Business-centric data solutions for safeguarding animal agriculture in the United States of America.

Business-centric solutions to data-related problems often yield the greatest positive impacts and improvements for private enterprises but are challenging to design and implement at scale within government agencies. The core mission of the Veterinary Services of the United States Department of Agriculture (USDA) Animal Plant Health Inspection Service is to safeguard animal agriculture in the United States of America, and effective data management underpins these efforts. As this agency works to assist data-driven decision-making in animal health management, it continues to use a blend of best practices from Federal Data Strategy initiatives and the International Data Management Association framework. This paper describes three case studies that focus on strategies to improve animal health data collection, integration, reporting and governance for animal health authorities. These strategies have enhanced the way USDA's Veterinary Services execute their mission and core operational activities for prevention, detection and early response to support disease containment and control.

S'agissant des problèmes en lien avec les données, les solutions centrées sur l'activité sont souvent celles qui génèrent le plus d'effets positifs et d'améliorations pour les entreprises du secteur privé, mais elles sont difficiles à concevoir et à mettre en oeuvre à grande échelle au sein des agences gouvernementales. Les Services vétérinaires du Service d'inspection de la santé animale et végétale du département américain de l'Agriculture (USDA) ont pour mission centrale de préserver les productions animales états-uniennes ; une gestion efficace des données vient soutenir cet effort. Dans leur action d'appui aux processus décisionnels de gestion de la santé animale fondés sur les données, ces Services recourent à une combinaison de bonnes pratiques mises en oeuvre aussi bien par les initiatives de la Stratégie fédérale sur les données que dans le cadre de l'Association internationale de gestion des données. Les auteurs décrivent trois études de cas sur des stratégies visant à améliorer la collecte, l'intégration, la notification et la gouvernance des données de santé animale afin de répondre aux besoins des autorités compétentes dans ce domaine. Ces stratégies ont permis aux Services vétérinaires de l'USDA de mieux s'acquitter de leur mission et d'améliorer leurs activités opérationnelles de prévention, de détection et de réaction rapide afin d'endiguer et contrôler les maladies.

Las soluciones eminentemente empresariales a problemas relacionados con los datos deparan con frecuencia los mejores frutos y resultados a la empresa privada, pero son difíciles de diseñar y aplicar a escala dentro de las administraciones públicas. Los Servicios Veterinarios adscritos al Servicio de Inspección Sanitaria de Animales y Plantas del Departamento de Agricultura de los Estados Unidos (USDA) tienen por principal cometido salvaguardar la producción animal estadounidense, labor que pasa en parte por una eficaz gestión de los datos. En su función de apoyo a la adopción de decisiones de gestión zoosanitaria basadas en los datos, este organismo sigue empleando una combinación de prácticas óptimas tomadas de iniciativas de la Estrategia Federal de Datos y de las pautas marcadas por la Asociación Internacional de Gestión de Datos. Los autores presentan y analizan tres ejemplos de estrategias para mejorar la obtención, integración, notificación y administración de datos zoosanitarios para las autoridades del ramo. Estas estrategias han conferido mayor eficacia a los Servicios Veterinarios del USDA en el cumplimiento de su misión y en la ejecución de sus principales actividades operativas de prevención, detección y pronta respuesta para ayudar a contener y combatir enfermedades.

Small molecules demonstrate the role of dynamin as a bi-directional regulator of the exocytosis fusion pore and vesicle release.

