracellularly from the day one post infection up till 40th day, while HCV core protein was detected from the second day up to 40th day consistently. In culture media supernatant HCV RNA was also actively detected conferring its ability to infect the naive Huh-7 cells. Furthermore, core specific siRNA showed significant inhibition at 24th hour post transfection both at RNA and protein level with progressive increase in the expression of core gene after 3rd day. It clearly depicts that the Huh-7 successfully retained the HCV replication after degradation of siRNA. Conclusion Finally, we report that our persistent infection cell culture model consistently replicate HCV genotype 3a for more than 1 month.

Abstract Background and Aims HCV infection may lead to hepatic fibrosis. In this study, we tried to determine whether there is any correlation of HCV genotypes and viral load to the clinical parameters such as ALT, AST, ALP, bilirubin, Hb level, patient's age and gender; and then correlated this association with disease progression in liver biopsy samples. Methods In cross-sectional and observational study, 6048 serum HCV RNA positive patients were chosen. The study consists of 53 months from March 2006 to September 2010. Patients were divided into three cohorts to validate our data. Statistical analysis and correlation of lab parameters with viral factors was determined by using SPSS version 16. Results The most prevalent genotype was 3 (70.9%) followed by 1 (13.3%) and 4 (7.4%), collectively. During Univariate analysis, in all cohorts; serum bilirubin, ALP, ALT and AAR showed significant correlation with genotypes, however multivariate analysis showed that all genotypes except 4a have no association with host biochemical markers. Disease progression was also independent of all genotypes. Serum ALP, ALT, bilirubin and viremea levels were significantly elevated in patients with genotype 4a. Viral load showed negative association with serum bilirubin (r = -0.112, P = 0.000) and ALP levels (r = -0.098, P = 0.000). We observed positive correlation of ALP and bilirubin levels, while negative associations of viral load with HCV liver disease progression. Conclusion Disease progression seems independent of the genotypes. Relationship between ALP and bilirubin with viral load may be an attractive marker to guess disease progression in patients with hepatitis C.

Abstract Background The 9.6 kb long RNA genome of Hepatitis C virus (HCV) is under the control of RNA dependent RNA polymerase, an error-prone enzyme, for its transcription and replication. A high rate of mutation has been found to be associated with RNA viruses like HCV. Based on genetic variability, HCV has been classified into 6 different major genotypes and 11 different subtypes. However this classification system does not provide significant information about the origin of the virus, primarily due to high mutation rate at nucleotide level. HCV genome codes for a single polyprotein of about 3011 amino acids which is processed into structural and non-structural proteins inside host cell by viral and cellular proteases. Results We have identified a conserved NS4A protein sequence for HCV genotype 3a reported from four different continents of the world i.e. Europe, America, Australia and Asia. We investigated 346 sequences and compared amino acid composition of NS4A protein of different HCV genotypes through Multiple Sequence Alignment and observed amino acid substitutions C22, V29, V30, V38, Q46 and Q47 in NS4A protein of genotype 1b. Furthermore, we observed C22 and V30 as more consistent members of NS4A protein of genotype 1a. Similarly Q46 and Q47 in genotype 5, V29, V30, Q46 and Q47 in genotype 4, C22, Q46 and Q47 in genotype 6, C22, V38, Q46 and Q47 in genotype 3 and C22 in genotype 2 as more consistent members of NS4A protein of these genotypes. So the different amino acids that were introduced as substitutions in NS4A protein of genotype 1 subtype 1b have been retained as consistent members of the NS4A protein of other known genotypes. Conclusion These observations indicate that NS4A protein of different HCV genotypes originally evolved from NS4A protein of genotype 1 subtype 1b, which in turn indicate that HCV genotype 1 subtype 1b established itself earlier in human population and all other known genotypes evolved later as a result of mutations in HCV genotype 1b. These results were further confirmed through phylogenetic analysis by constructing phylogenetic tree using NS4A protein as a phylogenetic marker.

