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Abstract
Objective—To determine and compare the number, type, location, and distribution of apoptotic epidermal cells in the laminae of clinically normal horses and horses with laminitis.
Sample Population—Formalin-fixed samples of digital lamellar tissue from 47 horses (including clinically normal horses [controls; n = 7], horses with acute [4] and chronic [7] naturally acquired laminitis, and horses with black walnut extract-induced [11] or carbohydrate overload-induced [18] laminitis).
Procedure—Blocks of paraffin-embedded lamellar tissues were stained for DNA fragmentation with the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique. Differential immunohistochemical staining for caspases 3 and 14 were used to confirm apoptosis.
Results—The number of TUNEL-positive epidermal cells per 0.1 mm of primary laminae was significantly greater in the acute laminitis group than in the other groups. In the acute laminitis group, there were 17 and 1,025 times as many TUNEL-positive basal layer cells and keratinocytes, respectively, compared with the control group. Apoptosis of TUNEL-positive basal layer cells was confirmed by results of caspase 3 immunohistochemical staining. The TUNEL-positive keratinocytes did not stain for caspases 3 or 14.
Conclusions and Clinical Relevance—The large number of apoptotic basal layer cells detected in the lamellar tissue of horses with acute naturally acquired laminitis suggests that apoptosis may be important in the development of acute laminitis. The role of the large number of TUNEL-positive keratinocytes detected in the interface of primary and secondary epidermal laminae of horses with acute laminitis remains to be elucidated. ( Am J Vet Res 2004;65:578–585)
Abstract
Objective—To characterize the in vitro response of equine cecal longitudinal smooth muscle (CLSM) to endothelin (ET)-1 and assess the role of ETA and ETB receptors in those ET-1–induced responses.
Animals—36 horses without gastrointestinal tract disease.
Procedure—To determine cumulative concentrationresponse relationships, CLSM strips were suspended in tissue baths containing graded concentrations of ET-1 (10–9 to 10–6M) with or without BQ-123 (ETA receptor antagonist); with or without IRL-1038 (ETB receptor antagonist); or with both antagonists at concentrations of 10–9, 10–7, and 10–5M. To determine the percentage change in baseline tension of CLSM, the areas under the curve during the 3-minute periods before and after addition of each dose were compared . Also, the effects of ET-1 and a combination of selective ETA and ETB receptor antagonists on electrically evoked contractions were studied.
Results—ET-1 caused sustained increases in CLSM tension in a concentration-dependent manner. Contractile responses to ET-1 were not significantly inhibited by either BQ-123 or IRL-1038 alone at any concentration; however, responses were significantly inhibited by exposure to the antagonists together at a concentration of 10–5M. Electrical field stimulation did not change the spontaneous contractile activity of CLSM and did not significantly alter the tissue response to ET-1, BQ-123, or IRL-1038.
Conclusions and Clinical Relevance—Results indicated that ET-1 has a contractile effect on equine CLSM that is mediated via ETA and ETB receptors. In vitro spontaneous contractions of equine CLSM apparently originate in the smooth muscle and not the enteric nervous system. (Am J Vet Res 2005;66:1202–1208)
Abstract
Objective—To identify differentially expressed genes in pulmonary tissues of horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD), which is a form of recurrent airway obstruction (RAO), compared with those of unaffected horses.
Animals—6 horses with SPAOPD-RAO and 6 unaffected (healthy) horses.
Procedures—Horses were assigned to 2 groups on the basis of medical history, clinical score, and transpleural pressure. Total RNA from each of the 5 lung lobes of each of the 6 SPAOPD-RAO–affected horses was extracted and pooled. Similarly, total RNA from unaffected horses was pooled. Differential display (DD) PCR assay was performed, and differentially expressed bands were purified and cloned into a plasmid vector. Plasmids were extracted from recombinant colonies, and purified DNA was sequenced. Genes of interest for RAO pathogenesis were identified. Real-time PCR assay was performed to confirm findings for the DD PCR assay.
Results—18 differentially expressed genes (17 upregulated and 1 downregulated) were identified. Three genes of particular interest were found to be altered (2 upregulated and 1 downregulated) in horses with SPAOPD-RAO by use of real-time PCR assay, and these findings matched the differential expression found by use of the DD PCR assay.
Conclusions and Clinical Relevance—SPAOPD-RAO in horses is a multifactorial, complex disease involving several genes. Upregulated genes, particularly β2-microglobulin, and the downregulated secretoglobin gene can serve as marker genes that may help to identify SPAOPD-RAO at an early age.
Abstract
OBJECTIVE
To evaluate the effects of a cognitive-behavioral skills building program (ie, MINDSTRONG; The Ohio State University) on the mental health outcomes and healthy lifestyle beliefs and behaviors of Doctor of Veterinary Medicine (DVM) students.
Sample
DVM students (n = 62) before beginning their program at a large public Midwest land-grant university.
Procedures
All 171 incoming DVM students (class of 2024) were required to take the cognitive-behavioral skills building program (7 weeks in length) before starting their 2020 school year. Students were given the option to consent to the study portion of the program. Consenting participants completed a pre- and postsurvey containing demographic questions and 5 valid and reliable scales, including the Patient Health Questionnaire-9 that assesses depressive symptoms, the Generalized Anxiety Disorder-7 that evaluates anxiety, the Brief Inventory of Perceived Stress that measures stress, and the Healthy Lifestyle Beliefs and Healthy Lifestyle Behaviors scales. Descriptive statistics described sample characteristics, paired t tests assessed changes over time in the outcomes Personal Wellness Assessment, and Cohen’s d determined effect sizes.
