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Effects of calcitriol on apoptosis, toll-like receptor 4 expression, and cytokine production of endotoxin-primed canine leukocytes Texte intégral
2018
Jaffey, Jared A. | Amorim, Juliana | DeClue, Amy E.
OBJECTIVE To determine the in vitro effect of calcitriol on indicators of immune system function in endotoxin-primed blood samples from healthy dogs. SAMPLE Blood samples from 6 healthy adult dogs. PROCEDURES Leukocytes were primed by incubation of blood samples with lipopolysaccharide (LPS; endotoxin) or PBS solution (unprimed control group) for 1 hour. Following priming, blood samples were incubated with calcitriol (2 × 10(−7)M) or ethanol (control substance) for 24 hours. After sample incubation, LPS-stimulated leukocyte production of tumor necrosis factor (TNF) and interleukin-10 (IL10) was measured with a canine-specific multiplex assay, and apoptosis and toll-like receptor 4 (TLR4) expression were evaluated via flow cytometry. RESULTS LPS stimulation of unprimed leukocytes but not endotoxin-primed leukocytes resulted in a significant increase in TNF and IL10 production, confirming the presence of endotoxin tolerance in dogs in vitro. Endotoxin priming significantly increased neutrophil viability with no effect on lymphocyte viability or TLR4 expression by neutrophils and monocytes. Calcitriol exposure significantly decreased LPS-stimulated production of TNF by unprimed and endotoxin-primed leukocytes. Conversely, calcitriol exposure had no effect on IL10 production by unprimed leukocytes but did significantly increase IL10 production by endotoxin-primed leukocytes. Calcitriol had no significant effect on the degree of neutrophil or lymphocyte apoptosis, nor was neutrophil and monocyte TLR4 expression affected in unprimed or endotoxin-primed leukocytes. CONCLUSIONS AND CLINICAL RELEVANCE These data indicated that calcitriol induced an anti-inflammatory shift in unprimed and endotoxin-primed canine leukocytes in vitro, without compromising neutrophil and monocyte TLR4 expression or altering the viability of neutrophils and lymphocytes in canine blood samples.
Afficher plus [+] Moins [-]Effects of two fractions of inspired oxygen during anesthesia on early postanesthesia oxygenation in healthy dogs Texte intégral
2018
Martin-Flores, Manuel | Tseng, Chia T. | Robillard, Steven D. | Abrams, Brittany E. | Campoy, Luis | Harvey, Jay | Gleed, Robin D.
OBJECTIVE To evaluate the effects of 2 fractions of inspired oxygen (Fio2s) during anesthesia on postanesthesia Pao2 and other measures of oxygen exchange. ANIMALS 22 healthy adult sexually intact female dogs undergoing ovariohysterectomy by ventral midline celiotomy. PROCEDURES Dogs were randomly assigned to receive either oxygen (Fio2 > 0.9 [100% oxygen]; n = 11; control group) or a mixture of nitrogen and oxygen (Fio2 = 0.4; 11; 40% oxygen group) as the carrier gas for isoflurane while anesthetized. All dogs were allowed to breathe spontaneously while anesthetized. For each dog, the Pao2, Paco2, other indices of oxygenation, and extent of sedation were monitored at predetermined times during and for 1 hour after anesthesia. Measured variables were compared between the 2 treatment groups and over time within each treatment group. RESULTS None of the measured variables differed significantly between the control and 40% oxygen groups at any time during the postanesthesia period. Within each treatment group, the Paco2 and extent of sedation decreased over time during the postanesthesia period. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that indices of oxygenation did not differ significantly between healthy dogs in which the Fio2 was maintained at > 0.9 and those in which the Fio2 was maintained at 0.4 while anesthetized for ovariohysterectomy. Thus, the addition of nitrogen to the carrier gas for an inhalant anesthetic conferred neither an advantage nor disadvantage in regard to oxygenation during the first hour of anesthesia recovery.
