Effect of desmopressin on plasma factor VIII and von Willebrand factor concentrations in Greyhounds

Objective To determine the effect of 1-Deamino-8-D-argi-nine vasopressin on plasma concentrations of von Willebrand factor and factor VIII in Greyhound blood donors, and to compare the response of 1-Deamino-8-D-arginine vaso-pressin injection on plasma concentrations of von Willebrand factor between groups with different resting plasma concentrations of von Willebrand factor.
Animals Fifteen Greyhound blood donors were used. Dogs were grouped into three categories depending on their von Willebrand factor concentrations.
Procedure Desmopressin was administered subcuta-neously at 1 mg/kg to all dogs. Plasma von Willebrand factor and factor VIII concentrations were measured before and 10, 20, 30, 45, 60, 90 and 120 min after desmopressin injection.
Results The von Willebrand factor and factor VIII concentrations in all dogs increased significantly and remained higher than base-line throughout the 2 h period.
Conclusion Desmopressin is useful in increasing von Willebrand factor concentrations in Greyhound blood donors, including those with low resting concentrations.     

Effect of Desmopressin Acetate on Bleeding Times and Plasma von Willebrand Factor in Doberman Pinscher Dogs with von Willebrand''s Disease

Effects of desmopressin acetate (1-desamino-8-D-arginine vasopressin [DDAVP]) on plasma von Willebrand factor (vWf) were studied in 12 purebred Doberman pinschers confirmed to have von Willebrand's disease (vWd) (plasma vWf antigen [vWf:Ag] concentrations, less than 30 U/dl). Twelve dogs had subnormal plasma botrocetin cofactor (BCf) activity and 11 dogs had prolonged buccal mucosa bleeding times. Tranquilization of three dogs with lenperone and three dogs with xylazine did not induce significant changes in mean plasma vWf:Ag concentrations or mean BCf activities. Thirty and 120 minutes after administration of DDAVP (1 micrograms/kg subcutaneously), there was significant shortening of the mean buccal mucosa bleeding time. Ten dogs responded to DDAVP with increases in BCf activity which exceeded 10 U/dl at 30 or 120 minutes, or both, after the drug was administered. At the same time, increases in plasma vWf:Ag concentrations were smaller than the increases in BCf activity. It was shown by multimeric analysis that primarily the higher molecular weight forms of vWf increased in plasma in response to DDAVP.     

The effects of desmopressin on plasma factor VIII/von Willebrand factor activity in dogs with von Willebrand''s disease.

Eight unanesthetized normal dogs and seven dogs with von Willebrand's disease (vWD) were given desmopressin (0.6 micrograms/kg, IV) in order to determine the effects of this drug on plasma Factor VIII/vWF activity. Seven of the normal dogs and four of the vWD dogs were administered an equal volume of saline (control infusion) on another occasion. The other three vWD dogs underwent major surgery after treatment with desmopressin. Plasma FVIII coagulant activity (FVIII:C), von Willebrand factor antigen (vWF:Ag), and FVIII-ristocetin co-factor activity (FVIII:RC) were quantitated before infusion and at 60 minutes postinfusion. Activities were expressed as a percentage of the activity of a pooled canine plasma (12 dogs) arbitrarily designated as having 100% FVIII:C, vWF:Ag, and FVIII:RC activity. Plasma FVIII:C activity increased by 28% in the normal dogs and by 37% in the dogs with vWD. Plasma vWF:Ag increased more than twofold in normal dogs after desmopressin treatment. In the vWD dogs the average increase was also twofold, however there was much greater variability between dogs with increases ranging from 1.2 fold to 2.4 fold. Plasma FVIII:RC activity almost doubled in normal dogs, however like vWF:Ag, the increases in vWD dogs were more variable. One vWD dog had no increase in FVIII:RC while in the remaining six dogs FVIII:RC increases ranged from 1.8 to 2.9 fold. The results of this study indicate that a single intravenous dose of desmopressin (0.6 micrograms/kg) causes a significant elevation in plasma vWF:Ag and FVIII:RC activity and a much lesser increase in FVIII:C activity in normal unanesthetized dogs.(ABSTRACT TRUNCATED AT 250 WORDS)     

