Photoreceptor cell dysplasia in two Tippler pigeons.

Two 12-week-old Tippler pigeons were evaluated for ocular abnormalities associated with congenital blindness. The pigeons were emaciated and blind. Biomicroscopy and direct and indirect ophthalmoscopy findings of the Tippler pigeons were normal with the exception of partially dilated pupils at rest. Scotopic (blue stimuli) and photopic monocular electroretinograms were extinguished in the blind Tippler pigeons. Histological and electron microscopy studies revealed reduced numbers of rods and cones, and an absence of the double cone complex. The photoreceptor cells' outer segments were absent, and the inner segments were short and broad. The number of cell nuclei in the outer and inner nuclear layers was decreased, and the internal and external plexiform layers were reduced in width. Photoreceptor cell endfeet with developing synaptic ribbons were present in the external plexiform layer. Inflammatory cell and subretinal debris was not seen. The electroretinographic, histopathological, and ultrastructural findings of the blind Tippler pigeons support the diagnosis of a photoreceptor cell dysplasia.

Chromogenic assays for equine coagulation factors VII, VIII:C, IX, and X, and C1-esterase inhibitor.

OBJECTIVES: To adapt manual human chromogenic assays for coagulation factors VII (F.VII), VIII:coagulant (F.VIII:C), IX (F.IX), and X (F.X), and C1-esterase inhibitor (C1-INH) for use with an automated analyzer, and to measure the activity of these proteins in horses. ANIMALS: 10 healthy horses were used to determine ranges for the assays. Pooled plasma for standards was collected from an additional 20 healthy horses. PROCEDURE: A computer-assisted analyzer was programmed from the manual method for commercially available human F.VII, F.VIII:C, F.IX, F.X, and C1-INH chromogenic assay kits. Standards were prepared from pooled citrated equine plasma for the F.VII, F.VIII:C, and F.X assays, and from commercial pooled citrated human plasma for F.IX and C1-INH assays. RESULTS: Mean +/- SD activities in citrated equine plasma from 10 horses were 226 +/- 19% for F.VII; 209 +/- 31% for F.VIII:C; 149 +/- 38% for F.IX; 88 +/- 12% for F.X; and 18.4 +/- 8.4% for C1-INH. Intra-assay coefficients of variation (CV) were 5.3% for F.VII; 2.1% for F.VIII:C; and 3.0% for C1-INH. Interassay CV were 5.7% for F.VII; 7.4% for F.VIII:C; 3.8% for F.IX; 14.4% for F.X; and 22.0% for C1-INH. CONCLUSIONS: Human chromogenic assay kits can be automated and used to measure F.VII, F.VIII:C, F.IX, F.X, and C1-INH activities in citrated equine plasma. CLINICAL RELEVANCE: Human chromogenic assays can be routinely used to measure F.VII, F.VIII:C, F.IX, F.X, and C1-INH in horses, and may be useful in evaluation of horses with disorders of hemostasis.

Analysis of coagulation proteins as acute-phase reactants in horses with colic.

OBJECTIVES: To measure coagulation factor VIII:coagulant (F.VIII:C) and C1-esterase inhibitor (C1-INH), hemostasis-associated acute-phase reactant proteins and coagulation factors VII (F.VII), IX (F.IX), and X (F.X), hemostasis proteins not associated with an acute-phase response, in a select population of horses with colic and hemostasis abnormalities, and presumed to have acute-phase changes. To compare these values and other routine hemostasis test results in the horses with colic with values for a population of healthy horses. To correlate the values of known equine acute-phase reactants, F.VIII:C and fibrinogen, to those of other tests of hemostasis. To identify hemostasis-associated acute-phase reactant proteins and gain insights into the effects the acute-phase response has on hemostatic abnormalities in horses with colic syndrome. SAMPLE POPULATION: 54 plasma samples from horses with colic attributable to inflammatory (n = 39) or strangulating (n = 15) intestinal disorders. PROCEDURE: Plasma samples were evaluated for activities of F.VII, F.VIII:C, F.IX, F.X, C1-INH, antithrombin III, protein C, plasminogen, and alpha 2-antiplasmin (alpha 2AP); fibrinogen concentration; and prothrombin (PT) and activated partial thromboplastin (APTT) times. RESULTS: Horses with colic had significantly higher fibrinogen concentration, greater alpha 2AP and protein C activities, and longer PT and APTT than did healthy horses. Horses with colic also had significantly lower mean F.VII activity than did healthy horses. Significant positive correlations between fibrinogen concentration and F.VIII:C, C1-INH, and alpha 2AP values, and between F.VIII:C activity and fibrinogen, C1-INH, alpha 2AP, and plasminogen values were identified. CONCLUSIONS: An acute-phase response contributes to changes observed in coagulation proteins in horses with colic attributable to inflammatory and strangulating intestinal disorders. The data suggest that plasminogen, alpha 2AP, and C1-INH, should be considered equine acute-phase proteins.

