Male Beagle Dogs Research Articles

Nonclinical Safety Pharmacology Study of the Herbal Product HAD-B1.

HAD-B1 is a Korean herbal formula designed to treat solid tumors, and through cell experiments, it has proven to have an anticancer effect. The current study aims to test the safety of HAD-B1. This experiment is under the regulation of ICH. In order to find if HAD-B1 has any effect on the CNS, 0, 500, 1000, and 2000 mg/kg/day of HAD-B1 were orally administered to male and female rats once. To discover any effect on the respiratory system, 0, 500, 1000, and 2000 mg/kg/day of HAD-B1 were orally given to male rats followed by measuring the respiratory rate, tidal volume, and minute respiratory volume. To assess the possibility of a delayed QT period as a result of the drug administration, hERG analysis was conducted at 0, 0.1, 0.3, and 1 μg/ml. To assess any effect on the cardiovascular system, 0, 500, 1000, and 2000 mg/kg/day of HAD-B1 were orally given to male beagle dogs once followed by temperature, blood pressure, ECG, and heart rate analyses. There were no clinically significant changes in both male and female rats on assessing any effects on the CNS. There were no clinically significant changes in male rats' respiratory assessment. There were no clinically significant changes in hERG analysis results. There were no clinically significant changes in the cardiovascular system of male beagle dogs. Our results demonstrate that HAD-B1 is a safe herbal formula that does not have a clinically significant effect on the CNS, respiratory, and cardiovascular systems.

Dietary ω-3 fatty acids reduced atrial fibrillation vulnerability via attenuating myocardial endoplasmic reticulum stress and inflammation in a canine model of atrial fibrillation

BackgroundDietary consumption of ω-3 fatty acids is correlated with a reduced incidence of cardiovascular events. Here, we investigated the effect of dietary ω-3 fatty acids on atrial fibrillation (AF) vulnerability in a canine model of AF and explored the related mechanisms. MethodsTwenty four male beagle dogs (weight, 8-10 kg) were randomly divided into four groups: (a) sham-operated group (normal chow); (b) AF+FO [AF and normal chow supplemented with fish oil (FO): 0.6 g n-3 polyunsaturated fatty acids (ω-3 PUFA) /kg/day]; (c) AF group (normal chow); (d) sham-operated FO group (chow supplemented with FO: 0.6 g ω-3 PUFA/kg/day). AF was induced by rapid atrial pacing (RAP: 400 bpm for 4 weeks). Daily oral administration of FO was initiated 1 week before surgery and continued for 4 weeks post operation. ResultsAtrial electric remodeling was significantly attenuated and AF vulnerability were significantly reduced in AF+FO group compared to AF group. Endoplasmic reticulum (ER) stress-related protein expression levels of glucose-regulated protein78, C/EBP homologous protein, cleaved-Caspase12, and phosphorylation of protein kinase R-like ER kinase as well as inflammatory cytokines interleukin-1β, interleukin-6, tumor necrosis factor-α in left atrium (LA) were significantly downregulated in AF+FO group than in AF group (all p<0.05). In addition, Masson staining revealed lower extent of LA interstitial fibrosis in AF+FO group than in AF group (p<0.01). Myocardial apoptosis was also significantly reduced in AF+FO group than in AF group (p<0.05). ConclusionsDietary ω-3 fatty acids could significantly reduce RAP-induced AF vulnerability, possibly via attenuating myocardial ER stress, inflammation, and apoptosis in this canine model of AF.

Effect of fentanyl, with or without treatment of bradycardia, on the minimum alveolar concentration of isoflurane and cardiovascular function in dogs