Afficher plus [+] Moins [-]Cyclooxygenase-2 expression in the eyes of cats with and without uveitis Texte intégral
2018
Sim, Zhi Hui | Pinard, Chantale L. | Plattner, Brandon L. | Bienzle, Dorothee
OBJECTIVE To characterize the distribution and intensity of cyclooxygenase (COX)-2 expression in the eyes of cats with and without uveitis and to determine whether COX-2 expression is correlated with severity of inflammation. SAMPLES Archived ocular tissue specimens from 51 cats with and 10 cats without ocular disease. PROCEDURES Specimens from only 1 eye were evaluated for each cat. Specimens were stained with H&E stain or immunohistochemical stain for detection of COX-2 and reviewed. For each eye, the type, severity, and distribution of inflammation and the distribution and intensity of COX-2 expression were determined for the uvea and other ocular tissues. Correlation between COX-2 expression and inflammation severity was also assessed. RESULTS COX-2 was not expressed in any nondiseased eye. Of the 51 diseased eyes, 20 had histologic evidence of lymphocytic-plasmacytic uveitis, 13 had neutrophilic uveitis, 11 had diffuse iris melanoma with uveitis, and 7 had diffuse iris melanoma without uveitis. Of the 44 eyes with uveitis, COX-2 was detected in the uvea of 16, including 11 eyes with lymphocytic-plasmacytic uveitis, 4 with neutrophilic uveitis, and 1 with diffuse iris melanoma–induced uveitis. Inflammation was severe, moderate, or mild in 10, 5, and 1 of those eyes, respectively. Cyclooxygenase-2 was detected in the cornea of 21 eyes with uveitis and 1 eye with diffuse iris melanoma without uveitis. Uveitis severity was positively correlated with COX-2 expression in both the uvea and cornea. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that COX-2 is an inflammatory mediator in feline uveitis but not diffuse iris melanoma.
Afficher plus [+] Moins [-]Venous blood gases, plasma biochemistry, and hematology of wild-caught common chameleons (Chamaeleo chamaeleon) Texte intégral
2018
Eshar, D. | Ammersbach, M. | Shacham, B. | Katzir, G. | Beaufrere, H.
The purpose of this study was to determine a wide range of selected hematologic, venous blood gases, and plasma biochemistry analytes in common chameleons (Chamaeleo chamaeleon). Blood samples were collected from the ventral tail vein of 41 common chameleons to determine reference intervals for 30 different blood analytes. The calcium-to-phosphorus ratio, packed cell volume (PCV), refractometric total solids (TS), blood cell counts, and differentials were also determined. The microscopic evaluation of blood smears revealed inclusion bodies in monocytes in 7 of the samples. Females showed significantly higher values of plasma proteins and calcium and cholesterol concentrations and males showed significantly higher values of aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) plasma concentrations. Significant differences were found between similar analytes determined by different testing methodologies in the PCV/hematocrit, electrolytes (sodium, potassium), and plasma proteins [TS, total protein (TP) and albumin]. Blood analytes determined in this study can provide baseline data that may be useful when evaluating the health status of common chameleons, taking into consideration the potential effects of gender and the type of analyzer used.
Afficher plus [+] Moins [-]Effects of prostaglandin-mediated and cholinergic-mediated miosis on morphology of the ciliary cleft region in dogs Texte intégral
2018
Park, Sangwan | Kang, Seonmi | Lim, Jaegook | Park, Eunjin | Nam, Taekjin | Jeong, Seowoo | Seo, Kangmoon
OBJECTIVE To compare morphology of the ciliary cleft (CC) region in dogs after topical administration of latanoprost, pilocarpine, or a combination of latanoprost and pilocarpine. ANIMALS 6 Beagles. PROCEDURES A prospective 4-phase crossover study with washout periods was performed. Latanoprost (phase L), pilocarpine (phase P), pilocarpine followed by latanoprost (phase PL), and latanoprost followed by pilocarpine (phase LP) were administered to the right eye. Artificial tears were administered to the left eye (control eye). For each phase, pupil diameter and intraocular pressure (IOP) were measured and ultrasonographic biomicroscopy was performed 2 hours after topical treatment. Angle opening distance (AOD), ciliary cleft width (CCW), ciliary cleft length (CCL), and ciliary cleft area (CCA) were evaluated. ESULTS All treated eyes had marked miosis without significant differences in pupil diameter among phases. Significant IOP reductions were detected for all phases, except phase P. The AOD and CCA were significantly increased in all phases for treated eyes, compared with results for control eyes. The CCW was significantly increased in phases P, PL, and LP; CCL was significantly increased in phases PL and LP. Comparison of treated eyes among phases revealed that CCW differed significantly between phases L and P and between phases L and PL. CONCLUSIONS AND CLINICAL RELEVANCE Prostaglandin-mediated and cholinergic-mediated miosis caused variations in CC configurations. When latanoprost and pilocarpine were used in combination, the first drug administered determined the cleft morphology, which was not fully reversed by the second drug. The CC morphology did not fully explain IOP reductions.
Afficher plus [+] Moins [-]Effect of trotting speed on kinematic variables measured by use of extremity-mounted inertial measurement units in nonlame horses performing controlled treadmill exercise Texte intégral
2018
Cruz, Antonio M. | Vidondo, Beatriz | Ramseyer, Alessandra A. | Maninchedda, Ugo E.
