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  • Author or Editor: Yoel Berhane x
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Objective—To determine in vitro vasoactive potency of monoamines formed in the cecum and found in the systemic circulation of horses.

Sample Population—Segments of digital blood vessels obtained from 6 healthy mixed-breed horses and ponies euthanatized at an abattoir and platelets isolated from 4 healthy ponies.

Procedure—Paired rings of digital artery and vein from the same horse were examined, and isometric tension was recorded. Concentration-response curves for tryptamine (TRP), tyramine (TYR), phenylethylamine (PEA), isoamylamine (IAA), and isobutylamine (IBA) were obtained. Vasoconstrictor mechanisms were investigated for TRP and TYR by the use of antagonists. Washed platelets loaded with [3H]-5-hydroxytryptamine (5-HT) were incubated with monoamines; the amount of radioactivity displaced after 30 minutes was estimated.

Results—TRP, TYR, and PEA were potent constrictors of arteries and veins, with TRP and TYR being more potent in veins than arteries. Constrictions induced by TYR were inhibited by benextramine (α-antagonist) and nisoxetine (neuronal-uptake blocker), whereas TRP responses were inhibited by ketanserin (5-HT receptor antagonist). All 5 amines displaced 5-HT from platelets with the order of potency being TYR > TRP > PEA > IAA > IBA.

Conclusions and Clinical Relevance—Amines from the equine cecum cause digital vasoconstriction. The most potent (TRP and TYR) cause selective venoconstriction. Tyrosine activates predominantly α-adrenoceptors through the release of neuronal norepinephrine, whereas TRP activates 5-HT receptors. All amines tested released 5-HT from platelets. Amines formed in the cecum and released into the systemic circulation warrant additional investigation as trigger factors for digital ischemia and subsequent laminitis. (Am J Vet Res 2003;64:1124–1131)

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in American Journal of Veterinary Research


Objective—To determine the effect of endotoxin (lipopolysaccharide [LPS]) on vasoactive mediator production by cultured equine digital vein endothelial cells (EDVECs).

Sample Population—EDVECs obtained from forelimb digital veins of 7 healthy adult horses.

Procedures—EDVECs were incubated with or without LPS (1 μg/mL) for 0, 2, 4, 6, 22, and 24 hours. The EDVECs were incubated for 18 hours with LPS (10 pg/mL to 1 μg/mL) with or without ibuprofen, cycloheximide, or L-nitroarginine methyl ester. Medium concentrations of prostacyclin, cyclic guanosine monophosphate, endothelin-1, and thromboxane A2 were determined. Changes in inducible nitric oxide synthase and cyclooxygenase-2 expression were determined.

Results—LPS stimulated mean 4.2- and 14.1-fold increases in EDVEC prostacyclin and cyclic guanosine monophosphate production, respectively, after 22 hours. These effects were LPS concentration–dependent (LPS concentrations that induced a response halfway between the maximum response and baseline of 1.50 and 1.22 ng/mL, respectively). The LPS-induced cyclic guanosine monophosphate production was significantly inhibited (to basal concentrations) by L-nitroarginine methyl ester, and prostacyclin production was inhibited by cycloheximide and ibuprofen. Production of thromboxane A2 by EDVECs was not detected. Endothelin-1 accumulated in the medium, but LPS did not enhance its production. Inducible nitric oxide synthase expression in EDVECs was not detected with the available antibodies, whereas LPS stimulated cyclooxygenase-2 expression in a time- and concentration-dependent manner.

Conclusions and Clinical Relevance—LPS stimulated vasoactive mediator production by equine endothelial cells, which may play a role in LPS-induced digital hypoperfusion.

Full access
in American Journal of Veterinary Research


Objective—To measure concentrations of amines formed in the cecum of clinically normal ponies, determine amine concentrations in plasma samples collected in spring and winter, and compare concentrations of amines and serotonin in plasma samples obtained from clinically normal ponies and ponies predisposed to laminitis.

Sample Population—Cecal contents obtained from 10 ponies euthanatized at an abattoir and blood samples obtained from 42 adult ponies.

Procedure—Cecal contents were assayed for amines by high-performance liquid chromatography (HPLC). Blood samples were collected at various times of the year from 20 ponies predisposed to acute laminitis and 22 clinically normal ponies. Plasma serotonin concentration was measured by HPLC, and tryptamine (TRP), tyramine (TYR), phenylethylamine (PEA), and isoamylamine (IAA) were measured by liquid chromatography- mass spectrometry.

Results—15 amines were identified in cecal contents. Plasma TRP, TYR, PEA, and IAA concentrations ranged from 10pM to 100nM in both groups of ponies. Plasma concentrations of serotonin or other amines did not differ between clinically normal ponies and those predisposed to laminitis; however, significantly higher concentrations of TRP, PEA, and IAA were found in samples obtained in the spring, compared with winter samples.

Conclusions and Clinical Relevance—Various amines are found in the cecum of ponies, several of which can be detected in the plasma. Concentrations increase significantly in the spring and may reach concentrations close to the threshold for causing vasoconstriction. Release of amines from the cecum into the systemic circulation may contribute to hemodynamic disturbances in horses and ponies with acute laminitis. (Am J Vet Res 2003;64:1132–1138)

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in American Journal of Veterinary Research


Objective—To determine whether pasture, and specifically the addition of fructan carbohydrate to the diet, induces exaggerated changes in serum insulin concentration in laminitispredisposed (LP) ponies, compared with ponies with no history of the condition, and also to determine insulin responses to the dexamethasone suppression test.

Design—Prospective study.

Animals—10 LP and 11 control adult nonobese mixed-breed ponies.

Procedures—Insulin-modified IV glucose tolerance tests were performed (5 ponies/group). In diet studies, ponies were kept on pasture and then changed to a hay diet (10 ponies/group). Second, ponies were maintained on a basal hay diet (4 weeks) before being fed a hay diet supplemented with inulin (3 g/kg/d [1.4 g/lb/d]). Serum insulin and plasma glucose concentrations were analyzed before and after dietary changes. Serum cortisol and insulin concentrations were also measured in a standard dexamethasone suppression test.

Results—The LP ponies were insulin resistant (median insulin sensitivity of 0.27 × 104 L•min−1•mU−1 in LP ponies, compared with 0.64 × 104 L•min−1•mU−1 in control ponies). Median insulin concentration in LP ponies was significantly greater than that in control ponies at pasture, decreased in response to feeding hay, and was markedly increased (5.5fold) following the feeding of inulin with hay. The LP ponies had a greater increase in serum insulin concentration at 19 hours after dexamethasone administration (median, 222.9 mU/L), compared with control ponies (45.6 mU/L).

Conclusions and Clinical Relevance—Nonobese ponies predisposed to develop laminitis had compensated insulin resistance, and this phenotype was revealed by feeding plant fructan carbohydrate or by dexamethasone administration.

Full access
in Journal of the American Veterinary Medical Association