Novel Mode of nanoLuciferase Packaging in SARS-CoV-2 Virions and VLPs Provides Versatile Reporters for Virus Production.

SARS-CoV-2 is a positive-strand RNA virus in the Coronaviridae family that is responsible for morbidity and mortality worldwide. To better understand the molecular pathways leading to SARS-CoV-2 virus assembly, we examined a virus-like particle (VLP) system co-expressing all structural proteins together with an mRNA reporter encoding nanoLuciferase (herein nLuc). Surprisingly, the 19 kDa nLuc protein itself was encapsidated into VLPs, providing a better reporter than nLuc mRNA itself.

Acyl-Coa Thioesterases: A Rheostat That Controls Activated Fatty Acids Modulates Dengue Virus Serotype 2 Replication.

During infection with dengue viruses (DENVs), the lipid landscape within host cells is significantly altered to assemble membrane platforms that support viral replication and particle assembly. Fatty acyl-CoAs are key intermediates in the biosynthesis of complex lipids that form these membranes. They also function as key signaling lipids in the cell.

Experimental Zika virus infection of Jamaican fruit bats (Artibeus jamaicensis) and possible entry of virus into brain via activated microglial cells.

The emergence of Zika virus (ZIKV) in the New World has led to more than 200,000 human infections. Perinatal infection can cause severe neurological complications, including fetal and neonatal microcephaly, and in adults there is an association with Guillain-Barré syndrome (GBS). ZIKV is transmitted to humans by Aedes sp.

Stearoly-CoA desaturase 1 differentiates early and advanced dengue virus infections and determines virus particle infectivity.

Positive strand RNA viruses, such as dengue virus type 2 (DENV2) expand and structurally alter ER membranes to optimize cellular communication pathways that promote viral replicative needs. These complex rearrangements require significant protein scaffolding as well as changes to the ER chemical composition to support these structures. We have previously shown that the lipid abundance and repertoire of host cells are significantly altered during infection with these viruses.

Dengue viruses cleave STING in humans but not in nonhuman primates, their presumed natural reservoir.

Human dengue viruses emerged from primate reservoirs, yet paradoxically dengue does not reach high titers in primate models. This presents a unique opportunity to examine the genetics of spillover versus reservoir hosts. The dengue virus 2 (DENV2) - encoded protease cleaves human STING, reducing type I interferon production and boosting viral titers in humans.

Murine Efficacy and Pharmacokinetic Evaluation of the Flaviviral NS5 Capping Enzyme 2-Thioxothiazolidin-4-One Inhibitor BG-323.

Arthropod-borne flavivirus infection continues to cause significant morbidity and mortality worldwide. Identification of drug targets and novel antiflaviviral compounds to treat these diseases has become a global health imperative. A previous screen of 235,456 commercially available small molecules identified the 2-thioxothiazolidin-4-one family of compounds as inhibitors of the flaviviral NS5 capping enzyme, a promising target for antiviral drug development.

Oxidative stress influences positive strand RNA virus genome synthesis and capping.

Flaviviruses are 5' capped positive-stranded RNA viruses that replicate their genomes within endoplasmic reticulum-derived vesicles. Flaviviruses are well known to induce oxidative stress late in infection but it is unknown if oxidative stress plays a positive role in the viral RNA replication cycle. We therefore examined how oxidation affects flavivirus RNA replication.

The accuracy of handheld pre-arrest breath test instruments as a predictor of the evidential breath alcohol test results.

Drunk driving is a serious threat to public safety. All available and appropriate tools for curbing this threat should be employed to their full extent. The handheld pre-arrest breath test instrument (PBT) is one tool for identifying the alcohol-impaired driver and enforcing drunk driving legislation.

Estimating the measurement uncertainty in forensic blood alcohol analysis.

For many reasons, forensic toxicologists are being asked to determine and report their measurement uncertainty in blood alcohol analysis. While understood conceptually, the elements and computations involved in determining measurement uncertainty are generally foreign to most forensic toxicologists. Several established and well-documented methods are available to determine and report the uncertainty in blood alcohol measurement.