ObjectiveTo determine the effect of fentanyl on the minimum alveolar concentration of isoflurane (MACISO) and cardiovascular variables in dogs, and how the treatment of bradycardia affects them. Study designProspective, randomized crossover-controlled trial. AnimalsA total of six male Beagle dogs weighing 9.9 ± 0.7 kg (mean ± standard deviation) and aged 13 months. MethodsTo each dog, two treatments were assigned on different days: fentanyl (FENTA) or fentanyl plus glycopyrrolate (FENTAglyco) to maintain heart rate (HR) between 100 and 132 beats minute−1. Determinations of MACISO were performed with 10 plasma fentanyl target concentrations ([Fenta]Target (0, 0.16, 0.32, 0.64, 1.25, 2.5, 5.0, 10.0, 20.0 and 40.0 ng mL−1) for FENTA and 5 [Fenta]Target (0, 1.25, 2.5, 5.0, 10.0 ng mL−1)) for FENTAglyco. During each MACISO determination, cardiovascular variables [mean arterial pressure (MAP), HR and cardiac index (CI)] were measured, and systemic vascular resistance index (SVRI) calculated. Pharmacodynamic models were used to describe the plasma fentanyl concentration [Fenta]–response relationship for the effect on MACISO and cardiovascular variables. A mixed-model analysis of variance followed by Dunnett’s or Tukey’s test, and the Bonferroni adjustment were used for comparisons within and between each treatment, respectively. Significance was set as p < 0.05. ResultsFentanyl decreased MACISO by a maximum of 84%. The [Fenta] producing 50% decrease in MAC, HR and CI were 2.64, 3.65 and 4.30 ng mL−1 (typical values of population model), respectively. The prevention of fentanyl-mediated bradycardia caused no significant effect on MACISO, but increased HR, MAP and CI, and decreased SVRI when compared with isoflurane alone. Conclusions and clinical relevanceFentanyl caused a plasma concentration-dependent decrease in MACISO, HR and CI and an increase in SVRI. Cardiovascular improvements associated with fentanyl in isoflurane-anesthetized dogs only occurred when the fentanyl-mediated bradycardia was prevented.

Effects of bronchial thermoplasty and cryoablation on airway smooth muscle.

Background:The effectiveness of bronchial thermoplasty (BT) has been reported in patients with severe asthma. This study compared the effects of BT and cryoballoon ablation (CBA) therapy on the airway smooth muscle (ASM).Methods:Eight healthy male beagle dogs were included in this experiment. In the preliminary experiment, one dog received BT treatment for both lower lobe bronchus, another dog received CBA treatment for 7 s on the upper and lower lobe of right bronchus, and 30 s on the left upper and lower lobe. The treatments were performed twice at an interval of 1 month. In subsequent experiments, the right lower lobe bronchus was treated with BT, and the left lower lobe bronchus was treated with CBA. The effects of treatment were observed after 1 (n = 3) month and 6 months (n = 3). Hematoxylin-eosin staining, Masson trichrome staining, and immunohistochemical staining were used to compare the effects of BT and CBA therapy on the ASM thickness, collagen fibers synthesis, and M3 receptor expression after treatment. One-way analysis of variance with Dunnett post hoc test was used to analyze the differences among groups.Results:In the preliminary experiment, the ASM ablation effect of 30-s CBA was equivalent to that of 7-s CBA (ASM thickness: 30.52 ± 7.75 μm vs. 17.57 ± 15.20 μm, P = 0.128), but the bronchial mucociliary epithelium did not recover, and large numbers of inflammatory cells had infiltrated the mucosal epithelium at 1-month post-CBA with 30-s freezing. Therefore, we chose 7 s as the CBA treatment time in our follow-up experiments. Compared with the control group (35.81 ± 11.02 μm), BT group and CBA group (13.41 ± 4.40 μm and 4.81 ± 4.44 μm, respectively) had significantly decreased ASM thickness after 1 month (P < 0.001). Furthermore, the ASM thickness was significantly lower in the 1-month post-CBA group than in the 1-month post-BT group (P = 0.015). There was no significant difference in ASM thickness between the BT and CBA groups after six months (9.92 ± 4.42 μm vs. 7.41 ± 7.20 μm, P = 0.540). Compared with the control group (0.161 ± 0.013), the average optical density of the ASM M3 receptor was significantly decreased in 6-month post-BT, 1-month post-CBA, and 6-month post-CBA groups (0.070 ± 0.022, 0.072 ± 0.012, 0.074 ± 0.008, respectively; all P < 0.001). There was no significant difference in the average optical density of ASM M3 receptor between the BT and CBA therapy groups after six months (P = 0.613).Conclusions:CBA therapy effectively ablates the ASM, and its ablation effect is equivalent to that of BT with a shorter onset time. A neural mechanism is involved in both BT and CBA therapy.

Pre-Clinical Pharmacokinetic Characterization, Tissue Distribution, and Excretion Studies of Novel Edaravone Oral Prodrug, TEJ-1704.

Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) is a free radical scavenger approved for the treatment of amyotrophic lateral sclerosis, a fatal neuromuscular disease. Edaravone is administered as an intravenous infusion over 60 min for several treatment cycles. To ease the burden of patients and caregivers, the oral formulation of edaravone has been developed. The purpose of this study was to evaluate pharmacokinetics and tissue distribution of TEJ-1704, an edaravone oral prodrug, in male Sprague Dawley rats and beagle dogs. Animal experiments were conducted using Sprague Dawley rats and beagle dogs to evaluate pharmacokinetics, tissue distribution, and excretion of TEJ-1704. Blood, tissues, cerebrospinal fluid, urine, and feces samples were collected at designated sampling time after intravenous (IV) or oral (PO) administration of edaravone or TEJ-1704. A modified bioanalysis method was developed to quantify edaravone in samples including plasma, tissues, cerebrospinal fluid, urine, and feces. The bioanalysis method was validated and successfully applied to pharmacokinetics, tissue distribution, and excretion studies of the novel edaravone prodrug. Although plasma Cmax of TEJ-1704 was low, groups administered with TEJ-1704 had high AUCinf, suggesting continuous metabolism of TEJ-1704 into edaravone. Groups treated with TEJ-1704 also showed lower CSF distribution than the control groups. After the administration of TEJ-1704, the majority of edaravone was distributed to the heart, lung, and kidney. It was excreted equally via urine and feces. The pharmacokinetics, tissue distribution, and excretion of TEJ-1704, a novel edaravone oral prodrug, were successfully characterized. Additional studies are needed to fully understand the difference between TEJ-1704 and edaravone and determine the potency of TEJ-1704.

Buccal Bone Remodeling Around Immediate Implants in STZ-Induced Diabetic Dogs: A Histologic and Microcomputed Tomographic Analysis.

Buccal bone remodeling around immediate implants placed in animals with streptozotocin (STZ)-induced diabetes has not been investigated. The present histologic and microcomputed tomography (μCT) in vivo experiment assessed the buccal bone remodeling around immediate implants, extraction socket healing, and bone-to-implant contact (BIC) in dogs with and without STZ-induced diabetes. Three male beagle dogs with STZ-induced diabetes and three healthy dogs (controls) were included. Fasting blood glucose levels were measured using a glucometer. Under general anesthesia, all animals underwent atraumatic tooth extraction of bilateral maxillary and mandibular second premolar teeth using Piezosurgery and immediate implant placement in the distal root socket. Primary closure was achieved for all implants and adjacent socket sites. After 11 months, all animals were sacrificed, and buccal bone thickness (BBT), marginal bone loss (MBL), BIC, and mesial extraction socket bone volume were assessed in control and diabetic animals using histologic and μCT examination. High-resolution μCT analyses were performed to identify the percentage of osteocytes and blood vessels in bone specimens harvested from a mesial extraction socket in each group. BBT and BIC were significantly higher in control dogs (P < .05), and diabetic dogs demonstrated significantly more MBL than control dogs (P < .05). Compared to healthy subjects, the sockets of induced diabetic dogs lost over 50% of bone horizontally and vertically. Control group had more significant osteocytes (38.85%) and blood vessels (37.87%), whereas the same values for STZ-induced diabetic dogs were 27.92% and 27.76%, respectively. Buccal bone loss and MBL were significantly higher around immediate implants placed in dogs with STZ-induced diabetes and were associated with multiple implant buccal thread exposure. A large percentage of socket space in diabetic dogs healed with nonosseous structure following tooth extraction.

Effect of Injectable-Platelet Rich Fibrin on marginal adaptation of Bioactive Materials Used as Direct Pulp Capping; An Experimental Animal Study

ABSTRACTIntroduction: Vital pulp therapy has been known as one of the treatment optionsto preserve pulp vitality after being exposed by trauma or caries. Aim: This experimentexplored the effect of injectable-Platelet Rich Fibrin on marginal adaptation of two pulp capping agents (Mineral Trioxide Aggregate and Bioactive Bone Graft). Materials andmethods: A total of 64 teeth were used out of 8 healthy male beagle dogs. The teethwere randomly assigned into four groups, they were exposed and capped with differentcapping agents. Group A; capped with Mineral Trioxide Aggregate (MTA), Group B;capped with MTA+ i-PRF, Group C; capped with Bioactive Bone Graft (BBG), Group D; capped with BBG+i-PRF. Finally the access cavity was restored with IntermediateRestorative Material (IRM). At each predetermined interval, the dogs were sacrificed(1 month, and 3 months). The samples were then prepared for electron microscopicscanning evaluation. To compare between the gap percentage of four groups at eachinterval, Kruskal-wallis test; was used. Mann-Whitney U test; was used to pair-wisecomparison when Kruskal-wallis test is significant. Bonferroni’s correction was utilizedfor the pair-wise comparisons. Statistical significance was considered at P area between the capping materials and dentin, followed by group C (BBG), with theleast value recorded in group A (MTA). Conclusion: the findings from the current studysuggested that i-PRF provided a better marginal adaptation of either MTA or BBG tothe pulp and dentin, which improved with time from one month to three months.