Abstract Background Several factors have been proposed to assess the clinical outcome of HCV infection. The correlation of HCV genotypes to possible serum markers in clinical prediction is still controversial. The main objective of this study was to determine the existence of any correlation between HCV genotypes to viral load and different clinical serum markers. Methods We performed a prospective cross-sectional and observational study. About 3160 serum HCV RNA positive patients were chosen from 4020 randomly selected anti-HCV positive patients. Statistical analysis was performed using the SPSS 16 software package. ROC (receiver operating characteristics) curves were used to compare diagnostic values of serum markers to predict genotypes. Results The most prevalent genotype was 3a (73.9%) followed by 1a (10.7%), 4a (6.4%) and 3b (6.1%) in Pakistani population. No correlation was found between viral load and serum markers for genotype 3a in a large no. of sample (n = 2336). While significant correlation was observed between viral load and AST in genotype 3b, ALP with viral load and ALT for genotype 1a. Patients with genotype 4a showed a significant inverse correlation with viral load and Hb level and AST with ALP. For genotype 4a, AUC (area under the curve) of ALT, ALP, AST, bilirubin, Hb level and viral load was 0.790, 0.763, 0.454, 0.664, 0.458 and 0.872 respectively. Conclusions In conclusion, there was a significant variable response of HCV genotypes with serum markers. Severity of disease is independent of serum marker level in genotype 3a, while the liver damage in genotype 4a may associate with viral cytopathic effect as well as the immune-mediated process. An index using six serum markers may correctly predict genotype 4a in patients with ≥75% accuracy.

Abstract Hepatocellular carcinoma is mainly associated with viral hepatitis B and C. Activation of cell growth stimulator IGF-II gene is observed in tumor formation especially in viral associated hepatocellular carcinoma. Elevated IGF-II levels are indicator of increased risk for cholangiocellular and hepatocellular carcinomas through over saturation of IGF-II binding capacities with IGF receptors leading to cellular dedifferentiation. In HCV, core protein is believed to trans-activate host IGF-II receptor through PKC pathway and the inhibition of tumor cell growth can be achieved by blocking IGF-II pathway either at transcriptional level or increasing its binding with IGFBPs (Insulin like growth factor proteins) at C-terminal, so that it is not available in free form. IGFBP-6 is a specific inhibitor of IGF-II actions. Affinity of IGFBPs with IGFs is controlled by post-translational modifications. Phosphorylation of IGFBPs inhibits IGFs action on target cells while O-glycosylation prevents binding of IGFBP-6 to glycosaminoglycans and cell membranes and resulting in a 10-fold higher affinity for IGF-II. O-glycosylation and phosphorylation operate the functional expression of cellular proteins, this switching on and off the protein expression is difficult to monitor in vivo. By using neural network based prediction methods, we propose that alternate O-β-GlcNAc modification and phosphorylation on Ser 204 control the binding of IGFBP-6 with IGF-II. This information may be used for developing new therapies by regulating IGFBP-6 assembly with IGF-II to minimize the risk of viral associated hepatocellular carcinoma. We can conclude that during HCV/HBV infection, O-β-GlcNAc of IGFBP-6 at Ser 204 diminish their binding with IGF-II, increase IGF-II cellular expression and promote cancer progression which can lead to hepatocellular carcinoma. Furthermore, this site can be used for developing new therapies to control the IGF-II actions during viral infection to minimize the risk of hepatocellular carcinoma.