Hemostatic and fibrinolytic indices in neonatal foals with presumed septicemia.

Thirteen coagulation tests evaluating hemostatic and fibrinolytic indices and serum cytokine and plasma endotoxin concentrations were obtained in 34 foals with a positive sepsis score (septic group) and 46 age-matched healthy foals. Compared to healthy foals, the prothrombin, activated partial thromboplastin, and whole blood recalcification times were significantly longer in septic foals. The fibrinogen and fibrin degradation products concentrations, percent plasminogen, alpha-2 antiplasmin, and plasminogen activator inhibitor activities, and tumor necrosis factor and interleukin-6 activities were greater in septic foals. Protein C antigen and antithrombin III activity were significantly lower in septic foals. Blood cultures were positive for growth and endotoxin was detected in 19 of 29 and 15 of 30 septic foals, respectively. In septicemic foals with detectable endotoxin in the plasma, the prothrombin and activated partial thromboplastin times were significantly longer and the plasminogen and antithrombin III activities were significantly less than in septic foals in which endotoxin was not detected. Twenty-three of the 34 septic foals did not survive. Septic foals that did not survive were most likely to have a positive blood culture in which a gram-negative organism was isolated. Histopathologic evidence of hemorrhage was evident in 11 foals at postmortem examination and thrombosis was identified in 2 foals. The prothrombin time was significantly longer in foals that had multisite hemorrhage at postmortem examination. The results of this study indicate that clinically relevant alternations in hemostatic and fibrinolytic indices occur in neonatal foals with septicemia and that derangements can be correlated with the presence of endotoxin in plasma. Derangements in hemostatic or fibrinolytic indices were helpful in identification of septic foals with increased risk of coagulopathy, but were not helpful in predicting hemorrhage as compared to thrombus formation. Survival of septicemic foals was correlated with gram-negative bacteremia, but not with the presence of endotoxin or coagulopathy.

Enzyme-linked immunosorbent assay for thrombin-antithrombin III complexes in horses.

OBJECTIVES: To adapt and characterize a human ELISA kit to quantify thrombin-antithrombin III (TAT) complexes in horses, and to evaluate TAT as a marker for hypercoagulation in horses. ANIMALS: 29 clinically normal horses used as controls, and 4 ill horses used to evaluate assay for known causes of hypercoagulation. PROCEDURE: A commercially available human sandwich-type ELISA kit with 2 antibodies against human thrombin and antithrombin III that bind selectively to their corresponding TAT antigenic sites was used. Equine TAT standards were made from purified equine thrombin and antithrombin III. Proteins diluted in a phosphate-buffered saline solution containing 0.1% Tween and 1 U of heparin/ ml were used to establish standard curves. Reference intervals for TAT concentration in citrated equine plasma, and intra- and interassay coefficients of variation were determined. RESULTS: Mean +/- SD values were 3.95 +/- 1.93 micrograms/L, with median of 3.18 micrograms/L and range of 1.95 to 9.03 micrograms/ L. One horse with cecal perforation had TAT concentration of 174.30 micrograms/L, and a horse infused IV with endotoxin had TAT concentration of 62.98 micrograms/L 12 hours after infusion. CONCLUSIONS: The data suggest that human TAT ELISA kits can be used to measure TAT concentration in citrated equine plasma, and that TAT is a marker for hypercoagulation in horses. CLINICAL RELEVANCE: Assays for equine TAT many help to further characterize the hypercoagulable state in horses.