Safety and toxicology assessment of sodium nitrite administered by intramuscular injection

Intramuscular (IM) injection of nitrite (1-10 mg/kg) confers survival benefit and protects against lung injury after exposure to chlorine gas in preclinical models. Herein, we evaluated safety/toxicity parameters after single, and repeated (once daily for 7 days) IM injection of nitrite in male and female Sprague Dawley rats and Beagle dogs. The repeat dose studies were performed in compliance with the Federal Drug Administration's (FDA) Good Laboratory Practices Code of Federal Regulations (21 CFR Part 58). Parameters evaluated consisted of survival, clinical observations, body weights, clinical pathology, plasma drug levels, methemoglobin and macroscopic and microscopic pathology. In rats and dogs, single doses of ≥100 mg/kg and 60 mg/kg resulted in death and moribundity, while repeated administration of ≤30 or ≤ 10 mg/kg/day, respectively, was well tolerated. Therefore, the maximum tolerated dose following repeated administration in rats and dogs were determined to be 30 mg/kg/day and 10 mg/kg/day, respectively. Effects at doses below the maximum tolerated dose (MTD) were limited to emesis (in dogs only) and methemoglobinemia (in both species) with clinical signs (e.g. blue discoloration of lips) being dose-dependent, transient and reversible. These signs were not considered adverse, therefore the No Observed Adverse Effect Level (NOAEL) for both rats and dogs was 10 mg/kg/day in males (highest dose tested for dogs), and 3 mg/kg/day in females. Toxicokinetic assessment of plasma nitrite showed no difference between male and females, with Cmax occurring between 5 mins and 0.5 h (rats) or 0.25 h (dogs). In summary, IM nitrite was well tolerated in rats and dogs at doses previously shown to confer protection against chlorine gas toxicity.

Effects of Interval Exercise Training on Serum Biochemistry and Bone Mineral Density in Dogs.

Simple SummaryIn this study, six male beagle dogs underwent 12 weeks of interval exercise following the Frequency, Intensity, Time/duration, Type, Volume, and Progression (FITT-VP) training principle. The heart rate (HR) response was measured during the entire exercise period, and changes in bone mineral density (BMD), muscle volume (MV), and hematology and serum biomarkers were evaluated at the pre-exercise training period and post-exercise training period. We showed that exercise training increased BMD in the femur and serum total alkaline phosphatase (TALP), aspartate aminotransferase, and creatine kinase levels. In addition, our data suggest a positive correlation between BMD and TALP, demonstrating that increased TALP might be an important contributing factor for enhancing BMD with physical training in dogs.Exercise has been suggested as a powerful intervention for health care and fitness management in humans; however, few studies have demonstrated the benefits of exercise training in dogs. The purpose of this study was to examine the effects of exercise training on heart rate (HR), bone mineral density (BMD), muscle volume (MV), and hematological and serum biomarkers in dogs. Six healthy beagles completed the interval treadmill exercise, developed on the basis of the FITT principle, two times a week for 12 weeks. To evaluate the physiological parameters, the HR values were analyzed using the Polar H10 system during the entire exercise period. At pre-and post-exercise, quantitative computed tomography and hematological and serum biochemical parameters were analyzed. The interval exercise resulted in a normal HR response and no adverse behavioral or physiological effects on the dogs. We showed that exercise improved BMD in the femur (541.6 ± 16.7 vs. 610.2 ± 27.8 HA, p < 0.01) and increased serum total alkaline phosphatase (TALP; 68.6 ± 9.2 vs. 81.3 ± 17.2, p < 0.01), aspartate aminotransferase (23.5 ± 1.0 vs. 33.5 ± 1.6, p < 0.01), and creatine kinase (114.8 ± 5.3 vs. 214.0 ± 20.8, p < 0.01) levels. There was a positive relationship between BMD and TALP (femur: r = 0.760, p = 0.004; vertebrae: r = 0.637; p = 0.025). Our findings suggest that interval exercise training is beneficial to increase BMD in the femur, and an increased TALP level would be a concomitant mechanism for enhancing BMD with exercise in dogs.

