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Abstract Hepatitis C virus causes acute and chronic hepatitis and can lead to permanent liver damage and hepatocellular carcinoma (HCC) in a significant number of patients via oxidative stress, insulin resistance (IR), fibrosis, liver cirrhosis and HCV induced steatosis. HCV induced steatosis and oxidative stress causes steato-hepatitis and these pathways lead to liver injury or HCC in chronic HCV infection. Steatosis and oxidative stress crosstalk play an important role in liver damage in HCV infection. This Review illustrates viral and host factors which induce Oxidative stress, steatosis and leads toward HCC. It also expresses Molecular cascade which leads oxidative stress and steatosis to HCC.

Abstract Background Hepatitis C virus (HCV) Core protein regulates multiple signaling pathways and alters cellular genes expression responsible for HCV induced pathogenesis leading to hepatocellular carcinoma (HCC). Prevalence of HCV genotype 3a associated HCC is higher in Pakistan as compare to the rest of world; however the molecular mechanism behind this is still unclear. This study has been designed to evaluate the effect of HCV core 3a on apoptosis and cell proliferation which are involved in HCC Methodology We examined the in vitro effect of HCV Core protein of genotype 3a and 1a on cellular genes involved in apoptosis by Real time PCR in liver cell line (Huh-7). We analyzed the effect of HCV core of genotype 1a and 3a on cell proliferation by MTT assay and on phosphrylation of Akt by western blotting in Huh-7 cells. Results The HCV 3a Core down regulates the gene expression of Caspases (3, 8, 9 and 10), Cyto C and p53 which are involved in apoptosis. Moreover, HCV 3a Core gene showed stronger effect in regulating protein level of p-Akt as compared to HCV 1a Core accompanied by enhanced cell proliferation in Huh-7 cell line. Conclusion From the current study it has been concluded that reduced expression of cellular genes involved in apoptosis, increased p-Akt (cell survival gene) and enhanced cell proliferation in response to HCV 3a core confirms anti apoptotic effect of HCV 3a Core gene in Huh-7 that may lead to HCC.

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/Aim Hepatitis C virus (HCV) is a major threat as almost 3% of the world's population (350 million individual) and 10% of the Pakistani population is chronically infected with this virus. RNA interference (RNAi), a sequence-specific degradation process of RNA, has potential to be used as a powerful alternative molecular therapeutic approach in spite of the current therapy of interferon-α and ribavirin against HCV which has limited efficiency. HCV structural gene E2 is mainly involved in viral cell entry via attachment with the host cell surface receptors i.e., CD81 tetraspanin, low density lipoprotein receptor (LDLR), scavenger receptor class B type 1 (SR-B1), and Claudin1 (CLDN1). Considering the importance of HCV E2 gene and cellular receptors in virus infection and silencing effects of RNAi, the current study was designed to target the cellular and viral factors as new therapeutic options in limiting HCV infection. Results In this study the potential of siRNAs to inhibit HCV-3a replication in serum-infected Huh-7 cells was investigated by combined treatment of siRNAs against the HCV E2 gene and HCV cellular receptors (CD81 and LDLR), which resulted in a significant decrease in HCV viral copy number. Conclusion From the current study it is concluded that the combined RNAi-mediated silencing of HCV E2 and HCV receptors is important for the development of effective siRNA-based therapeutic option against HCV-3a.

Abstract Background Hepatitis C virus (HCV) is a major causative agent of liver associated diseases leading to the development of hepatocellular carcinoma (HCC) all over the world and genotype-3a responsible for most of the cases in Pakistan. Due to the limited efficiency of current chemotherapy of interferon-α (IFN-α) and ribavirin against HCV infection alternative options are desperately needed out of which the recently discovered RNAi represent a powerful silencing approach for molecular therapeutics through a sequence-specific RNA degradation process to silence virus infection or replication. HCV translation is mediated by a highly conserved internal ribosome entry site (IRES) within the 5'UTR region making it a relevant target for new drug development. Materials and methods The present study was proposed to assess and explore the possibility of HCV silencing using siRNA targeting 5'UTR. For this analysis full length HCV 5'UTR of HCV-3a (pCR3.1/5'UTR) was tagged with GFP protein for in vitro analysis in Huh-7 cells. siRNA targeting 5'UTR were designed, and tested against constructed vector in Huh-7 cell line both at RNA and Protein levels. Furthermore, the effect of these siRNAs was confirmed in HCV-3a serum infected Huh-7 cell line. Results The expression of 5'UTR-GFP was dramatically reduced both at mRNA and protein levels as compared with Mock transfected and control siRNAs treated cells using siRNAs against IRES of HCV-3a genotype. The potential of siRNAs specificity to inhibit HCV-3a replication in serum-infected Huh-7 cells was also investigated; upon treatment with siRNAs a significant decrease in HCV viral copy number and protein expression was observed. Conclusions Overall, the present work of siRNAs against HCV 5'UTR inhibits HCV-3a expression and represents effective future therapeutic opportunities against HCV-3a genotype.