Abstract HCV is a leading cause of hepatocellular carcinoma and cirrhosis all over the world. Claudins belong to family of tight junction's proteins that are responsible for establishing barriers for controlling the flow of molecules around cells. For therapeutic strategies, regulation of viral entry into the host cells holds a lot of promise. During HCV infection claudin-1 is highly expressed in liver and believed to be associated with HCV virus entry after HCV binding with or without co-receptor CD81. The claudin-1 assembly with tight junctions is regulated by post translational modifications. During claudins assembly and disassembly with tight junctions, phosphorylation is required at C-terminal tail. In cellular proteins, interplay between phosphorylation and O-β-GlcNAc modification is believed to be functional switch, but it is very difficult to monitor these functional and vibrant changes in vivo. Netphos 2.0 and Disphos 1.3 programs were used for potential phosphorylation; NetPhosK 1.0 and KinasePhos for kinase prediction; and YinOYang 1.2 and OGPET to predict possible O-glycosylation sites. We also identified Yin Yang sites that may have potential for O-β-GlcNAc and phosphorylation interplay at same Ser/Thr residues. We for the first time proposed that alternate phosphorylation and O-β-GlcNAc modification on Ser 192, Ser 205, Ser 206; and Thr 191 may provide an on/off switch to regulate assembly of claudin-1 at tight junctions. In addition these phosphorylation sites may be targeted by novel chemotherapeutic agents to prevent phosphorylation lead by HCV viral entry complex.

Abstract HIV (Human immunodeficiency virus) transmission has been reduced by protected sex and screening of blood products and other body fluids in the developed countries. It has been reported that Pakistan is at high risk of HIV/AIDS infection but presently the prevalence rate is considerably low. The number of reported cases of HIV/AIDS in Pakistan has been continuously increasing since 1987. By 2010 the total number of registered cases has reached to 6000 and this figure is on the rise with the passage of time. Some serious strategies must be implemented to control this deadly disease.

Abstract Background Hepatitis C can lead to liver fibrosis and cirrhosis. We compared readily available non-invasive fibrosis indexes for the fibrosis progression discrimination to find a better combination of existing non-invasive markers. Methods We studied 157 HCV infected patients who underwent liver biopsy. In order to differentiate HCV fibrosis progression, readily available AAR, APRI, FI and FIB-4 serum indexes were tested in the patients. We derived a new fibrosis-cirrhosis index (FCI) comprised of ALP, bilirubin, serum albumin and platelet count. FCI = [(ALP × Bilirubin) / (Albumin × Platelet count)]. Results Already established serum indexes AAR, APRI, FI and FIB-4 were able to stage liver fibrosis with correlation coefficient indexes 0.130, 0.444, 0.578 and 0.494, respectively. Our new fibrosis cirrhosis index FCI significantly correlated with the histological fibrosis stages F0-F1, F2-F3 and F4 (r = 0.818, p < 0.05) with AUROCs 0.932 and 0.996, respectively. The sensitivity and PPV of FCI at a cutoff value < 0.130 for predicting fibrosis stage F0-F1 was 81% and 82%, respectively with AUROC 0.932. Corresponding value of FCI at a cutoff value ≥1.25 for the prediction of cirrhosis was 86% and 100%. Conclusions The fibrosis-cirrhosis index (FCI) accurately predicted fibrosis stages in HCV infected patients and seems more efficient than frequently used serum indexes.

Abstract Background Chronic HCV is one of the major causes of morbidity and mortality in the present day world. The assessment of disease progression not only provides useful information for diagnosis and therapeutic supervision judgment but also for monitoring disease. Different invasive and non invasive methods are applied to diagnose the disease from initial to end stage (mild fibrosis to cirrhosis). Although, liver biopsy is still considered as gold standard to identify liver histological stages, an assessment of the disease development based on non-invasive clinical findings is also emerging and this may replace the need of biopsy in near future. This review gives brief insight on non-invasive methods currently available for predicting liver fibrosis in HCV with their current pros and cons to make easier for a clinician to choose better marker to assess liver fibrosis in HCV infected patients. Methods More than 200 studies regarding invasive and noninvasive markers available for HCV liver disease diagnosis were thoroughly reviewed. We examined year wise results of these markers based on their sensitivity, specificity, PPV, NPV and AUROCs. Results We found that in all non-invasive serum markers for HCV, FibroTest, Forn's Index, Fibrometer and HepaScore have high five-year predictive value but with low AUROCs (0.60~0.85) and are not comparable to liver biopsy (AUROC = 0.97). Even though from its beginning, Fibroscan is proved to be best with high AUROCs (> 0.90) in all studies, no single noninvasive marker is able to differentiate all fibrosis stages from end stage cirrhosis. Meanwhile, specific genetic markers may not only discriminate fibrotic and cirrhotic liver but also differentiate individual fibrosis stages. Conclusions There is a need of marker which accurately determines the stage based on simplest routine laboratory test. Genetic marker in combination of imaging technique may be the better non invasive diagnostic method in future.