DDAVP-induced increases in coagulation factor VIII and von Willebrand factor in the plasma of conscious dogs

Infusion of the vasopressin analogue DDAVP into five normal dogs at doses of 0.1-2.0 micrograms DDAVP per kg body weight induced dose-dependent increases in the plasma content of coagulation factor VIII and von Willebrand factor. Plasma concentrations of von Willebrand factor (determined antigenically as factor VIII-related antigen and functionally as coagglutinin cofactor activity) and coagulation factor VIII were measured immediately before and at 10, 30, and 120 min after 10-min intravenous infusions of DDAVP. The greatest increases in coagulation factor VIII were produced with the 2.0 micrograms/kg dose. Ten minutes after infusion the mean increase in coagulation factor VIII was 32 units/dl (concentrations of all indices were reported relative to concentrations in a standard canine plasma pool, arbitrarily assigned a concentration of 100 units/dl) and this increase did not change significantly throughout the duration of the experiment. At 10 min post-infusion, the mean factor VIII-related antigen concentration increased 81 units/dl (dose = 2.0 micrograms/kg) and did not change significantly for the duration of the experiment. The maximum mean increase in coagglutinin cofactor activity, 141 units/dl, occurred 10 min after infusion (dose = 1.0 microgram/kg). Coagglutinin cofactor activity decreased significantly from peak activity by 120 min post-infusion.     

Stability of canine factor VIII activity and von Willebrand factor antigen concentration in vitro

The in vitro stability of canine factor VIII activity, von Willebrand factor antigen concentration and the ratio of these two factors was studied. Samples were stored for up to 48 hours, either as plasma or as whole blood, at 4 degrees, 20 degrees and 37 degrees C. Factor VIII activity was generally stable in both plasma and whole blood samples for up to 48 hours at 4 degrees or 20 degrees C. The concentration of von Willebrand factor antigen was more stable in samples stored as plasma than whole blood, and for a shorter time than factor VIII activity. Consequently, the stability of the ratio of these two factors was relatively poor in vitro.     

The effects of desmopressin on hemostatic parameters in the normal dog.

Seven normal unanesthetized dogs were infused with desmopressin in saline (doses 0.2, 0.4 and 0.6 micrograms/kg i.v.) or saline alone, on separate occasions in order to determine the effects of desmopressin on circulating factor VIII coagulant and factor VIII-related antigen activities, on partial thromboplastin times and prothrombin times, on blood platelet count and packed cell volume, and on serum osmolarity. Desmopressin caused a rapid dose-dependent elevation in both factor VIII activities peaking at 30-60 minutes postinfusion. At a dosage of 0.6 microgram/kg desmopressin induced rises in factor VIII coagulant activity and factor VIII-related antigen of 150% and 198% of the preinfusion levels respectively, while the half disappearance times were approximately five and six hours respectively. The other parameters were not significantly affected by even the highest dose of desmopressin used. These results indicate that normal dogs respond to desmopressin in a manner similar to man. The major differences are in the dosage required to produce comparable effects (higher in dogs), and in the fact that the factor VIII-related antigen activity consistently responds to a greater degree than the coagulant activity. It is concluded that desmopressin may have particular value in stimulating elevations in plasma factor VIII-related antigen in dogs deficient in this plasma protein. Six dogs were pretreated with the endorphin inhibitor, naloxone, (0.04 mg/kg I.V.) or sterile saline, three minutes prior to infusion with desmopressin at a dose of 0.6 microgram/kg i.v.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of acepromazine, xylazine and thiopentone on factor VIII activity and von Willebrand factor antigen concentration in dogs

