The present study examined the food value, techno-functional properties, and potential metabolites of Agave sisalana Perrine (Ram kandmool, RKM) flour. The proximate analysis revealed that RKM flour has a moisture content (10.74 %), fat content (1.12 %), protein content (5.42 %), and crude fiber content (11.35 %). The total carbohydrate content was 15.11 %, with an energy value of 385.76 kJ 100 g-1, and total sugar content of 36.61 mg glucose g-1. Additionally, the high total phenolic content (310.79 mg GAE g-1) and total flavonoid content (51.32 mg QE g-1) highlight its significant antioxidant properties, as evidenced by DPPH* and ABTS scavenging activities of 71.22 % and 93.68 %, respectively. Functional properties such as solubility index increase with temperature, indicating good solubility in hot food preparations, while the swelling ratio peaks at 70 °C, making it a valuable thickening agent. FTIR, SEM, LCMS, and amino acid analysis revealed a complex chemical composition, high surface area due to the rough, porous surface of RKM fibers, and significant health-benefiting compounds, respectively. Overall, RKM flour's diverse nutritional and functional properties, high mineral content, and significant antioxidant properties make it a valuable ingredient for food formulations aimed at improving health outcomes.

The present study aims to characterize the nutritional and potential applications of Dioscorea esculenta (Suthni) flour (SF) in comparison to potato flour (PF). Results revealed similarities in mineral content and crystalline structures, while the higher fat content in SF presents potential advantages in mouthfeel and flavor. SF exhibits a notably shorter peak viscosity (506.6 m.Pa) compared to PF (616 m.Pa), indicative of its rapid gelatinization kinetics and making it advantageous in rapid thickening products such as instant sauces or soups. The irregular and agglomerated structure of SF, compared to the uniform oval shape of PF, was revealed by SEM. Similar functional groups were observed in both tubers, as revealed by FTIR. Amino acid profiles highlight distinctions in protein quality, with SF showing higher a glutamic acid content and limited sulfur-containing amino acids and aromatic amino acids. The results indicated that a combination of 60

This study compares Arenga obtusifolia (tassey) flour with potato flour, focusing on nutritional, techno-functional, and metabolite profiles. Tassey flour has higher fat (2.50 %), fiber (7.11 %), and carbohydrate (75.58 %) contents, while potato flour offers more protein (21.33 %) and ash (3.44 %), making it a better protein source. Tassey flour exhibits strong nutraceutical potential, with a total phenolic content of 439.58 mg GAE g- 1 and flavonoid content of 16.60 mg QE g-1, resulting in high antioxidant activity (ABTS inhibition: 79.56 %, DPPH: 72.23 %) than potato flour. Tassey's peak viscosity (1092.3 mPa s) and pasting temperature (70.76 degrees C) suit it for baked goods requiring moisture retention, while potato flour's higher amylose (13.64 %) and starch content (64.49 %) lend structural benefits. Tassey flour also excels in water absorption (40.59 %) and oil holding capacity (12.20 %) compared to potato flour, making it ideal for snacks and bakery applications needing enhanced moisture. FTIR spectrum reveals the presence of various functional compounds, while SEM images show that the flour granules were smooth, uniform, and ranged from 10 to 50 mu m. Amino acid analysis shows valuable sulfurcontaining and aromatic amino acids, though potato flour has higher in-vitro protein digestibility (79.03 %). HRLCMS analysis further reveals a diverse array of nutraceutical compounds promoting metabolic health and reducing oxidative stress. Sensory testing of cookies reveals strong acceptability for tassey-enhanced variants, particularly Variant D. Tassey's unique attributes make it an excellent alternative to potato flour in healthfocused and nutrient-enriched food products.