Use of enzyme-linked immunosorbent assay to measure thrombin-antithrombin III complexes in horses with colic.

OBJECTIVES: To evaluate new ELISA for measurement of thrombin-antithrombin III (TAT) concentration, and to correlate the values to other tests of hemostasis in horses with colic. DESIGN: Plasma TAT concentration and 8 other hemostasis analytes were measured in horses with colic at hospital admission and during the next 4 days. Retrospectively, data were analyzed by outcome, broad-category diagnosis, and clinical management, and for correlation between TAT and other assays. ANIMALS: 100 horses with colic. PROCEDURE: Plasma samples were evaluated for TAT, fibrinogen, and fibrin degradation products concentrations; antithrombin III (ATIII), protein C, alpha 2-antiplasmin, and plasminogen activities; prothrombin time (PT); and activated partial thromboplastin time. RESULTS: Changes were indicative of a hypercoagulable state, most severe in nonsurviving horses, characterized by increased TAT concentration; decreased ATIII, protein C, and plasminogen activities; and increased PT. Nonsurvivors had significantly increased TAT concentration compared with that in survivors, without regard to sample collection time; however, compared over time, TAT was significantly increased only at admission. Highest TAT concentration was in nonsurvivors with inflammatory intestinal lesions. There was significant negative correlation between TAT and ATIII, protein C, alpha 2-antiplasmin, and plasminogen values, and significant positive correlation between TAT and PT, and fibrin degradation products values. CONCLUSIONS: Plasma TAT reflects the current state of coagulation system activation and is a good assay for early diagnosis of the hypercoagulable state in horses with the most severe forms of colic. CLINICAL RELEVANCE: Measurement of equine TAT provides further information to characterize the hypercoagulable state in horses to aid in case management.

Surgical creation of portacaval shunts during temporary intestinal arterial and venous occlusion in dogs.

OBJECTIVE: To surgically create complete portacaval shunts in dogs during temporary arrest of intestinal arterial and portal venous blood flow. DESIGN: Complete portacaval anastomoses were surgically created, and liver function was evaluated for 14 to 18 weeks after surgery. ANIMALS: 32 adult mixed-breed dogs of either sex. PROCEDURE: Administration of deferoxamine and temporary intestinal arterial occlusion were used to minimize the intestinal cellular damage resulting from the complete, temporary arrest of portal venous blood flow during creation of the portacaval anastomosis. Side-to-side, appositional anastomoses ( > 2 cm diameter) were formed between the portal vein and caudal vena cava. Dogs were observed daily for signs of hepatic encephalopathy, and food intake was recorded. Body weight was recorded weekly. Preprandial plasma ammonia, serum urea nitrogen, and glucose concentrations and sulfobromophthalein retention were measured monthly. The dogs were euthanatized, and necropsy was performed 14 to 18 weeks after surgery. RESULTS: 30 of 32 dogs recovered without complications. Complete portosystemic shunting was documented by increased plasma ammonia concentration, decreased serum urea nitrogen and glucose concentrations, prolonged sulfobromophthalein retention (P < 0.01), and inspection at necropsy. CONCLUSION: This method of providing temporary, complete arrest of portal venous blood flow was helpful in allowing accurate, appositional portacaval anastomoses to be created that remained patent for 14 to 18 weeks. CLINICAL RELEVANCE: This method of providing temporary, complete arrest of portal venous blood flow may prove useful in clinical surgery when temporary arrest of portal blood flow is desired.

Accuracy of interpretation of microscopic images of cytologic, hematologic, and histologic specimens using a low-resolution desktop video conferencing system.

