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Evaluation of high–molecular weight adiponectin in horses
2012
Wooldridge, Anne A. | Edwards, Heather Gray | Plaisance, Eric P. | Applegate, Rory | Taylor, Debra R. | Taintor, Jennifer | Zhong, Qiao | Judd, Robert L.
Objective: To characterize adiponectin protein complexes in lean and obese horses. Animals: 26 lean horses and 18 obese horses. Procedures: Body condition score (BCS) and serum insulin activity were measured for each horse. Denaturing and native western blot analyses were used to evaluate adiponectin complexes in serum. A human ELISA kit was validated and used to quantify high–molecular weight (HMW) complexes. Correlations between variables were made, and HMW values were compared between groups. Results: Adiponectin was present as a multimer consisting of HMW (> 720-kDa), low-molecular weight (180-kDa), and trimeric (90-kDa) complexes in serum. All complexes were qualitatively reduced in obese horses versus lean horses, but the percentage of complexes < 250 kDa was higher in obese versus lean horses. High–molecular weight adiponectin concentration measured via ELISA was negatively correlated with serum insulin activity and BCS and was lower in obese horses (mean ± SD, 3.6 ± 3.9 μg/mL), compared with lean horses (8.0 ± 4.6 μg/mL). Conclusions and Clinical Relevance: HMW adiponectin is measurable via ELISA, and concentration is negatively correlated with BCS and serum insulin activity in horses. A greater understanding of the role of adiponectin in equine metabolism will provide insight into the pathophysiology of metabolic disease conditions.
Show more [+] Less [-]Evaluation of ultrasonography for measurement of skin thickness in Shar-Peis
2012
Objective: To determine whether high-frequency diagnostic ultrasonography is useful for assessment of skin thickness in Shar-Peis. Animals: 10 healthy Shar-Peis and 10 healthy Beagles used as controls. Procedures: Ultrasonographic examination of the skin was performed on 4 cutaneous sites by use of a 13-MHz linear-array transducer, and the mean of 3 measurements was calculated. Ultrasonography results were compared with histologic findings of skin specimens stained with H&E, Alcian blue at a pH of 2.5, and Masson trichrome stains, with histometric measurements of skin thickness made by use of a microscope, and with measurements of skin thickness made by use of a plicometer. Ultrasonograpy results were also compared via age and sex of selected animals. Results: A clear correlation was detected between ultrasonography results and results of histologic and histometric analysis in both groups. In Shar-Peis, no correlation was found between ultrasonography results and age and sex, whereas in Beagles, a weak positive correlation was found only between skin thickness in dorsal cervical and frontal (on the rostral margins of the supraorbital processes) regions and age. A positive overall correlation was found in Shar-Peis between measurements made via ultrasonography and plicometery. Conclusions and Clinical Relevance: Ultrasonography was a useful tool to assess skin thickness, and in Shar-Peis, it might be considered a valid alternative to invasive methods such as histologic examination to objectively estimate the severity of hereditary cutaneous hyaluronosis.
Show more [+] Less [-]Estimates of the pharmacokinetics of famciclovir and its active metabolite penciclovir in young Asian elephants (Elephas maximus)
2012
Brock, A Paige | Isaza, Ramiro | Hunter, Robert P. | Richman, Laura K. | Koch, David E. | Schmitt, Dennia L. | Montali, Richard J. | Lindsay, William A.
Objective: To determine plasma pharmacokinetics of penciclovir following oral and rectal administration of famciclovir to young Asian elephants (Elephas maximus). Animals: 6 healthy Asian elephants (5 females and 1 male), 4.5 to 9 years old and weighing 1,646 to 2,438 kg. Procedures: Famciclovir was administered orally or rectally in accordance with an incomplete crossover design. Three treatment groups, each comprising 4 elephants, received single doses of famciclovir (5 mg/kg, PO, or 5 or 15 mg/kg, rectally); there was a minimum 12-week washout period between subsequent famciclovir administrations. Serial blood samples were collected after each administration. Samples were analyzed for famciclovir and penciclovir with a validated liquid chromatography–mass spectroscopy assay. Results: Famciclovir was tolerated well for both routes of administration and underwent complete biotransformation to the active metabolite, penciclovir. Mean maximum plasma concentration of penciclovir was 1.3 μg/mL at 1.1 hours after oral administration of 5 mg/kg. Similar results were detected after rectal administration of 5 mg/kg. Mean maximum plasma concentration was 3.6 μg/mL at 0.66 hours after rectal administration of 15 mg/kg; this concentration was similar to results reported for humans receiving 7 mg/kg orally. Conclusions and Clinical Relevance: Juvenile Asian elephants are susceptible to elephant endotheliotropic herpesvirus. Although most infections are fatal, case reports indicate administration of famciclovir has been associated with survival of 3 elephants. In Asian elephants, a dose of 8 to 15 mg of famciclovir/kg given orally or rectally at least every 8 hours may result in penciclovir concentrations that are considered therapeutic in humans.
