Full-length sequence analysis of hepatitis C virus genotype 3b strains and development of an in vivo infectious 3b cDNA clone.

IMPORTANCE: HCV genotype 3b is a difficult-to-treat subtype, associated with accelerated progression of liver disease and resistance to antivirals. Moreover, its prevalence has significantly increased among persons who inject drugs posing a serious risk of transmission in the general population. Thus, more genetic information and antiviral testing systems are required to develop novel therapeutic options for this genotype 3 subtype. We determined the complete genomic sequence and complexity of three genotype 3b isolates, which will be beneficial to study its biology and evolution. Furthermore, we developed a full-length in vivo infectious cDNA clone of genotype 3b and showed its robustness and genetic stability in human-liver chimeric mice. This is, to our knowledge the first reported infectious cDNA clone of HCV genotype 3b and will provide a valuable tool to evaluate antivirals and neutralizing antibodies in vivo, as well as in the development of infectious cell culture systems required for further research.

Emerging Anti-Inflammatory Attributes of Curcumin: A Novel Paradigm and Ameliorative Attributes for the Treatment of Osteoarthritis.

Strong oxidant, curcumin, is diferuloyl methane; a member of the class of phenols known as curcuminoids that give Indian medicinal plants their characteristic turmeric-yellow hue. Over 5000 years ago, curcumin was first employed in the traditional Indian medical system. A growing amount of investigation reveals that curcumin has several pharmacological characteristics, including anticancer, hepatoprotective, anti-inflammatory, antioxidant, and antibacterial properties. Clinical tests revealed no harm, but some participants did have moderate nausea or diarrhea. A degenerative joint condition known as osteoarthritis affects elderly populations all over the world. It has a chronic inflammatory etiology, which contributes to chondrocyte loss that results in a thinner surface of cartilage at the affected joints. Curcumin has been scientifically demonstrated to exhibit medicinal benefits for osteoarthritis (OA), and further research is being conducted on the numerous ways through which it suppresses inflammation and slows the progression of ailments. Clinical and preclinical studies suggest the potential efficacy of curcumin in managing osteoarthritis, warranting further investigation. With emphasis on the mechanisms associated, this review seeks to completely outline the clinical effectiveness of curcumin using data from a variety of scientific studies that have been published so far.

Signaling pathways and molecular process of natural polyphenols in the amelioration of inflammatory bowel disease: A privileged scaffold in new drug discovery.

BACKGROUND: GIT is seriously affected by inflammatory bowel disease (IBD), which is characterized by extreme inflammation and an imbalance in a person's healthy life span. The frequency of occurrence of such chronic diseases as IBD would continue to increase. In the past decade, increasing attention has been paid to polyphenols from natural sources have been shown to serve as successful therapeutic agents for altering the signalling pathways linked to IBD and oxidative stress. METHODS: We conducted a structured search for peer-reviewed research articles using the various keywords in bibliographic databases. By using common tools and a deductive qualitative content analysis technique, the quality of the retrieved papers and the distinctive findings of the articles included in the study were evaluated. RESULTS: Notably, experimental and clinical evidence has proved that natural polyphenols could act as a targeted modulator to play a key role in the prevention or treatment of IBD. Polyphenol phytochemicals have shown noticeable alleviative effects by acting on the TLR/NLR, and NF-κB signaling pathway in intestinal inflammation. CONCLUSION: This study examines the potential of polyphenols for treating IBD, with an emphasis on modulating cellular signalling mechanisms, regulating the balance of gut microbiota, and restoring the epithelial barrier. The available evidence concluded that the utilization of polyphenol-rich sources could control inflammation, mucosal healing, and positive benefits with minimal side effects. Even though additional study is required in this area, particularly that which focuses on the interactions, connections, and precise mechanisms of action linking polyphenols and IBD.

Oxazoline/amide derivatives against M. tuberculosis: experimental, biological and computational investigations.