OBJECTIVE: To determine the accuracy of interpretation of microscopic images for pathologic study transmitted over Switched-56 lines using a desktop interactive video conferencing system. MATERIALS AND METHODS: In subjective studies, two systems were connected using null-modem cables, which allowed evaluation of different bandwidths from 56 kbps to 384 kbps. Objective studies were done with two systems connected at distant sites via paired Switched-56 lines that produced an effective bandwidth of 112 kbps. A video camera mounted on a microscope was attached to the sending system. The resolution of the video image on the video conferencing system was 352 x 288 lines. Cases for cytology, hematology, and histopathology studies were selected from archives; one pathologist transmitted microscopic images, and a second pathologist made interpretations. The three pathologists were Board certified with similar experience that ranged from 20 to 35 years. Categories of interpretations or observations were predetermined for each study to allow the data on agreement between the direct microscopic interpretation or observation and that recorded by the receiving pathologist to be corrected for agreement attributable to chance alone. The results were analyzed using the kappa statistic. RESULTS: In the subjective studies, image degradation prevented interpretation while the microscope stage was moved. This problem occurred at all bandwidths tested. Image quality limited microscopic details. Organisms < 1 micron in diameter could not be seen reliably. In objective cytologic studies, overall agreement was recorded on 89 of 99 observations. In the four categories of specimens, observed agreement ranged from 0.778 to 0.958, and kappa was 0.704 to 0.948. For hematology specimens, overall agreement was found on 69 of 80 observations; observed agreement on eight types of nucleated blood cells ranged from 0.5 to 1.0, and kappa was 0.429 to 1.0. Poorer color definition and image quality prevented accurate identification of lymphoblasts and eosinophils in particular. For histologic specimens, overall agreement was obtained on 56 of 66 observations, observed agreement on four categories of histologic change ranged from 0.73 to 0.93, and kappa was 0.47 to 0.9. CONCLUSIONS: The desktop interactive video conferencing system, as configured in this study, was unsuitable for making definitive diagnoses from transmitted microscopic images.

Intravascular and peritoneal coagulation and fibrinolysis in horses with acute gastrointestinal tract diseases.

Components of the coagulation and fibrinolytic cascades, prothrombin and activated partial thromboplastin times, endotoxin activity, and albumin concentration were measured in blood and peritoneal fluid from 20 healthy horses and from 153 horses with acute gastrointestinal tract diseases at admission. Overall, 77% (117/153) of affected horses survived to discharge from the hospital, and 85% (82/97) of horses discharged were reported to be normal 9 to 14 months later. Significant differences in hemostatic factors were more common in peritoneal fluid than in blood. Tissue plasminogen activator, plasminogen, protein C, antithrombin III, and alpha 2-antiplasmin activities and concentrations of fibrinogen and fibrin degradation products were significantly (P < 0.05) greater in peritoneal fluid from horses with colic, and, with the exception of fibrinogen concentration, were associated with detection of endotoxin. Higher values for these variables, except tissue plasminogen activator activity, were significantly (P < 0.05) associated with survival. Plasminogen, antithrombin III, and alpha 2-antiplasmin activities were significantly (P < 0.05) greater in peritoneal fluid from horses with inflammatory or strangulating lesions, compared with those in horses with simple colic. Plasminogen-activator inhibitor type 1 activity, fibrin degradation products concentration, and prothrombin time were significantly (P < 0.05) greater in the blood of horses with colic. Survival was inversely associated with significantly (P < 0.05) greater intravascular concentrations of fibrin degradation products and fibrinogen and prothrombin time. This study revealed marked contrasts between peritoneal and intravascular coagulation and fibrinolysis in horses with colic, indicating that inferences regarding the peritoneal environment, particularly with respect to fibrinolytic capacity, should not be made on the basis of factors measured in blood.

Hemostatic indices in healthy foals from birth to one month of age.

Hemostatic indices were determined in 45 healthy light breed foals, from birth to 1 month of age, and in 20 healthy adult (> 2 years of age) light breed horses. Blood samples were obtained from each foal at 4 ages: 1) < 24 hours, 2) 4-7 days, 3) 10-14 days, and 4) 25-30 days. The following hemostatic indices were determined: platelet count; prothrombin and activated partial thromboplastin times; activity concentrations of protein C, antithrombin III, plasminogen, alpha-2 antiplasmin, tissue plasminogen activator, and plasminogen activator inhibitor-1; plasma protein C antigen and fibrinogen concentrations; and serum fibrin degradation products concentration. Prothrombin and activated partial thromboplastin times were significantly longer at birth than in older foals. The plasma concentrations of the following were significantly lower at birth than in older foals: antithrombin III, plasminogen and tissue plasminogen activator activities, protein C antigen, and fibrinogen. Concentrations of the following were significantly higher at birth than in older foals: protein C and plasminogen activator inhibitor-1 activities and fibrin degradation products. These results indicate that hemostatic indices of neonatal foals differ significantly from those of older foals and adults. With the exceptions of antithrombin III and tissue plasminogen activator activities, all hemostatic indices measured in foals at 1 month of age were equivalent to adult values.

