Frequency of respiratory pathogens and SARS-CoV-2 in canine and feline samples submitted for respiratory testing in early 2020.

OBJECTIVES: The emergence of the 2019 novel coronavirus (SARS-CoV-2) has necessitated evaluation of the potential for SARS-CoV-2 infection in dogs and cats. Using a large data set, we evaluated the frequency of SARS-CoV-2 and other respiratory pathogens in samples submitted for respiratory testing from mid-February to mid-April 2020. MATERIALS AND METHODS: A SARS-CoV-2 real-time PCR was developed and validated. A subset of canine and feline samples submitted for respiratory pathogen panel testing to reference laboratories in Asia, Europe, and North America were also tested for SARS-CoV-2. The frequency of respiratory pathogens was compared for the February-April period of 2020 and 2019. RESULTS: Samples from 4616 patients were included in the study and 44% of canine and 69% of feline samples were PCR positive with Mycoplasma cynos and Bordetella bronchiseptica and Mycoplasma felis and feline calicivirus, respectively. No SARS-CoV-2 infections were identified. Positive results for respiratory samples were similar between years. CLINICAL SIGNIFICANCE: The data in this study suggest that during the emergence of the SARS-CoV-2 pandemic in early 2020, respiratory diseases in tested pet cats and dogs were caused by common veterinary pathogens and that SARS-CoV-2 infections in dogs and cats are rare.

Development of a selective androgen receptor modulator for transdermal use in hypogonadal patients.

We have identified a non-steroidal selective androgen receptor modulator (SARM), termed LY305, that is bioavailable through a transdermal route of administration while highly cleared via hepatic metabolism to limit parent compound exposure in the liver. Selection of this compound and its transdermal formulation was based on the optimization of skin absorption properties using both in vitro and in vivo skin models that supported PBPK modeling for human PK predictions. This molecule is an agonist in perineal muscle while being a weak partial agonist in the androgenic tissues such as prostate. When LY305 was tested in animal models of skeletal atrophy it restored the skeletal muscle mass through accelerated repair. In a bone fracture model, LY305 remained osteoprotective in the regenerating tissue and void of deleterious effects. Finally, in a small cohort of healthy volunteers, we assessed the safety and tolerability of LY305 when administered transdermally. LY305 showed a dose-dependent increase in serum exposure and was well tolerated with minimal adverse effects. Notably, there were no statistically significant changes to hematocrit or HDL after 4-week treatment period. Collectively, LY305 represents a first of its kind de novo development of a non-steroidal transdermal SARM with unique properties which could find clinical utility in hypogonadal men.

The effects on cow performance and calf birth and weaning weight of replacing grass silage with brewers grains in a barley straw diet offered to pregnant beef cows of two different breeds.

The effects on cow and calf performance of replacing grass silage with brewers grains in diets based on barley straw and fed to pregnant beef cows are reported. Using a 2 × 2 factorial arrangement of breed and diet, cows pregnant by artificial insemination (n = 34) of two breeds (cross-bred Limousin, n = 19 and pure-bred Luing, n = 15) were fed diets ad libitum which consisted of either (g/kg dry matter) barley straw (664) and grass silage (325; GS) or barley straw (783) and brewers grains (206, BG) and offered as total mixed rations. From gestation day (GD) 168 until 266, individual daily feed intakes were recorded and cow body weight (BW) and body condition score (BCS) measured weekly. Calving date, calf sex, birth and weaning BW, and calf age at weaning were also recorded. Between GD 168 and 266, cross-bred Limousin cows gained more weight than Luing cows (p < 0.05) and cows offered BG gained more weight than cows offered GS (p < 0.001). Luing cows lost more BCS than cross-bred Limousin cows (p < 0.05), but diet did not affect BCS. There were no differences in dry matter intake as a result of breed or diet. Calf birth BW, however, was greater for cows fed BG than GS (44 vs. 38 kg, SEM 1.0, p < 0.001) with no difference between breeds. At weaning, calves born to BG-fed cows were heavier than those born to GS-fed cows (330 vs. 286 kg, SEM 9.3, p < 0.01). In conclusion, replacement of grass silage with brewers grains improved the performance of beef cows and increased calf birth and weaning BW. Further analysis indicated that the superior performance of cows offered the BG diet was most likely due to increases in protein supply which may have improved both energy and protein supply to the foetus.