Abstract Background Hepatitis C Virus (HCV) has two envelop proteins E1 and E2 which is highly glycosylated and play an important role in cell entry. Inhibition of virus at entry step is an important target to find antiviral drugs against HCV. Glanthus Nivalis Agglutinin (GNA) is a mannose binding lectin which has tendency for specific recognition and reversible binding to the sugar moieties of a wide variety of glycoproteins of enveloped viruses. Results In the present study, HCV pseudoparticles (HCVpp) for genotype 3a were produced to investigate the ability of GNA to block the HCV entry. The results demonstrated that GNA inhibit the infectivity of HCVpp and HCV infected serum in a dose-dependent manner and resulted in 50% reduction of virus at 1 ± 2 μg concentration. Molecular docking of GNA and HCV glycoproteins (E1 and E2) showed that GNA inhibit HCV entry by binding N-linked glycans. Conclusion These results demonstrated that targeting the HCV glycans is a new approach to develop antiviral drugs against HCV.

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 Hepatitis C virus (HCV) Core protein is thought to trigger activation of multiple signaling pathways and play a significant role in the alteration of cellular gene expression responsible for HCV pathogenesis leading to hepatocellular carcinoma (HCC). However, the exact molecular mechanism of HCV genome specific pathogenesis remains unclear. We examined the in vitro effects of HCV Core protein of HCV genotype 3a and 1a on the cellular genes involved in oxidative stress and angiogenesis. We also studied the ability of HCV Core and Cox-2 siRNA either alone or in combination to inhibit viral replication and cell proliferation in HCV serum infected Huh-7 cells. Results Over expression of Core gene of HCV 3a genotype showed stronger effect in regulating RNA and protein levels of Cox-2, iNOS, VEGF, p-Akt as compared to HCV-1a Core in hepatocellular carcinoma cell line Huh-7 accompanied by enhanced PGE2 release and cell proliferation. We also observed higher expression levels of above genes in HCV 3a patient's blood and biopsy samples. Interestingly, the Core and Cox-2-specific siRNAs down regulated the Core 3a-enhanced expression of Cox-2, iNOS, VEGF, p-Akt. Furthermore, the combined siRNA treatment also showed a dramatic reduction in viral titer and expression of these genes in HCV serum-infected Huh-7 cells. Taken together, these results demonstrated a differential response by HCV 3a genotype in HCV-induced pathogenesis, which may be due to Core and host factor Cox-2 individually or in combination. Conclusions Collectively, these studies not only suggest a genotype-specific interaction between key players of HCV pathogenesis but also may represent combined viral and host gene silencing as a potential therapeutic strategy.

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.

Effervescent formulation helps in faster and better absorption of drugs, especially those that are rapidly soluble in water. However, these tablets require special packaging in order to prevent them from absorbing moisture, hence increasing cost. We compared an effervescent tablet prepared using an in-house developed method (multi-layer tablet with acid and base part separated by an inert layer) to a European effervescent tablet (Efferalgan®) in a single-center, randomized cross-over study among twelve healthy volunteers. Blood samples were collected for 8 h and analyzed for paracetamol concentration using HPLC. Our results showed that both the products have similar pharmacokinetic profiles with no significant difference observed for C last , T half , K elim , and MRT (p-value > 0.05). Moreover, to assess bioequivalence we did not find any significant difference (p-value > 0.05) in AUC (27.12 ± 6.02 vs. 27.29 ± 2.64 µg.h/ml), C max (7.42 ± 1.06 vs. 7.83 ± 1.19 µg/ml) and t max (0.85 ± 0.22 vs. 0.83 ± 0.25 h). The TR ratios for AUC, C max , and t max were 0.99, 0.95, and 1.02 respectively, and were all within the specified FDA limits i.e., 0.8-1.25. We found our test tablet to be bioequivalent to that of Efferalgan®., (© 2024. The Author(s).)