Hormones and neurotransmitters are stored in specialised vesicles and released from excitable cells through exocytosis. During vesicle fusion with the plasma membrane, a transient fusion pore is created that enables transmitter release. The protein dynamin is known to regulate fusion pore expansion (FPE). The mechanism is unknown, but requires its oligomerisation-stimulated GTPase activity. We used a palette of small molecule dynamin modulators to reveal bi-directional regulation of FPE by dynamin and vesicle release in chromaffin cells. The dynamin inhibitors Dynole 34-2 and Dyngo 4a and the dynamin activator Ryngo 1-23 reduced or increased catecholamine released from single vesicles, respectively. Total internal reflection fluorescence (TIRF) microscopy demonstrated that dynamin stimulation with Ryngo 1-23 reduced the number of neuropeptide Y (NPY) kiss-and-run events, but not full fusion events, and slowed full fusion release kinetics. Amperometric stand-alone foot signals, representing transient kiss-and-run events, were less frequent but were of longer duration, similarly to full amperometric spikes and pre-spike foot signals. These effects are not due to alterations in vesicle size. Ryngo 1-23 action was blocked by inhibitors of actin polymerisation or myosin II. Therefore, we demonstrate using a novel pharmacological approach that dynamin not only controls FPE during exocytosis, but is a bi-directional modulator of the fusion pore that increases or decreases the amount released from a vesicle during exocytosis if it is activated or inhibited, respectively. As such, dynamin has the ability to exquisitely fine-tune transmitter release.

Damaging legacy: maternal cigarette smoking has long-term consequences for male offspring fertility.

STUDY QUESTION: What are the effects on fertility of cigarette smoke-induced toxicity on male offspring exposed during the gestational/weaning period? SUMMARY ANSWER: Maternal cigarette smoke exposure during the gestational/weaning period causes long-term defects in male offspring fertility. WHAT IS KNOWN ALREADY: Cigarette smoke is a well-known reproductive toxicant which is particularly harmful to both fetal and neonatal germ cells. However, recent studies suggest a significant portion of young mothers in the developed world still smoke during pregnancy. In the context of male reproductive health, our understanding of the effects of in utero exposure on offspring fertility is limited. STUDY DESIGN, SIZE, DURATION: In this study, 27 C57BL/6 5-week-old female mice were exposed via the nose-only to cigarette smoke (treatment) or 27 were exposed to room air (control) for 6 weeks before being housed with stud males to produce litters. In the treatment group, smoke exposure continued throughout mating, pregnancy and lactation until weaning of pups at 21 days post birth. Male offspring were examined at post-natal days 3, 6, 12, 21 and 98 (adult). PARTICIPANTS/MATERIALS, SETTING, METHODS: Approximately 108 maternal smoke-exposed C57BL/6 offspring and controls were examined. Spermatogenesis was examined using testicular histology and apoptosis/DNA damage was assessed using caspase immunohistochemistry and TUNEL. Sertoli cell morphology and fluctuations in the spermatogonial stem cell population were also examined using immunohistochemistry. Microarray and QPCR analysis were performed on adult testes to examine specific long-term transcriptomic alteration as a consequence of maternal smoke exposure. Sperm counts and motility, zona/oolemma binding assays, COMET analysis and mitochondrial genomic sequencing were also performed on spermatozoa obtained from adult treated and control mice. Fertility trials using exposed adult male offspring were also performed. MAIN RESULTS AND THE ROLE OF CHANCE: Maternal cigarette smoke exposure caused increased gonocyte and meiotic spermatocyte apoptosis (P < 0.01) as well as germ cell depletion in the seminiferous tubules of neonatal and juvenile offspring. Aberrant testicular development characterized by abnormal Sertoli and germ cell organization, a depleted spermatogonial stem cell population (P < 0.01), atrophic seminiferous tubules and increased germ cell DNA damage (P < 0.01) persisted in adult offspring 11 weeks after exposure. Microarray analysis of adult offspring testes associated these defects with meiotic germ cell development, sex hormone metabolism, oxidative stress and Sertoli cell signalling. Next generation sequencing also revealed a high mitochondrial DNA mutational load in the testes of adult offspring (P < 0.01). Adult maternal smoke-exposed offspring also had reduced sperm counts with spermatozoa exhibiting morphological abnormalities (P < 0.01), affecting motility and fertilization potential. Odf2, a spermatozoa flagellum component required for coordinated ciliary beating, was also significantly down-regulated (P < 0.01) in maternal smoke-exposed adult offspring, with aberrant localization along the spermatozoa flagellum. Adult maternal smoke-exposed offspring took significantly longer to impregnate control females and had a slight but significant (P < 0.01) reduction in litter size. LIMITATIONS, REASONS FOR CAUTION: This study examined only one species (mouse) using a smoking model which only simulates human cigarette smoke exposure. WIDER IMPLICATIONS OF THE FINDINGS: This study represents the first comprehensive animal model of maternal smoking on male offspring reproductive function, suggesting that exposure during the gestational/weaning period causes long-term defects in male offspring fertility. This is due to a compromised spermatogonial stem cell population resulting from gonocyte apoptosis and impaired spermatogenic development. This results in significant germ cell damage and Sertoli cell dysfunction, impacting germ cell number, tubule organization, DNA damage and spermatozoa in adult offspring. This study strengthens the current literature suggesting that maternal exposure impairs male offspring fertility, which is currently debated due to conflicting studies. STUDY FUNDING/COMPETING INTERESTS: This study was funded by the Australian Research Council, Hunter Medical Research Institute, National Health and Medical Research Council of Australia and the Newcastle Permanent Building Society Charitable Trust. The authors declare no conflict of interest.