Results
62 DVM students completed both surveys. Postintervention, students had significant improvements in depressive symptoms, anxiety, and healthy lifestyle beliefs and behaviors.
Clinical Relevance
Although this study used a small convenience sample of DVM students from a single university, a cognitive-behavioral skills building program demonstrated the ability to decrease rates of depression, anxiety, and suicidal ideation and improve healthy lifestyle beliefs and behaviors. Requiring DVM students to participate in such programming could provide benefit during their professional education and throughout their careers.
Abstract
Objective—To characterize alterations in systemic and local colonic hemodynamic variables associated with IV infusion of ATP-MgCl2 in healthy anesthetized horses.
Animals—12 adult horses.
Procedure—Six horses were given ATP-MgCl2, IV, beginning at a rate of 0.1 mg of ATP/kg of body weight/min with incremental increases until a rate of 1.0 mg/kg/min was achieved. The remaining 6 horses were given an equivalent volume of saline (0.9% NaCl) solution over the same time period. Colonic and systemic hemodynamic variables and colonic plasma nitric oxide (NO) concentrations were determined before, during, and after infusion.
Results—Infusion of ATP-MgCl2 caused a rate-dependent decrease in systemic and colonic vascular resistance, principally via its vasodilatory effects. A rate of 0.3 mg of ATP/kg/min caused a significant decrease in systemic and colonic arterial pressure and colonic vascular resistance without a significant corresponding decrease in colonic arterial blood flow. Consistent alterations in NO concentrations of plasma obtained from colonic vasculature were not detected, despite profound vasodilatation of the colonic arterial vasculature.
Conclusions and Clinical Relevance—Results revealed that IV infusion of ATP-MgCl2 may be beneficial in maintaining colonic perfusion in horses with ischemia of the gastrointestinal tract, provided a sufficient pressure gradient exists to maintain blood flow. (Am J Vet Res 2001;62:1240–1249)
Abstract
Objective—To determine concentrations of nitric oxide (NO) in plasma and bronchoalveolar lavage fluid (BALF) and localize nitric oxide synthesis in the lungs of horses with summer pasture-associated obstructive pulmonary disease (SPAOPD).
Animals—7 adult horses with SPAOPD and 6 clinically normal adult horses.
Procedure—Severity of SPAOPD was determined by use of clinical scores, change in intrapleural pressure (ΔPpl) during tidal breathing, cytologic analysis of BALF, and histologic evaluation of lung specimens obtained during necropsy. Nitric oxide concentrations in plasma, BALF, and epithelial lining fluid (ELF) were determined by use of a chemiluminescent method. Inducible nitric oxide synthase (iNOS) and nitrotyrosine (NT) were localized in formalin-fixed lung specimens by use of immunohistochemical staining, and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) activity was localized in cryopreserved specimens by use of histochemical staining.
Results—Plasma concentration of NO in affected horses was slightly but not significantly greater than concentration in nonaffected horses. Nitric oxide concentrations in BALF or ELF did not differ between groups. Immunoreactivity of iNOS in bronchial epithelial cells of 3 of 5 lung lobes was greater in horses with SPAOPD, compared with nonaffected horses. However, staining for NT and NADPHd activity did not differ between groups.
Conclusions and Clinical Relevance—Expression of iNOS was greater in bronchial epithelial cells of horses with SPAOPD, compared with nonaffected horses, suggesting that NO may play a role in amplifying the inflammatory process in the airways of horses with this disease. (Am J Vet Res 2001;62:1381–1386)
Abstract
Objective—To evaluate the in vitro effects of adenosine tryphosphate (ATP) on vasomotor tone of equine colonic vasculature.
Sample Population—Arteries and veins from the left ventral colon of 14 mixed-breed horses euthanatized for reasons unrelated to cardiovascular or gastrointestinal tract disease.
Procedures—Endothelium-intact and -denuded arterial and venous rings were precontracted with 10–7 and 1.8 × 10–8 M endothelin-1, respectively. In 1 trial, endothelium-intact rings were also incubated with 10–4 M Nω-nitro-L-arginine methyl ester (L-NAME) to inhibit nitric oxide (NO) production. Adenosine triphosphate (10–8 to 10–3 M) was added in a noncumulative manner, and relaxation percentage versus time curves were generated. Areas under the curves (ie, percentage of relaxation time) were calculated.
Results—Relaxation response of arterial and venous rings to ATP was dose-dependent. Percentage of relaxation time in response to 10–4 and 10–3 M ATP was significantly greater, compared with that for rings not treated with ATP. Removal of endothelium attenuated but did not eliminate the relaxation response. Addition of L-NAME did not attenuate the relaxation response in arteries. At higher concentrations, the vascular response to ATP was biphasic.
Conclusions and Clinical Relevance—ATP applied to equine colonic arterial and venous rings with and without intact endothelium induced a biphasic response characterized by transient contraction followed by slow, substantial, and sustained relaxation. This ATP-induced response is possibly mediated by a mechanism other than NO. Adenosine triphosphate may be a useful treatment to modulate colonic vasomotor tone in horses with strangulating volvulus of the ascending colon. (Am J Vet Res 2001;62:1928–1933)
