Computer-aided identification of dengue virus NS2B/NS3 protease inhibitors: an integrated molecular modelling approach for screening of phytochemicals.

Globally, dengue (DENV) fever has appeared as the most widespread vector-borne disease, affecting more than 100 million individuals annually. No approved anti-DENV therapy or preventive vaccine is available yet. DENV NS3 protein is associated with protease activity and is essential for viral replication process within the host cell. NS2B is linked with NS3 protein as a cofactor. Hence, NS3/NS2B is a potential druggable target for developing inhibitors against dengue virus. In the present study, a dataset of Beta vulgaris L.-based natural compounds was developed. Virtual ligand screening of 30 phytochemicals was carried out to find novel inhibitors against the NS2B/NS3 protein. Spatial affinity, drug-likeness, and binding behaviors of selected phytochemicals were analyzed. Post-simulation analysis, including Principal Component Analysis (PCA), MMGBSA, and Co-relation analysis, was also performed to provide deep insight for elucidating protein-ligand complexes. This computer-aided screening scrutinized four potent phytochemicals, including betavulgaroside II, vitexin xyloside, epicatechin, and isovitexin2-O-xyloside inhibitors exhibiting optimal binding with viral NS3/NS2B protein. Our study brings novel scaffolds against DENV NS2B/NS3 of serotype-2 to act as lead molecules for further biological optimization. In future, this study will prompt the exploration and development of adjuvant anti-DENV therapy based on natural compounds.Communicated by Ramaswamy H. Sarma.

SARS-CoV-2 Detection and COVID-19 Diagnosis: A Bird's Eye View.

The battle against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) associated coronavirus disease 2019 (COVID-19) is continued worldwide by administering firsttime emergency authorized novel mRNA-based and conventional vector-antigen-based anti- COVID-19 vaccines to prevent further transmission of the virus as well as to reduce the severe respiratory complications of the infection in infected individuals. However; the emergence of numerous SARS-CoV-2 variants is of concern, and the identification of certain breakthrough and reinfection cases in vaccinated individuals as well as new cases soaring in some low-to-middle income countries (LMICs) and even in some resource-replete nations have raised concerns that only vaccine jabs would not be sufficient to control and vanquishing the pandemic. Lack of screening for asymptomatic COVID-19-infected subjects and inefficient management of diagnosed COVID-19 infections also pose some concerns and the need to fill the gaps among policies and strategies to reduce the pandemic in hospitals, healthcare services, and the general community. For this purpose, the development and deployment of rapid screening and diagnostic procedures are prerequisites in premises with high infection rates as well as to screen mass unaffected COVID-19 populations. Novel methods of variant identification and genome surveillance studies would be an asset to minimize virus transmission and infection severity. The proposition of this pragmatic review explores current paradigms for the screening of SARS-CoV-2 variants, identification, and diagnosis of COVID-19 infection, and insights into the late-stage development of new methods to better understand virus super spread variants and genome surveillance studies to predict pandemic trajectories.

Synthesis, structural characterization and in vitro pharmacological properties of betanin-encapsulated chitosan nanoparticles.

Betanin, a natural food color and the only betalain, is approved for use in pharmaceutical and food industries as natural antioxidative and preservative agent, respectively. However, the antioxidant power and health-promoting properties of betanin have been disregarded due to its low stability in physiological conditions. Therefore, this study is designed to synthesize and evaluate in vitro pharmacological characteristics of betanin-encapsulated chitosan nanoparticles (ChBetNPs). ChBetNPs were synthesized by ionic gelation method and characterized by DLS, UV, FTIR, SEM and zeta potential analysis. The encapsulation efficiency (EE) and in vitro release kinetics were analyzed using spectrophotometric technique for quantifying the encapsulated amount of betanin in ChBetNPs as a function of time. The antioxidant activity of ChBetNPs was analyzed by DPPH and H2O2 radical scavenging assays, anti-inflammatory activity by protein denaturation and human RBCs stabilization assays, and anti-acetylcholinesterase activity using standard protocol with minor modifications. Unloaded chitosan nanoparticles (CSNPs) were found to be sized at 161.4 ± 5.75 nm while an increase in the size to 270.3 ± 8.50 nm was noticed upon encapsulating betanin. EE of ChBetNPs was measured to be ∼87.5%. The IC50 of ChBetNPs depicted significant free radical scavenging activities as compared to CSNPs. Similarly, a strong anti-inflammatory activity of ChBetNPs was noted. Significant decrease in acetylcholinesterase activity by ChBetNPs was measured (IC50 0.5255 µg/mL vs. control 26.09 µg/mL). The vegetables coated with 3% ChBetNPs showed decreased weight loss as compared to uncoated control. ChBetNPs was shown to exhibit strong antioxidant, anti-inflammatory and anti-acetylcholinesterase activities thus making it a significant therapeutic agent for the management of Alzheimer's disease.