Background: Treatment cost and high prevalence of Poly Cystic Ovarian Syndrome (PCOS) is a very challenging issue globally. Due to this reason; current study was conducted to determine pharmaco-economy of conventional and non-conventional treatments for the management of PCOS., Methods: Prospective Cross-Sectional study was conducted in the metropolitan city of Karachi from January - December 2019. Primary data of 200 PCOS patients were collected from different hospitals and clinics. An instrument was used to collect data pertaining to the direct and indirect cost associated with the disease management. Collected data was analyzed by the tools for cost analysis and software called Statistical Package of Social Sciences (SPSS) - 22., Results: In Cost Minimization Analysis (CMA); Allopathic treatment [Mean cost/month: PKR:4479.32 ± 350.95 (USD:27.46 ± 2.15)], Herbal treatment [Mean cost/month: PKR:1527.78 ± 78.15 (USD:9.37 ± 0.48)], Combination treatment [Mean cost/month: PKR:2803.09 ± 654.22 (USD:17.18 ± 4.01)], and Homoeopathic treatment [Mean cost/month: PKR:976.95 ± 46.19 (USD:5.99 ± 0.28)]. Incremental cost/month for Allopathic treatment is 358%, Herbal treatment is 56%, Combination treatment is 187%. In Cost Effectiveness Analysis (CEA); Allopathic treatment (Incremental cost-effectiveness ratio/month: 1334.24), Herbal treatment (Incremental cost-effectiveness ratio/month: 936.41), Combination treatment (Incremental cost-effectiveness ratio/month: 1017.09). Due to lowest cost of Homeopathic treatment, cost of Homeopathic treatment was considered as a threshold value. In-direct cost/month of Allopathic treatment is PKR:593.33 ± 24.00 (USD:3.64 ± 0.15), Herbal treatment is PKR:307.84 ± 26.69 (USD:1.89 ± 0.16), Combination treatment is PKR:409.09 ± 45.63 (USD:2.51 ± 0.28) and Homoeopathic treatment is PKR:300.00 ± 26.39 (USD:1.84 ± 0.16)., Conclusion: The most cost-effective is treatment is Homeopathic; Herbal treatment is second most cost-effective option for the treatment of PCOS. Lowest direct and indirect costs and short treatment duration collaboratively lessen the %incremental cost per year and incremental cost effectiveness ratio per year., (© 2024. The Author(s).)

Scope: This study investigates the impact of extracts derived from Antarctic fish species, Trematomus newnesi and Trematomus bernacchii, on the migration of human placental trophoblast JEG-3 cells, which is a crucial aspect of successful pregnancy., Methods and Results: The extracts, obtained from the muscles of these fish, significantly enhance the migration and invasion of JEG-3 cells in in vitro wound healing, Transwell, and collagen invasion assays. These effects are accompanied by an increase in matrix metalloproteinase (MMP) 9 activity, as demonstrated by zymography. Furthermore, the extracts activated Akt and protein phosphatase 1, resulting in the dephosphorylation of β-catenin at Ser33/37/Thr41, as confirmed by western blot analysis. Consequently, MMP9 is upregulated, while metallopeptidase inhibitor 1/3 is downregulated, as verified by western blot and qRT-PCR analyses. Finally, utilizing ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis, followed by matching with the Global Natural Product Social Molecular Networking library, the study annotates the compound responsible for the observed migratory activity as taurocholic acid. Importantly, the study confirms that taurocholic acid enhances cell migration in JEG-3 cells., Conclusion: The results of this study emphasize the potential of Antarctic fish extracts in promoting extravillous trophoblast migration and invasion, which are critical for successful pregnancy., (© 2024 The Author(s). Molecular Nutrition & Food Research published by Wiley‐VCH GmbH.)

Melissa officinalis and Panax ginseng extracts were investigated to determine combinatorial effects on cognitive behaviors' of albino-rats. The study was prospective-experimental; lasted from June-2022 to March-2023. Learning and memory measurements were done by animal-models. Data analyzed by 22nd version of SPSS. In Passive-avoidance-test both doses of Melissa officinalis and Panax ginseng (100/100mg/kg and 200/200mg/kg) showed significant differences in number of acquisition-trial between groups (p<0.001); drug treated groups showed longer latency-period compared to control and scopolamine (p<0.001). In time-spent-in-dark-chamber treated groups spent less-time in dark-chamber as compared to control and scopolamine (p<0.001). In Morris-water-maze-task treatment groups (100/100mg/kg and 200/200mg/kg) showed significant (p<0.001) decrease in escape-latency compared with control and scopolamine. Spatial-memory-probe showed significant interaction between drugs and days (p<0.001); time-spent in platform region is significantly increased (p<0.001) in both treatment groups compared with control and scopolamine. 8-arm-radial-maze-test showed the significant increase (p<0.05) in total number of correct responses in treatment groups (100/100mg/kg and 200/200mg/kg) compared to control and scopolamine. In-vitro studies revealed acetyl-choline-esterase inhibition by 36.40% from Melissa officinalis and Panax ginseng combination. Study concluded that combination of M. officinalis and P. ginseng extracts may significantly improve the effects on memory and cognition.