Abstract Background Hepatitis C virus (HCV) is a major causative agent of liver associated diseases throughout the world, with genotype 3a responsible for most of the cases in Pakistan. Due to the limited efficiency of current therapy, RNA interference (RNAi) a novel regulatory and powerful silencing approach for molecular therapeutics through a sequence-specific RNA degradation process represents an alternative option. Results The current study was purposed to assess and explore the possibility of RNAi to silence the HCV-3a Core gene expression, which play complex role in regulation of cell growth and host genes expression essential for infectivity and disease progression. To identify the potent siRNA target sites, 5 small interfering RNAs (siRNAs) against Core gene were designed and in vitro transcribed after consensus sequence analysis of different HCV-3a isolates. Antiviral effects of siRNAs showed upto 80% inhibition of Core gene expression by different siRNAs into Huh-7 cells as compared with Mock transfected and control siRNAs treated cells. For long lasting effect of siRNAs, vector based short hairpin siRNAs (shRNAs) were designed and tested against HCV-3a Core which resulted in a similar pattern of inhibition on RNA and protein expression of HCV Core as synthetic siRNAs. Furthermore, the efficacy of cell culture tested siRNA and shRNA, were evaluated for inhibition of HCV replication in HCV infected serum inoculated Huh-7 cells and a significant decrease in HCV viral copy number was observed. Conclusions Our results support the possibility of using consensus siRNA and shRNA-based molecular therapy as a promising strategy in effective inhibition of HCV-3a genotype.

We have developed an in situ bioprinting method that allows the printing of cells under true physiological conditions by applying self-assembling ultrashort peptides as bioinks. This method avoids cell stressing methods, such as UV-treatment, chemical crosslinking and viscous bioink printing methods. We further demonstrate that different nanomaterials can easily be synthesized or incorporated in the 3D bioprinted peptide scaffolds which opens up the possibility of functionalized 3D scaffolds. [ABSTRACT FROM AUTHOR]

The endosymbiont Wolbachia is known to block replication of several important arboviruses, including dengue virus (DENV), in the mosquito vector Aedes aegypti. So far, the exact mechanism of this viral inhibition is not fully understood. A recent study in Drosophila melanogaster has demonstrated an interaction between the pelo gene and Drosophila C virus. In this study, we explored the possible involvement of the pelo protein, that is involved in protein translation, in Wolbachia-mediated antiviral response and mosquito-DENV interaction. We found that pelo is upregulated during DENV replication and its silencing leads to reduced DENV virion production suggesting that it facilities DENV replication. However, in the presence of Wolbachia, specifically in female mosquitoes, the pelo protein is downregulated and its subcellular localization is altered, which could contribute to reduction in DENV replication in Ae. aegypti. In addition, we show that the microRNA aae-miR-2940-5p, whose abundance is highly enriched in Wolbachia-infected mosquitoes, might mediate regulation of pelo. Our data reveals identification of pelo as a host factor that is positively involved in DENV replication, and its suppression in the presence of Wolbachia may contribute to virus blocking exhibited by the endosymbiont. [ABSTRACT FROM AUTHOR]