Sustainable finance, natural resource abundance, and energy poverty trap: the environmental challenges in the era of COVID-19.

Energy poverty is a global challenge and the scarcity has been emerging as a global issue. Therefore, the relationship among energy scarcity, sustainable finance, and carbon emissions is analyzed with the help of global data from 40 developing countries until the beginning of the COVID-19 era. For empirical results' estimation, the study analyzed a panel data ranging from 2000 to 2019. To measure the energy poverty, some part of population that has no access to energy is considered, and empirical analysis based on augmented mean group (AMG) regression method was carried out. The findings of the study suggest the inverse relation among energy poverty and carbon emissions. Moreover, a negative relationship was also observed between sustainable finance and carbon emissions. These findings highlight that alleviation of energy poverty can intensify environmental pollution. While improvement in access to clean energy will benefit society by alleviating energy poverty and controlling carbon emissions. Moreover, improvement in the share of sustainable finance in total investment may improve the environment quality by reducing carbon emissions. Therefore, it is suggested that regional plans along with sustainable finance are required on a priority basis for the promotion of clean energy to control carbon emissions and alleviate energy poverty at the household level.

Effectiveness of health education intervention on diabetes mellitus among the teachers working in public sector schools of Pakistan.

BACKGROUND: Diabetes Mellitus (DM) is considered as one of the major public health problems globally. Health education strategies can help in managing blood glucose level and complications among DM patients. Health education intervention is effective to manage and control the blood glucose levels among diabetic patients. This study explored the effectiveness of health education intervention on DM among school teachers in public sector schools of Pakistan. METHODS: This was quasi-experimental study where baseline & end line assessments were conducted on teachers of public sector schools of Sindh province, Pakistan, from October to December 2019. Pretested structured questionnaire was used in this study. Participants (n = 136). were randomly selected from the list of government schools registered with district education department An intervention comprised of health education sessions with DM patients was undertaken after conducting baseline assessment followed by end line assessment. The institutional review board of Health Services Academy Pakistan ethically approved this study. RESULTS: All the respondents completed post-test with mean ± SD age of participants being 39.2 ± 1.34 years. Female teachers comprised 65% out of which 70% were living in rural areas. Knowledge on DM pre-test score was 20.03 ± 3.31 that increased in post-test to 49.11 ± 2.21 (p < 0.05). Mean score of information on symptoms and causes of DM was 1.98 ± 0.21 for pre-test whereas for post-test it was 4.78 ± 0.12 (p < 0.05). The effect of intervention was significant on diabetes related complications (p < 0.05), symptoms (p < 0.05), overall score (p < 0.05) and preventive practices (p < 0.05). CONCLUSIONS: The study provides evidence of the importance and effectiveness of health education intervention related to diabetes among school teachers, which has a positive impact on the knowledge and practices. We concluded that the health education session sensitized the teachers and they can bring cogent changes to enhance their knowledge about diabetes and its risks.

Alleviation of cadmium stress in rice by inoculation of Bacillus cereus.

Heavy metal resistant bacteria are of great importance because they play a crucial role in bioremediation. In the present study, 11 bacterial strains isolated from industrial waste were screened under different concentrations of cadmium (Cd) (100 µM and 200 µM). Among 11 strains, the Cd tolerant Bacillus cereus (S6D1-105) strain was selected for in vitro and in vivo studies. B. cereus was able to solubilize potassium, and phosphate as well as produce protease and siderophores during plate essays. Moreover, we observed the response of hydroponically grown rice plants, inoculated with B. cereus which was able to promote plant growth, by increasing plant biomass, chlorophyll contents, relative water content, different antioxidant enzymatic activity such as catalase, superoxide dismutase, ascorbate peroxidase, polyphenol oxidase and phenylalanine ammonia-lyase and reducing malondialdehyde content in both roots and leaves of rice plants under Cd stress. Our results showed that the B. cereus can be used as a biofertilizer which might be beneficial for rice cultivation in Cd contaminated soils.