Materials and Methods: The cross-sectional survey was conducted; primary data were collected from asthmatic patients in different hospitals and clinics of allopathic, homeopathic, and herbal practitioners in Karachi, Pakistan. The study duration was from January 2020 to December 2020. Asthmatic patients aged over 13 years were selected for the study. A written informed consent was taken from the patients before the interview. Collected data were analyzed by the Statistical Package of Social Sciences (SPSS) 22., Result: Among 255 asthmatic patients; 51.4% ( n  = 131) were male and 48.6% ( n  = 124) were female. For control of acute attacks of asthma 88.2% ( p = 0.0001) of patients significantly preferred allopathic treatment while 6.3% ( p = 0.008) used homeopathic treatment and 5.5% chose herbal treatment. For maintenance of asthma, 78.8% ( p = 0.0001) patients used allopathic treatment while 12.4% ( p = 0.0001) homeopathic and 8.8% ( p = 0.0001) patients used herbal treatment. About 63.4% ( p = 0.0001) of the asthmatic patients used short-acting β -2 agonists for managing acute asthmatic episodes while long-acting β -2 agonists ( p = 0.0001) and inhaled corticosteroids ( p = 0.0001) were found to be the preferred medicines for maintenance therapy. Effectiveness of treatment ( p = 0.004) and cost effectiveness ( p = 0.0001) significantly act as contributing factors for the selection of the treatment. The majority of the patients were satisfied with their chosen treatments for control of asthmatic symptoms., Conclusion: Most asthmatic patients preferred allopathic treatment for the management of acute episodes and control of asthmatic symptoms. It was found that the major factors for selecting a specific treatment include effectiveness, cost, and minimal side effects., Competing Interests: The authors declare that there are no conflicts of interest., (Copyright © 2022 Javeria Farooq et al.)

The aim of study was to find effective treatment option which reduces the risk of complications among patients of polycystic-ovarian-syndrome. A cross-sectional study was conducted from January-2019 to December-2019. Data was collected from 200 patients that have visited hospitals and clinics located in Karachi. A questionnaire was used in the survey. Collected data was analyzed with SPSS-22. Hormonal-imbalance (p=0.0001), polycystic-ovaries (p=0.008), irregular-menstruation (p=0.0001), obesity (p=0.0001), diabetes (p=0.0001) and infertility (p=0.014) significantly treated by allopathic-medications. Hormonal-imbalance (p=0.025), polycystic-ovaries (p=0.0001), irregular-menstruation (p=0.0001), obesity (p=0.046), diabetes (p=0.001), acne (p=0.046), anxiety (p=0.014), depression (p=0.014) and eating disorder (p=0.046) significantly treated by homeopathic-medications. Polycystic-ovaries (p=0.0001), irregular-menstruation (p=0.0001), obesity (p=0.014), diabetes (p=0.0001) and acne (p=0.014) significantly treated by herbal-medications. Allopathic treatment was found effective in reducing risk of complication associated with PCOS; hormonal-imbalance (59%), hirsutism (42%), obesity (89%), diabetes (90%), hypertension (17%), infertility (60%) and anxiety (75%). Risk reduction of complications by Homeopathic treatment; polycystic-ovaries (54%), irregular menstruation (91%) and depression (43%). Combination treatment was found effective in reducing the risk of acne (43%) and eating disorder (100%). Allopathic treatment is effective in reducing the majority of risks of complications and the complications of polycystic-ovaries-syndrome can be significantly controlled with the homeopathic mode of treatment.

Guanine nucleotide exchange factors (GEFs) activate GTPases by stimulating the release of guanosine diphosphate to permit the binding of guanosine triphosphate. ARHGEF3 or XPLN (exchange factor found in platelets, leukemic, and neuronal tissues) is a selective guanine nucleotide exchange factor for Rho GTPases (RhoGEFs) that activates RhoA and RhoB but not RhoC, RhoG, Rac1, or Cdc42. ARHGEF3 contains the diffuse B-cell lymphoma homology and pleckstrin homology domains but lacks similarity with other known functional domains. ARHGEF3 also binds the mammalian target of rapamycin complex 2 (mTORC2) and subsequently inhibits mTORC2 and Akt. In vivo investigation has also indicated the communication between ARHGEF3 and autophagy-related muscle pathologies. Moreover, studies on genetic variation in ARHGEF3 and genome-wide association studies have predicted exciting novel roles of ARHGEF3 in controlling bone mineral density, platelet formation and differentiation, and Hirschsprung disease. In conclusion, we hypothesized that additional biochemical and functional studies are required to elucidate the detailed mechanism of ARHGEF3-related pathologies and therapeutics., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Khaliq, Umair and Yoon.)