Detailed knowledge of the three-dimensional (3D) structure of a protein is essential for the proper understanding of its function(s) that could be modified through posttranslational modifications (PTMs). Among these PTMs, alterations of serine/threonine residues of a protein through phosphorylation and O-glycosylation are extremely dynamic and could modulate the functions of a protein by affecting their 3D structure. Potential of a protein for certain PTMs could be evaluated through computer-based methods. Erythropoietin (EPO) is a multifunctional protein that primarily regulates red blood cell production and is also involved in other nonhematopoietic functions; for example, EPO also has cardioprotective and neuroprotective effects. In this study, multifunctional EPO behavior has been revealed based on transient modifications of its receptor. In this study, PTMs of erythropoietin receptor (EPO-R) were predicted using neural network tools, and the possible effects of these modifications are suggested. Phosphorylation and O-glycosylation at serine 380 and 444 of the cytoplasmic domain of EPO-R seem to have an antagonistic role in controlling signaling events induced by EPO. O-glycosylation at threonine 423 might hinder β-TrCP (a ubiquitin ligase) binding, which ubiquitinates at K 428, and ultimately results in the recycling of EPO-R, thus increasing EPO sensitivity. In contrast, the phosphorylated form of the same residue inhibits the recycling of EPO-R and thereby decreases the EPO sensitivity. Additionally, the interplay of O-glycosylation modification at serine 478 and phosphorylation at tyrosine 479 might help in controlling the duration of EPO-induced signaling. [ABSTRACT FROM AUTHOR]

Long intergenic non-coding RNAs (lincRNAs) are appearing as an important class of regulatory RNAs with a variety of biological functions. The aim of this study was to identify the lincRNA profile in the dengue vector Aedes aegypti and evaluate their potential role in host-pathogen interaction. The majority of previous RNA-Seq transcriptome studies in Ae. aegypti have focused on the expression pattern of annotated protein coding genes under different biological conditions. Here, we used 35 publically available RNA-Seq datasets with relatively high depth to screen the Ae. aegypti genome for lincRNA discovery. This led to the identification of 3,482 putative lincRNAs. These lincRNA genes displayed a slightly lower GC content and shorter transcript lengths compared to protein-encoding genes. Ae. aegypti lincRNAs also demonstrate low evolutionary sequence conservation even among closely related species such as Culex quinquefasciatus and Anopheles gambiae. We examined their expression in dengue virus serotype 2 (DENV-2) and Wolbachia infected and non-infected adult mosquitoes and Aa20 cells. The results revealed that DENV-2 infection increased the abundance of a number of host lincRNAs, from which some suppress viral replication in mosquito cells. RNAi-mediated silencing of lincRNA_1317 led to enhancement in viral replication, which possibly indicates its potential involvement in the host anti-viral defense. A number of lincRNAs were also differentially expressed in Wolbachia-infected mosquitoes. The results will facilitate future studies to unravel the function of lncRNAs in insects and may prove to be beneficial in developing new ways to control vectors or inhibit replication of viruses in them. [ABSTRACT FROM AUTHOR]

Background: Several factors have been proposed to assess the clinical outcome of HCV infection. The correlation of HCV genotypes to possible serum markers in clinical prediction is still controversial. The main objective of this study was to determine the existence of any correlation between HCV genotypes to viral load and different clinical serum markers. Methods: We performed a prospective cross-sectional and observational study. About 3160 serum HCV RNA positive patients were chosen from 4020 randomly selected anti-HCV positive patients. Statistical analysis was performed using the SPSS 16 software package. ROC (receiver operating characteristics) curves were used to compare diagnostic values of serum markers to predict genotypes. Results: The most prevalent genotype was 3a (73.9%) followed by 1a (10.7%), 4a (6.4%) and 3b (6.1%) in Pakistani population. No correlation was found between viral load and serum markers for genotype 3a in a large no. of sample (n = 2336). While significant correlation was observed between viral load and AST in genotype 3b, ALP with viral load and ALT for genotype 1a. Patients with genotype 4a showed a significant inverse correlation with viral load and Hb level and AST with ALP. For genotype 4a, AUC (area under the curve) of ALT, ALP, AST, bilirubin, Hb level and viral load was 0.790, 0.763, 0.454, 0.664, 0.458 and 0.872 respectively. Conclusions: In conclusion, there was a significant variable response of HCV genotypes with serum markers. Severity of disease is independent of serum marker level in genotype 3a, while the liver damage in genotype 4a may associate with viral cytopathic effect as well as the immune-mediated process. An index using six serum markers may correctly predict genotype 4a in patients with ≥75% accuracy. [ABSTRACT FROM AUTHOR]

