Analysis of Transmission of Hepatitis C Viruses: Dental vs. Surgical Procedures

Objectives: The primary purpose of the present study is to evaluate the cause of HCV transmission either by dental or surgical procedures in Pakistan. It will reveal the standards of current practices and knowledge of surgical instrument sterilization in Pakistan. Methods:The contemporary study design was cross-sectional. A total of 7097 patients were included in the study that was the confirmed cases of Hepatitis C from four districts of Punjab, Pakistan, during the period of three years from Jul 1, 2018, to Jun 30, 2021. The ANN model simulates a discrete Pareto distribution; descriptive statistics were applied by using SPSS 23.Results:In age distribution, most patients were aged 26-37 from the 2020- 2021 year of study with 45.6%age. The total mean ± SD was 2.42 ± .935 with 0.000P-values. Results show that high-risk variables are strongly associated with the surgical history, such as sector analysis (public and private) with 0.000 P-values. Analysis of these variables indicated that inquiry about surgical instruments sterilization and age groups(100%)were the most vital variable in the ANN model. Pareto distribution values of surgical procedures that are transmitting hepatitis C viruses more than the dental procedures. Conclusion: Surgical procedures are high transmission source of hepatitis C viruses than dental procedures. Clinical Significance: More care should be taken, and more severe sterilization measures should be taken to prevent the transmission of hepatitis C infection, especially in dental and general surgery procedures in Pakistan.

Recapitulating Trafficking of Nucleosides Into the Active Site of Polymerases of RNA Viruses: The Challenge and the Prize

Nucleoside analogs are very effective antiviral agents with currently over 25 compounds approved for the therapy of viral infections. Still, their successful use against RNA viruses is very recent, despite RNA viruses comprising some of the most damaging human pathogens (e.g., Coronaviruses, Influenza viruses, or Flaviviridae such as dengue, Zika and hepatitis C viruses). The breakthrough came in 2013–2014, when the nucleoside analog Sofosbuvir became one of the cornerstones of current curative treatments for hepatitis C virus (HCV). An analog designed on the same principles, Remdesivir, has been the first approved compound against SARS-CoV-2, the coronavirus that causes the current COVID-19 pandemic. Both of these nucleoside analogs target the RNA-dependent RNA polymerase (RdRp) (NS5B for HCV, nsp12 for SARS-CoV-2). RdRps of RNA viruses display a peculiar elaboration of the classical polymerase architecture that leads to their active site being caged. Thus, triphosphate nucleosides and their analogs must access this active site in several steps along a narrow and dynamic tunnel. This makes straightforward computational approaches such as docking unsuitable for getting atomic-level details of this process. Here we give an account of ribose-modified nucleoside analogs as inhibitors of viral RdRps and of why taking into account the dynamics of these polymerases is necessary to understand nucleotide selection by RdRps. As a case study we use a computational protocol we recently described to examine the approach of the NTP tunnel of HCV NS5B by cellular metabolites of Sofosbuvir. We find major differences with natural nucleotides even at this early stage of nucleotide entry.

An assessment of knowledge and awareness on hepatitis B and hepatitis C viruses among police trainees attending police training college, Ongole, Prakasam District, Andhra Pradesh, India

Background: HBV and HCV are blood borne viral diseases, and knowledge about modes of transmission and preventive aspects can help in reduce the risk of getting liver diseases and mainly transmitted through transfusion of contaminated blood and blood products. The knowledge regarding HBV and HCV infection studied in many professional groups, but few surveys are available among the police professionals. The study was conducted to assess the knowledge and awareness regarding Hepatitis B and C viruses’ transmission among these police trainees, and to find out the awareness regarding symptoms, signs, complications and importance of Hepatitis B vaccine.Methods: A cross sectional observationalstudywasconductedamongthepolicetraineesattendingdistrictpolicetraining college, Ongole, Prakasam district, Andhra Pradesh. Information about socio demographic characteristics, basic knowledge about hepatitis B and C viruses, its modes of transmission, prevention, symptoms and signs and also complications, Hepatitis B vaccination were also assessed from all these police trainees. The data were collected, tabulated, and statistically analyzed.Results: Among total police trainees 61.8% belong to upper middle class, 62.4% were degree holders and most of them belonging to Krishna (32.2%) district. About 59% of police trainees aware about hepatitis B virus infection and 67% know about hepatitis C virus infection. 67.2% were listen about availability of hepatitis B vaccination, none of the participants were aware about hepatitis B vaccine zero dose schedule.Conclusions: Efficient health programme managing departments must take the responsibility for HBV education, transmission, testing, vaccination accessibility and availability.