The Insight of In Silico and In Vitro evaluation of Beta vulgaris phytochemicals against Alzheimer's disease targeting acetylcholinesterase.

B. vulgaris extracts possess antioxidant, anti-inflammatory along with its role in improving memory disorders. Subsequently, in vitro and in silico studies of its purified phytochemicals may expand complementary and alternative Alzheimer's therapeutic option. Super activation of acetylcholinesterase enzyme is associated explicitly with Alzheimer's disease (AD) ultimately resulting in senile dementia. Hence, acetylcholinesterase enzyme inhibition is employed as a promising approach for AD treatment. Many FDA approved drugs are unable to cure the disease progression completely. The Present study was devised to explore the potential bioactive phytochemicals of B. vulgaris as alternative therapeutic agents against AD by conducting in vitro and in silico studies. To achieve this, chemical structures of phytochemicals were recruited from PubChem. Further, these compounds were analyzed for their binding affinities towards acetylcholinesterase (AChE) enzyme. Pharmacophoric ligand-based models showed major characteristics like, HBA, HBD, hydrophobicity, aromaticity and positively ionizable surface morphology for receptor binding. Virtual screening identified three hit compounds including betanin, myricetin and folic acid with least binding score compared to the reference drug, donepezil (-17 kcal/mol). Further, in vitro studies for anti-acetylcholinesterase activity of betanin and glycine betaine were performed. Dose response analysis showed 1.271 µM and 1.203 µM 50% inhibitory concentration (IC50) values for betanin and glycine betaine compounds respectively. Our findings indicate that phytoconstituents of B. vulgaris can be implicated as an alternative therapeutic drug candidate for cognitive disorders like Alzheimer's disease.

Phytochemical Screening and Protective Effects of Prunus persica Seeds Extract on Carbon Tetrachloride-Induced Hepatic Injury in Rats.

BACKGROUND AND PURPOSE: Carbon tetrachloride (CCl4) is a dynamic environmental toxin released from chemical factories and its concentration in the atmosphere is accelerating at an alarming proportion. The potential presence of CCl4 in the human body causes liver injury via free radical stimulated inflammatory responses. OBJECTIVES: In this study, protective effects of hydromethanolic seeds extract of Prunus persica (PPHM) were evaluated for free radical scavenging potential in CCl4 mediated acute liver toxicity in the murine model. EXPERIMENTAL APPROACH: Followed by acute oral toxicity analysis, liver cells of Sprague-Dawley (SD) rats were treated with CCl4 and subsequently, the chemoprophylactic effect of extract (400 mg/Kg dose) was evaluated using in vivo studies including, silymarin as the positive control. Biochemical parameters, staining (hematoxylin and eosin (H & E) and Masson's Trichome) and quantitative gene expression analysis via real-time PCR were used to evaluate hepatic damage control. RESULTS: The results illustrated that PPHM extract exhibit strong anti-oxidant activity, comparable to the positive control, gallic acid. Research study results also demonstrated that the extract treatment at 400 mg/Kg concentration is highly effective in protecting liver damage due to CCl4 exposure. Mechanistic investigations indicated that the therapeutic action of PPHM was correlated with the increase in Nrf2, NQO-1 and decrease in collagen III mRNA genes expression compared to CCl4 treated group. CONCLUSIONS AND IMPLICATIONS: Accordingly, our research study indicated that PPHM alleviated CCl4-mediated oxidative stress through Nrf2/NQO-1 pathway, thereby protecting liver damage against environmental toxins. Our findings provide supportive evidence to suggest PPHM as a novel nontoxic hepatoprotective agent.

Hepatitis C Diagnosis: Simplified Solutions, Predictive Barriers, and Future Promises.