Main aim of current study was to determine the anxiolytic and antidepressant potential of Bougainvillea glabra Extract (BVE). The effects were investigated by using Open-Field-Test (OFT), Light-and-Dark Model (LD), Hole-Board (HB) and Forced-Swimming-Test (FST). Different doses for BVE were given to Wistar-Rats and compared with Control and Diazepam. Data has been collected by simple observations of animal behaviors in mentioned models. Collected data was analyzed by SPSS-22 version. In OFT (number of squares travelled), significant differences noted between Control and BV100mg/kg (p=0.001), Diazepam and BV100mg/kg (p=0.0001), Diazepam and BV200mg/kg (p=0.015), Diazepam and BV300 mg/kg (p=0.002). In LD-Test, significant differences were noted between Control and BV100mg/kg, BV200mg/kg and BV300mg/kg (p=0.0001), Diazepam and BV100mg/kg, 200mg/kg (p=0.0001), Diazepam and BV300mg/kg (p=0.028). In HB-Test by head dips, significant differences noted between control group and BV100mg/kg and 200mg/kg (p=0.0001), Control group and BV300mg/kg (p=0.005). For number of head dips, significant differences noted between Diazepam and BV100mg/kg, 200mg/kg and 300mg/kg (p=0.0001). In FST, significant differences were observed between Control group and BV100mg/kg, BV200mg/kg and BV300mg/kg (p=0.0001), Fluoxetine and BV100mg/kg, BV200mg/kg and BV300mg/kg (p=0.0001). It is observed that MAO-A and MAO-B are inhibited by BVE. Study demonstrates that BV flowers have anxiolytic and antidepressant activities.

Osteoarthritis is the most common form of arthritis worldwide and progression of arthritis may lead to secondary complications. Osteoarthritis is the degradation of joint tissues from the effects of injury, grinding sensation and inflammation. There are three main systems of medicine including allopathic, homeopathic and herbal system for the treatment of osteoarthritis. Plants are the basis for traditional medicine which is still widely practiced today. The objective of the present study is to find out the complaints and secondary complications in patients of osteoarthritis on allopathic, homeopathic and herbal system of medicine. This study was conducted in Karachi from January 2019 to January 2020. Allopathic, Homeopathic and Herbal practitioners were consulted in the study (n=600, with= 200 in each study group). Data was analyzed by SPSS 22 version. Finding of the study suggest that after the treatment with allopathic medicines orthopaedician, family physician or rheumatologist / internal medicine specialist, there was no significant difference in complaints (Chi-square =0.598, p=0.742). Moreover, there was no significant difference in complications (Chi-square = 0.039, p=0.981). While treatment with homeopathic medicines either by orthologist or family physician significant difference in complaints were observed (Chi-square =4.86, p=0.027). The patient visiting family physicians have higher number of complications than orthologists. While treatment with herbal medicines either by orthologists or family physician there was significant difference in complaints. (Chi-square =5.404, p=0.020). The study reveals that there is no significant difference of complaints and complications in allopathic systems of medicine when treating osteoarthritis, while it is significant for homeopathic and herbal medicine.

Decidualization refers to the functional differentiation of endometrial stromal cells and plays a significant role in embryo implantation and pregnancy. C-peptide is excreted in equimolar concentrations as that of insulin during the metabolism of proinsulin in pancreatic beta-cells. High levels of C-peptide are correlated with hyperinsulinemia and polycystic ovarian syndrome, which show a defect in decidualization. However, the role of C-peptide in decidualization has not yet been studied. Here, we identified C-peptide as an endogenous antideciduogenic factor. This inhibitory function was confirmed by the reduced expression of decidual markers, including prolactin, insulin-like growth factor-binding protein-1, and Forkhead box protein O1 as well as by the fibroblastic morphological change in the presence of C-peptide. C-peptide also enhanced cellular senescence and decreased the proportion of apoptotic cells during decidualization. In addition, C-peptide potentiated the inhibitory effects of both insulin and palmitic acid in an AKT- and autophagy-independent manner, respectively. Furthermore, C-peptide augmented protein phosphatase 1 (PP1) activity, leading to a reduction in the inhibitory phosphorylation of glycogen synthase kinase (GSK)3β, which resulted in enhanced cellular senescence and decreased apoptosis during decidualization. Taken together, our findings suggest that C-peptide is an antideciduogenic factor acting via the regulation between PP1 and GSK3β in patients with hyperinsulinemia., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Khaliq, Baek, Cho, Chon and Yoon.)

Sarcopenia is the degenerative loss of skeletal muscle mass and function associated with aging and occurs in the absence of any underlying disease or condition. A comparison of the age-related molecular signaling signatures of different muscles has not previously been reported. In this study, we compared the age-related molecular signaling signatures of the intercostal muscles, the diaphragm, and the gastrocnemii using 6-month and 20-month-old rats. The phosphorylation of Akt, ribosomal S6, and Forkhead box protein O1 (FoxO1) in diaphragms significantly increased with age, but remained unchanged in the intercostal and gastrocnemius muscles. In addition, ubiquitin-proteasome degradation, characterized by the levels of MuRF1 and Atrogin-1, did not change with age in all rat muscles. Interestingly, an increase in LC3BII and p62 levels marked substantial blockage of autophagy in aged gastrocnemii but not in aged respiratory muscles. These changes in LC3BII and p62 levels were also associated with a decrease in markers of mitochondrial quality control. Therefore, our results suggest that the age-related signaling events in respiratory muscles differ from those in the gastrocnemii, most likely to preserve the vital functions played by the respiratory muscles.