HCV is a leading cause of hepatocellular carcinoma and cirrhosis all over the world. Claudins belong to family of tight junction's proteins that are responsible for establishing barriers for controlling the flow of molecules around cells. For therapeutic strategies, regulation of viral entry into the host cells holds a lot of promise. During HCV infection claudin-1 is highly expressed in liver and believed to be associated with HCV virus entry after HCV binding with or without co-receptor CD81. The claudin-1 assembly with tight junctions is regulated by post translational modifications. During claudins assembly and disassembly with tight junctions, phosphorylation is required at C-terminal tail. In cellular proteins, interplay between phosphorylation and O-β-GlcNAc modification is believed to be functional switch, but it is very difficult to monitor these functional and vibrant changes in vivo. Netphos 2.0 and Disphos 1.3 programs were used for potential phosphorylation; NetPhosK 1.0 and KinasePhos for kinase prediction; and YinOYang 1.2 and OGPET to predict possible O-glycosylation sites. We also identified Yin Yang sites that may have potential for O-β-GlcNAc and phosphorylation interplay at same Ser/Thr residues. We for the first time proposed that alternate phosphorylation and O-β-GlcNAc modification on Ser 192, Ser 205, Ser 206; and Thr 191 may provide an on/off switch to regulate assembly of claudin-1 at tight junctions. In addition these phosphorylation sites may be targeted by novel chemotherapeutic agents to prevent phosphorylation lead by HCV viral entry complex. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma is mainly associated with viral hepatitis B and C. Activation of cell growth stimulator IGF-II gene is observed in tumor formation especially in viral associated hepatocellular carcinoma. Elevated IGF-II levels are indicator of increased risk for cholangiocellular and hepatocellular carcinomas through over saturation of IGF-II binding capacities with IGF receptors leading to cellular dedifferentiation. In HCV, core protein is believed to transactivate host IGF-II receptor through PKC pathway and the inhibition of tumor cell growth can be achieved by blocking IGF-II pathway either at transcriptional level or increasing its binding with IGFBPs (Insulin like growth factor proteins) at C-terminal, so that it is not available in free form. IGFBP-6 is a specific inhibitor of IGF-II actions. Affinity of IGFBPs with IGFs is controlled by post-translational modifications. Phosphorylation of IGFBPs inhibits IGFs action on target cells while O-glycosylation prevents binding of IGFBP-6 to glycosaminoglycans and cell membranes and resulting in a 10-fold higher affinity for IGF-II. O-glycosylation and phosphorylation operate the functional expression of cellular proteins, this switching on and off the protein expression is difficult to monitor in vivo. By using neural network based prediction methods, we propose that alternate O-β-GlcNAc modification and phosphorylation on Ser 204 control the binding of IGFBP-6 with IGF-II. This information may be used for developing new therapies by regulating IGFBP-6 assembly with IGF-II to minimize the risk of viral associated hepatocellular carcinoma. We can conclude that during HCV/HBV infection, O-β-GlcNAc of IGFBP-6 at Ser 204 diminish their binding with IGF-II, increase IGF-II cellular expression and promote cancer progression which can lead to hepatocellular carcinoma. Furthermore, this site can be used for developing new therapies to control the IGF-II actions during viral infection to minimize the risk of hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