The simplification of current hepatitis C diagnostic algorithms and the emergence of digital diagnostic devices will be very crucial to achieving the WHO's set goals of hepatitis C diagnosis (i.e., 90%) by 2030. From the last decade, hepatitis C diagnosis has been revolutionized by the advent and approval of state-of-the-art HCV diagnostic platforms which have been efficiently implemented in high-risk HCV populations in developed nations as well as in some low-to-middle income countries (LMICs) to identify millions of undiagnosed hepatitis C-infected individuals. Point-of-care (POC) rapid diagnostic tests (RDTs; POC-RDTs), RNA reflex testing, hepatitis C self-test assays, and dried blood spot (DBS) sample analysis have been proven their diagnostic worth in real-world clinical experiences both at centralized and decentralized diagnostic settings, in mass hepatitis C screening campaigns, and hard-to-reach aboriginal hepatitis C populations in remote areas. The present review article overviews the significance of current and emerging hepatitis C diagnostic packages to subvert the public health care burden of this 'silent epidemic' worldwide. We also highlight the challenges that remain to be met about the affordability, accessibility, and health system-related barriers to overcome while modulating the hepatitis C care cascade to adopt a 'test and treat' strategy for every hepatitis C-affected individual. We also elaborate some key measures and strategies in terms of policy and progress to be part of hepatitis C care plans to effectively link diagnosis to care cascade for rapid treatment uptake and, consequently, hepatitis C cure.

Sodium nitroprusside application improves morphological and physiological attributes of soybean (Glycine max L.) under salinity stress.

Salinity is among the major abiotic stresses negatively affecting the growth and productivity of crop plants. Sodium nitroprusside (SNP) -an external nitric oxide (NO) donor- has been found effective to impart salinity tolerance to plants. Soybean (Glycine max L.) is widely cultivated around the world; however, salinity stress hampers its growth and productivity. Therefore, the current study evaluated the role of SNP in improving morphological, physiological and biochemical attributes of soybean under salinity stress. Data relating to biomass, chlorophyll and malondialdehyde (MDA) contents, activities of various antioxidant enzymes, ion content and ultrastructural analysis were collected. The SNP application ameliorated the negative effects of salinity stress to significant extent by regulating antioxidant mechanism. Root and shoot length, fresh and dry weight, chlorophyll contents, activities of various antioxidant enzymes, i.e., catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) were improved with SNP application under salinity stress compared to control treatment. Similarly, plants treated with SNP observed less damage to cell organelles of roots and leaves under salinity stress. The results revealed pivotal functions of SNP in salinity tolerance of soybean, including cell wall repair, sequestration of sodium ion in the vacuole and maintenance of normal chloroplasts with no swelling of thylakoids. Minor distortions of cell membrane and large number of starch grains indicates an increase in the photosynthetic activity. Therefore, SNP can be used as a regulator to improve the salinity tolerance of soybean in salt affected soils.

Tectona grandis leaf extract ameliorates hepatic fibrosis: Modulation of TGF- ß /Smad signaling pathway and upregulating MMP3/TIMP1 ratio.

ETHNOBOTANICAL RELEVANCE: Tectona grandis L.f (or syn: Jatus grandis (L.f.) Kuntze Revis), from family Lamiaceae, also known as Teak, is widely recognized in ayurvedic system of medicine and confer curative potential against inflammation, liver disorders, biliousness, diabetes, bronchitis, leprosy and dysentery. Its leaves are rich source of edible food colorant and reported nontoxic for liver and various organs. AIM OF STUDY: Hepatic injury progression to liver cirrhosis and cancer is a serious health issue across the world. Currently, anti-fibrotic therapeutic options are limited and expensive with no FDA approved direct anti-hepato-fibrotic drug validated in clinic. Thus, the aim of this study was to understand ameliorative effect of Tectona grandis L.f, leaves in early liver fibrosis. METHOD AND RESULTS: C57BL/6 mice suffering from CCl4 induced liver injury, were orally administered at three different doses (50, 100 & 200 mg/kg) of Tectona grandis L.f, leaf extract, thrice a week, up to 4 and 8 weeks. Anti-fibrotic effect was evaluated through animal body/liver weight measurements, serological tests (AST, ALT, GSH, MDA and LDH assays), tissue hydroxyproline content, and histochemical analysis (H&E, Masson trichrome, Sirius red and αSMA localization). Moreover, transcriptional and post-transcriptional expression of fibrosis associated biomarkers and TGF-ß/Smad cascade were analyzed. It was observed that 100 mg/kg dose optimally downregulated TGF-ß1/Smad2 with upregulation of Smad7 and regulated αSMA, Col 1, PDGF, TIMP1 and MMP3 expression, post 8 weeks of treatment. In addition, MMP3/TIMP1 ratio was upregulated to 0.7, 2.5 and 1.7 fold at 50 mg/kg, 100 mg/kg & 200 mg/kg treatments respectively, in comparison to untreated liver fibrosis models. The extract contains gallic acid, caffeic acid, sinapinic acid and myricetin when analyzed through high performance liquid chromatography. CONCLUSION: Tectona grandis L.f, leaves have potential to ameliorate liver fibrosis induced by CCl4 in mice via modulation of TGF-ß1/Smad pathway and upregulated MMP3/TIMP1 ratio.

