A Validatable Droplet Digital Polymerase Chain Reaction Assay for the Detection of Adeno-Associated Viral Vectors in Bioshedding Studies of Tears.

The use of viral vectors to treat genetic diseases has increased substantially in recent years, with over 2,000 studies registered to date. Adeno-associated viral (AAV) vectors have found particular success in the treatment of eye related diseases, as exemplified by the approval of voretigene neparvovec-rzyl. To bring new therapies to market, regulatory agencies typically request qualified or validated bioshedding studies to evaluate release of the vector into the environment.

Assessment of naloxone as a therapeutic for inhaled carfentanil in the ferret.

Carfentanil is a powerful synthetic opioid that is approximately 100 times more potent than fentanyl and 10,000 times more potent than morphine. Carfentanil was originally intended to be used as a sedative for big game animals in a veterinary setting, but it is becoming increasingly recognized as a public health concern. We set out to investigate the effectiveness of naloxone against a potentially lethal dose of inhaled carfentanil in male ferrets.

Symptomatic treatment of botulism with a clinically approved small molecule.

Botulinum neurotoxins (BoNTs) are potent neuroparalytic toxins that cause mortality through respiratory paralysis. The approved medical countermeasure for BoNT poisoning is infusion of antitoxin immunoglobulins. However, antitoxins have poor therapeutic efficacy in symptomatic patients; thus, there is an urgent need for treatments that reduce the need for artificial ventilation.

Disparate Entry of Adenoviruses Dictates Differential Innate Immune Responses on the Ocular Surface.

Human adenovirus infection of the ocular surface is associated with severe keratoconjunctivitis and the formation of subepithelial corneal infiltrates, which may persist and impair vision for months to years following infection. Long term pathology persists well beyond the resolution of viral replication, indicating that the prolonged immune response is not virus-mediated. However, it is not clear how these responses are sustained or even initiated following infection.

A Randomized Cross-Over Trial Comparing the Effect of Intramuscular Versus Intranasal Naloxone Reversal of Intravenous Fentanyl on Odor Detection in Working Dogs.

Fentanyl is a potent opioid used clinically as a pain medication and anesthetic but has recently seen a sharp rise as an illicit street drug. The potency of fentanyl means mucous membrane exposure to a small amount of the drug can expose first responders, including working canines, to accidental overdose. Naloxone, a fast-acting opioid antagonist administered intranasally (IN) or intramuscularly (IM) is currently carried by emergency personnel in the case of accidental exposure in both humans and canines.

Effects of orally administered raltegravir in cats with experimentally induced ocular and respiratory feline herpesvirus-1 infection.

Objective: To determine the effects of orally administered raltegravir in cats with experimentally induced ocular and respiratory feline herpesvirus-1 (FHV-1) infection.

Animals: 14 healthy 6-month-old unvaccinated specific pathogen-free cats.

Procedures: On day 0, all cats were experimentally inoculated by topical application of 0.

In vitro efficacy of povidone iodine and hydroxyethyl cellulose, alone and in combination, against common feline ocular pathogens.

Infectious ocular disease, such as conjunctivitis, is common in cats and can be caused by several viruses and bacteria, either as a single infection or as co-infections. In this study, povidone-iodine (PVP-I), alone or compounded with hydroxyethyl cellulose (HEC), was investigated for its efficacy against these pathogens in vitro. Whilst PVP-I alone was effective at inhibiting feline herpesvirus type 1 (FHV-1), Chlamydia felis, and Mycoplasma felis, PVP-I with HEC exerted a synergistic inhibitory effect against FHV-1 and C.

The HIV Integrase Inhibitor Raltegravir Inhibits Felid Alphaherpesvirus 1 Replication by Targeting both DNA Replication and Late Gene Expression.

Alphaherpesvirus-associated ocular infections in humans caused by human alphaherpesvirus 1 (HHV-1) remain challenging to treat due to the frequency of drug application required and the potential for the selection of drug-resistant viruses. Repurposing on-the-market drugs is a viable strategy to accelerate the pace of drug development. It has been reported that the human immunodeficiency virus (HIV) integrase inhibitor raltegravir inhibits HHV-1 replication by targeting the DNA polymerase accessory factor and limits terminase-mediated genome cleavage of human betaherpesvirus 5 (HHV-5).