Regulation of equine fibrinolysis in blood and peritoneal fluid based on a study of colic cases and induced endotoxaemia.

Much of the pathophysiology associated with equine gastrointestinal diseases is attributed to the effects of endotoxin on haemostasis. Because little is known about the responses of the equine fibrinolytic system to endotoxin, regulation of the system was investigated. Tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor type-1 (PAI-1) were identified as the primary plasminogen activator and plasminogen activator inhibitor, respectively, in equine blood. Under experimental conditions, the equine fibrinolytic system responded to endotoxin in a manner similar to that reported in man, with an early, transient increase in t-PA activity followed by an overwhelming and prolonged increase in activity of PAI-1. To investigate the response of the equine fibrinolytic system to clinical endotoxaemia, endotoxin concentrations were measured in plasma and peritoneal fluid, and activities of t-PA and PAI-1 were compared between healthy horses (n = 38) and horses with naturally occurring gastrointestinal diseases (n = 150). It was observed that plasma PAI-1 and peritoneal t-PA were increased concurrently in abnormal horses; and that these increases were associated with the presence of endotoxin. The results of this study suggest that 1) fibrinolysis is regulated in horses in a manner similar to that in man; 2) regulation of fibrinolysis is altered in endotoxaemic horses with gastrointestinal diseases; 3) events occurring in the vascular system may not reflect those in the peritoneal cavity; and 4) t-PA activity is increased in the peritoneal fluid of endotoxaemic horses with gastrointestinal diseases.

Microcytosis and iron status in dogs with surgically induced portosystemic shunts.

Microcytosis is common in dogs with congenital portosystemic shunts (PSS) and acquired liver disease. The objective of this study was to determine if microcytosis could be induced in normal dogs by surgical creation of PSS, and to characterize the changes in hematology and iron status. Hematocrit, mean cell volume, mean cell hemoglobin, and mean cell hemoglobin concentration decreased linearly from 45.5%, 69.1 fL, 22.8 g/dL and 33.1% to 39.5%, 55.9 fL, 17.8 g/dL and 31.9%, respectively, 18 weeks after creation of PSS. The erythrocyte count did not change, but red cell distribution widths indicated a shift to a heterogenous population with decreased volume. Mean cell volume and mean cell hemoglobin decreased rapidly after induction of PSS and were significantly (P < .05) different from presurgery values within 2 weeks. Serum iron and copper concentrations and total iron binding capacity were decreased in dogs with PSS. Liver iron concentration doubled after creation of PSS, with the majority of stainable iron located in Kupffer cells. The changes in erythrocyte indices and measures of iron status in dogs with surgically induced PSS were similar to those in dogs with congenital PSS. Microcystosis developed rapidly in dogs after induction of PSS. These results indicate that iron deficiency was not the cause of microcytosis in these dogs.

Analysis of hemostasis in horses with colic.

Eight tests of hemostasis were measured in 233 horses with colic. Blood samples were obtained at admission and for 4 consecutive days of hospitalization. Data were analyzed retrospectively by outcome, by broad-category diagnosis group, by small intestinal disorder, and by smaller categories for comparing specific diseases. Nonsurviving horses and horses with the most severe forms of intestinal ischemia had changes interpreted as hypercoagulative, the intensity of which was increased on the first and second mornings (sample times 2 and 3) after admission, when most significant differences for results of specific tests were detected. Nonsurvivors had decreased antithrombin III activity and prolonged prothrombin and activated partial thromboplastin times; those with strangulating obstructions also had decreased protein C and plasminogen activities. During hospitalization and with survival, these changes tended to reverse. In most horses, regardless of diagnosis or outcome, concentration of fibrin degradation products and fibrinogen, and alpha 2-antiplasmin activity increased over time. Whether these changes reflected specific effects of colic or of the acute-phase response was not determined. In comparisons of small intestinal disorders (proximal enteritis, strangulations, and impactions), diagnostically distinguishing features were not found. Likewise, in comparisons of specific diseases (small vs large intestinal impaction, proximal enteritis vs colitis, small vs large intestinal obstruction), diagnostically distinguishing features were not found.