The pharmacokinetics and pharmacokinetic/pharmacodynamic relationships of evacetrapib administered as monotherapy or in combination with statins.

Evacetrapib is a novel cholesteryl ester transfer protein (CETP) inhibitor currently being evaluated in a late-stage cardiovascular outcome trial. Using population-based models, we analyzed evacetrapib concentration data along with high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) data from a 12-week study in dyslipidemic patients treated with evacetrapib alone or in combination with atorvastatin, simvastatin, or rosuvastatin. Evacetrapib pharmacokinetics were characterized using a two-compartment model with first-order absorption. Evacetrapib exposure increased in a less than dose-proportional manner, similar to other CETP inhibitors. No patient factors had a clinically relevant impact on evacetrapib pharmacokinetics. The relationships between evacetrapib exposure and HDL-C and LDL-C were characterized using Emax models. The theoretical maximal mean HDL-C increase and LDL-C decrease relative to baseline were 177 and 44.1%, respectively. HDL-C change from baseline was found to be negatively correlated with baseline HDL-C. A pharmacologically independent LDL-C reduction was found when evacetrapib was coadministered with statins.CPT Pharmacometrics Syst. Pharmacol. (2014) 3, e94; doi:10.1038/psp.2013.70; published online 22 January 2014.

Evaluation of various static in vitro-in vivo extrapolation models for risk assessment of the CYP3A inhibition potential of an investigational drug.

Nine static models (seven basic and two mechanistic) and their respective cutoff values used for predicting cytochrome P450 3A (CYP3A) inhibition, as recommended by the US Food and Drug Administration and the European Medicines Agency, were evaluated using data from 119 clinical studies with orally administered midazolam as a substrate. Positive predictive error (PPE) and negative predictive error (NPE) rates were used to assess model performance, based on a cutoff of 1.25-fold change in midazolam area under the curve (AUC) by inhibitor. For reversible inhibition, basic models using total or unbound systemic inhibitor concentration [I] had high NPE rates (46-47%), whereas those using intestinal luminal ([I]gut) values had no NPE but a higher PPE. All basic models for time-dependent inhibition had no NPE and reasonable PPE rates (15-18%). Mechanistic static models that incorporate all interaction mechanisms and organ specific [I] values (enterocyte and hepatic inlet) provided a higher predictive precision, a slightly increased NPE, and a reasonable PPE. Various cutoffs for predicting the likelihood of CYP3A inhibition were evaluated for mechanistic models, and a cutoff of 1.25-fold change in midazolam AUC appears appropriate.

Optimal design criteria for discrimination and estimation in nonlinear models.

Nonlinear models are common in pharmacokinetics and pharmacodynamics. To date, most work in design in this area has concentrated on parameter estimation. Here, we introduce the idea of optimization of both estimation and model selection. However, experimental designs that provide powerful discrimination between a pair of competing model structures are rarely efficient in terms of estimating the parameters under each model. Conversely, designs which are efficient for parameter estimation may not provide suitable power to discriminate between the models. Several different methods of addressing both of these objectives simultaneously are introduced in this paper and are compared to an existing optimality criterion.

Optimal crossover designs for logistic regression models in pharmacodynamics.

Pharmacodynamics (PD) is the study of the biochemical and physiological effects of drugs. The construction of optimal designs for dose-ranging trials with multiple periods is considered in this paper, where the outcome of the trial (the effect of the drug) is considered to be a binary response: the success or failure of a drug to bring about a particular change in the subject after a given amount of time. The carryover effect of each dose from one period to the next is assumed to be proportional to the direct effect. It is shown for a logistic regression model that the efficiency of optimal parallel (single-period) or crossover (two-period) design is substantially greater than a balanced design. The optimal designs are also shown to be robust to misspecification of the value of the parameters. Finally, the parallel and crossover designs are combined to provide the experimenter with greater flexibility.