Progression of diabetes mellitus may leads to secondary complications. Management of such complication is a major challenge for diabetologists. Objective of current study was to evaluate the development of chronic complications in patients of type-II diabetes by comparing their treatment options. This cross sectional study was conducted in outpatient facilities of Karachi from July 2017 to July 2018. 201 type -II DM patients were enrolled in the study. Study was divided into four arms of treatment; herbal, homeopathic, allopathic and combination. Outcome measures were development of acute and chronic complications. Data was analyzed by SPSS 22 version. Finding of study reveals that relative risk reduction of diabetic coma is 50% (p=0.0001) with combination treatment, relative risk reduction of retinopathy is 52% (p=0001) with herbal treatment, relative risk reduction of nephropathy is 58% (p=0.0001) with combination treatment, relative risk reduction of hypertension is 49% (p=0.0001) with herbal treatment, relative risk reduction of myocardial infarction is 15% (p=0.0001) with herbal treatment and relative risk reduction of neuropathies is 27% (p=0.0001) with combination treatment and relative risk reduction of other complications is 12% with allopathic treatment (p=0.20). Finding of study reveals that risk of acute and chronic complications in type-II DM patients is low if they use either herbal system of medicine or combination of different systems of medicine.

Decidualization is characterized by the differentiation of endometrial stromal cells (eSCs), which is critical for embryo implantation and maintenance of pregnancy. In the present study, we investigated the possible effect of simulated microgravity (SM) on the process of proliferation and in vitro decidualization using primary human eSCs. Exposure to SM for 36 h decreased the proliferation and migration of eSCs significantly, without inducing cell death and changes in cell cycle progression. The phosphorylation of Akt decreased under SM conditions in human eSCs, accompanied by a simultaneous decrease in the level of matrix metalloproteinase (MMP)-2 and FOXO3a. Treatment with Akti, an Akt inhibitor, decreased MMP-2 expression, but not FOXO3a expression. The decreased level of FOXO3a under SM conditions impeded autophagic flux by reducing the levels of autophagy-related genes. In addition, pre-exposure of eSCs to SM significantly inhibited 8-Br-cAMP induced decidualization, whereas restoration of the growth status under SM conditions by removing 8-Br-cAMP remained unchanged. Treatment of human eSCs with SC-79, an Akt activator, restored the reduced migration of eSCs and decidualization under SM conditions. In conclusion, exposure to SM inhibited decidualization in eSCs by decreasing proliferation and migration through Akt/MMP and FOXO3a/autophagic flux.

Purpose: To compare the outcomes of infants with a right (RCDH) versus a left-sided (LCDH) congenital diaphragmatic hernia (CDH) and whether these differed according to whether the infants had undergone fetoscopic tracheal occlusion (FETO)., Methods: Demographics, the type of surgical repair, preoperative and postoperative courses and respiratory, gastrointestinal, surgical and skeletal morbidities at follow-up were compared between infants with a RCDH or LCDH. A sub-analysis was undertaken in those who had undergone FETO., Results: During the study period, there were 167 infants with a LCDH and 24 with a RCDH; 106 underwent FETO (15 RCDH). Overall, the need for inhaled nitric oxide (p = 0.036) was higher in the RCDH group and, at follow-up, infants with RCDH were more likely to have a hernia recurrence (p = 0.043), pectus deformity (p = 0.019), scoliosis (p = 0.029) and suffer chronic respiratory morbidity (p = 0.001). There were, however, no significant differences in short term or long term outcomes (hernia recurrence (p = 0.237), pectus deformity (p = 0.322), scoliosis (p = 0.0174) or chronic respiratory morbidity (p = 0.326)) between infants with a right or left sided CDH who had undergone FETO., Conclusion: Overall, infants with a RCDH compared to those with a LCDH had greater long-term morbidity, but not if they had undergone FETO.

Background  Chikungunya fever (CHIKF) is an infectious illness spread by the bite of mosquitoes and caused by an arbovirus known as Chikungunya virus (CHIKV). This disease has become an epidemic in Karachi and various other cities of Pakistan, affecting a large population, especially those from poor, socioeconomically underdeveloped areas. It is proving to be a severe and alarming cause of debility due to its prolonged detrimental effects on the joints. A significant number of cases are reported daily in different hospitals of Karachi, with Civil Hospital being one of the major tertiary care hospitals. The aim of this study is to determine the frequency and symptoms of chikungunya as well as to assess the participants' awareness about the spread and preventive measures of this disease. Methods This is a cross-sectional study that was carried out in Civil Hospital Karachi by approaching patients in the out-patient department (OPD) and the emergency department with complaints of fever and joint pain. All the data was collected via a pre-coded questionnaire during May-June 2017 by taking prior informed verbal consent and were analyzed through Statistical Package for the Social Sciences (SPSS) version 22 software. Results The age group most affected by this disease was the 21-30 years range, which represented almost a quarter of the cases (n=83, 32.17%). Majority of the respondents (n=214, 82.95%) had heard of the disease, mainly due to their own prior experience with it (n=100, 38.76%). Lethargy (n=219, 84.88%), difficulty in walking (n=213, 82.56%), and headache (n=209, 81.01%) were the major symptoms reported apart from fever (n=258, 100.00%) and arthralgia (n=258, 100.00%). Conclusion CHIKF is proving to be a great threat to people as it impairs their quality of life to a great extent. The recent outbreak of chikungunya has victimized a considerable population of Karachi. This study mainly assessed the severity of the disease and its symptoms as well as the lack of awareness among patients. Proper and effective preventive measures can further help to eradicate this disease on a large scale and prevent future epidemics., Competing Interests: The authors have declared that no competing interests exist.