Scrambled and fried: cigarette smoke exposure causes antral follicle destruction and oocyte dysfunction through oxidative stress.

Cigarette smoke is a reproductive hazard associated with pre-mature reproductive senescence and reduced clinical pregnancy rates in female smokers. Despite an increased awareness of the adverse effects of cigarette smoke exposure on systemic health, many women remain unaware of the adverse effects of cigarette smoke on female fertility. This issue is compounded by our limited understanding of the molecular mechanisms behind cigarette smoke induced infertility. In this study we used a direct nasal exposure mouse model of cigarette smoke-induced chronic obstructive pulmonary disease to characterise mechanisms of cigarette-smoke induced ovotoxicity. Cigarette smoke exposure caused increased levels of primordial follicle depletion, antral follicle oocyte apoptosis and oxidative stress in exposed ovaries, resulting in fewer follicles available for ovulation. Evidence of oxidative stress also persisted in ovulated oocytes which escaped destruction, with increased levels of mitochondrial ROS and lipid peroxidation resulting in reduced fertilisation potential. Microarray analysis of ovarian tissue correlated these insults with a complex mechanism of ovotoxicity involving genes associated with detoxification, inflammation, follicular activation, immune cell mediated apoptosis and membrane organisation. In particular, the phase I detoxifying enzyme cyp2e1 was found to be significantly up-regulated in developing oocytes; an enzyme known to cause molecular bioactivation resulting in oxidative stress. Our results provide a preliminary model of cigarette smoke induced sub-fertility through cyp2e1 bioactivation and oxidative stress, resulting in developing follicle depletion and oocyte dysfunction.

A novel naphthalimide that selectively targets breast cancer via the arylhydrocarbon receptor pathway.

We report that the naphthalimide analogue 2-(2-aminophenyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (NAP-6) is a highly potent and selective breast cancer targeting molecule. These effects are mediated via the aryl hydrocarbon receptor (AHR) pathway and the subsequent induction of CYP1 metabolising monooxygenases in breast cancer cell line models. Indeed the triple negative breast cancer cell line MDA-MB-468 with a GI50 value of 100 nM is greater than 500-fold more sensitive to NAP-6 compared with other tumour derived cell models. Within 1 h exposure of these cells to NAP-6, CYP1A1 expression increases 25-fold, rising to 250-fold by 24 h. A smaller concurrent increase in CYP1A2 and CYP1B1 is also observed. Within 24 h these cells present with DNA damage as evident by enhanced H2AXγ expression, cell cycle checkpoint activation via increased CHK2 expression, S-phase cell cycle arrest and cell death. Specific small molecule inhibitors of the AHR and CYP1 family ameliorate these events. A positive luciferase reporter assay for NAP-6 induced XRE binding further confirms the role of the AHR in this phenomenon. Non-sensitive cell lines fail to show these biological effects. For the first time we identify 2-(2-aminophenyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione as a new AHR ligand that selectively targets breast cancer.

