A detailed model and Monte Carlo simulation for predicting DIP genome length distribution in baculovirus infection of insect cells

Baculoviruses have enormous potential for use as biopesticides to control insect pest populations without the adverse environmental effects posed by the widespread use of chemical pesticides. However, continuous baculovirus production is susceptible to DNA mutation and the subsequent production of defective interfering particles (DIPs). The amount of DIPs produced and their genome length distribution are of great interest not only for baculoviruses but for many other DNA and RNA viruses. In this study, we elucidate this aspect of virus replication using baculovirus as an example system and both experimental and modeling studies. The existing mathematical models for the virus replication process consider DIPs as a lumped quantity and do not consider the genome length distribution of the DIPs. In this study, a detailed population balance model for the cell‐virus culture is presented, which predicts the genome length distribution of the DIP population along with their relative proportion. The model is simulated using the kinetic Monte Carlo algorithm, and the results agree well with the experimental results. Using this model, a practical strategy to maintain the DIP fraction to near to its maximum and minimum limits has been demonstrated.     

江苏省300例艾滋病人群联合检测 HCV、TP、HIV的结果分析

摘要 目的：探究江苏省2017年至2019年度艾滋病哨点监测人群丙型肝炎病毒（Hepatitis C virus,HCV）、梅毒螺旋体（Treponema Pallidum,TP）以及人类免疫缺陷病毒（Treponema Pallidum,HIV）感染状况,以期为制定对应的疾患干预手段提供临床数据支撑。方法：严格按照《全国艾滋病哨点监测实施方案操作手册（2012版）》对入组的900例个体开展相关问卷调查,并对入组对象进行HCV、TP以及HIV感染血清学检测,分别统计3年总体感染情况、年度感染情况、不同年龄段人群感染情况以及合并感染情况。结果：（1）性工作者HCV、TP以及HIV感染率分别为3.00 %、4.00 %和9.00 %,吸毒人群感染率分别为13.00 %、73.00 %和13.00 %,孕产妇感染率分别为1.00 %、0.67 %和1.33 %,比较显示,吸毒人群HCV、TP以及HIV感染率明显高于其余两类人群（P<0.05）;（2）就各年度不同人群HCV、TP以及HIV感染率开展年度横向比较显示,不同年度不同人群的HCV、TP以及HIV感染率差异无统计学意义（P>0.05）;（3）针对不同年龄段不同人群的HCV、TP以及HIV感染率开展比较显示,吸毒人群中≥50岁HCV和HIV感染率明显高于其他年龄段（P<0.05）,性工作者中18~20岁HIV感染率明显高于其他年龄段（P<0.05）,其余差异无统计学意义（P>0.05）;（4）合并感染情况分析显示合并TP以及HIV感染率要高于其他合并感染。结论：吸毒人群是江苏省HCV、TP以及HIV感染高发群体,以≥50岁年龄段感染率最高,总体来看HCV、TP以及HIV感染率呈现逐年降低趋势,但仍应该加强对上述疾患的防控工作。     

Virological surveillance,molecular phylogeny,and evolutionary dynamics of hepatitis C virus subtypes 1a and 4a isolates in patients from Saudi Arabia

Hepatitis C virus (HCV) subtypes are pre-requisite to predict endemicity, epidemiology, clinical pathogenesis, diagnosis, and treatment of chronic hepatitis C infection. HCV genotypes 4 and 1 are the most prevalent in Saudi Arabia, however; less consensus data exist on circulating HCV subtypes in infected individuals. This study was aimed to demonstrate the virological surveillance, phylogenetic analysis, and evolutionary relationship of HCV genotypes 4 and 1 subtypes in the Saudi population with the rest of the world. Fifty-five clinical specimens from different parts of the country were analyzed based on 5′ untranslated region (5′ UTR) amplification, direct sequencing, and for molecular evolutionary genetic analysis. Pair-wise comparison and multiple sequence alignment were performed to determine the nucleotide conservation, nucleotide variation, and positional mutations within the sequenced isolates. The evolutionary relationship of sequenced HCV isolates with referenced HCV strains from the rest of the world was established by computing pairwise genetic distances and generating phylogenetic trees. Twelve new sequences were submitted to GenBank, NCBI database. The results revealed that HCV subtype 4a is more prevalent preceded by 1a in the Saudi population. Molecular phylogeny predicts the descendants’ relationship of subtype 4a isolates very close to Egyptian prototype HCV strains, while 1a isolates were homogeneous and clustering to the European and North American genetic lineages. The implications of this study highlight the importance of HCV subtyping as an indispensable tool to monitor the distribution of viral strains, to determine the risk factors of infection prevalence, and to investigate clinical differences of treatment outcomes among intergenotypic and intragenotypic isolates in the treated population.     