Microencapsulated phytogenic in dog feed modulates immune responses, oxidative status and reduces bacterial (Salmonella and Escherichia coli) counts in feces

Improving the health and immunity of pets is a concern of the guardians, while maintaining the health of the animals directly impacts the owners' health and public health. The objective of this study was to produce a dog feed containing a microencapsulated phytogenic blend, as well as determine the impact of the additive on animal health and its intestinal microbiota. The composition included microencapsulated thymol, carvacrol, and cinnamaldehyde, at 300 mg/kg of feed. Ten male beagle dogs were divided into two groups, identified as follows: the control group (C; ingested the feed without the additive) and the treated group (T; consumed feed containing the phytogenic blend). The dogs received 300 g of feed/day divided into two meals. Greater neutrophil counts in group T and lymphocyte counts were observed at the end of the experiment, as well as levels of α2-globulins and γ-globulin, while β1 and β2-globulins levels were lower in group T. Serum nitrogen oxide levels were higher in group T. Levels of reactive oxygen species were lower in group T at days 30 and 45, unlike activity of glutathione peroxidase that was higher. We found less bacterial contamination in the feces of dogs in group T, i.e., total bacterial count, total coliform counts, and counts of Salmonella and Escherichia coli were lower on days 30 and 45. Phytogenic blend intake reduces bacterial counts in stool and improves antioxidant/oxidative status and immune responses.

Peri-implant tissue augmentation by volume-stable collagen matrix transplantation: a study of dog mandibles.

The aim of this study was to investigate histologically the amount of peri-implant tissue augmentation after volume-stable porcine collagen matrix transplantation. Six male beagle dogs were used in the experiment. P2, P4, and M1 distal roots were extracted under general anesthesia. After 6months, implants were placed in the same sites, and volume-stable porcine collagen matrix transplantation was performed. Impressions were taken at 1 and 2weeks and at 1, 2, and 3months after transplantation. The dogs were euthanized at 3months, and their mandibles were removed and scanned using micro-computed tomography. Standard Triangulated Language data were also obtained. Using preoperative models as a reference, the data for all time points were compared, and changes in the thickness of the cross-section of the implant sites were measured. The model created at 3months was then compared with the mandible data, and the thickness of collected peri-implant soft tissue was measured under optical microscopy. Increased thickness was found at some of the sites on the buccal side. Regarding the peri-implant soft tissue, the thickness of the measured sites on the buccal side was significantly increased at 3months in the experimental group. Histological observations of the internal structures of the tissue in the experimental group revealed irregular collagen fibers and a remnant collagen matrix. Endogenous tissue was observed within the collagen matrix, indicating good fusion with the surrounding autologous tissue. These results suggest that volume-stable porcine collagen matrix transplantation promotes peri-implant tissue augmentation on the buccal side.

Influence of long-stay jugular catheters on hemostatic variables in healthy dogs.

To compare hemostatic variables performed on blood samples obtained from indwelling jugular catheters or direct venipuncture over a 72-hour period. Prospective experimental study. University research laboratory. Five healthy neutered male purpose-bred Beagle dogs. Each dog was sedated to facilitate placement of a long-stay 20-Ga polyurethane IV catheter into the jugular vein. Blood samples were obtained from the preplaced catheters at 4 time points corresponding to 0, 24, 48, and 72hours relative to placement. Blood samples were also obtained by direct venipuncture of a peripheral vein using a 21-Ga butterfly catheter and evacuated blood tubes at the same time points. Platelet count, platelet closure time, prothrombin time, activated partial thromboplastin time, fibrinogen, and kaolin-activated thromboelastography were performed on these paired samples at each time point. The patency of the indwelling catheters was maintained by flushing every 6hours with heparinized saline. No significant differences were identified in any of the hemostatic variables obtained by either blood collection technique at any time point during the study (P>0.05). There was also no significant day-to-day variation in any catheter-derived hemostatic variable obtained from individual dogs identified over the course of the study. These data suggest that accurate hemostatic variables may be obtained using blood collected from indwelling jugular catheters, maintained with heparinized saline for at least 72hours, in healthy dogs.