Risk factors associated with clinical signs of vesicular stomatitis and seroconversion without clinical disease in Colorado horses during the 2014 outbreak.

Vesicular stomatitis (VS) is caused by a contagious rhabdovirus that affects horses, cattle, and swine. Clinical signs of vesicular stomatitis virus (VSV) infection in pigs and cattle are indistinguishable from foot-and-mouth disease (FMD), a foreign animal disease and reportable disease in the United States (Rodriguez et al., 2000). A VS epidemic occurred in the Rocky Mountain region in 2014-15. A study was conducted in Colorado to evaluate horse- and management-level factors associated with VS. For a horse to be considered a clinical VS horse, there were two requirements. First, clinical VS horses had to have clinical signs consistent with VS, including one or more of the following: vesicles, ulcers, erosions or crusting on the muzzle, nares, lips, oral or nasal mucosa, ears, ventrum, udder or penile sheath, or coronary band lesions. Second, clinical VS horses had to have laboratory confirmation of VSV exposure via virus isolation from lesions or a positive complement fixation test performed on sera. All non-clinical horses residing on VSV-affected premises enrolled in the study were evaluated for exposure (i.e., seroconversion) to VSV. Overall, management and housing data were collected from 334 horses on 48 premises in Colorado. Approximately one-third (31.4%) of enrolled horses were clinical cases and two-thirds (68.6%) were controls. Three premises-matched logistic regression models were constructed in SAS using backward elimination (P-value < 0.05) after univariate screening of a priori-selected variables (P-value < 0.20). Model outcomes included differences in characteristics and management of 1) clinical and nonclinical horses, 2) exposed and unexposed horses, and 3) exposed nonclinical and unexposed nonclinical horses. Overall, factors most strongly associated with risk of being a VS clinical horse were access to pasture (P-value = 0.002), and pregnancy status (P-value = 0.001). Factors most strongly associated with VSV exposure among horses were access to pasture (P-value = 0.003) and lack of any insect control (P-value = 0.001). The only factor associated with VSV-exposed nonclinical horses compared with unexposed VSV horses was contact with clinical horses (P-value = 0.013). There were no associations identified regarding clinical horses compared with exposed nonclinical horses. With regard to severity of lesions (severe vs. moderate or mild), no variables met the criteria for inclusion in the multivariable model. Results of this study provide evidence that pasture access and fly control are important factors associated with VSV exposure.

Targeted delivery of an ADP-ribosylating bacterial toxin into cancer cells.

The actin cytoskeleton is an attractive target for bacterial toxins. The ADP-ribosyltransferase TccC3 from the insect bacterial pathogen Photorhabdus luminescence modifies actin to force its aggregation. We intended to transport the catalytic part of this toxin preferentially into cancer cells using a toxin transporter (Protective antigen, PA) which was redirected to Epidermal Growth Factor Receptors (EGFR) or to human EGF receptors 2 (HER2), which are overexpressed in several cancer cells. Protective antigen of anthrax toxin forms a pore through which the two catalytic parts (lethal factor and edema factor) or other proteins can be transported into mammalian cells. Here, we used PA as a double mutant (N682A, D683A; mPA) which cannot bind to the two natural anthrax receptors. Each mutated monomer is fused either to EGF or to an affibody directed against the human EGF receptor 2 (HER2). We established a cellular model system composed of two cell lines representing HER2 overexpressing esophageal adenocarcinomas (EACs) and EGFR overexpressing esophageal squamous cell carcinomas (ESCCs). We studied the specificity and efficiency of the re-directed anthrax pore for transport of TccC3 toxin and established Photorhabdus luminescence TccC3 as a toxin suitable for the development of a targeted toxin selectively killing cancer cells.

Small molecule inhibitors of protein kinases in cancer- how to overcome resistance.