Biochemical Profiling and Elucidation of Biological Activities of Beta vulgaris L. Leaves and Roots Extracts.

RESEARCH BACKGROUND: Red beet (Beta vulgaris L.) is commercially grown in Punjab and Khyber Pakhtunkhwa (KPK) regions while cultivated as vegetable in Baluchistan and Sindh regions of Pakistan. It is well known for its health-promoting role in several clinical and pathologic outcome due to abundance of betalains and other bioactive compounds. The purpose of study was to interpret bioactivity of of B. vulgaris leaves (BvLM) and roots (BvRM) extracts for finding natural cure of neurodegenerative diseases (NDs). EXPERIMENTAL APPROACH: BvLM and BvRM extracts were evaluated for phytochemical composition, antioxidant, anti-inflammatory and anticholinesterase potential using standard protocols with modifications. RESULTS: Phytochemicals analysis of BvLM and BvRM extracts depicted the presence of flavonoids, phenols, alkaloids, saponins and glycosides. The IC50 values for free radical scavenging activity for BvRM and BvLM showed that BvLM (DPPH: 2.20 ± 0.72 µg/mL, H 2 O 2 : 0.0519 ± 1.02 µg/mL) is more significant as compared to BvRM (DPPH: 2.312 ± 0.72 µg/mL, H 2 O 2 : 2.668 ± 0.49 µg/mL). BvLM showed significant protection against heat induced hemolysis of HRBCs and protein denaturation (2.322 ± 0.1 > 2.324 ± 0.06 µg/mL) as compared to BvRM (8.572 ± 0.2 > 50.18 ± 1.0 µg/mL). Both extracts found to exhibit strong inhibitory potential against acetylcholinesterase enzyme. DISCUSSION/CONCLUSION: Our study highlighted B. vulagris extracts as rich and nutritious source of antioxidants, anti-inflammatory and cholinesterase inhibitors that could be helpful in preventing and treating neurodegenerative disorders (NDs). In consideration of multifactorial and complex etiology of NDs, BvLM and BvRM extracts would be apt candidates for development of therapeutic strategy for management of multiple cognitive disorders.

Phytochemical analysis of Berberis lyceum methanolic extract and its antiviral activity through the restoration of MAPK signaling pathway modulated by HCV NS5A

Objective: To evaluate the antiviral activity and phytochemicals of selected plant extracts and their effect on the mitogen-activated protein kinase (MAPK) signaling pathway modulated by hepatitis C virus (HCV) nonstructural protein 5A (NS5A). Methods: A total of ten plant extracts were initially screened for their toxicities against HepG2 cells. The non-toxic plants were tested for their inhibitory effect on the expression of HCV NS5A at both mRNA and protein levels using real-time PCR and Western blotting assays, respectively. The differential expression of the genes associated with MAPK pathway in the presence of NS5A gene and plant extract was measured through real-time PCR. Subsequently, the identification of secondary metabolites was carried out by phytochemical and HPLC analysis. Results: The phytochemical profiling of Berberis lyceum revealed the presence of alkaloids, phenols, saponins, tannins, flavonoids, carbohydrates, terpenoids, steroids, and glycosides. Similarly, quercetin, myricetin, gallic acid, caffeic acid, and ferulic acid were identified through HPLC analysis. The methanolic extract of Berberis lyceum strongly inhibited HCV RNA replication with an IC50 of 11.44 μg/mL. RT-PCR and Western blotting assays showed that the extract reduced the expression of HCV NS5A in a dose-dependent manner. Berberis lyceum extract also attenuated NS5A-induced dysregulation of the MAPK signaling pathway. Conclusions: Our findings suggest that Berberis lyceum extract strongly inhibits HCV propagation by reducing HCV NS5A-induced perturbation of MAPK signaling.

