A Remote Assay for Measuring Canine Platelet Activation and the Inhibitory Effects of Antiplatelet Agents

Background

Antiplatelet medications are increasingly used in dogs. Remote analysis of platelet activity is challenging, limiting assessment of antiplatelet drug efficacy.

Hypothesis/Objectives

To evaluate a method used in humans for stimulation and remote analysis of canine platelet activity.

Animals

Forty‐five dogs of various ages without a coagulopathy or thrombocytopenia. Six were receiving antiplatelet medication.

Methods

Prospective observational study. Platelets were stimulated with combinations of arachidonic acid (AA) and epinephrine (Epi) or adenosine diphosphate (ADP) and the thromboxane A₂‐mimetic U46619 (U4). PAMFix was added to the blood samples to facilitate delayed analysis of platelet activity. Activity was assessed by flow cytometric measurement of surface P‐selectin (CD62P) expression.

Results

Canine platelets could be stimulated with both AA/Epi and ADP/U4. The levels of P‐selectin were significantly greater than paired, unstimulated samples (P < 0.001). Inhibition of P‐selectin expression occurred after this stimulation by adding antiplatelet drugs in vitro. The efficacy of antiplatelet drugs in samples from treated dogs was also measurable ex vivo using this method. Delayed analysis of platelet activity at time points up to 22 days demonstrated excellent correlation between respective mf values at each time point (r² = 0.92, P < 0.0001).

Conclusions and Clinical Importance

This study evaluated a new method to remotely assess canine platelet activity. It shows that PAMFix can be used for this purpose. This provides opportunities to interrogate the inhibitory action of antiplatelet drugs in clinical settings.     

Assessment of the red blood cell proteome in a dog with unexplained hemolytic anemia

A 7‐year‐old female neutered Jack Russell Terrier was presented to Langford Vets, the University of Bristol, with a history of chronic intermittent lethargy. Investigations and clinical course were compatible with hereditary hemolysis due to a red blood cell membrane defect. Proteomics was used to explore protein alterations in the presence of a hypothesized red blood cell membrane protein deficiency. Proteomic analysis revealed downregulation of the band 3, and alpha‐ and beta‐adducin proteins, and alterations in the red blood cell proteome consistent with previous reports of changes due to the presence of reticulocytosis and ongoing hemolysis. The spectrum of protein alterations identified in the affected dog may be homologous to a band 3 protein deficiency secondary to hereditary spherocytosis, as described in people.     

Crop evapotranspiration of processing tomato in the San Joaquin Valley of California, USA

Yield of processing tomato has increased by 53% over the past 35 years. Thus, concerns exist about the current seasonal crop evapotranspiration requirements of processing tomato compared to the past published requirements, which were about 645 mm. Also, the mid-season crop coefficient for processing tomato developed 35 years ago with sprinkler irrigation was 1.25, while a mid-season coefficient developed 20 years ago with subsurface drip irrigation was 1.05. Because of the age and variability of crop coefficients and the long-term yield increase, a study was conducted to determine the seasonal crop evapotranspiration and crop coefficients of processing tomatoes using the Bowen ratio energy balance method in eight commercial fields from 2001 to 2004. Measurements were made in both furrow- and drip-irrigated fields. Results showed seasonal crop evapotranspiration to range from 528 to 752 mm with an average of 648 mm. No statistical differences were found between furrow and drip irrigation. Mid-season crop coefficients varied between 0.96 and 1.09 with statistically similar values between furrow- and drip-irrigation for a given year. Current evapotranspiration rates were similar to those of the early 1970s, indicating that the water use efficiency of processing tomato increased substantially with time during the past 35 years.     

The effect of drip line placement on yield and quality of drip-irrigated processing tomatoes

Subsurface drip irrigation of processing tomatoes is increasing in California. The common design approach is to bury drip lines 0.2–0.36 m deep in the middle of the plant row, which places drip lines directly beneath plant rows. This design limits the use of the drip irrigation system to only those crops compatible with this drip line and bed spacing, and thus, other design approaches are being investigated to increase the flexibility of the drip systems. These approaches are installing drip lines in alternate furrows and installing drip lines in every furrow, both of which place drip lines midway between plant rows. The furrows are the result of the cultural practices used to form beds for planting.This study investigated the effect of the different drip line placements on crop yield and quality. Results showed that the highest yields occurred for the buried placement and the smallest yields for the alternate furrow placement. For the buried placement, soil water content and root density were concentrated around the drip lines, directly beneath the plant rows, while for the furrow placements, zones of high soil water content and root density did not coincide with the plant rows. However, some growers have found the furrow placement to reduce some of the disease problems normally experienced with the traditional furrow irrigation methods.     