This review delves into the utilization of non-conventional starch sources through a specific cultural lens, focusing on the practices and significance of Arenga obtusifolia Griff., commonly referred to as the sugar palm, and its derivative product called Tassey. The tassey is widely popular in the Nyishi tribe of Arunachal Pradesh, India, a region renowned for its biodiversity and abundant palm tree varieties, including the sugar palm. Highlighting the multifaceted dimensions of the sugar palm, this review expounds upon Tassey's creation, consumption, and versatile applications. Tassey, a substance extracted from the sugar palm's stem, emerges as a pivotal element of the Nyishi tribe's cultural heritage and sustenance strategies, especially during times of scarcity and natural calamities. This review sheds light on the integral role played by Nyishi women in the cultivation and preparation of tassey, underscoring their vital contribution to upholding this time-honoured practice. Furthermore, the economic ramifications of tassey production and its commercial viability are explored, elucidating its role in bolstering the tribe's livelihood. Extending beyond its dietary role, tassey exhibits a diverse array of applications, spanning from human consumption to its incorporation in the crafting of traditional alcoholic beverages and even as a therapeutic resource for livestock. By offering insights into the profound relationship between the Nyishi tribe and the sugar palm, this review enriches our comprehension of traditional ecological wisdom and sustainable exploitation of resources within a culturally vibrant backdrop. Ultimately, this comprehensive exploration contributes to a nuanced appreciation of the intricate interplay between heritage, environment, and innovation in this distinctive region.

Cereals provide nutrients such as lipids, carbs, proteins, minerals, and vitamins and make up the majority of human nutrition. Some cereals also contain a significant quantity of dietary fiber (soluble and insoluble). Cereals, when consumed as a whole or in coloured varieties, are the source of bioactive components with functional properties. Dietary fiber in whole grains has a unique blend of bioactive components such as resistant starch, vitamins, minerals, phytochemicals, and antioxidants that provide different health benefits. Traditionally, whole grains were consumed, but currently, most foods are derived from refined fractions of cereal and pulse crops. Minimal processing of cereals increases both nutrient bioavailability and health benefits. The bioactive compounds present in whole cereals ultimately fight against diseases and prevent or control some diseases in the body, such as cardiovascular risk, reducing the risk of cancer, type-2 diabetes, hypertension, and high blood pressure, improving gastrointestinal health, etc. The consumer demands minimally processed foods, which are essential for maintaining health and improving metabolism. Therefore, developing products using whole cereals or millet to improve the health and safety of humans is of great interest. The present review discusses the nutraceutical potential of cereals and their products, their impact on gut microbiota, and consumer acceptability. The potential for future research in the processing of such healthy cereals in combination with other pseudocereals is identified and discussed.

The uptake or expression of hepatitis B virus (HBV) proteins by dendritic cells (DCs) is considered important for disease outcome. Differential expression of microRNA (miRNA) may have a role in viral persistence and hepatocellular injury. The miRNA expression was investigated by microarray in DCs from different stages of HBV infection and liver disease namely, immune active (IA; n = 20); low replicative (LR; n = 20); HBeAg negative (n = 20); acute viral hepatitis (AVH, n = 20) and healthy controls (n = 20). miRNA levels were analyzed by unsupervised hierarchical clustering and principal component analyses and validated by quantitative polymerase Chain Reaction (qPCR). The miRNA-messenger RNA (mRNA)regulatory networks identified 19 miRNAs and 12 target gene interactions in major histocompatibility complex and other immune pathways. miR-2278, miR-615-3p, and miR-3681-3p were downregulated in the IA group compared to healthy control, miR-152-3p and miR-3613-3p in the LR group compared to IA group and miR-152-3p and miR-503-3p in HBe negative compared to LR group. However, miR-7-1-1-3p, miR-192-5p, miR-195-5p, and miR-32-5p in LR, miR-342-3p, and miR-940 in HBe negative, and miR-34a-5p, miR-130b-3p, miR-221-3p, miR-320a, miR-324-5p, and miR-484 in AVH were upregulated. Further, qPCR confirmed changes in miRNA levels and their target genes associated with antigen processing and presentation. Thus, a deregulated network of miRNAs-mRNAs in DCs seems responsible for an impaired immune response during HBV pathogenesis.