Protective effect of liposome encapsulation on paclitaxel developmental toxicity in the rat.

Paclitaxel is an anticancer drug that has demonstrated severe embryotoxicity in chicks. This embryotoxicity is reduced by liposome encapsulation of the drug. The current study was designed to evaluate the potential of liposome encapsulation for reducing paclitaxel embryotoxicity in rats. Wistar rats were treated with paclitaxel on day 8 of pregnancy (plug = day 0) at doses of 0.67, 2.0, or 10.0 mg/kg intravenously. The same doses of paclitaxel encapsulated in liposomes were administered intravenously to other groups of animals. Control animals were given blank liposomes. Free paclitaxel produced maternal and embryotoxicity at 10.0 mg/kg with three of seven dams dying and resorption of all embryos in surviving dams. Liposome encapsulation at 10.0 mg/kg was not associated with maternal death and there were live fetuses on evaluation at term, although litter size was reduced and malformations occurred in surviving fetuses. At 2.0 mg/kg free paclitaxel, fetal weight was decreased and resorptions increased. Liposome encapsulation at 2.0 mg/kg produced litter results similar to those obtained in control animals given empty liposomes. Malformations were prominent at 2.0 mg/kg free paclitaxel and at 10.0 mg/kg paclitaxel in liposomes and included exencephaly/anencephaly, ventral wall defects, facial clefts, anophthalmia, diaphragmatic hernia, and defects of the kidney, cardiovascular system, and tail. Liposome encapsulation appeared to shift the developmental response to paclitaxel such that 10 mg/kg encapsulated drug produced effects similar to 2.0 mg/kg free drug. These results may have implications for drug delivery of therapeutic agents used during human pregnancy.

Structural anomalies of the reproductive tract.

Structural anomalies of the female reproductive tract may be divided into disorders of lateral fusion and disorders of vertical fusion of the Müllerian duct system. In the past, these disorders were diagnosed at or after menarche or later in life during evaluation of various forms of reproductive tract failure, such as infertility and pregnancy wastage. Newer techniques including pelvic ultrasound and magnetic resonance imaging have allowed diagnosis in the pediatric patient and on occasion, even in utero. Surgical correction of lateral-fusion defects (didelphic, bicornuate, septate, and unicornuate uteri) have remained essentially unchanged except for surgical reconstruction of the septate uterus. Surgical correction of vertical defects (vaginal septi and cervical agenesis or dysgenesis) has received considerable recent interest with the development of newer techniques that may be more effective than the reconstructive procedures of the past.

Changes in mitochondrial and microsomal 3 beta-hydroxysteroid dehydrogenase activity in mouse ovary over the course of the estrous cycle.

3 beta-Hydroxysteroid dehydrogenase (HSD) is located in the endoplasmic reticulum and mitochondria. To determine whether the separate enzymes play different roles in steroidogenesis, the specific activity (SA) of both were measured at four different stages of the mouse estrous cycle. Microsomal HSD activity changed little throughout, averaging 8.7 +/- 0.7 nmol progesterone/min/mg protein. In contrast, mitochondrial HSD activity changed dramatically at diestrus, increasing to 14.4 nmol progesterone/min/mg protein. When measured at proestrus, estrus, and metestrus, mitochondrial HSD activity was 5.5, 7.4, and 4.5 nmol progesterone/min/mg protein, respectively. To ascertain whether the increase in mitochondrial HSD activity at diestrus could be due to a preferential induction of enzyme, its SA and the SA of a mitochondrial inner membrane enzyme, cytochrome C oxidase, were compared to the SA of a mitochondrial outer membrane enzyme, rotenone-insensitive NADH cytochrome C reductase. The SA of all three enzymes changed proportionally at diestrus, suggesting that the increase in mitochondrial HSD activity was not due to its preferential induction. Rather, we believe that the HSD activity in the mitochondrial fraction, as measured at the four stages of the estrous cycle, is a reflection of the combined contributions from an ever changing population of ovarian cells. Mitochondria from luteal cells have the highest HSD activity, and are very likely responsible for the major synthesis of progesterone during the luteal phase.