In the year 2003 to 2005 a prospective study was conducted to find out the predominance of Staphylococcus (Staphylococcus aureus) resistance pattern in opposition to five life saving antibiotics as these are the sole agents to treat critically ill patients in hospitals. During the period of two years almost 2500 samples of bacterial culture were taken from different pathological laboratories and hospitals in Karachi. Among these 1500 were Gram positive cocci and 1000 samples were identified as Staphylococcus aureus. Life saving antibiotics were taken from five different groups and by mean of disk diffusion technique antibiogram of Staphylococcus aureus against these antibiotic were determined. During the course of study imipenem showed 11%, amikacin exhibited 58%, cefipime showed 31%, vancomycin and piperacillin/tazobactam displayed 24% resistance against Staphylococcus aureus. Imipenem was found to be most effective against Staphylococcus aureus.Resistance to other antibiotics developed quickly in Staphylococcus aureus collected from clinical areas where these antimicrobial agents are extensively used.

Retrospective study of Cancer types in different ethnic groups & genders determines the pattern of cancers in different ethnic groups & genders during the last eight years reported in Oncology wards of hospitals of Karachi, Pakistan. Every single one male & female case with histologically and cytologically established cancer was enrolled from January 2003 to December 2010. Data for all patients were collected retrospectively by patient's file & charts, which represents the population of Karachi, Interior Sindh & Balochistan. 5134 patients (Male = 2432 / Female = 2702) investigated for their diagnosis of cancer type, ethnicity, age & gender. Classification of malignancy was done according to the International Classification of Disease coding system by W.H.O (ICD-10). The statistical analysis was performed for mean, standard error & proportions for ethnic groups & genders. Proportionately 47.37% males and among which major ethnic groups 17% Sindhi, 17% Immigrant, 4% Baloch, 3% Pukhtoon, ≈ 4% Punjabi, 1% Siraiki, 2% Minorities and 52.62% females, in which 16% Sindhi, 21% Immigrant, 4% Baloch 3% Pukhtoon, 5% Punjabi, 1% Siraiki, 3% Minorities. Mean age of males = 45.75 years, SE ± 0.227 and for females = 44.07, SE ± 0.183. The three most occurring tumors in all cancers of male were found Head & Neck, Adenoma/Carcinoma of Glands & Body cavity membranes, GIT, and females Breast, Head & Neck, Adenoma/Carcinoma of Glands & Body cavity membranes, GIT. The analysis of data indicates Head & Neck is most common cancer among male, in the similar way Breast cancer is the most common malignancy among female.

The present study was conducted to determine the antibiotic susceptibility pattern of Pseudomonas aeruginosa from sputum samples of lower respiratory tract infection patients admitted to different hospitals of Karachi. Most of the hospitals are hampered with high frequency of nosocomial infections generally caused by multiresistant nosocomial pathogen. Among Gram-negative pathogens Pseudomonas aeruginosa considered as most challenging pathogen. The objective of the study was to determine frequency of Pseudomonas aeruginosa from sputum samples and to find out susceptibility pattern against four antibiotics widely used for treatment. The sputum samples from 498 patients were collected consecutively between January 2010 and March 2011 and were cultured and identified. According to CLSI (Clinical Laboratory Standards Institute) guidelines antimicrobial susceptibility testing was performed by disc diffusion method. Pseudomonas aeruginosa were isolated from 24% (120/498) of the lower respiratory tract patient. A higher resistance to Pseudomonas aeruginosa isolate was observed with piperacillin/tazobactam and cefipime i.e. 42% and 40% respectively. Imipenem was found to be most effective antibiotic against Pseudomonas aeruginosa (76% sensitivity) but amikacin resistance was continuously increasing. In conclusion the frequency of Pseudomonas aeruginosa was also higher among lower respiratory tract infection patients with alarmingly high rate of resistance among widely used antibiotics. These findings focused on careful consideration for monitoring and optimization of antimicrobial use in order to reduce occurrence and spread of antimicrobial resistant pathogen.