Foamy viruses (FVs) or spumaviruses are retroviruses that are explored as vectors for gene therapy. The good feature of foamy viruses is its broad tropism; however, their infections result in non-targeted gene expression. Here, we attempted to design the liver targeted viral gene delivery by employing liver specific gene promoters like albumin (ALB), transthyretin (TTR) and hepatitis B virus (HBV) promoters. We compared the relative gene expression of liver specific promoters versus the U3 promoter in liver cell line (HepG2) and non-liver cell lines: human fibrosarcoma cell line (HT1080), baby hamster kidney cell line (BHK), human embryonic kidney cell line (HEK 293T) and cervical cancer cell line (HeLa). We have found that the promoter exchange didn't affect viral assembly. The ability to drive gene expression was best with TTR promoter which was followed by HBV and ALB promoter. The use of TTR, HBV and ALB promoters are helpful in achieving liver specific gene expression.

Hepatocellular carcinoma (HCC) is an aggressive tumor with limited systemic and locoregional modalities of treatment. Although microRNA (miRNA) based therapies have significant potential, their targeted delivery remains a major challenge. miR-199a-3p functions as an important tumor suppressor in HCC, which regulates various cellular processes. Recently, peptide-based nanoparticles (NPs) have been developed to deliver oligonucleotides including miRNA. Here, we describe the synthesis and characterization of arginine ,-dehydrophenylalanine (RF) nanoparticles for the selective delivery of miR-199a-3p to restore dysregulated gene expression in HCC. Targeted delivery was achieved by conjugating lactobionic acid (LA) with RF NPs (RF-LA NPs), a ligand for the asialoglycoprotein receptor known to be overexpressed in HCC cell lines. RF-LA NPs condensed miR-199a-3p had an average size of approximate to 60nm and a zeta potential of approximate to+2.54 mV. RF-LA/miR NPs were found to be stable in serum as well as against RNase attack. RF-LA/miR NPs showed an enhanced cellular uptake and an efficient delivery of miR-199a-3p leading to a significant increase in miR-199a-3p levels (over 500 fold). The increased miR-199a-3p levels remarkably suppressed cell proliferation and migration as well as induced cellular apoptosis and downregulation of the specific target gene (mTOR) in vitro. RF-LA/miR NPs showed high tumor/ low organ ratios after intravenous injection into HCC tumor bearing nude mice. RF-LA/miR NPs treated mice demonstrated>50% decline in tumor growth, which also corresponded well with suppression of mTOR protein expression, tumor cell proliferation and increased survival rate (P<0.05). Conclusion: RF-LA/miR NPs showed significantly enhanced delivery of the miRNA which underscores their potential for further development as a therapeutic approach for HCC. (Hepatology 2018;67:1392-1407)

Hepatitis B virus (HBV) can manipulate the microRNA (miRNA) regulatory networks in infected cells to create a permissive environment for viral replication, cellular injury, disease onset, and its progression. The aim of the present study was to understand the miRNA networks and their target genes in the liver of hepatitis B patients involved in HBV replication, liver injury, and liver fibrosis. We investigated differentially expressed miRNAs by microarray in liver biopsy samples from different stages of HBV infection and liver disease (immune‐tolerant [n = 8], acute viral hepatitis [n = 8], no fibrosis [n = 16], early [F1+F2, n = 19] or late [F3+F4, n = 14] fibrosis, and healthy controls [n = 7]). miRNA expression levels were analyzed by unsupervised principal component analysis and hierarchical clustering. Analysis of miRNA–mRNA regulatory networks identified 17 miRNAs and 18 target gene interactions with four distinct nodes, each representing a stage‐specific gene regulation during disease progression. The immune‐tolerant group showed elevated miR‐199a‐5p, miR‐221‐3p, and Let‐7a‐3p levels, which could target genes involved in innate immune response and viral replication. In the acute viral hepatitis group, miR‐125b‐5p and miR‐3613‐3p were up, whereas miR‐940 was down, which might affect cell proliferation through the signal transducer and activator of transcription 3 pathway. In early fibrosis, miR‐34b‐3p, miR‐1224‐3p, and miR‐1227‐3p were up, while miR‐499a‐5p was down, which together possibly mediate chronic inflammation. In advanced fibrosis, miR‐1, miR‐10b‐5p, miR‐96‐5p, miR‐133b, and miR‐671‐5p were up, while miR‐20b‐5p and miR‐455‐3p were down, possibly allowing chronic disease progression. Interestingly, only 8 of 17 liver‐specific miRNAs exhibited a similar expression pattern in patient sera. Conclusion: miRNA signatures identified in this study corroborate previous findings and provide fresh insight into the understanding of HBV‐associated liver diseases which may be helpful in developing early‐stage disease diagnostics and targeted therapeutics. (Hepatology 2018;67:1695‐1709)