Evaluation of topical ophthalmic ganciclovir gel for the treatment of dogs with experimentally induced ocular canine herpesvirus-1 infection.

OBJECTIVE To determine the in vitro half maximal effective concentration (EC) of ganciclovir for canine herpesvirus-1 (CHV-1) and to evaluate the efficacy of ganciclovir ophthalmic gel in dogs with experimentally induced ocular CHV-1 infection. ANIMALS 10 specific pathogen-free adult Beagles. PROCEDURES Cytotoxicity and EC of ganciclovir for CHV-1 were determined during in vitro experiments.

Transcriptome profiling of alphaherpesvirus-infected cells treated with the HIV-integrase inhibitor raltegravir reveals profound and specific alterations in host transcription.

Anti-microbial compounds typically exert their action by directly interfering with one or more stages of the pathogen's life cycle. However, some compounds also have secondary effects on the host that aid in pathogen clearance. Raltegravir is a human immunodeficiency virus (HIV)-integrase inhibitor that has been shown to alter the host immune response to HIV in addition to its direct antiviral effect.

Plasma Concentration of Meloxicam in Pediatric Rats.

In this study, we compared the plasma concentrations of meloxicam in pediatric rat pups (ages: 7, 14, 21, and 28 d) with those of young adult rats. Adult rats received 1.34 mg/kg SC meloxicam to determine the target peak plasma concentration (Cmax) for comparison with the pediatric animals.

New Paradigms for the Study of Ocular Alphaherpesvirus Infections: Insights into the Use of Non-Traditional Host Model Systems.

Ocular herpesviruses, most notably human alphaherpesvirus 1 (HSV-1), canid alphaherpesvirus 1 (CHV-1) and felid alphaherpesvirus 1 (FHV-1), infect and cause severe disease that may lead to blindness. CHV-1 and FHV-1 have a pathogenesis and induce clinical disease in their hosts that is similar to HSV-1 ocular infections in humans, suggesting that infection of dogs and cats with CHV-1 and FHV-1, respectively, can be used as a comparative natural host model of herpesvirus-induced ocular disease. In this review, we discuss both strengths and limitations of the various available model systems to study ocular herpesvirus infection, with a focus on the use of these non-traditional virus-natural host models.

Electric Cell-Substrate Impedance Sensing To Monitor Viral Growth and Study Cellular Responses to Infection with Alphaherpesviruses in Real Time.

Electric cell-substrate impedance sensing (ECIS) measures changes in an electrical circuit formed in a culture dish. As cells grow over a gold electrode, they block the flow of electricity and this is read as an increase in electrical impedance in the circuit. ECIS has previously been used in a variety of applications to study cell growth, migration, and behavior in response to stimuli in real time and without the need for cellular labels.

Towards Development of Small Molecule Lipid II Inhibitors as Novel Antibiotics.

Recently we described a novel di-benzene-pyrylium-indolene (BAS00127538) inhibitor of Lipid II. BAS00127538 (1-Methyl-2,4-diphenyl-6-((1E,3E)-3-(1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)pyryl-1-ium) tetrafluoroborate is the first small molecule Lipid II inhibitor and is structurally distinct from natural agents that bind Lipid II, such as vancomycin. Here, we describe the synthesis and biological evaluation of 50 new analogs of BAS00127538 designed to explore the structure-activity relationships of the scaffold.

Phase Separation on Bicontinuous Cubic Membranes: Symmetry Breaking, Reentrant, and Domain Faceting.

We study the phase separation of binary lipid mixtures that form bicontinuous cubic phases. The competition between the nonuniform Gaussian membrane curvature and line tension leads to a very rich phase diagram, where we observe symmetry breaking of the membrane morphologies and reentrant phenomena due to the formation of bridges between segregated domains. Upon increasing the line tension contribution, we also find faceting of lipid domains that we explain using a simple argument based on the symmetry of the underlying surface and topology.

Equine Mesenchymal Stromal Cells from Different Sources Efficiently Differentiate into Hepatocyte-Like Cells.