A 2-year retrospective study of viral and host-associated risk factors in Pakistani hepatocellular carcinoma patients.

OBJECTIVE: Persistent chronic hepatitis C (CHC) infection is associated strongly with serious complications such as hepatitis C virus-associated liver cirrhosis (HCV-LC) and hepatitis C virus-associated hepatocellular carcinoma (HCV-HCC). The aim of this study was to assess the distribution of hepatitis C virus (HCV) genotypes among HCV-positive patients and examine the potential associations between viral and host-associated factors with the risk of developing HCV-HCC. PATIENTS AND METHODS: HCV-positive patients (n = 300) were enrolled and divided into three groups: CHC (n = 171), HCV-LC (n = 51), and HCV-HCC (n = 78). RESULTS: HCV genotype 3a showed the highest prevalence among HCV-positive individuals (66% of patients), followed by genotype 1a (15% of patients). The proportion of individuals infected with mixed HCV genotypes was higher among HCV-HCC patients. Interestingly, there were a significantly higher proportion of women (54/78; 69.2%) among HCV-HCC patients compared with CHC patients (89/171 or 52%; χ = 6.47; P=1 × 10). Women with HCV had two-fold higher odds of developing HCV-HCC (odds ratio = 2.07, 95% confidence interval: 1.18-3.71). In comparison with CHC patients, significantly more HCV-HCC patients were 50 years of age or older (59/78 or 75.6% of HCV-HCC patients and 61/171 or 35.7% of CHC patients; χ = 34.27; P < 0.0001), suggesting that HCV-positive patients aged 50 years or older had an ~five-fold higher risk of developing HCV-HCC (odds ratio = 5.6, 95% confidence interval: 3.02-10.01). CONCLUSION: In summary, HCV genotype 3a had the highest prevalence in the studied HCV-positive population, and women and older patients were at a higher risk of developing HCV-LC and HCV-HCC following CHC infections.

Anti-hepatitis C virus activity and synergistic effect of Nymphaea alba extracts and bioactive constituents in liver infected cells.

BACKGROUND: Without an effective vaccine, hepatitis C virus (HCV) remains a global threat, inflicting 170-300 million carriers worldwide at risk of cirrhosis and hepatocellular carcinoma (HCC). Though various direct acting antivirals have been redeemed the hepatitis C treatment, a few restraints persist including possible side effects, viral resistance emergence, excessive cost which restricts its availability to a common person. HYPOTHESIS: There is no preventive HCV vaccine available today so the discovery of potent antiviral natural flora and their bioactive constituents may help to develop preventive cures against HCV infection. STUDY DESIGN: In current study, we aim to clarify anti-HCV activity of methanol and acetone extracts along with the purified fractions of Pakistani local plant, Nymphaea alba L (N. alba) using Huh-7 cell line as transfection model. Synergistic study of purified fractions with interferon was performed using MDBK cell line (expressing interferon receptors) as transfection model. MATERIALS AND METHODS: Recent study by our research group has observed potent anti-HCV NS3 protease activity of methanol and acetone extracts of N. alba. Effect of N. alba extracts, its fractions precisely, the N1 and N8 fractions on HCV replication was demonstrated by analyzing viral gene expression using in vitro transfection model. Considering NS3 protease as a dynamic drug target, fourteen phytochemicals of N. alba were selected as ligands for interaction with NS3 protein using Molecular Operating Environment (MOE) software. Boceprevir, FDA approved NS3 protease inhibitor, was used as standard for comparative study in docking screening. RESULTS: Herein we report 84% and 94% reduction of 3a genotype of HCV NS3/4A gene expression at mRNA level at non-toxic concentration. Specifically, two fractions 'N1' & 'N8' isolated from acetone extract suppressed HCV NS3 gene expression in transfected target cells with an EC50 value of 37 ±â€¯0.03 µg/ml and 20 ±â€¯0.02 µg/ml respectively. Similarly, viral genotype 1a replication is strongly suppressed in target cells by N. alba flower extracts and purified fractions. Moreover, combination of fractions with standard antiviral drug displayed synergistic effects for inhibition of HCV replication. Phytochemicals including Isoquercetin, Hyperoside, Quercetin, Reynoutrin, Apigenin and Isokaempferide displayed minimum binding energies as compared to standard protease inhibitor. CONCLUSION: N. alba and its purified phytochemicals with new scaffolds might significantly serve as valuable and alternative regimen against HCV either alone or in combination with other potential anti-HCV agents.