1 Department of Molecular and cellular medicine, Institute of Liver & Biliary Sciences, New Delhi, India, 2 International centre for genetic engineering and biotechnology, New Delhi, India, 3 Institute of Nano Science and Technology, Mohali, India 4 Department of Pathology Institute of Liver & Biliary Sciences, New Delhi, India, 5 Amity Institute of Molecular Medicine and Stem Cell Research, AUUP, Noida, India. 6 Department of Hepatology, Institute of Liver & Biliary Sciences, New Delhi, India *Shared Corresponding Authors: Virander Singh Chauhan, International Centre for Genetic Engineering & Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India Telephone: +91-11-26742316, Ex-480 Fax: +91-11-26123504 Email: viranderschauhan@gmail.com or Shiv Kumar Sarin, Institute of Liver & Biliary Sciences (ILBS), D-1 Vasant Kunj, New Delhi 10070, India Telephone: +91-11-46300000;Fax: +91-11-26123504; Email:shivsarin@gmail.com

Persistent or chronic infection with the hepatitis B virus (HBV) represents one of the most common viral diseases in humans. The hepatitis B virus deploys the hepatitis B virus X protein (HBx) as a suppressor of host defenses consisting of RNAi-based silencing of viral genes. Because of its critical role in countering host defenses, HBx represents an attractive target for antiviral drugs. Here, we developed and optimized a loss-of-function screening procedure, which identified a potential pharmacophore that abrogated HBx RNAi suppression activity. In a survey of 14,400 compounds in the Maybridge Screening Collection, we prioritized candidate compounds via high-throughput screening based on reversal of green fluorescent protein (GFP)-reported, RNAi-mediated silencing in a HepG2/GFP-shRNA RNAi sensor line. The screening yielded a pharmacologically active compound, N-(2,4-difluorophenyl)-N-[3-(1H-imidazol-1-yl) propyl] thiourea (IR415), which blocked HBx-mediated RNAi suppression indicated by the GFP reporter assay. We also found that IR415 reversed the inhibitory effect of HBx protein on activity of the Dicer endoribonuclease. We further confirmed the results of the primary screen in IR415-treated, HBV-infected HepG2 cells, which exhibited a marked depletion of HBV core protein synthesis and down-regulation of pre-genomic HBV RNA. Using a molecular interaction analysis system, we confirmed that IR415 selectively targets HBx in a concentration-dependent manner. The screening assay presented here allows rapid and improved detection of small-molecule inhibitors of HBx and related viral proteins. The assay may therefore potentiate the development of next-generation RNAi pathway-based therapeutics and promises to accelerate our search for novel and effective drugs in antiviral research.

Antiviral therapy for chronic hepatitis B (CHB) is often required for prolonged periods. We investigated the instance of one HBV genotype switching to another during tenofovir therapy. Of the 67 patients, genotype A was present in 6 (8.9%), D in 43 (65.6%), C in 1 (1.5%), and mixed in 17 (23.8%) patients. Genotype changes were detected in 51 (76.1%) patients on therapy during a follow-up of 192 (range 52-312) weeks. Inter-genotype changes were seen in 17 (33.3%) and intra-genotype in 28 (55%) and both inter-and intra-genotype in 6 of 51 (11.7%) patients. The distribution of genotypes in patients achieving complete virological response was genotype D, 32/43 (74.4%); genotype A, 6/6 (100%); and mixed genotypes, 13/17 (76.47%). The cumulative time of genotype switch among genotype A was 12 months (range 6-18), in genotype D, 12 months (range 6-48), and mixed genotype, 18 months (range 6-24). The type of intergenotype switch most frequently detected among genotype A1 was from A1 to D1 5/6 (83.3%), followed by mixed to genotype D3 7/13 (54%) and among intra-genotype changes, from D1 to D3 in 14/20 (70%). Pretreatment HBV genotype was the only factor predicting inter-genotype switches with genotype A or mixed genotypes more likely to undergo intergenotype switches as compared to genotype D patients (OR 66.6 [13.6-327.0, P<0.001]). Compared to genotype D, genotype A, and mixed genotypes are more inclined to switch while on tenofovir therapy. Genotypes tend to switch and select to a particular type possibly due to constant antiviral drug pressure. (C) 2016 Wiley Periodicals, Inc.