The effect of acepromazine maleate, xylazine and thiopentone on the packed cell volume, plasma protein content, factor VIII activity and von Willebrand factor antigen concentration of blood was studied in normal dogs. The same variables were measured in dogs with haemophilia A given acepromazine maleate and thiopentone. Both the packed cell volume and plasma protein content decreased after the administration of either acepromazine maleate or xylazine. Values were not changed further after administration of thiopentone. Changes in the haemostatic variables measured were generally small. Consequently, blood samples collected from dogs under the influence of premedicant doses of acepromazine maleate or xylazine, and when subsequently anaesthetised with thiopentone, are adequate for the assay of factor VIII activity and von Willebrand factor antigen concentration for establishing an animal's haemophilia A and von Willebrand's disease status.     

Von Willebrand's disease in dogs in the United Kingdom

An assay for the measurement of von Willebrand factor antigen has been established. In a period of 18 months, 13 dogs have been identified as suffering from von Willebrand's disease. The affected animals had levels of von Willebrand factor antigen which ranged from undetectable to 43 per cent of normal. Factor VIII levels were also reduced. Haemorrhagic episodes were usually associated with trauma or surgery, and often required transfusion with fresh blood or plasma to arrest haemorrhage.     

Plasma von Willebrand factor concentration and thyroid function in dogs

Plasma von Willebrand factor antigen concentration was determined in 15 dogs with suspected hypothyroidism, in 1 dog with hyperthyroidism, and in 14 euthyroid dogs. The mean +/- SEM von Willebrand factor:antigen concentration in hypothyroid dogs (47.1% +/- 12.6%) was significantly decreased (P less than 0.0005), compared with that in euthyroid dogs (94.7 +/- 5.6%). Four hypothyroid dogs were given thyroxine for 1 month and all 4 had an increase in von Willebrand factor:antigen concentration. The plasma von Willebrand factor:antigen concentration was 200% in the hyperthyroid dog. Seemingly, reduced concentrations of plasma von Willebrand factor:antigen can be found in dogs in association with congenital von Willebrand disease or with von Willebrand disease acquired through hypothyroidism.     

von Willebrand disease phenotype and von Willebrand factor marker genotype in Doberman Pinschers

OBJECTIVE: To define the relationship between clinical expression of a type-1 von Willebrand disease phenotype and genotype at 2 von Willebrand factor marker loci in Doberman Pinschers. ANIMALS: 102 client-owned Doberman Pinschers. PROCEDURES: Dogs were recruited on the basis of plasma von Willebrand factor concentration, clinical history, and pedigree. Blood samples and response to a history questionnaire were obtained for each dog. Plasma von Willebrand factor concentration was measured by use of an ELISA, and genotyping was performed via polymerase chain reaction for 1 intragenic and 1 extragenic von Willebrand factor marker. Amplification product size was determined by use of polyacrylamide gel electrophoresis (intragenic marker) or automated sequence analysis (extragenic marker). Western blots were prepared from a subset of dogs with low plasma von Willebrand factor concentration to evaluate multimer distribution. RESULTS: Strong associations were detected between plasma von Willebrand factor concentration and von Willebrand factor marker genotype. Twenty-five dogs had substantial reduction in plasma von Willebrand factor concentration and multiple hemorrhagic events. All were homozygous for a 157-base-pair intragenic marker allele and homozygous or compound heterozygous for 1 of 4 extragenic marker alleles. These marker genotypes were exclusively detected in dogs with low plasma von Willebrand factor concentration, although some dogs with these genotypes did not have abnormal bleeding. CONCLUSIONS AND CLINICAL RELEVANCE: Type-1 von Willebrand disease in Doberman Pinschers is associated with the von Willebrand factor gene locus; however, the expression pattern in this breed appears more complex than that of a simple recessive trait.     