Tuberculosis (TB) is a treatable contagious disease that continuously kills approximately 2 million people yearly. Different oxazoline/amide derivatives were synthesized, and their anti-tuberculosis activity was performed against different strains of Mtb. This study designed the anti-Mtb compounds based on amide and oxazoline, two different structural moieties. The compounds were further synthesized and characterized by spectral techniques. Their anti-Tb activity was evaluated against strain (M. tuberculosis: H37Rv). Selectivity and binding affinity of all synthesized compounds (2a-2e, 3a-3e) against PanK in Mtb were investigated through molecular docking. Molecular dynamics simulation studies for the promising compounds 2d and 3e were performed for 100 ns. The stability of these complexes was assessed by calculating the root mean square deviation, solvent-accessible surface area, and gyration radius relative to their parent structures. Additionally, free energy of binding calculations were performed. Among all synthesized compounds, 2d and 3e had comparable antitubercular activity against standard drug, validated by their computational and biological study.

Novel hepatitis B virus reverse transcriptase mutations in patients with sustained viremia despite long-term tenofovir treatment.

BACKGROUND: Chronic hepatitis B virus (HBV) treatment consists of nucleos(t)ide analogues to suppress viral replication. The HBV inhibitor tenofovir has a high barrier to resistance, however, evidence of virus-escape is emerging. This study investigates HBV evolution in patients undergoing tenofovir treatment with the primary aim to assess the emergence of putative resistance mutations. METHODS: HBV DNA was extracted from blood samples of two patients with HBeAg-positive chronic HBV infection and persistent viremia despite tenofovir treatment, and subsequently amplified by PCR before full-length HBV genomes were assembled by deep sequencing. The mutation linkage within the viral population was evaluated by clonal analysis of amplicons. RESULTS: Sequence analysis of HBV, derived from 11 samples collected 2010-2020 from one patient, identified 12 non-synonymous single-nucleotide polymorphisms (SNPs) emerging during a tenofovir treatment interruption from 2014 to 2017. Two of the SNPs were in the reverse transcriptase (RT; H35Q and D263E). The two RT mutations were linked and persisted despite restarting tenofovir treatment in 2017. For the second patient, we analyzed HBV derived from six samples collected 2014-2020 following 10 years of tenofovir treatment, and identified five non-synonymous SNPs, that confer resistance towards entecavir and/or lamivudine. Two RT mutations (H35N and P237T) emerged during subsequent 5-year entecavir treatment. H35N was maintained during final tenofovir treatment. CONCLUSIONS: Our findings indicate that changes at the conserved residue 35 (H35N/Q) in the HBV RT may be associated with tenofovir resistance. These variants have not previously been described, and further studies are warranted to assess resistance in vitro and in vivo.

Novel HCV Genotype 4d Infectious Systems and Assessment of Direct-Acting Antivirals and Antibody Neutralization.

Hepatitis C virus (HCV) genotype 4 is highly prevalent in the Middle East and parts of Africa. Subtype 4d has recently spread among high-risk groups in Europe. However, 4d infectious culture systems are not available, hampering studies of drugs, as well as neutralizing antibodies relevant for HCV vaccine development. We determined the consensus 4d sequence from a chronic hepatitis C patient by next-generation sequencing, generated a full-length clone thereof (pDH13), and demonstrated that pDH13 RNA-transcripts were viable in the human-liver chimeric mouse model, but not in Huh7.5 cells. However, a JFH1-based DH13 Core-NS5A 4d clone encoding A1671S, T1785V, and D2411G was viable in Huh7.5 cells, with efficient growth after inclusion of 10 additional substitutions [4d(C5A)-13m]. The efficacies of NS3/4A protease- and NS5A- inhibitors against genotypes 4a and 4d were similar, except for ledipasvir, which is less potent against 4d. Compared to 4a, the 4d(C5A)-13m virus was more sensitive to neutralizing monoclonal antibodies AR3A and AR5A, as well as 4a and 4d patient plasma antibodies. In conclusion, we developed the first genotype 4d infectious culture system enabling DAA efficacy testing and antibody neutralization assessment critical to optimization of DAA treatments in the clinic and for vaccine design to combat the HCV epidemic.