Adult equine hepatocytes have proven challenging to culture long term in vitro as they rapidly lose their morphology and functionality, thus limiting studies on liver function and response to disease. In this study, we describe for the first time the differentiation of equine mesenchymal stromal cells (MSC) from a variety of sources into functional hepatocyte-like cells (HLC). First, we differentiated equine umbilical cord blood (UCB)-derived MSC into HLC and found that these cells exhibited a distinct polygonal morphology, stored glycogen as visualized by periodic acid Schiff's reagent staining, and were positive for albumin and other hepatocyte-specific genes.

A novel corneal explant model system to evaluate antiviral drugs against feline herpesvirus type 1 (FHV-1).

Feline herpesvirus type-1 (FHV-1) is the most common viral cause of ocular surface disease in cats. Many antiviral drugs are used to treat FHV-1, but require frequent topical application and most lack well-controlled in vivo studies to justify their clinical use. Therefore, better validation of current and novel treatment options are urgently needed.

In Vitro and In Vivo Evaluation of Cidofovir as a Topical Ophthalmic Antiviral for Ocular Canine Herpesvirus-1 Infections in Dogs.

Purpose: The effects of cidofovir were investigated against canine herpesvirus-1 (CHV-1) in vitro and in dogs with experimentally induced recurrent ocular CHV-1 infection, a host-adapted pathogen animal model of ocular herpes simplex virus-1 (HSV-1) infection.

Methods: The cidofovir EC50 was determined for CHV-1 and HSV-1. A randomized, masked vehicle-controlled trial was performed using beagles with experimentally induced recurrent ocular CHV-1 infection.

Silicon is a frequent component of atmospheric nanoparticles.

Nanoparticles are the largest fraction of aerosol loading by number. Knowledge of the chemical components present in nanoparticulate matter is needed to understand nanoparticle health and climatic impacts. In this work, we present field measurements using the Nano Aerosol Mass Spectrometer (NAMS), which provides quantitative elemental composition of nanoparticles around 20 nm diameter.

Selective detection and characterization of nanoparticles from motor vehicles.

Numerous studies have shown that exposure to motor vehicle emissions increases the probability of heart attacks, asthma attacks, and hospital visits among at-risk individuals. However, while many studies have focused on measurements of ambient nanoparticles near highways, they have not focused on specific road-level domains, such as intersections near population centers. At these locations, very intense spikes in particle number concentration have been observed.

Molecular transformations accompanying the aging of laboratory secondary organic aerosol.

The aging of fresh secondary organic aerosol (SOA), formed in a flow tube reactor by α-pinene ozonolysis, was studied by passing the fresh SOA into a second chamber for reaction with high levels of the hydroxyl radical. Two types of experiments were performed: (1) injection of a short plug of fresh SOA into the second chamber, where the particle mass and average O/C mole ratio were measured as a function of time after injection, and (2) injection of a continuous stream of fresh SOA into the second chamber, where particles were collected on a filter over a period of time for off line analysis by high performance mass spectrometry. These setups allowed the chemistry of SOA aging to be elucidated.

Quantitative and time-resolved nanoparticle composition measurements during new particle formation.

The chemical composition of 20 nm diameter particles was measured with the Nano Aerosol Mass Spectrometer (NAMS) in a rural/coastal environment during days when new particle formation (NPF) occurred and days when NPF did not occur. NAMS provides a quantitative measure of nanoparticle elemental composition with high time resolution. These measurements show that nanoparticle chemical composition is dynamic on both types of days and that changes in nanoparticle chemical composition do not necessarily correlate with changes in aerosol mass or number concentration.

Quantitative assessment of the sulfuric acid contribution to new particle growth.

The Nano Aerosol Mass Spectrometer (NAMS) was deployed to rural/coastal and urban sites to measure the composition of 20-25 nm diameter nanoparticles during new particle formation (NPF). NAMS provides a quantitative measure of the elemental composition of individual, size-selected nanoparticles. In both environments, particles analyzed during NPF were found to be enhanced in elements associated with inorganic species (nitrogen, sulfur) relative to that associated with organic species (carbon).