Multimeric analysis of von Willebrand factor before and after desmopressin acetate (DDAVP) administration intravenously and subcutaneously in male beagle dogs

Eight adult male Beagles were given 1 microgram of 1-deamino-8-D-arginine vasopressin (DDAVP)/kg of body weight, SC or by slow IV infusion on separate occasions. Both routes of administration induced highly significant increases (P less than 0.0001) in plasma concentrations of von Willebrand factor (vWf) antigen (Ag) and botrocetin cofactor (BCf) activity, an indication of platelet-associated vWf activity. In most instances, increases in plasma vWf:Ag and BCf values induced by SC injection were as large as or larger than those induced by slow IV infusion. With both routes of administration, BCf activity increased more than the vWf:Ag concentration, so that high BCf-to-vWf:Ag ratios were found in plasma after DDAVP administration. In plasma samples obtained before DDAVP administration, sodium dodecyl sulfate-electrophoresis resolved the vWf into a series of multimeric proteins with molecular weights similar to those of human vWf. Sodium dodecyl sulfate-electrophoresis of plasma samples obtained after DDAVP administration revealed mainly the larger molecules of vWf that were increased by DDAVP administration.     

Changes in factor VIII activity and von Willebrand factor antigen concentration with age in dogs.

The factor VIII activity of 38 German shepherd puppies, 6-12 weeks old, submitted for diagnosis of haemophilia A was measured. Eight of these puppies had values higher than would be expected for haemophiliacs, but less than the reference range for adult dogs. A further sequential study of 21 puppies (6-26 weeks of age) indicated that the factor VIII activity of puppies is generally less than that of adult dogs until about 14 weeks of age. Changes in the concentration of von Willebrand factor antigen in the puppies were irregular. These variations are probably not sufficient to interfere with accurate diagnosis of haemophilia A in most affected young dogs, but may interfere with the detection of heterozygotes in young bitches.     

Effect of levothyroxine administration on hemostatic analytes in Doberman Pinschers with von Willebrand disease

Levothyroxine administration has been suggested to be an effective treatment for canine von Willebrand disease (vWd), but evidence supporting this treatment is lacking. Effects of levothyroxine administration were evaluated in 8 euthyroid Doberman Pinschers with plasma von Willebrand factor (vWf) concentrations < 15%, characteristic of type 1 vWd. Levothyroxine (0.04 mg/kg PO q12h) and placebo were administered for 30 days in a 2-period, 2-treatment, double-blinded, crossover design with a 30-day washout period between treatments. Buccal mucosal bleeding time (BMBT), plasma vWf concentration (vWf: Ag), vWf collagen binding activity (vWf:CBA), factor VIII coagulant activity (FVIII:C), and serum concentrations of total thyroxine (T4), free thyroxine (fT4), 3,5,3'-triiodothyronine (T3), and thyroid-stimulating hormone (TSH) were measured on days 0, 2, and 30 of each treatment period. The 8 dogs (1 male, 7 females) had markedly low plasma vWf:Ag (mean, 8.9%; reference range, 70-180%) and vWf:CBA (mean, 11.1%; reference range, >70%). Response to placebo versus levothyroxine treatment was not significantly different between groups at day 0, 2, or 30 for BMBT, vWf:Ag, vWf:CBA, and FVIII:C. Serum T4, fT4, and T3 concentrations were significantly higher and serum TSH significantly lower in the levothyroxine-treated group than in the placebo group at days 2 and 30. Administration of levothyroxine at 0.04 mg/kg caused laboratory evidence of hyperthyroidism but did not affect plasma FVIII:C and vWf:Ag concentrations or vWf-dependent collagen binding and BMBT. The results of this study failed to identify a direct action of levothyroxine supplementation on plasma vWf concentration or activity in euthyroid Doberman Pinschers with vWd.     