Replacement of a 5-cm Intrathoracic Trachea With a Tissue-Engineered Prosthesis in a Canine Model

BackgroundCritical obstacles must be addressed before clinical application of artificial tracheas. The major complications of long tracheal replacement include anastomotic dehiscence and stenosis owing to poor vascularity and incomplete reepithelialization. The objective of this report was to clarify whether preincubation of the prosthesis in the omentum could be applicable for reconstruction of a long segment of the intrathoracic trachea in a canine model. MethodsThe framework of an artificial trachea was fabricated from a polypropylene mesh tube and coated with 1% neutral atelocollagen inside and outside the lumen. The prosthesis was placed in the omentum of 9 healthy male beagle dogs for 3 weeks. Then, the pedicled prosthesis was used to replace a 50-mm-long section of intrathoracic trachea. Results were evaluated bronchoscopically, macroscopically, and histologically. ResultsAfter 3 weeks of abdominal incubation, the prostheses were incorporated into the host tissue. None of the dogs showed dehiscence of the anastomosis or infection of the prostheses during the postoperative period. Seven of the 9 dogs survived for more than 1 year. One dog died of a bowel obstruction resulting from a diaphragmatic hernia 3 months after replacement, and another died due to reasons unrelated to the prosthesis at 6 months. Bronchoscopic examination revealed no stenosis or dehiscence, and microscopic examination of all dogs showed that the luminal surface was covered by newly regenerated connective tissue and respiratory epithelium. ConclusionsPedicled omentum-prosthesis complexes may allow successful reconstruction of a long segment of the intrathoracic trachea.

Safety Pharmacology Study of ET-26 Hydrochloride, a Potential Drug for Intravenous General Anesthesia, in Rats and Beagle Dogs.

Background: ET-26 hydrochloride (ET-26HCl), a class 1 new drug, was developed to reserve the advantages of etomidate with a mild adrenocortical inhibition. Purpose: this study was to evaluate the potential adverse effects on the cardiovascular system of beagle dogs and the respiratory and central nervous systems of rats. Methods: three established methods, the whole-body plethysmography for respiratory function, the prototype telemetry transmitter for cardiovascular function, and the standardized functional observational battery for central nervous system function, were accomplished with Good Laboratory Practice standards. Results: no significant difference in the tidal volume, but the respiratory rate and minute ventilation were reduced. The degree of inhibition was the most serious in the first 15 min after dosing and function fully recovered after 1 h. For male rats, the respiratory rate of male rats was reduced significantly at 15 min after injection with ET-26HCl (4 mg/kg, 28.6%, p ≤ 0.01; 8 mg/kg, 24.5%, p ≤ 0.01; 16 mg/kg, 44.5%, p ≤ 0.001), and the minute ventilation at 15 min was decreased by 20.1% (4 mg/kg, p = 0.034), 22.2% (8 mg/kg, p = 0.019), and 44.6% (16 mg/kg, p ≤ 0.001) as compared to control group. As with male rats, the respiratory rate of the female rats was reduced significantly at 15 min (4 mg/kg, 23.3%, p ≤ 0.01; 8 mg/kg, 29.2%, p ≤ 0.001; 16 mg/kg, 44.1%, p ≤ 0.001), and the minute ventilation was decreased by 25.2% (4 mg/kg, p ≤ 0.001), 23.0% (8 mg/kg, p ≤ 0.01), and 47.6% (16 mg/kg, p ≤ 0.001). Then, all the variations in cardiovascular functions were within the expected range for normal biological variation, we concluded that ET-26HCl, even at 10-fold ED50, still does not exert toxicological effects on the cardiovascular system. For male beagle dogs, the systolic blood pressure after 24 h following administration of vehicle control or 8, 12, or 16 mg/kg ET-26HCl was 137.80 ± 5.55, 131.76 ± 10.03, 139.88 ± 8.35, and 141.28 ± 8.75 mmHg, respectively. The diastolic blood pressure was 71.16 ± 4.84, 66.52 ± 8.50, 73.64 ± 8.51, and 74.24 ± 8.68 mmHg, respectively. For female beagle dogs, the systolic blood pressure after 24 h following administration of vehicle control or 8, 12, or 16 mg/kg ET-26HCl was 128.28 ± 5.22, 124.76 ± 7.29, 134.88 ± 5.56, and 135.36 ± 8.72 mmHg, respectively. The diastolic blood pressure was 67.00 ± 4.10, 62.12 ± 7.87, 69.44 ± 6.40, and 70.20 ± 8.42 mmHg, respectively. In central nervous system function experiment, all the changes observed in the functional observational battery tests, including motor activity, behavior, coordination, and sensory and motor reflex responses, and reduced body temperature, were resulted in general anesthesia effect of ET-26HCl. Conclusion: ET-26HCl exerts mild, reversible effects on respiratory, cardiovascular, and central nervous system function as verified by standard in vivo animal models.