Show more [+] Less [-]Analysis of the transport of and cytotoxic effects for nalbuphine solution in corneal cells
2012
Spatola, Ronald A. | Thangavelu, Mirunalni | Upadhyayula, Vijayasaradhi | Lee, Seungsoo | Phelps, Mitch A. | Chandler, Heather L.
Objective: To assess the in vitro effects of various nalbuphine concentrations on viability and wound healing ability of corneal cells and potential drug transport through the corneal epithelium. Sample: Cultured canine and human corneal epithelial cells (CECs) and cultured canine corneal stromal fibroblasts. Procedures: CECs and stromal fibroblasts were exposed to nalbuphine (concentration of solutions ranged from 0% to 1.2%) for up to 30 minutes, and viability was assessed with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A standard scratch test technique was used. Wound healing of CECs and stromal fibroblasts was evaluated following treatment with nalbuphine solutions < 0.1%. Liquid chromatography–mass spectrometry–mass spectrometry analysis was used to evaluate drug transport across a monolayer and a multilayer of human CECs. Results: A progressive decrease in viability was detected in canine CECs for all nalbuphine treatment groups, whereas treatment with only 0.5% or 1.2% nalbuphine significantly reduced corneal stromal fibroblast viability, compared with results for control cells. Within 24 hours, treatment with 0.1% nalbuphine solution significantly altered the healing rate of both canine CECs and stromal fibroblasts. Continuous increases in transport rates of nalbuphine were detected with time for both the monolayer and multilayer of human CECs. Conclusions and Clinical Relevance: In vitro, nalbuphine potentially could penetrate through corneal tissue, but it may cause damage to the corneal epithelium and stromal fibroblasts. Therefore, nalbuphine potentially may impair corneal wound healing.
Show more [+] Less [-]Investigation of hydration processes of the equine hoof via nuclear magnetic resonance microscopy
2012
Sugimoto, Miki | Kuwano, Atsutoshi | Ikeda, Shuntaro | Kume, Shin-ichi | Yoshihara, Eiru | Wada, Shinya
Objective: To examine the distribution of water in hoof wall specimens of horses via nuclear magnetic resonance (NMR) microscopy and determine changes in water distribution during hydration. Sample: 4 hoof wall specimens (2 obtained from the dorsum and 1 each obtained from the lateral quarter and lateral heel regions) of the stratum medium of healthy hooves of 1 horse. Procedures: Equine hoof wall specimens were examined via NMR microscopy. Proton density–weighted 3-D images were acquired. Changes during water absorption were assessed on sequential images. Results: The inner zone of the stratum medium had higher signals than did the outer zone. Areas of high signal intensity were evident in transverse images; these corresponded to the distribution of horn tubules. During water absorption, the increase in signal intensity started at the bottom of a specimen and extended to the upper region; it maintained the localization pattern observed before hydration. The relationship between the local maximal signals in areas corresponding to the horn tubules and minimal signal intensities in areas corresponding to the intertubular horn was similar and maintained approximately a linear distribution. Conclusions and Clinical Relevance: Based on the premise that signal intensity reflects water content, hydration in the equine hoof wall during water absorption occurred concurrently in the tubules and intertubular horn, and there was maintenance of the original water gradients. This technique can be applied for the assessment of pathophysiologic changes in the hoof wall on the basis of its hydration properties.
Show more [+] Less [-]Respiratory mechanics and results of cytologic examination of bronchoalveolar lavage fluid in healthy adult alpacas
2012
Pacheco, Ana P. | Bedenice, Daniela | Mazan, Melissa R. | Hoffman, Andrew M.