Small molecule protein kinase inhibitors show great promise as anti-cancer agents, however, de novo and acquired resistance present problems. These are reviewed and illustrated using the receptor tyrosine kinase, KIT, as an example. Emerging solutions are presented, such as targeting active kinase conformations.

Experimental targeted radioiodide therapy following transfection of the sodium iodide symporter gene: effect on clonogenicity in both two-and three-dimensional models.

To evaluate the potential of the expression of the sodium/iodide symporter (NIS) as a means of targeting radioiodine to tumor cells, we have employed plasmid-mediated transfection of the NIS gene into a range of mammalian cell hosts. We observed perchlorate-inhibitable iodide uptake up to 41-fold over control in all NIS-transfected cells. We assessed the effect of NIS expression followed by exposure to 131I- on the clonogenic survival of UVW glioma cells. After exposure of two-dimensional monolayer cultures of UVW-NIS cells to 131I- at a radioactive concentration of 4 MBq/mL, clonogenic survival was reduced to 21%. Similar treatment of UVW-NIS cells in three-dimensional spheroid cultures resulted in a reduction of clonogenic survival to 2.5%. This increase in sensitivity to 131I- exposure is likely to be due to a radiological bystander effect. These results are very encouraging for the development of a novel cytotoxic gene-therapy strategy in which a radiological bystander effect plays a significant role in tumor cell sterilization.

Pharmacophore development for corticotropin-releasing hormone: new insights into inhibitor activity.

Corticotropin-releasing hormone (CRH) is an endogenous 41-amino acid peptide involved in a wide ranging series of systems including the brain, the coordination of the body's overall response to stress, and more recently as a crucial initiator in the onset of labor, also known as the placental clock. Although more physiological data on CRH is emerging shedding more light on the processes involved and their integration, the mode of action of the hormone and the postulated binding site(s) remain unknown. Recently, a number of small-molecular-weight ligands have emerged as potent antagonists but, as therapeutics, suffer from a lack of solubility. Additionally, despite a number of exhaustively large patents, the lack of structural diversity with these antagonists has enabled little scope for comprehensive and wide ranging studies into the structure of the binding sites of this hormone. As part of a program investigating new, structurally diverse antagonists and agonists of CRH, we have developed a preliminary pharmacophore based on the known small-molecular-weight ligands as an initial step in our program. This pharmacophore was validated by comparison with some of the compounds we postulated to be active.

Small molecule inhibitors of serine/threonine protein phosphatases.

Serine/threonine protein phosphatases have long been ignored as potential therapeutic targets for two reasons, one the biochemical significance of these proteins has not been appreciated and two, many natural protein phosphatase inhibitors are potent toxins and are considered unsuitable for clinical use. This review outlines the biochemical role of this protein family in cancer, cystic fibrosis, immunosuppression and, cardiac and neurological disorders. Particular emphasis is also given to the synthesis of selective small molecule inhibitors and their clinical exploitation.

The prevention of preterm labour -- corticotropin releasing hormone type 1 receptors as a target for drug design and development.

The role of the corticotropin releasing hormone in the onset of labour and the subsequent medicinal chemistry implications of CRH antagonists for the prevention of premature birth, and identification of the CRH type 1 receptor as the target for this drug design, are reviewed here.

Effect of rescuer fatigue on performance of continuous external chest compressions over 3 min.