Factors contributing to the variability observed in duplicate forensic breath alcohol measurement.

Several factors contribute to the variability observed among repeated measurements of breath alcohol concentration. Identifying these factors and the magnitude of their contribution is the focus of this study. Large breath alcohol data sets consisting of duplicate test results from drivers arrested for driving while intoxicated were obtained from four jurisdictions: Sweden, Alabama, New Jersey and Washington State.

Differentiating new cannabis use from residual urinary cannabinoid excretion in chronic, daily cannabis users.

Aims: To develop and validate empirically a mathematical model for identifying new cannabis use in chronic, daily cannabis smokers.

Design: Models were based on urinary creatinine-normalized (CN) cannabinoid excretion in chronic cannabis smokers.

Setting: For model development, participants resided on a secure research unit for 30 days.

Employing components-of-variance to evaluate forensic breath test instruments.

The evaluation of breath alcohol instruments for forensic suitability generally includes the assessment of accuracy, precision, linearity, blood/breath comparisons, etc. Although relevant and important, these methods fail to evaluate other important analytical and biological components related to measurement variability. An experimental design comparing different instruments measuring replicate breath samples from several subjects is presented here.

New Zealand's breath and blood alcohol testing programs: further data analysis and forensic implications.

Paired blood and breath alcohol concentrations (BAC, in g/dL, and BrAC, in g/210 L), were determined for 11,837 drivers apprehended by the New Zealand Police. For each driver, duplicate BAC measurements were made using headspace gas chromatography and duplicate BrAC measurements were made with either Intoxilyzer 5000, Seres 679T or Seres 679ENZ Ethylometre infrared analysers. The variability of differences between duplicate results is described in detail, as well as the variability of differences between the paired BrAC and BAC results.

Estimating the uncertainty associated with Widmark's equation as commonly applied in forensic toxicology.

No computation is performed more frequently by forensic toxicologists than that involving Widmark's equation. The equation is employed to estimate either the number of drinks consumed or the corresponding blood or breath alcohol concentration. Despite the wide use of Widmark's equation, rarely is an uncertainty estimate also provided.

Results of a proposed breath alcohol proficiency test program.

Although proficiency test programs have long been used in both clinical and forensic laboratories, they have not found uniform application in forensic breath alcohol programs. An initial effort to develop a proficiency test program appropriate to forensic breath alcohol analysis is described herein. A total of 11 jurisdictions participated in which 27 modern instruments were evaluated.

Factors associated with breath test refusals in drunken driving arrests.

Objective: Jurisdictions with per se breath alcohol legislation rely heavily on breath test evidence in prosecuting drunk driving cases. Depending on other legal considerations, where subjects refuse the breath test, prosecution may be more difficult. The objective was to identify factors significantly associated with the risk of test refusal.

Common Legal Challenges and Responses in Forensic Breath Alcohol Determination.

No other single criminal/civil offense connects the forensic science community together with the legal community more than driving while under the influence of alcohol (DUI). Statutory changes along with political pressure have caused DUI to be a very serious and expensive offense. The result has been an enormous increase in DUI defense challenges in recent years with the primary focus on breath alcohol measurement.

Breath alcohol measurement variability associated with different instrumentation and protocols.

Breath alcohol measurement has variability resulting from instrumental, procedural and biological components. Reliable estimates of the standard deviation (S.D.

Breath alcohol analysis in one subject with gastroesophageal reflux disease.

A large number of people suffer from the heartburn symptoms associated with gastroesophageal reflux disease (GERD). Relatively little has been published on its potential for biasing a breath alcohol measurement. The present case describes an individual (white male, aged 23) who experimentally consumed 1.

Enforcement of Zero Tolerance in the State of Washington - Evidence from Breath-Test Records.

Several studies have demonstrated the effectiveness of zero tolerance (ZT) laws that make it illegal for drivers younger than 21 years to have any measurable alcohol in their bodies when operating a vehicle. All 50 states in the United States now have such laws. However, there is some question as to the extent they are being enforced.