Comparative stability of canine and feline hemostatic proteins in freeze‐thaw‐cycled fresh frozen plasma

Objective – To evaluate the stability of canine and feline hemostatic proteins in freeze‐thaw‐cycled (FTC) fresh frozen plasma (FFP). Design – Prospective study. Setting – Veterinary Teaching Hospital. Animals – Nine blood donor dogs and 10 blood donor cats. Interventions – Whole blood was collected and separated into packed RBC and plasma units according to standard methods. Each unit of plasma was divided into 2 equal aliquots and frozen (?41°C). One aliquot from each donor (FTC) was then thawed and then refrozen (?41°C) until time of analysis. The second aliquot (nonfreeze‐thaw‐cycled; NFTC) remained frozen until time of analysis. The hemostatic proteins assessed included coagulation factors, anticoagulant factors (antithrombin and Protein C), and adhesive proteins (fibrinogen and von Willebrand Factor). The coagulant activities of factors II, VII, VIII, IX, X, XI, and XII were measured in modified one‐stage activated partial thromboplastin time or prothrombin time assays. Antithrombin and Protein C activities were measured in chromogenic substrate assays. Clottable fibrinogen was measured via the Clauss method, and von Willebrand Factor concentration (vWF:Ag) was measured in an ELISA. A paired t‐test was utilized to identify differences in factor activity or concentration between FTC FFP and NFTC FFP. Measurements and Main results – No clinically or statistically significant differences (all P>0.05) were identified between FTC FFP and NFTC FFP. Conclusions – Refreezing FFP within 1 hour of initial thawing appeared to have no deleterious effects on the hemostatic protein activity or content of that unit. Transfusion of FTC FFP is expected to provide the recipient with comparable replacement of hemostatic proteins as FFP that has remained frozen.     

Hemostatic defects associated with two infusion rates of dextran 70 in dogs.

We investigated changes in hemostatic function after infusion of 6% dextran 70 (high molecular weight dextran) at 2 rates. Six healthy dogs underwent 3 regimens: 20 ml of dextran/kg of body weight administered in 1 hour (trial A), 20 ml of dextran/kg administered in 30 minutes (trial B), and 0.9% sodium chloride solution as a control administered over 1 hour to achieve hemodilution equivalent to that for 20 ml of dextran/kg (trial C). Before and at 2, 4, 8, and 24 hours after the start of trials A and B, we measured PCV, total solids (TS) concentration, amount of von Willebrand factor antigen (vWf:Ag), factor VIII coagulant activity (VIII:C), prothrombin time, activated partial thromboplastin time (APTT), platelet retention in a glass bead column, and buccal mucosa bleeding time (BMBT). Values were not obtained at 8 and 24 hours for trial C. Saline-induced changes in hemostasis were significant (P less than 0.05) from baseline throughout the sample collection period. Significant differences (P less than 0.05) between trial A and control were observed for vWf:Ag, VIII:C, BMBT, APTT, TS, and PCV values at 2 hours, and for VIII:C at 4 hours. Significant differences (P less than 0.05) between trial B and control were observed for APTT, TS, and PCV values at 2 hours, and for vWf:Ag, VIII:C, BMBT, APTT, TS, and PCV values at 4 hours. During trials A and B, mean values of analytes infrequently deviated from reference intervals, and clinical signs of bleeding were not observed in any dog.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression of von Willebrand factor in plasma and platelets of cats

Immunochemical methods that are used to assess von Willebrand factor in human beings and dogs were used to assess von Willebrand factor in 3 cat species. Our findings indicated that the expression and multimeric composition of von Willebrand factor in plasma and platelets of cats were similar to those reported in human beings and dogs. We suggest that these methods may be used to evaluate von Willebrand disease in members of the cat family used in this study.     

Clearance of nasal mucus in nonanesthetized and anesthetized dogs

Clearance rates for nasal mucus in the maxillary turbinate region were measured in 8 Beagle dogs. 99mTc Macroaggregated albumin (10 microliters) was instilled in the nasal maxillary region of dogs under general anesthesia. A gamma camera was used to detect movement of the 99mTc macroaggregated albumin in the nose for 1 hour after it was instilled. Velocity of mucus was measured in the 8 dogs each under 3 conditions of anesthesia: anesthesia with pentobarbital given IV (20 mg/kg of body weight), anesthesia with halothane gas, and no anesthesia. Mean velocities (+/- SD) were 3.7 +/- 1.4 mm/min in dogs anesthetized with pentobarbital, 4.3 +/- 2.5 mm/min in dogs anesthetized with halothane, and 3.4 +/- 1.7 mm/min in awake dogs. The differences between the 3 anesthetic conditions were not significant at the P less than 0.05 level. Use of anesthesia at a light surgical plane provides a controlled method for measurement of clearance of nasal mucus with minimal alterations from the nonanesthetized state.     