Guidelines for the performance of cardiopulmonary resuscitation (CPR) have been revised recently and now advocate that chest compressions are performed without interruption for 3 min in patients during asystole and pulseless electrical activity. The aim of the present study was to determine if rescuer fatigue occurs during 3 min of chest compressions and if so, the effects on the rate and quality of compressions. Forty subjects competent in basic life support (BLS) were studied. They performed continuous chest compressions on a Laerdal Skillmeter Resusci-Anne manikin for two consecutive periods of 3 min separated by 30 s. The total number of compressions attempted was well maintained at approximately 100 min(-1) throughout the period of study. However, the number of satisfactory chest compressions performed decreased progressively during resuscitation (P < 0.001) as follows: first min, 82 min(-1); second, 68 min(-1); third, 52 min(-1); fourth, 70 min(-1); fifth, 44 min(-1); sixth, 27 min(-1). We observed significant correlations between the number of satisfactory compressions performed and both height and weight of the rescuer. Female subjects achieved significantly fewer satisfactory compressions compared with males (P = 0.03). Seven subjects (five female, two male) were unable to complete the second 3-min period because of exhaustion. We conclude that rescuer fatigue adversely affects the quality of chest compressions when performed without interruption over a 3-min period and that this effect may be greater in females due to their smaller stature. Consideration should be given to rotating the rescuer performing chest compressions after 1 min intervals.

Dose-response relationship for radiation-induced mutations at micro- and minisatellite loci in human somatic cells in culture.

PURPOSE: The study was designed to determine the dose-response relationship for radiation induction of mutations at mini- and microsatellite loci in human somatic cells. Mutations induced by graded doses of gamma-irradiation were quantified by screening clones derived from single irradiated cells for micro- and minisatellite alterations following irradiation with 1, 2 or 3 Gy. MATERIALS AND METHODS: After irradiation, the moderately radioresistant glioma cell line UVW was seeded at low density into Petri dishes to allow formation of discrete colonies, 100 of which were examined at each dose. All the cells within a colony were presumed to have arisen from a single irradiated cell. Radiation-induced microsatellite alterations were determined at 16 different loci, by PCR amplification and visualization on polyacrylamide gels. Minisatellite alterations were identified at four different minisatellite loci by restriction enzyme digestion and Southern blotting. RESULTS: A dose-response curve for mutation frequency was obtained by analysis of 100 clones, yielding a minisatellite mutation rate of 5.5x10(-3) mutations/locus/Gy/cell and a microsatellite mutation rate of 8.75x10(-4) mutations/locus/ Gy/cell. At microsatellite loci, alterations were predominantly simple loss or gain of repeat units and loss of heterozygosity (LOH). The mutations in minisatellite loci resulted predominantly in LOH and variation in repeat number. The background instability at each locus was determined by analysis of non-irradiated clones. Only 2% and 1% of the micro-and minisatellite loci respectively showed altered bands. CONCLUSIONS: This is the first report of a dose-response relationship for radiation-induced micro- and minisatellite mutations in human somatic cells. Described is a sensitive method for analysis of low-dose radiation mutagenesis in somatic cells that may prove to be a useful tool for radiation protection and dosimetry.

Case-control study of a multistate equine herpesvirus myeloencephalopathy outbreak.

BACKGROUND: A large multistate outbreak of equine herpesvirus myeloencephalopathy (EHM) occurred in May 2011 among horses that participated in a competitive event. OBJECTIVE: To identify EHM risk factors among horses with a common exposure venue. ANIMALS: A total of 123 horses: 19 horses with EHM, 14 equine herpesvirus-1 cases with no reported neurologic signs, and 90 control horses. METHODS: EHM case survey data were compared with data from EHV-1 cases with no neurologic signs and healthy controls using univariable and multivariable methods. RESULTS: Significant factors associated with higher risk for EHM compared with EHV-1 cases with no neurologic signs were (1) greater number of biosecurity risks at the event, (2) female sex, (3) increasing number of classes competed in at the event, and (4) an interaction between sex and number of classes competed in. In the EHM versus controls comparison, in addition to sex and biosecurity risks, factors associated with higher EHM risk included EHV-1 vaccination in the 5 weeks before the event and increasing number of events attended in April 2011; zinc dietary supplementation was associated with decreased risk. An interaction between sex and the number of events attended in April 2011 also was significant. CONCLUSIONS AND CLINICAL IMPORTANCE: Findings from this study suggest that dietary zinc supplementation may be associated with decreased risk of EHM. Several factors were associated with increased risk of EHM. Additional investigations of factors associated with risk of EHM are warranted to evaluate the importance of these factors in this complex disease of horses.