Phytochemical profiling and antiviral activity of Ajuga bracteosa, Ajuga parviflora, Berberis lycium and Citrus lemon against Hepatitis C Virus.

Hepatitis C is a serious health issue and cause liver disorders in millions of people. Available therapeutic agents require long term administration with numerous side effects. Therefore, there is a dire need to find alternative treatment options for this disease. Since ancient times, medicinal plants are widely used to cure various diseases with no or less harmful effects. Therefore, this study was designed to find out phytochemicals and investigate antiviral activity of methanol extract of Ajuga bracteosa, Ajuga parviflora, Berberis lycium and Citrus lemon against Hepatitis C Virus (HCV infection). Phytochemical analysis of the plant extract was performed using various chemical tests. Toxicity of the plant extract was determined against using trypan blue exclusion method. Antiviral activity of the selected plant extract was find out against HCV infected HepG2 cells. For this purpose, HepG2 cells were seeded with HCV positive and negative serum and nontoxic doses of plant extract for 24 and 48 h. After this RNA was extracted and viral load was determined using Real-time PCR. Phytochemical analysis showed the presence of flavonoids and phenols in all plant extracts while amino acids, alkaloids and tannins were present in B. lycium and saponins were detected in C. lemon. Toxicity assay showed that all plant extracts were nontoxic at maximum concentration of 200 µg/ml except B. lycium, which showed mild toxicity at 40 µg/ml and were extremely toxic at 60 µg/ml and above doses. Real-time PCR quantitation result revealed that after 24 h treatments A. parviflora showed highest antiviral activity, followed by A. bracteosa, while B. lycium extract had low (35%) and C. lemon has no antiviral effects. The 48 h treatments showed an increase antiviral activity by A. bracteosa followed by A. parviflora and B. lycium while C. lemon showed negative effect. Our results depicted that mentioned plants might be used as an alternative therapeutic regime or in combination with existing treatments against HCV.

Comparison of Anti-HCV Activity of Multiple Punica granatum Extracts and Fractions in Virus-infected Human Hepatocytes.

BACKGROUND: Plants extracts and their bioactive constituents can provide an alternative approach for new treatment. Pakistani flora reveals a huge, largely untapped source of potential antiviral constituents. OBJECTIVE: High-cost concerns of direct-acting anti-HCV drugs limit their employment specifically in developing countries like Pakistan. Therefore, discovery of inexpensive and nontoxic agents is needed for HCV treatment. METHODOLOGY: In this study, we used plasmid constructs of pCR3.1/FLAGtag/HCV NS3/4A (genotype 1a & 3a) and Punica granatum extracts (PK AV 003) and semi-purified fractions (P1-P11) were evaluated for their anti-HCV activity. Acetone extract along with two fractions (P4 & P11) revealed a useful therapeutic index. RESULTS: The fractions P4 (IC50=28.5±0.02 µg/ml; IC25=16±0.02 µg/ml) and P11 (IC50=72±0.02 µg/ml; IC25=41±0.03 µg/ml) dramatically suppressed HCV replication as measured by quantitative PCR (qPCR) and HCV NS3 protein expression level in transient transfection model. Consistent with suppression in genome replication, inhibition of HCV infectious particles by PK AV 003 extract was also judged in an infectious model system. This data is the first description of Pakistani indigenous cultivated fruit-producing plant, Punica granatum, possessing anti-HCV activity. Further analyses are being performed for investigating the mechanistic studies and structural characterization of purified fractions of PK AV 003. CONCLUSION: Our findings suggest that PK AV 003 fruit extract might be useful as an add-on therapeutic candidate for treating HCV infection.