Pharmacokinetics of a high-concentration formulation of buprenorphine (Simbadol) in male dogs

ObjectiveTo describe the pharmacokinetics of buprenorphine in dogs following administration of a high-concentration formulation of buprenorphine. Study designProspective, randomized, crossover study. AnimalsA total of six healthy male intact Beagle dogs, aged 9–13 months and weighing 10.3 ± 1.4 kg (mean ± standard deviation). MethodsDogs were randomized to be administered buprenorphine (0.12 mg kg–1; Simbadol, 1.8 mg mL–1) via the intravenous (lateral saphenous) or subcutaneous (dorsal interscapular) route followed by the alternative route of administration after a 14 day interval. Blood was sampled before administration and at set times up to 72 hours after injection. Plasma buprenorphine concentration was measured using liquid chromatography–tandem mass spectrometry. ResultsA three-compartment model with zero or biphasic rapid and slow first-order input in (intravenous or subcutaneous data, respectively) and first-order elimination from the central compartment best fitted the data. The rapid first-order input accounted for 63% of the dosage absorption. Typical values (% interindividual variability) for the three compartment volumes were 900 (33), 2425 (not estimated) and 6360 (28) mL kg–1. The metabolic and two distribution clearances were 25.7 (21), 107.5 (74) and 5.7 (61) mL minute–1 kg–1. The absorption half-life for the fast absorption phase was 8.9 minutes with a 0.7 (103) minute delay. The absorption half-life for the slow absorption phase was 347 minutes with a 226 (42) minute delay. Median (range) bioavailability calculated from noncompartmental analysis was 143 (80–239)%. Calculated terminal half-life was 963 minutes. Conclusions and clinical relevanceThe high-concentration formulation of buprenorphine administered subcutaneously had a large volume of distribution and a rapid absorption phase followed by slower, delayed absorption. The high estimate of bioavailability should be interpreted with caution as values above 100% are most commonly related to experimental issues.

Non-viral derivation of a transgene-free induced pluripotent stem cell line from a male beagle dog

We previously reported the non-viral derivation of transgene-free induced pluripotent stem cells (iPSCs) from somatic fibroblasts of a female beagle dog using an optimized induction medium and integration-free episomal vectors. Here, we report novel derivation of a male canine iPSC line OF35Y-iPS, which showed standard characteristics of pluripotency such as a strong gene expression profile of pluripotency markers, differentiation potential into all three germ layers, and normal karyotype (78XY). Furthermore, we demonstrated targeted integration of 2A-EGFP into the canine NANOS3 locus. The novel iPSC line would be a useful resource for stem cell research and regenerative veterinary medicine.

Melt extrusion deposition (MED™) 3D printing technology – A paradigm shift in design and development of modified release drug products

Three-dimensional printing (3DP) technology offers unique advantages for pharmaceutical applications. However, most of current 3D printing methods and instrumentations are not specifically designed and developed for pharmaceutical applications. To meet the needs in pharmaceutical applications for precision, compatibility with a wide range of pharmaceutical excipients and drug materials without additional processing, high throughput and GMP compliance, an extrusion-based 3D printer based on Melt Extrusion Deposition (MED™) 3D printing technology was developed in this study. This technology can process powder pharmaceutical excipients and drugs directly without the need of preparing filament as required by FDM 3D printing. Six different tablet designs based on compartment models were used to demonstrate the precision and reproducibility of this technology. The designed tablets were fabricated using the GMP-compliant MED™ 3D printer and were evaluated in vitro for drug release and in vivo for selected designs using male beagle dogs. Tablet designs with one or more compartments showed versatile release characteristics in modulating the release onset time, release kinetics, duration of release and mode of release. Multiple drugs or formulations were fabricated into a single tablet to achieve independent release kinetics for each drug or to fine-tune the pharmacokinetic profile of a drug. Building upon the theoretical analysis of models, precision and reproducibility of MED™ 3D printing technology, a novel product development approach, 3D printing formulation by design (3DPFbD®) was developed to provide an efficient tool for fast and efficient pharmaceutical product development. The MED™ 3D printing represents a novel and promising technology platform encompassing design and development of modified drug release products and has potential to impact the drug delivery and pharmaceutical product development.