Influence of Diesel Soot Particles and Sulfite on Functions of Polymorphonuclear Leukocytes

Aqueous suspensions of diesel soot particles in combination with sulfite influence certain functions of human polymorphonuclear neutrophils in vitro. Chemiluminescence, generated after activation by opsonized zymosan as well as oxygen uptake were decreased, whereas phagocytosis was increased. An enhancement of degranulation could not be observed. The single substances show little or no effects on the above properties. The results indicate that combinations of air pollutants such as diesel soot and sulfite may modulate vital functions of activated leukocytes in vivo.     

Characterization of Indoor Environmental Quality in Primary Schools in Maia: A Portuguese Case Study

Scientific evidence associates indoor environment pollutants with respiratory effects (asthma and allergies), and children constitute one of most sensitive groups. Indoor air quality (IAQ) in schools may indeed be a significant health factor for children, with effects on school attendance and performance. Our aim was to characterize IAQ of classrooms in Maia County (north of Portugal) for which there was no information available. The study was conducted in 21 of the 40 primary schools, selected by stratified random sampling. Depending on the dimension, one or two classrooms were tested at each school. Walkthrough surveys of school grounds, buildings, and individual classrooms were done. Continuous measurements were taken of temperature, relative humidity, airborne respirable particles, total volatile organic compounds, and carbon dioxide, whereas bioaerosols were counted on Plate Count Agar during regular school activities. The indoor arithmetic mean for PM₁₀, CO₂, TCOV, and bioaerosol concentrations were 0.14 mg/m³, 999 ppm, 0.41 mg/m³, and 4140 UCF/m³, respectively. The values of PM₁₀ and CO₂ were close to their acceptable maximum values, but bioaerosols were much higher. This study supports previous studies conducted in school environments and emphasizes the need for proactive indoor air quality audits in school buildings.     

Anti-cancer and anti-hepatitis C virus NS5B polymerase activity of etodolac 1,2,4-triazoles

Abstract

Arachidonic acid is an unsaturated fatty acid liberated from phospholipids of cell membranes. NSAIDs are known as targets of cyclooxygenase enzyme (COX-1, COX-2 and COX-3) in arachidonic acid metabolism. This mechanism of COX-2 in carcinogenesis causes cancer. In addition, COX-2 plays a role in the early stages of hepatocarcinogenesis. Hepatitis C virus (HCV) infection is cause of liver cirrhosis and hepatocellular carcinoma (HCC). The aim of our study was to improve effective agents against HCV. A novel series of new etodolac 1,2,4-triazoles derivatives (4a–h) have been synthesized and investigated for their activity against HCV NS5B polymerase. Compound 4a was found to be the most active with IC₅₀ value of 14.8?µM. In accordance with these results, compound 4a was screened for anti-cancer activity on liver cancer cell lines (Huh7, Mahlavu, HepG2, FOCUS). Compound 4a showed anti-cancer activity against Huh7 human hepatoma cell line with IC₅₀ value of 4.29?µM. Therefore, compound 4a could be considered as a new anti-cancer and anti-HCV lead compound.     