Objective-To evaluate respiratory mechanical function and bronchoalveolar lavage (BAL) cytologic results in healthy alpacas. Animals-16 client-owned adult alpacas. Procedures-Measurements of pulmonary function were performed, including functional residual capacity (FRC) via helium dilution, respiratory system resistance via forced oscillatory technique (FOT), and assessment of breathing pattern by use of respiratory inductive plethysmography (RIP) in standing and sternally recumbent alpacas. Bronchoalveolar lavage was performed orotracheally during short-term anesthesia. Results-Mean +/- SD measurements of respiratory function were obtained in standing alpacas for FRC (3.19 +/- 0.53 L), tidal volume (0.8 +/- 0.13 L), and respiratory system resistance at 1 Hz (2.70 +/- 0.88 cm H2O/L/s), 2 Hz (2.98 +/- 0.70 cm H2O/L/s), 3 Hz (3.14 +/- 0.77 cm H2O/L/s), 5 Hz (3.45 +/- 0.91 cm H2O/L/s), and 7 Hz (3.84 +/- 0.93 cm H2O/L/s). Mean phase angle, as a measurement of thoracoabdominal asynchrony, was 19.59 +/- 10.06°, and mean difference between nasal and plethysmographic flow measurements was 0.18 +/- 0.07 L/s. Tidal volume, peak inspiratory flow, and peak expiratory flow were significantly higher in sternally recumbent alpacas than in standing alpacas. Cytologic examination of BAL fluid revealed 58.52 +/- 12.36% alveolar macrophages, 30.53 +/- 13.78% lymphocytes, 10.95 +/- 9.29% neutrophils, 0% mast cells, and several ciliated epithelial cells. Conclusions and Clinical Relevance-Pulmonary function testing was tolerated well in nonsedated untrained alpacas. Bronchoalveolar lavage in alpacas yielded samples with adequate cellularity that had a greater abundance of neutrophils than has been reported in horses.
Show more [+] Less [-]Cardiorespiratory, gastrointestinal, and analgesic effects of morphine sulfate in conscious healthy horses
2012
Figueiredo, Juliana P. | Muir, William W. | Sams, Richard
Objective: To compare the cardiorespiratory, gastrointestinal, analgesic, and behavioral effects between IV and IM administration of morphine in conscious horses with no signs of pain. Animals: 6 healthy adult horses. Procedures: Horses received saline (0.9% NaCl) solution (IM or IV) or morphine sulfate (0.05 and 0.1 mg/kg, IM or IV) in a randomized, masked crossover study design. The following variables were measured before and for 360 minutes after drug administration: heart and respiratory rates; systolic, diastolic, and mean arterial blood pressures; rectal temperature; arterial pH and blood gas variables; intestinal motility; and response to thermal and electrical noxious stimuli. Adverse effects and horse behavior were also recorded. Plasma concentrations of morphine, morphine-3-glucuronide, and morphine-6-glucuronide were measured via liquid chromatography–mass spectrometry. Results: No significant differences in any variable were evident after saline solution administration. Intravenous and IM administration of morphine resulted in minimal and short-term cardiorespiratory, intestinal motility, and behavioral changes. A decrease in gastrointestinal motility was detected 1 to 2 hours after IM administration of morphine at doses of 0.05 and 0.1 mg/kg and after IV administration of morphine at a dose of 0.1 mg/kg. Morphine administration yielded no change in any horse's response to noxious stimuli. Both morphine-3-glucuronide and morphine-6-glucuronide were detected in plasma after IV and IM administration of morphine. Conclusions and Clinical Relevance: Clinically relevant doses of morphine sulfate yielded minimal and short-term behavioral and intestinal motility effects in healthy horses with no signs of pain. Neither dose of morphine affected their response to a noxious stimulus.
Show more [+] Less [-]Evaluation of the diagnostic and prognostic utility of ultrasonography at first diagnosis of presumptive bovine respiratory disease
2012
Abutarbush, Sameeh M. | Pollock, Colleen M. | Wildman, Brian K. | Perrett, Tye | Schunicht, Oliver C. | Fenton, R Kent | Hannon, Sherry J. | Vogstad, Amanda R. | Jim, G Kee | Booker, Calvin W.
This project investigated the use of ultrasonography at first diagnosis of presumptive early bovine respiratory disease (BRD) in feedlot cattle from western Canada. One hundred seventy-four cattle (116 cases and 58 controls) at high risk of developing BRD were enrolled in a prospective longitudinal study over 2 y (2006-2007). Cattle with clinical signs relating to the respiratory system and assessed as sick at the time of feedlot arrival (arrival fever cases) or assessed as sick in the pen 3 to 30 d post-arrival (post-arrival fever cases, post-arrival no fevers cases) were eligible for enrollment. Control animals were identified at the time of case enrollments. Ultrasonography was done using a 3.5 sector transducer at enrollment and at 2, 4, and 6 wk post-enrollment. Lung lesions were identified at least 1 time over the course of the trial in 32/116 (28%) cases and 9/58 (16%) controls. At enrollment, lung lesions were identified in 20/115 (17%) cases and 2/55 (4%) controls (data unreadable n = 4). Post-arrival fever cases (14/48) were the most likely to have a lesion identified using ultrasound. In arrival fever cases, average daily gain (enrollment to last ultrasound, average 34 d) was improved (P = 0.007) in cattle identified with a lesion at enrollment using ultrasound compared with those not identified with a lesion at that time, potentially demonstrating the effects of gut fill at arrival weighing, as these sicker animals may have eaten less prior to arrival and, therefore, had more room for improvement in weight over time due to restoration of normal gut fill. None of the ultrasound time points explored (enrollment, 2, 4, or 6 wk post-enrollment) were associated with the animal health outcomes of interest (subsequent treatment, chronicity, wastage, or mortality) for cattle enrolled at arrival or post-arrival.