Laryngeal function in normal dogs administered isoflurane following partial clearance of alfaxalone or propofol

ObjectiveTo assess laryngeal function in normal dogs administered isoflurane following partial clearance of alfaxalone or propofol. Study designRandomized experimental crossover study. AnimalsA group of 12 purpose-bred, male Beagle dogs. MethodsDogs were randomly assigned to one of two treatments: alfaxalone–isoflurane (ALF-ISO) or propofol–isoflurane (PRO-ISO) and anesthetized for three video laryngoscopy examinations. The alternate treatment occurred after ≥ 14 days interval. Examinations were performed after induction of anesthesia (LS-A), after 20 minutes of breathing isoflurane via a facemask (LS-B) and after a further 20 minutes of isoflurane (LS-C). Parameters of objective laryngeal function included inspiratory rima glottidis surface area (RGSA-I), expiratory rima glottidis surface area (RGSA-E) and % RGSA increase, calculated from three consecutive respiratory cycles in the final 15 seconds of each video laryngoscopy examination. The % RGSA increase was calculated using [(RGSA-I – RGSA-E)/RGSA-E] × 100. Subjective laryngeal function was evaluated independently by two experienced surgeons blinded to treatment. ResultsThe % RGSA increase within each treatment was greater for LS-B and LS-C than for LS-A (ALF-ISO: p = 0.03, PRO-ISO: p = < 0.001). There was no difference within each treatment from LS-B compared with LS-C. RGSA-I increased within each treatment from LS-A to both LS-B and LS-C (ALF-ISO: p = 0.002) and to LS-C (PRO-ISO: p = 0.006). Subjective laryngeal function scores improved from LS-A to LS-C. Conclusions and clinical relevanceLaryngeal function improved from postinduction examination following either 20 or 40 minutes of anesthesia with isoflurane via facemask. This study demonstrates that isoflurane may have a lesser effect on arytenoid abduction activity compared with more commonly used intravenous induction anesthetics (alfaxalone and propofol).

Automated Blood Sampling in a Canine Telemetry Cardiovascular Model.

Successful implementation of automated blood sampling (ABS) into a telemetry instrumented canine cardiovascular model provides simultaneous cardiovascular assessment of novel compounds while collecting multiple blood samples for analysis of drug level, cytokines, and biomarkers. Purpose-bred male Beagle dogs (n = 36) were instrumented with a dual-pressure telemetry transmitter and vascular access port. Modifications to acclimation practices, surgical procedures, and housing were required for implementation of ABS in our established cardiovascular canine telemetry colony. These modifications have increased the use and reproducibility of the model by combining early pharmacokinetic and cardiovascular studies, thus achieving both refinement and reduction from a 3R perspective. In addition, the modified model can shorten timelines and reduce the compound requirement in early stages of drug development. This telemetry-ABS model provides an efficient means to quickly identify potential effects on key cardiovascular parameters in a large animal species and to obtain a more complete pharmacokinetic-pharmacodynamic profile for discovery compounds.

Entelon (vitis vinifera seed extract) reduces degenerative changes in bovine pericardium valve leaflet in a dog intravascular implant model.

Inflammation and calcification are major factors responsible for degeneration of bioprosthetic valve and other substitute heart valve implantations. The objective of this study was to evaluate the anti-inflammatory and anti-calcification effects of Entelon150® (consisting of grape-seed extract) in a beagle dog model of intravascular bovine pericardium implantation. In total, 8 healthy male beagle dogs were implanted with a bovine pericardium bilaterally in the external jugular veins and divided into two groups. Animals in the Entelon150® group (n = 4) were treated with 150 mg of Entelon150® twice daily for six weeks after surgery. The negative control (NC) group (n = 4) was treated with 5 ml of saline using the same method. After six weeks, we measured the calcium content, performed histological examination, and performed molecular analysis. The calcium content of implanted tissue in the Entelon150® group (0.56±0.14 mg/g) was significantly lower than that in the NC group (1.48±0.57 mg/g) (p < 0.05). Histopathological examination showed that infiltration of chronic inflammatory cells, such as fibroblasts and macrophages, occurred around the graft in all groups; however, the inflammation level of the implanted tissue in the Entelon150® group was s lower than that in the NC group. Both immunohistochemical and western blot analyses revealed that bone morphogenetic protein 2 expression was significantly attenuated in the Entelon150® group. Our results indicate that Entelon150® significantly attenuates post-implantation inflammation and degenerative calcification of the bovine pericardium in dogs. Therefore, Entelon150® may increase the longevity of the bovine pericardium after intravascular implantation.