Hypercoagulable state associated with a deficiency of protein C in a thoroughbred colt.

Protein C is a vitamin K-dependent serine protease with anticoagulant and profibrinolytic activity which is synthesized in the liver. Decreased protein C activity was detected in a Thoroughbred colt with clinical and histopathologic evidence of recurrent venous thrombosis. Although protein C activity was reduced, protein C antigen concentration was normal. Consumptive coagulopathies produce a decrease in both the functional and antigenic concentrations of protein C, thus a defect in protein C synthesis was suspected. Inhibition of gamma-carboxylation secondary to vitamin K antagonism results in the synthesis of a protein C molecule with antigenicity, but without biological activity. However, there was no evidence of vitamin K antagonism. The hypercoaguable state resulting from the reduced activity of protein C in this colt was associated with uncomplicated renal disease, rather than a protein C consumptive process such as endotoxemia. A primary hypercoagulable state due to a deficiency of protein C activity was diagnosed. Primary deficiencies of protein C activity have not been previously documented in horses.

Effects of exploratory laparotomy on plasma and peritoneal coagulation/fibrinolysis in horses.

Plasma and peritoneal fluid samples were collected before and after surgery from 6 horses undergoing a ventral midline exploratory laparotomy and from 6 anesthetized control horses. Coagulation/fibrinolytic components measured in the plasma and peritoneal fluid of these horses included the functional activity of antithrombin III, alpha-2 antiplasmin, plasminogen, and protein C, and the concentrations of fibrinogen and fibrin degradation products. Peritoneal fluid antithrombin III, fibrin degradation products, and plasminogen values were significantly increased after surgery (over time) in principal horses. Compared with control horses, postoperative peritoneal fluid from horses undergoing laparotomy had significantly increased antithrombin-III activity at 12 and 72 hours, alpha-2 antiplasmin activity at 24 hours, fibrin degradation product concentrations at 6, 12, 24, 72, 96, and 144 hours, plasminogen activity at 6, 12, 24, 48, 72 and 96 hours, and protein-C activity at 12, 24, 72, and 96 hours. There were no significant changes in the peritoneal fibrinogen concentration in principal horses. Plasma plasminogen activity was significantly decreased at 24 hours after surgery in principal horses, compared with controls. Changes were minimal in the remaining plasma coagulation/fibrinolytic components of horses undergoing laparotomy. Plasma and peritoneal fluid values of anesthetized control horses did not change.

Use of newly developed assays for protein C and plasminogen in horses with signs of colic.

Protein C content and plasminogen activity were measured in plasma from 100 horses with signs of colic. Data were analyzed by grouping horses 4 ways. Each horse was allotted to 1 of 2 outcome groups (survivors and nonsurvivors), 1 of 3 broad-category diagnosis groups (inflammatory disorders, strangulating obstructions, and all other gastrointestinal disorders), and 1 of 2 clinical management groups (medical and surgical). In a fourth grouping, all horses (although numbers of horses included in each subgroup were small) were assigned either to specific diagnostic groups that had high expectation for activated hemostasis (intestinal ischemia, endotoxemia, jugular thrombosis, peritoneal adhesions, and laminitis) or to a control group, in which active hemostasis was unlikely. Within 2 to 24 hours after admission, nonsurvivors developed lower protein C content than did survivors. Protein C content and plasminogen activity became low during hospitalization in horses with strangulating obstructions and in horses having surgery. The results from the grouping by specific diagnosis must be considered pilot data because the numbers of horses in each subgroup were small. Although not statistically significant, trends were noticed in protein C and plasminogen: (1) horses with intestinal ischemia and endotoxemia developed low protein C content and plasminogen activity, (2) protein C content became low in horses that developed peritoneal adhesions or laminitis, and (3) plasminogen activity became low in horses that developed jugular thrombosis. Low protein C content or low plasminogen activity, or both, may be useful as predictors for outcome and for these specific complications of equine colic.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of equine digital Starling forces and hemodynamics during early laminitis.