Show more [+] Less [-]Comparison of equine tendon- and bone marrow–derived cells cultured on tendon matrix with or without insulin-like growth factor-I supplementation
2012
Durgam, Sushmitha S. | Stewart, Allison A. | Pondenis, Holly C. | Gutierrez-Nibeyro, Santiago M. | Evans, Richard B. | Stewart, Matthew C.
Objective-To compare in vitro expansion, explant colonization, and matrix synthesis of equine tendon- and bone marrow-derived cells in response to insulin-like growth factor-I (IGF-I) supplementation. Sample-Cells isolated from 7 young adult horses. Procedures-Tendon- and bone marrow-derived progenitor cells were isolated, evaluated for yield, and cultured on autogenous cell-free tendon matrix for 7 days. Samples were analyzed for cell viability and expression of collagen type I, collagen type III, and cartilage oligomeric matrix protein mRNAs. Collagen and glycosaminoglycan syntheses were quantified over a 24-hour period. Results-Tendon- and bone marrow–derived cells required 17 to 19 days of monolayer culture to reach 2 passages. Mean +/- SE number of monolayer cells isolated was higher for tendon-derived cells (7.9 +/- 0.9 × 10(6)) than for bone marrow–derived cells (1.2 +/- 0.1 × 10(6)). Cell numbers after culture for 7 days on acellular tendon matrix were 1.6- to 2.8-fold higher for tendon-derived cells than for bone marrow-derived cells and 0.8- to 1.7-fold higher for IGF-I supplementation than for untreated cells. New collagen and glycosaminoglycan syntheses were significantly greater in tendon-derived cell groups and in IGF-I-supplemented groups. The mRNA concentrations of collagen type I, collagen type III, and cartilage oligomeric matrix protein were not significantly different between tendon- and bone marrow-derived groups. Conclusions and Clinical Relevance-In vitro results of this study suggested that tendon-derived cells supplemented with IGF-I may offer a useful resource for cell-based strategies in tendon healing.
Show more [+] Less [-]Evaluation of Clostridium novyi–NT spores in dogs with naturally occurring tumors
2012
Krick, Erika L. | Sorenmo, Karin U. | Rankin, Shelley C. | Cheong, Ian | Kobrin, Barry | Thornton, Katherine | Kinzler, Kenneth W. | Vogelstein, Bert | Zhou, Shibin | Diaz, Luis A. Jr
Objective-To establish the maximum tolerated dose of Clostridium novyi-NT spores in tumor-bearing dogs and evaluate spore germination within tumors and tumor response. Animals-6 client-owned dogs. Procedures-A standard dose-escalation study was planned, with maximum tolerated dose defined as the highest dose at which 0 or 1 of 6 dogs had dose-limiting toxicoses (DLT). Dogs received 1 dose of C novyi-NT spores IV. Toxicoses were graded and interventions performed according to specific guidelines. Grade 3 or higher toxicosis or any toxicosis combination that substantially affected patient status was considered DLT. Clinical response was measured by use of response evaluation criteria in solid tumors at 28 days. Results-The first 2 dogs had DLT. The dose was decreased. Two of the next 4 dogs had DLT; therefore, dose administration was stopped because the study endpoint had been reached. The most common toxicosis was fever (n = 6 dogs). Two dogs developed abscesses (1 within a nasal carcinoma and 1 splenic abscess) attributable to C novyi-NT infection; both required surgical intervention. Clostridium novyi-NT was cultured from 1 of 6 tumors. Five dogs were available for response assessment (4 had stable disease; 1 had progressive disease). Conclusions and Clinical Relevance-Results indicated that C novyi-NT can germinate within tumors of dogs. Toxicosis, although common and sometimes severe, was manageable with treatment. Further studies in dogs with superficial tumors may allow for continued dose escalation and provide information for use in clinical trials in veterinary and human oncology.
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