Ligand design,synthesis and biological anti-HCV evaluations for genotypes 1b and 4a of certain 4-(3- & 4-[3-(3,5-dibromo-4-hydroxyphenyl)-propylamino]phenyl) butyric acids and 3-(3,5-dibromo-4-hydroxyphenyl)-propylamino-acetamidobenzoic acid esters

4-(4-[N-1-carboxy-3-(3,5-dibromo-4-hydroxyphenyl)-3-oxo-propylamino]phenyl)-4-oxo-butyric acid (V), 4-(3- & 4-[N-1-carboxy-3-(3,5-dibromo-4-hydroxyphenyl)-3-oxo-propylaminophenyl]-2-aryl-4-oxo-butyric acids (Xa–e) and 4-(2-alkyl-2-[N-3-(3,5-dibromo-4-hydroxyphenyl)-1-carboxy-3-oxo-propylamino]acetamido) benzoate esters (XVa–e) were designed, synthesized and biologically evaluated as anti-HCV for genotypes 1b and 4a. The design was based on their docking scores with HCV NS3/4A protease-binding site of the genotype 1b (1W3C), which is conserved in the genotype 4a structure. The docking scores predicted that most of these molecules have higher affinity to the HCV NS3/4A enzyme more than Indoline lead. These compounds were synthesized and evaluated for their cytopathic inhibitory activity against RAW HCV cell cultures of genotype 4a and also examined against Huh 5–2 HCV cell culture of genotype 1b, utilizing Luciferase and MTS assays. Compounds Xa and Xb have 95 and 80% of the activity of Ribavirin against genotype 4a and compounds XVa, XVb and XVd exerted high percentage inhibitory activity against genotype 1b equal 87.7, 84.3 and 82.8%, respectively, with low EC₅₀ doses.     

Synthesis and Antiviral Evaluation of Ribavirin Congeners Containing a Hexitol Moiety

Abstract

Several ribavirin congeners containing a hexitol moiety were prepared via ring opening of two different epoxides with the methylcarboxylate ester of triazole and further elaboration. Unfortunately, none of the newly synthesized compounds displayed appreciable antiviral activity.     

Development Of A Nucleic Acid Lateral Flow Strip For Detection Of Hepatitis C Virus (Hcv) Core Antigen

The object of this study was to develop a simple, rapid, specific, and highly sensitive method to detect HCV core antigen. A nucleic acid aptamer was designed with the high specificity and sensitivity in a nucleic acid lateral flow strip to compete with HCV core antigen and DNA probes. The lower detection limit of the test strip was calculated to be 10 pg/mL with the scanner and 100 pg/mL with naked eyes. Results showed that there were no cross-interactions with other proteins such as HCV NS3, E1/E2 antigens, HIV p24 antigens, or BSA proteins (HCV unrelated protein). When the viral load exceeded 10⁴ copies/mL, the positive coincidence rates of ELISA and strip detection, when compared with the HCV RNA assay, were 98.44% and 97.28%, respectively. The results indicated that the ELISA detection and strip assay were in good agreement with the measured value. The results indicated that a nucleic acid lateral flow strip was a simple, rapid, specific, highly sensitive, and cost-effective field-based method for detecting HCV core antigen. The strip assay is an acceptable alternative to diagnose HCV core antigen and to investigate its epidemiology in clinical laboratories lacking specialized equipment and skills.     

Inhibition of HCV Replication in HCV Replicon by shRNAs

We show that the vector-derived long dsRNA specifically inhibits the replication of HCV RNA in HCV replicon. We designed a long dsRNA targeted to the full-length HCV IRES/core elements (1-to 377-nt). Our results revealed that the replication of HCV RNA was reduced to near background levels in a sequence-specific manner by the long dsRNAs in the HCV replicon. We also designed four shRNAs against several regions (120- to 139-nt, 260- to 279-nt, 330- to 349-nt, and 340- to 359-nt) of the HCV IRES/Core elements. The two HCV IRES/core-specific shRNAs, 330- to 349-nt and 340- to 359-nt, containing the AUG initiation codon sequence showed stronger HCV inhibitory effects than the other two shRNAs, 120- to 139-nt and 260- to 279-nt.