Haemophilia A in German shepherd dogs

Haemophilia A was diagnosed in 14 male German shepherd dogs. Factor VIII: coagulant (FVIII:C) activities ranged from 1.13% of a normal canine plasma pool. von Willebrand's factor antigen values were normal or increased in all 9 of these dogs which were tested. Twelve of these dogs had a common maternal grandsire. Five dogs had exhibited no tendency to bleed (when tested between 2 to 23 months of age). They were tested because of pedigree links with clinically affected animals. Common clinical signs in the latter dogs included: bleeding from the mouth, subcutaneous and intramuscular haematomas and lameness. Since these dogs usually had a mild to moderate deficiency of FVIII:C, they may survive to adulthood without exhibiting clinical signs severe enough to necessitate veterinary attention.     

Evaluation of the haemostatic profile in the pre- and post parturient mare, with particular focus on the perinatal period.

Various haemostatic analytes were systematically evaluated for four months pre-partum and five months post partum in 14 healthy mares. The plasma fibrinogen concentration and both Factor VIII:C and von Willebrand factor activity showed gradual increases from mid-gestation and reached maximal, or near maximal activity at parturition. These increases were paralleled by an increase in plasma fibronectin concentration, the appearance of fibrinogen degradation products, and a modest rise in antithrombin III concentration. In contrast, the activity of Factor VII and Factor IX, and the one-stage prothrombin (PT) time and the activated partial thromboplastin (APTT) time remained relatively constant throughout the pre- and post parturient period.     

Efficacy of Fresh-Frozen Plasma and Cryoprecipitate in Dogs with von Willebrand's Disease or Hemophilia A

Here we report the comparative efficacy of fresh-frozen plasma (FFP) and Cryoprecipitate in the treatment of 2 inherited bleeding disorders in dogs. The dogs were divided into 3 groups, consisting of 4 Doberman Pinschers with type I von Willebrand's disease (vWD) (group I), 1 Scottish Terrier with type III vWD (group 2), and 4 German Shepherd Dogs with hemophilia A (group 3). In vWD, therapeutic efficacy was determined by the ability of the products to increase von Willebrand factor antigen (vWf:Ag) concentrations above 35 canine units (CU)/dL and to correct the prolonged buccal mucosal bleeding time. Therapeutic efficacy in hemophilia A was assessed by the ability of the products to increase the factor VIII coagulant (FVIII:C) activity above 30 CU/dL. In both groups 1 and 2, higher increases in vWf:Ag were achieved with Cryoprecipitate than with FFP, despite a significantly smaller total amount of vWf:Ag (in CU) being infused with Cryoprecipitate. The maximum vWf:Ag attained after infusion in group 1 was dependent on both the baseline vWf:Ag concentration and on the type of infusion product. The dogs with vWD in both groups also displayed a delayed increase in FVIII:C activity after infusion of both plasma products, which is characteristic of the disease. In group 3, Cryoprecipitate achieved similar increases in FVIII:C activity compared to FFP, although a significantly lesser amount of FVIII:C (in CU) was delivered with Cryoprecipitate. Six of the 9 dogs treated with FFP experienced adverse effects ranging from mild pruritus to pallor and weakness, whereas none of the 9 dogs treated with Cryoprecipitate had any observable adverse reactions ( P = .009). Based on its efficacy and safety, we recommend Cryoprecipitate over FFP for treatment or prophylaxis of hemorrhagic episodes in dogs with vWD or hemophilia A.