Incidences of different types of cancer are increasing in Pakistan, among which cancer of Cervix and Respiratory pappilomatosis are of great concern because of their association with human Pappilomavirus (HPV). Cervical cancers typically distress women of middle age or older; however it may affect women in any age after the puberty. Two serotypes of HPV (16 & 18) accounts 70% of cervical cancer cases, while HPV (6 & 11) are considered low-risk viruses associated with genital warts (Condyloma acuminata) and Respiratory pappilomatosis in both gender. Generally, there is transient role of HPV in human body and are removed by immune system in or around 1 year. Data from different Pakistani hospitals provides sound evidence for increasing trends of cervical cancer, which is, being developing country imperative for us. As the cost of cancer management is increasing day by day with poor survival rate and its burden is borne by patient, their family or society in-large, so if screening or prevention is possible then there would be need to identify target population for screening and vaccination. By quality adjusted life year (QALY) measurement, the data from different sources indicates that adolescent age is the appropriate target population and is cost effective for vaccination. Two vaccines manufactured by recombinant DNA technology are licensed in some parts of the world for prevention of HPV related cancers, however both have certain advantage over another, as one of the vaccines contains viral like proteins of two HPV serotypes 16 & 18 and provide additional cross protection against HPV type 13 and 45 with 100% seroprotection, while the other vaccine, being quadrivalent offers protection against four serotypes 6, 11, 16 and 18. Both vaccines tolerability and safety profiles are similar and acceptable, however bivalent vaccine appears to provide long-lasting immunity by the development of memory B-cells hypothetically due to difference of adsorbing agent used by manufacturer, on the other hand, quadrivalent vaccine offers protection against cervical cancer but also offers additional protection against Condyloma acuminata and respiratory Pappilomatosis. As these vaccines are new in the market and initial trials indicate availability of antibodies for up to around 5 years i.e. why it is controversial at the moment that whether booster dose is recommended or not, however it is assumed that, there is no harm to have booster dose at 5th year of vaccination.

Paratuberculosis is a chronic bowel disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Its secretion through semen highlights the importance of paratuberculosis-free breeding bulls. The breeding and teaser bulls at three semen production units (SPUs) located in Punjab, Pakistan, were screened for the presence of antibodies against MAP. A total of 253 samples were collected from SPUs and a commercially available indirect screen ELISA (Is-ELISA) was applied. Is-ELISA detected antibodies in 20 (24.6%), 16 (22.8%), and 17 (16.6%) samples from SPU-I, SPU-II, and SPU-III, respectively. Collectively, seroprevalence of 20.0% (47/235) in breeding bulls and 33.3% (6/18) in teaser bulls was observed, and thus it poses a potential threat of disease spread to a high number of heifers and cows through artificial insemination. Therefore, this paper highlights the presence of the disease for the first time at SPUs and triggers attempts to ascertain the prevalence of paratuberculosis throughout the country.

Hepatitis C virus is a major cause of liver associated diseases all over the world. Irrespective of the significant advances in the current therapy, drugs and vaccines are restricted with many factors such as toxicity, complexity, cost and resistance. New technologies particularly RNA interference (RNAi) mediated by small interfering RNA (siRNA) have become more and more interesting and effective therapeutic entities to silence pathogenic gene products associated with disease, including cancer, viral infections and autoimmune disorders. RNAi works at a posttranscriptional level by targeting mRNA as a mean for inhibiting the synthesis of the encoded protein. Several reports have indicated the efficiency and specificity of synthetic and vector based siRNAs inhibiting HCV replication. In the present review, we focused on the recent development in the potential use and issues regarding siRNA as a therapy for HCV.

Demineralised bone matrix (DBM) products are complex mixtures of proteins known to influence bone growth, turnover, and repair. They are used extensively in orthopaedic surgery, and are bioassayed in vivo prior to being used in clinical applications. Many factors contribute to the osteogenic potency of DBM, but the relative contributions of these factors, as well as the possibility of interactive effects, are not completely defined. The "gold standard" measure of the therapeutic value of DBM, the in vivo assay for ectopic bone formation, is costly, time-consuming, and involves the use of numerous animal subjects. We have measured the levels of five growth factors released by the collagenase digestion of DBM, and statistically related these levels with osteogenic potency as determined by a standard in vivo model, in order to determine which value or combination of values of growth factors best predict osteogenic activity. We conclude that the level of BMP-2 is the best single predictor of osteogenic potency, and that adding the values of other growth factors only minimally increases the predictive power of the BMP-2 measurement. A small, but significant, interactive effect between BMP-2 and BMP-7 was demonstrated. We present a statistical model based on growth factor (e.g. BMP-2) analysis that best predicts the in vivo assay score for DBM. This model allows the investigator to predict which lots of DBM are likely to exhibit in vivo bioactivity and which are not, thus reducing the need to conduct in vivo testing of insufficiently active lots of DBM. This model uses cut-point analysis to allow the user to assign an estimate of acceptable uncertainty with respect to the "gold standard" test. This procedure will significantly reduce the number of animal subjects used to test DBM products.