To Assess the Prevalence Rate of Hepatitis C among the Solid Waste Handler in Selected Areas

Introduction: Solid waste handlers are subjected to a wide range of occupational hazards. Among these hazards is the infection from hepatitis A, B or C viruses (HAV, HBV or HCV). This relationship has been the study subject of many researchers around the world, given that the infection of hepatitis viruses is a significant cause of morbidity and a socio-economic burden.Viral hepatitis is a liver inflammation arising from viral infections. There are classes of viruses affecting the liver. The most popular forms are hepatitis B (HBV) and hepatitis C viruses (HCV). Objective: this study is planning to assess the prevalence rate of Hepatitis C among solid waste handlers in selected areas. Methodology: Cross-sectional study research designed will be used in this study conducted on a solid waste handler. The unlikely purposefully sampling technique was used to select a sample for this study to analyze the prevalence rate. The researcher's study included 100 population from the selected area and investigation of Hepatitis C will do at A.V.B.R.H Hospital Sawangi (M) Wardha. The population was selected according to inclusion and exclusion criteria. Expected Outcome: In this research study expected outcome is solid waste handlers are may be suffering from Hepatitis C because of their working pattern and environment. Conclusion: The conclusion will be drawn from the results.

Profiling of tumour-associated microbiota in human hepatocellular carcinoma

Liver cancer is the fourth leading cause of cancer-related death. Hepatocellular carcinoma (HCC) is a primary liver cancer that results from chronic hepatitis caused by multiple predisposing factors such as viral infection, alcohol consumption, and non-alcoholic fatty liver disease. Accumulating studies have indicated that dysfunction of the gut epithelial barrier and hepatic translocation of gut microbes may be implicated in the pathogenesis of HCC. However, the translocated bacteria in HCC patients remains unclear. Here, we characterised tumour-associated microbiota in patients with liver cancer and focused on HCC. We observed that the number of amplicon sequence variants in tumour-associated microbiota was significantly higher compared with that in non-tumour regions of the liver. The tumour-associated microbiota consisted of Bacteroidetes, Firmicutes, and Proteobacteria as the dominant phyla. We identified an unclassified genus that belonged to the Bacteroides, Romboutsia, uncultured bacterium of Lachnospiraceae as a signature taxon for primary liver cancer. Additionally, we identified Ruminococcus gnavus as a signature taxon for HCC patients infected with hepatitis B and/or hepatitis C viruses. This study suggests that tumour microbiota may contribute to the pathology of HCC.

EXPRESSION OF NUCLEOCAPSID VIRAL PROTEINS IN THE BACTERIAL SYSTEM OF Escherichia coli: THE INFLUENCE OF THE CODON COMPOSITION AND THE UNIFORMITY OF ITS DISTRIBUTION WITHIN GENE

A heterologous host has got a unique expression ability of each gene. Differences between the synonymous sequences play an important role in regulation of protein expression in organisms from Escherichia coli to human, and many details of this process remain unclear. The work was aimed to study the composition of codons, its distribution over the sequence and the effect of rare codons on the expression of viral nucleocapsid proteins and their fragments in the heterologous system of E.coli. The plasmid vector pJC 40 and the BL 21 (DE 3) E. coli strain were used for protein expression. The codon composition analysis was performed using the online resource (www.biologicscorp.com). 10 recombinant polypeptides were obtained encoding the complete nucleotide sequence of nucleocapsid proteins (West Nile and hepatitis C viruses) and the fragments including antigenic determinants (Lassa virus, Marburg, Ebola, Crimean-Congo hemorrhagic fever (CCHF), Puumaravala, Hantaan, and lymphocytic choriomeningitis (LHM)). Hybrid plasmid DNAs provide efficient production of these proteins in the prokaryotic system with the recombinant protein yield varying by a factor of 8: from 5 to 40 mg per 1 liter of bacterial culture. No correlation was found between the level of protein expression and the frequency of occurrence of rare codons in the cloned sequence: the maximum frequency of occurrence of rare codons per cloned sequence was observed for the West Nile virus (14.6%), the minimum was for the CCHF virus (6.6%), whereas the expression level for these proteins was 30 and 5 mg/L culture, respectively. The codon adaptation index (CAI) values, calculated on the basis of the codon composition in E. coli, for the cloned viral sequences were in the range from 0.50 to 0.58, which corresponded to the average expressed proteins. The analysis of the distribution profiles of CAI in the cloned sequences indicated the absence of clusters of rare codons that could create difficulties in translation. A statistically significant difference between the frequencies of the distribution of amino acids in the cloned sequences and their content in E. coli was observed for the nucleocapsid proteins of the Marburg, Ebola, West Nile, and hepatitis C viruses.