P0277 ENHANCED AND EFFECTIVE DELIVERY OF MICRORNA BY USING LIGAND MODIFIED SELF ASSEMBLED CATIONIC DIPEPTIDE NANOPARTICLES IN HEPATOCELLULAR CARCINOMA (HCC) A. Varshney, J.J. Panda, A.K. Singh, S.B. Rooge, S. Biswas, S.K. Sarin, V.S. Chauhan. Department of Research, Institute of Liver & Biliary Sciences, New Delhi, Institute of Nano Science and Technology, Mohali, Department of Hepatology & Research, Institute of Liver & Biliary Sciences, International Centre for Genetic Engineering and Biotechnology, New Delhi, India E-mail: virander@icgeb.res.in

The host-mediated RNAi pathways restrict replication of viruses in plant, invertebrate and vertebrate systems. However, comparatively little is known about the interplay between RNAi and various viral infections in mammalian hosts. We show in the present study that the siRNA-mediated silencing of Drosha, Dicer and Ago2 [argonaute RISC (RNA-induced silencing complex) catalytic component 2] transcripts in Huh7 cells resulted in elevated levels of HBV (hepatitis B virus)-specific RNAs and, conversely, we observed a decrease in mRNA and protein levels of same RNAi components in HepG2 cells infected with HBV. Similar reductions were also detectable in CHB (chronic hepatitis B) patients. Analysis of CHB liver biopsy samples, with high serum HBV DNA load (>log108 IU/ml), revealed a reduced mRNA and protein levels of Drosha, Dicer and Ago2. The low expression levels of key RNAi pathway components in CHB patient samples as well as hepatic cells established a link between HBV replication and RNAi components. The HBV proteins were also examined for RSS (RNA-silencing suppressor) properties. Using GFP-based reversion of silencing assays, in the present study we found that HBx is an RSS protein. Through a series of deletions and substitution mutants, we found that the full-length HBx protein is required for optimum RSS activity. The in vitro dicing assays revealed that the HBx protein inhibited the human Dicer-mediated processing of dsRNAs into siRNAs. Together, our results suggest that the HBx protein might function as RSS to manipulate host RNAi defence, in particular by abrogating the function of Dicer. The present study may have implications in the development of newer strategies to combat HBV infection.

BACKGROUND:Gene therapy is most effective when delivery is both efficient and safe. However, it has often proven difficult to find a balance between efficiency and safety in case of viral or polymeric vectors for gene therapy. Peptide based delivery systems may be attractive alternatives but their relative instability to proteolysis is a major concern in realizing their potential application in biomedical sciences. In this work we report gene delivery potential of nanoparticles (Nps) synthesized from cationic dipeptides containing a non-protein amino acid α, β-dehydrophenylalanine (∆Phe) residue.METHODS:Dipeptides were synthesized using solution phase peptide synthesis method. Nps were formed using self-assembly. Nps were characterized using light scattering, electron microscopy. Transfection efficiency was tested in hepatocellular carcinoma (HuH 7) cells.RESULTS:The cationic dipeptides condensed plasmid DNA into discrete vesicular nanostructures. Dipeptide Nps are non-cytotoxic, protected the condensed DNAs from enzymatic degradation and ferried them successfully inside different types of cells. GFP encoding plasmid DNA loaded dipeptide Nps showed positive transfection and gene expression in HuH 7 cells.CONCLUSIONS:The cationic dipeptide Nps can successfully deliver DNA without exerting any cytotoxic effect. Owing to their simple dipeptide origin, ease of synthesis, enhanced enzymatic stability as well unmatched biocompatibility, these could be successfully developed as vehicles for effective gene therapy.