OBJECTIVE To assess effects of speed on kinematic variables measured by use of extremity-mounted inertial measurement units (IMUs) in nonlame horses performing controlled exercise on a treadmill. ANIMALS 10 nonlame horses. PROCEDURES 6 IMUs were attached at predetermined locations on 10 nonlame Franches Montagnes horses. Data were collected in triplicate during trotting at 3.33 and 3.88 m/s on a high-speed treadmill. Thirty-three selected kinematic variables were analyzed. Repeated-measures ANOVA was used to assess the effect of speed. RESULTS Significant differences between the 2 speeds were detected for most temporal (11/14) and spatial (12/19) variables. The observed spatial and temporal changes would translate into a gait for the higher speed characterized by increased stride length, protraction and retraction, flexion and extension, mediolateral movement of the tibia, and symmetry, but with similar temporal variables and a reduction in stride duration. However, even though the tibia coronal range of motion was significantly different between speeds, the high degree of variability raised concerns about whether these changes were clinically relevant. For some variables, the lower trotting speed apparently was associated with more variability than was the higher trotting speed. CONCLUSIONS AND CLINICAL RELEVANCE At a higher trotting speed, horses moved in the same manner (eg, the temporal events investigated occurred at the same relative time within the stride). However, from a spatial perspective, horses moved with greater action of the segments evaluated. The detected changes in kinematic variables indicated that trotting speed should be controlled or kept constant during gait evaluation.
Afficher plus [+] Moins [-]Immunohistochemical expression of insulin, glucagon, and somatostatin in pancreatic islets of horses with and without insulin resistance Texte intégral
2018
Newkirk, Kim M. | Ehrensing, Gordon | Odoi, Agricola | Boston, Ray C. | Frank, Nicholas
OBJECTIVE To assess insulin, glucagon, and somatostatin expression within pancreatic islets of horses with and without insulin resistance. ANIMALS 10 insulin-resistant horses and 13 insulin-sensitive horses. PROCEDURES For each horse, food was withheld for at least 10 hours before a blood sample was collected for determination of serum insulin concentration. Horses with a serum insulin concentration < 20 μU/mL were assigned to the insulin-sensitive group, whereas horses with a serum insulin concentration > 20 μU/mL underwent a frequently sampled IV glucose tolerance test to determine sensitivity to insulin by minimal model analysis. Horses with a sensitivity to insulin < 1.0 × 10(−4) L•min−1•mU−1 were assigned to the insulin-resistant group. All horses were euthanized with a barbiturate overdose, and pancreatic specimens were harvested and immunohistochemically stained for determination of insulin, glucagon, and somatostatin expression in pancreatic islets. Islet hormone expression was compared between insulin-resistant and insulin-sensitive horses. RESULTS Cells expressing insulin, glucagon, and somatostatin made up approximately 62%, 12%, and 7%, respectively, of pancreatic islet cells in insulin-resistant horses and 64%, 18%, and 9%, respectively, of pancreatic islet cells in insulin-sensitive horses. Expression of insulin and somatostatin did not differ between insulin-resistant and insulin-sensitive horses, but the median percentage of glucagon-expressing cells in the islets of insulin-resistant horses was significantly less than that in insulin-sensitive horses. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that, in insulin-resistant horses, insulin secretion was not increased but glucagon production might be downregulated as a compensatory response to hyperinsulinemia.
Afficher plus [+] Moins [-]Genome-wide sequencing and quantification of circulating microRNAs for dogs with congestive heart failure secondary to myxomatous mitral valve degeneration Texte intégral
2018
Jung, SeungWoo | Bohan, Amy
OBJECTIVE To characterize expression profiles of circulating microRNAs via genome-wide sequencing for dogs with congestive heart failure (CHF) secondary to myxomatous mitral valve degeneration (MMVD). ANIMALS 9 healthy client-owned dogs and 8 age-matched client-owned dogs with CHF secondary to MMVD. PROCEDURES Blood samples were collected before administering cardiac medications for the management of CHF. Isolated microRNAs from plasma were classified into microRNA libraries and subjected to next-generation sequencing (NGS) for genome-wide sequencing analysis and quantification of circulating microRNAs. Quantitative reverse transcription PCR (qRT-PCR) assays were used to validate expression profiles of differentially expressed circulating microRNAs identified from NGS analysis of dogs with CHF. RESULTS 326 microRNAs were identified with NGS analysis. Hierarchical analysis revealed distinct expression patterns of circulating microRNAs between healthy dogs and dogs with CHF. Results of qRT-PCR assays confirmed upregulation of 4 microRNAs (miR-133, miR-1, miR-let-7e, and miR-125) and downregulation of 4 selected microRNAs (miR-30c, miR-128, miR-142, and miR-423). Results of qRT-PCR assays were highly correlated with NGS data and supported the specificity of circulating microRNA expression profiles in dogs with CHF secondary to MMVD. CONCLUSIONS AND CLINICAL RELEVANCE These results suggested that circulating microRNA expression patterns were unique and could serve as molecular biomarkers of CHF in dogs with MMVD.