Translational Perspective in Hepatocellular Carcinoma

The burden of liver cancer is higher in Hispanics, African Americans, and Asians. Viral hepatitis (Hepatitis B and Hepatitis C viruses), non-alcoholic steatohepatitis (NASH), and alcoholic liver disease (ALD) are the most common etiological/risk factors for liver cancer. Approximately 80–90% of hepatocellular carcinoma (HCC) occurs in patients with underlying liver cirrhosis. Individuals with advanced cirrhosis represent a high-risk group for liver cancer. To fill the increasing gap between basic science and clinical research, translational research has been developed as an emerging technology. Basic science attempts to unravel the mechanisms of disease using tools (e.g., culture systems and animal models) that allow for easy manipulation of biological processes. Further, culture systems and animal models are useful to derive causal associations, but they generally do not include an endpoint directly applicable to clinical practice. Hence, development of new tools for early detection, including the evaluation of liquid biopsy, identification of tissue biomarkers of treatment response, execution of precision and enhancement of patient stratification in patients at risk for HCC development to enable chemoprevention clinical trials becomes important. It was identified as translational research has begun as an effective approach to facilitate the development of novel molecular-based biomarkers and to accelerate the implementation of laboratory discoveries into clinically applicable tools. Despite great advancement in diagnosis and management of HCC, the exact biology of the tumor remains poorly understood generally limiting the clinical outcome. Comprehensive analysis and characterization of the molecular mechanisms and subsequently individual prediction of corresponding prognostic traits would transform both diagnosis and treatment of HCC and is the key goal of modern medicine. To overcome the challenge and to accelerate the progress, a collaborative effort from various clinical research groups and translational approach is needed.

Optimizing Consensus Generation Algorithms for Highly Variable Amino Acid Sequence Clusters

Producing a functional consensus sequence is a preliminary bioinformatics task, which is a necessity for many research purposes. However, the existence of hypervariable regions in the input multiple sequence alignment files causes complications in generating a useful consensus sequence. The current methods for consensus generation, Threshold, and majority algorithms, have several problems, which exclude them as applicable algorithms for such highly variable sequence clusters. Hence, we designed a novel alternative algorithm for the same purpose. The algorithm was explained both using a mathematical formula and a practical implementation in Python programming language. A sequence set from HCV genotype 1b E2 protein has been utilized as a practical example for evaluating the algorithm’s performance. A few in silico tests have been performed on the output sequence and the results have been compared to results from other algorithms. Epitope-mapping analysis indicates the functionality of this algorithm, by preserving the hotspot residues in the consensus sequence, and the antigenicity index shows significant antigenicity of the consensus sequence. Moreover, phylogenetic analysis shows no significant change in the placement of the new consensus sequence on the phylogenetic tree compared to other algorithms. This approach will have several implications in designing a new vaccine for highly variable viruses such as HIV-1, Influenza, and Hepatitis C Viruses (HCV).

EKSTRAKSI SENYAWA BIOAKTIF THEAFLAVIN TEH OOLONG (CAMELLIA SINESIS) DENGAN AIR SEBAGAI GREEN SOLVENT

Theaflavin, a polyphenolic compounds found in oolong tea and black tea, thought to have medicinal potency. Theaflavins and their derivatives such as theaflavin gallate have shown a broad spectrum of antiviral activity against several viruses, including influenza A, B and hepatitis C viruses. Previous research show that theaflavins could inhibit RdRp activity through blocking the active site in the catalytic pocket of RdRp in SARS‐CoV‐2, SARS‐CoV and MERS‐CoV.. This research aims to extract theaflavins in oolong tea with water solvent at a temperature of 100oC, pressure > 1 atm and varied extraction times (10-60 minutes). The effect of oolong tea extraction time towards the theaflavin content was observed. The results showed that with the longer extraction time, the theaflavin levels were reduced due to the degradation of thermal theaflavins into thearubigins. The model exponential equation obtained is y=14,91488 e-0,2631x + 0,11865, with the R2=0.99161. The best operating conditions was obtained at 10 minutes of extraction time, resulted in 1.19% of the total theaflavin content of oolong tea.