Background: Chronic HCV is one of the major causes of morbidity and mortality in the present day world. The assessment of disease progression not only provides useful information for diagnosis and therapeutic supervision judgment but also for monitoring disease. Different invasive and non invasive methods are applied to diagnose the disease from initial to end stage (mild fibrosis to cirrhosis). Although, liver biopsy is still considered as gold standard to identify liver histological stages, an assessment of the disease development based on non-invasive clinical findings is also emerging and this may replace the need of biopsy in near future. This review gives brief insight on non-invasive methods currently available for predicting liver fibrosis in HCV with their current pros and cons to make easier for a clinician to choose better marker to assess liver fibrosis in HCV infected patients. Methods: More than 200 studies regarding invasive and noninvasive markers available for HCV liver disease diagnosis were thoroughly reviewed. We examined year wise results of these markers based on their sensitivity, specificity, PPV, NPV and AUROCs. Results: We found that in all non-invasive serum markers for HCV, FibroTest, Forn's Index, Fibrometer and HepaScore have high five-year predictive value but with low AUROCs (0.60∼0.85) and are not comparable to liver biopsy (AUROC = 0.97). Even though from its beginning, Fibroscan is proved to be best with high AUROCs (> 0.90) in all studies, no single noninvasive marker is able to differentiate all fibrosis stages from end stage cirrhosis. Meanwhile, specific genetic markers may not only discriminate fibrotic and cirrhotic liver but also differentiate individual fibrosis stages. Conclusions: There is a need of marker which accurately determines the stage based on simplest routine laboratory test. Genetic marker in combination of imaging technique may be the better non invasive diagnostic method in future. [ABSTRACT FROM AUTHOR]

Background: Hepatitis C can lead to liver fibrosis and cirrhosis. We compared readily available non-invasive fibrosis indexes for the fibrosis progression discrimination to find a better combination of existing non-invasive markers.Methods: We studied 157 HCV infected patients who underwent liver biopsy. In order to differentiate HCV fibrosis progression, readily available AAR, APRI, FI and FIB-4 serum indexes were tested in the patients. We derived a new fibrosis-cirrhosis index (FCI) comprised of ALP, bilirubin, serum albumin and platelet count. FCI = [(ALP × Bilirubin) / (Albumin × Platelet count)].Results: Already established serum indexes AAR, APRI, FI and FIB-4 were able to stage liver fibrosis with correlation coefficient indexes 0.130, 0.444, 0.578 and 0.494, respectively. Our new fibrosis cirrhosis index FCI significantly correlated with the histological fibrosis stages F0-F1, F2-F3 and F4 (r = 0.818, p < 0.05) with AUROCs 0.932 and 0.996, respectively. The sensitivity and PPV of FCI at a cutoff value < 0.130 for predicting fibrosis stage F0-F1 was 81% and 82%, respectively with AUROC 0.932. Corresponding value of FCI at a cutoff value ≥1.25 for the prediction of cirrhosis was 86% and 100%.Conclusions: The fibrosis-cirrhosis index (FCI) accurately predicted fibrosis stages in HCV infected patients and seems more efficient than frequently used serum indexes. [ABSTRACT FROM AUTHOR]

Background: Hepatitis C virus (HCV) is a major causative agent of liver associated diseases throughout the world, with genotype 3a responsible for most of the cases in Pakistan. Due to the limited efficiency of current therapy, RNA interference (RNAi) a novel regulatory and powerful silencing approach for molecular therapeutics through a sequence-specific RNA degradation process represents an alternative option. Results: The current study was purposed to assess and explore the possibility of RNAi to silence the HCV-3a Core gene expression, which play complex role in regulation of cell growth and host genes expression essential for infectivity and disease progression. To identify the potent siRNA target sites, 5 small interfering RNAs (siRNAs) against Core gene were designed and in vitro transcribed after consensus sequence analysis of different HCV-3a isolates. Antiviral effects of siRNAs showed upto 80% inhibition of Core gene expression by different siRNAs into Huh-7 cells as compared with Mock transfected and control siRNAs treated cells. For long lasting effect of siRNAs, vector based short hairpin siRNAs (shRNAs) were designed and tested against HCV-3a Core which resulted in a similar pattern of inhibition on RNA and protein expression of HCV Core as synthetic siRNAs. Furthermore, the efficacy of cell culture tested siRNA and shRNA, were evaluated for inhibition of HCV replication in HCV infected serum inoculated Huh-7 cells and a significant decrease in HCV viral copy number was observed. Conclusions: Our results support the possibility of using consensus siRNA and shRNA-based molecular therapy as a promising strategy in effective inhibition of HCV-3a genotype. [ABSTRACT FROM AUTHOR]