A carbohydrate overload model was used in 8 horses to evaluate Starling forces and hemodynamics of the digit during the prodromal stage of acute laminitis. A pump-perfused extracorporeal digital preparation was used to evaluate blood flow, arterial pressure, venous pressure, capillary pressure, isogravimetric capillary filtration coefficient, osmotic reflection coefficient, and vascular compliance. From these data, pre- and postcapillary resistances and pre- to postcapillary resistance ratios were determined. Vascular and tissue oncotic pressures were estimated from plasma and lymph protein concentrations, respectively. The osmotic reflection coefficient, an estimation of capillary permeability, was determined by means of the lymph protein wash-down technique. Using the collected data, tissue pressure in the digit was calculated by use of the Starling equation. In the isolated digit, mean isogravimetric capillary pressure was 55.13 mm of Hg, mean plasma and lymph oncotic pressures were 22.29 mm of Hg and 7.2 mm of Hg, respectively, the mean osmotic reflection coefficient was 0.66, the mean capillary filtration coefficient was 0.003 ml/min/mm of Hg/100 g, and mean interstitial fluid pressure was 44.82 mm of Hg. The high capillary pressure appeared to be caused by high vascular resistance from the venous side, predisposing to enhanced capillary filtration and interstitial fluid accumulation.

Evaluation of coagulation and fibrinolysis during the prodromal stages of carbohydrate-induced acute laminitis in horses.

The balance of coagulation and fibrinolysis was studied in 15 horses during the prodromal stages of acute laminitis induced by carbohydrate overload. Progression of the disease was stopped 12 to 24 hours before the expected onset of lameness in trial 1 (8 horses) and at the onset of lameness in trial 2 (7 horses). The end points in each trial were identified by specific changes in blood pressures (trial 1) and by changes in pulse, rectal temperature, and arterial pressure (trial 2) that were anticipated on the basis of original description of the experimental model. Blood samples for hemostasis evaluation were collected before and after carbohydrate overload in trial 1 and after carbohydrate overload in trial 2. Significant changes were not detected in platelet count, mean platelet volume, prothrombin time, activated partial thromboplastin time, fibrinogen concentration, plasminogen concentration, alpha-2-antiplasmin, antithrombin III, protein C, thromboxane B2, or fibrin(ogen) degradation product concentration. We concluded that an imbalance in coagulation and fibrinolysis is not pathogenic in the onset of experimentally induced equine acute laminitis. Because several test methods used to evaluate hemostasis in these horses were new, reference values for 34 healthy adult horses were established.

Antigenic assay for protein C determination in horses.

An antigenic assay was developed for determination of protein C in horses. Protein C, a natural, vitamin K-dependent anticoagulant component in blood, was isolated from equine plasma, a specific antibody was produced in goats, and a rocket electroimmunophoresis assay was established. Tests were performed to verify the identity of the isolated protein C and to determine the purity of the antibody. Protein C antigen was measured in plasma from 34 clinically normal horses, and values were compared with amidolytic function values. The mean (+/- SD) values for the 2 test methods were similar (antigen content, 104.5 +/- 13.8%; amidolytic activity, 104.6 +/- 17.5%), but the correlation coefficient was 0.1. Four horses given Na coumarin had markedly decreased plasma protein C amidolytic activity and minimal decrease in protein C antigen content.

Chromogenic assay for equine plasminogen.

A functional assay for equine plasminogen was established, using urokinase as the activator, a synthetic chromogenic substrate, a computer-assisted centrifugal analyzer, and acidified/neutralized plasma. One documented effect of plasma acidification appears to be inactivation of alpha-2-antiplasmin. Intra- and interassay precision testing yielded coefficients of variation of 4.1% (n = 10) and 5.6% (n = 26), respectively. Plasminogen was stable in equine plasma stored up to 1 week at 4 C and up to 5 months at -70 C. Plasminogen in nonacidified equine plasma was not activated by urokinase, streptokinase, tissue plasminogen activator, or tissue plasminogen activator plus soluble fibrin. Streptokinase also failed to activate plasminogen in acidified/neutralized equine plasma.