Oxidative toxicity in diabetes and Alzheimer's disease: mechanisms behind ROS/ RNS generation.

Reactive oxidative species (ROS) toxicity remains an undisputed cause and link between Alzheimer's disease (AD) and Type-2 Diabetes Mellitus (T2DM). Patients with both AD and T2DM have damaged, oxidized DNA, RNA, protein and lipid products that can be used as possible disease progression markers. Although the oxidative stress has been anticipated as a main cause in promoting both AD and T2DM, multiple pathways could be involved in ROS production. The focus of this review is to summarize the mechanisms involved in ROS production and their possible association with AD and T2DM pathogenesis and progression. We have also highlighted the role of current treatments that can be linked with reduced oxidative stress and damage in AD and T2DM.

Therapeutic potential of Taraxacum officinale against HCV NS5B polymerase: In-vitro and In silico study.

Discovery of alternative and complementary regimens for HCV infection treatment is a need of time from clinical as well as economical point of views. Low cost of bioactive natural compounds production, high biochemical diversity and inexistent/milder side effects contribute to new therapies. Aim of this study is to clarify anti-HCV role of Taraxacum officinale, a natural habitat plant rich of flavonoids. In this study, methanol extract of T. officinale leaves was initially analyzed for its cytotoxic activity in human hepatoma (Huh-7) and CHO cell lines. Hepatoma cells were transfected with pCR3.1/Flagtag/HCV NS5B gene cloned vector (genotype 1a) along with T. officinale extract. Considering NS5B polymerase as potential therapeutic drug target, twelve phytochemicals of T. officinale were selected as ligands for molecular interaction with NS5B protein using Molecular Operating Environment (MOE) software. Sofosbuvir (Sovaldi: brand name) currently approved as new anti-HCV drug, was used as standard in current study for comparative analysis in computational docking screening. HCV NS5B polymerase as name indicates plays key role in viral genome replication. On the basis of which NS5B gene is targeted for determining antiviral role of T. officinale extract and 65% inhibition of NS5B expression was documented at nontoxic dose concentration (200µg/ml) using Real-time PCR. In addition, 57% inhibition of HCV replication was recorded when incubating Huh-7 cells with high titer serum of HCV infected patients along with leaves extract. Phytochemicals for instance d-glucopyranoside (-31.212 Kcal/mol), Quercetin (-29.222 Kcal/mol), Luteolin (-26.941 Kcal/mol) and some others displayed least binding energies as compared to standard drug Sofosbuvir (-21.0746 Kcal/mol). Results of our study strongly revealed that T. officinale leaves extract potentially blocked the viral replication and NS5B gene expression without posing any toxic effect on normal fibroblast cells of body.

Antiviral drugs against hepatitis C virus.

Hepatitis C virus (HCV) infection is a major worldwide problem causes acute and chronic HCV infection. Current treatment of HCV includes pegylated interferon-α (PEG IFN- α) plus ribavirin (RBV) which has significant side effects depending upon the type of genotype. Currently, there is a need to develop antiviral agents, both from synthetic chemistry and Herbal sources. In the last decade, various novel HCV replication, helicase and entry inhibitors have been synthesized and some of which have been entered in different phases of clinical trials. Successful results have been acquired by executing combinational therapy of compounds with standard regime in different HCV replicons. Even though, diverse groups of compounds have been described as antiviral targets against HCV via Specifically Targeted Antiviral Therapy for hepatitis C (STAT-C) approach (in which compounds are designed to directly block HCV or host proteins concerned in HCV replication), still there is a need to improve the properties of existing antiviral compounds. In this review, we sum up potent antiviral compounds against entry, unwinding and replication of HCV and discussed their activity in combination with standard therapy. Conclusively, further innovative research on chemical compounds will lead to consistent standard therapy with fewer side effects.