Afficher plus [+] Moins [-]Pharmacokinetics and pharmacodynamics after oral administration of tapentadol hydrochloride in dogs Texte intégral
2018
Howard, James | Aarnes, Turi K. | Dyce, Jonathan | Lerche, Phillip | Wulf, Larry W. | Coetzee, Johann F. | Lakritz, Jeffrey
OBJECTIVE To evaluate pharmacokinetic and pharmacodynamic characteristics of 3 doses of tapentadol hydrochloride orally administered in dogs. ANIMALS 6 healthy adult mixed-breed dogs. PROCEDURES In a prospective, randomized crossover study, dogs were assigned to receive each of 3 doses of tapentadol (10, 20, and 30 mg/kg, PO); there was a 1-week washout period between subsequent administrations. Plasma concentrations and physiologic variables were measured for 24 hours. Samples were analyzed by use of high-performance liquid chromatography–tandem mass spectrometry. RESULTS Tapentadol was rapidly absorbed after oral administration. Mean maximum plasma concentrations after 10, 20, and 30 mg/kg were 10.2, 19.7, and 31 ng/mL, respectively. Geometric mean plasma half-life of the terminal phase after tapentadol administration at 10, 20, and 30 mg/kg was 3.5 hours (range, 2.7 to 4.5 hours), 3.7 hours (range, 3.1 to 4.0 hours), and 3.7 hours (range, 2.8 to 6.5 hours), respectively. Tapentadol and its 3 quantified metabolites (tapentadol sulfate, tapentadol-O-glucuronide, and desmethyltapentadol) were detected in all dogs and constituted 0.16%, 2.8%, 97%, and 0.04% of the total area under the concentration-time curve (AUC), respectively. Plasma AUCs for tapentadol, tapentadol sulfate, and tapentadol-O-glucuronide increased in a dose-dependent manner. Desmethyltapentadol AUC did not increase in a linear manner at the 30-mg/kg dose. Sedation scores and heart and respiratory rates were not significantly affected by dose or time after administration. CONCLUSIONS AND CLINICAL RELEVANCE Oral administration of tapentadol was tolerated well, and the drug was rapidly absorbed. Adverse events were not apparent in any dogs at any doses in this study.
Afficher plus [+] Moins [-]Does antimicrobial therapy improve outcomes in horses with severe equine asthma and a positive tracheal wash bacterial culture? Texte intégral
2018
The objective of this study was to observe the outcomes of adding an antimicrobial treatment to a conventional treatment regime in horses with severe equine asthma in a clinical setting. Eleven client-owned horses with a history consistent with severe equine asthma, increased respiratory effort and nostril flaring, ≥ 20% neutrophils on bronchoalveolar lavage (BAL), and a positive tracheal wash (TW) bacterial culture were treated with environmental management, corticosteroids, and bronchodilators. Six horses were also treated with an antimicrobial (principal group), while the other 5 were administered saline as a placebo (control group). Treatment with antimicrobials significantly improved the post-treatment clinical score of the principal group compared with the pre-treatment score, whereas no significant difference occurred in the control group. The principal group also had significantly less neutrophil myeloperoxidase (MPO) activity post-treatment than pre-treatment, with a median difference of -0.39 units/[protein] in the principal group and a median difference of -0.21 units/[protein] in the controls. There was no difference in MPO activity pre- versus post-treatment in the control group. No differences were noted in the intra-group comparisons of pre- versus post-treatment BAL neutrophil counts, mucus scores, and concentrations of interleukin-8 (IL-8) or tumor necrosis factor-alpha (TNF-α) in bronchoalveolar lavage fluid (BALF) in either group. There were no differences found in the inter-group comparisons of the principal versus controls for each of the pre- and post-treatment time periods for BAL neutrophil count, mucus score, clinical scores, MPO activity, and IL-8 or TNF-α concentrations. The role of airway bacteria in horses with severe equine asthma requires further investigation as antimicrobial therapy improved post-treatment clinical scores and decreased MPO activity in the group of horses studied, but did not affect other measures of airway inflammation.
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