Minimally invasive cholecystostomy in the dog: evaluation of placement techniques and use in extrahepatic biliary obstruction

Objective— To evaluate 4 methods of cholecystostomy catheter placement and to report on laparoscopic (Lap) cholecystostomy for the management of extrahepatic biliary obstruction (EHBO) in 3 dogs.
Study Design— Experimental study and clinical report.
Animals— Cadaveric dogs (n=20); 2 canine and 1 feline patient.
Methods— Pigtail cholecystostomy catheters were inserted in 20 canine cadavers using ultrasound (US) or Lap guidance. Insertion routes were either transperitoneal or transhepatic. Methods studied included Lap-transperitoneal, US-transperitoneal, US-transhepatic, and US-Seldinger (n=5 dogs/group). Insertion success, pleural penetration, and insertion site leakage (Lap-transperitoneal group) were evaluated. Three clinical EHBO cases were treated by Lap-transperitoneal technique.
Results— Insertion success was 100% by Lap-transperitoneal but 0% with US-transperitoneal and US-Seldinger methods. US-transhepatic yielded 3 of 5 successful placements. The pleura was penetrated in all US-transhepatic and US-Seldinger insertions. Leakage pressure for Lap-transperitoneal catheters averaged 75 cm H₂O (±20 cm H₂O). Lap-transperitoneal cholecystostomy resulted in marked improvement in 2 dogs, but the catheter became obstructed in the cat. One dog spontaneously regained common bile duct patency and the remaining 2 animals had successful cholecystoenterostomy.
Conclusions— In cadaver testing, the Lap-transperitoneal cholecystostomy method was superior based on high insertion success with no pleural penetration. In 2 clinical cases, Lap-transperitoneal placement successfully provided biliary drainage for patient stabilization.
Clinical Relevance— The role for temporary cholecystostomy has yet to be established, but may aid patient stabilization and mortality reduction in EHBO.     

Solitary plasmacytoma of bone in two successfully treated cats

This is the first report of feline solitary plasmacytoma of bone. We describe the clinical, clinico-pathological, radiographic and pathological findings of two successfully treated cats with long-term follow-up. The first case presented with spinal pain and neurological deficits. Radiographs demonstrated sclerosis of lumbar vertebra L6 and a myelogram confirmed interference to flow of contrast in the L4-7 region. A biopsy of L6 revealed neoplastic plasma cell infiltration. There was no evidence of paraproteinaemia on serum protein electrophoresis. The cat underwent hypofractionated megavoltage radiotherapy. Clinical signs resolved completely and 4 years after diagnosis the cat remains well and has no electrophoretically detectable paraproteinaemia. The second case presented with neurological deficits of the tail and spinal radiographs revealed extensive osteolysis of the sacrum. A biopsy of sacral bone demonstrated neoplastic plasma cell infiltration. The animal was normoglobulinaemic. The cat improved clinically with induction chemotherapy (melphalan and methylprednisolone). The same chemotherapeutics were continued at maintenance doses for 4.3 years, at which time there was recurrence of neurological deficits and a palpable sacral mass. Cytological examination of a fine needle aspirate confirmed recurrence of plasma cell neoplasia. A low concentration monoclonal paraproteinaemia was detected. Vincristine was administered resulting in resolution of neurological deficits and a palpably smaller sacral mass. Eighteen months into vincristine therapy, there was recurrence of clinical signs and the cat was euthanased, more than 6 years after the initial diagnosis.     

Effect of different head–neck positions on physical and psychological stress parameters in the ridden horse

Different head–neck positions (HNPs) are used in equestrian sports and are regarded as desirable for training and competition by riders, judges and trainers. Even though some studies have been indicative of hyperflexion having negative effects on horses, this unnatural position is frequently used. In the present study, the influence of different HNPs on physical and psychological stress parameters in the ridden horse was investigated. Heart rate (HR), heart rate variability (HRV) and blood cortisol levels were measured in 18 horses. Low frequency (LF) and high frequency (HF) are power components in the frequency domain measurement of HRV which show the activity of the sympathetic and parasympathetic nervous system. Values were recorded at rest, while riding with a working HNP and while riding with hyperflexion of the horse's head, neck and poll. In addition, rideability and behaviour during the different investigation stages were evaluated by the rider and by an observer. Neither the HR nor the HRV showed a significant difference between working HNP (HR = 105 ± 22/min; LF/HF = 3.89 ± 5.68; LF = 37.28 ± 10.77%) and hyperflexion (HR = 110 ± 18; LF/HF = 1.94 ± 2.21; LF = 38.39 ± 13.01%). Blood cortisol levels revealed a significant increase comparing working HNP (158 ± 60 nm ) and hyperflexion (176 ± 64 nm , p = 0.01). The evaluation of rider and observer resulted in clear changes of rideability and behavioural changes for the worse in all parameters collected between a working HNP and hyperflexion. In conclusion, changes of the cortisol blood level as a physical parameter led to the assumption that hyperflexion of head, neck and poll effects a stress reaction in the horse, and observation of the behaviour illustrates adverse effects on the well‐being of horses during hyperflexion.     

Sex differences in the influence of social context,salient social stimulation and amphetamine on ultrasonic vocalizations in prairie voles

Prairie voles (Microtus ochrogaster) are a socially monogamous rodent species and their cooperative behaviors require extensive communication between conspecifics. Rodents use ultrasonic vocalizations (USVs) to communicate and because a prairie vole breeder pair must engage in extensive cooperation for successful reproduction, auditory communication may be critical for this species. Therefore, we sought to characterize USVs in adult male and female prairie voles, and to determine how these calls are influenced by social context, salient social stimuli and the psychostimulant drug of abuse amphetamine (AMPH). Here, we characterize prairie vole USVs by showing the range of frequencies of prairie vole USVs, the proportion of various call types, how these call types compare between males and females, and how they are influenced by social stimulation and AMPH. AMPH caused a robust increase in the number of USVs in both males and females and there was a dramatic sex difference in the complexity of call structures of AMPH‐induced USVs, with males emitting more elaborate calls. Moreover, we show that novel (i.e. salient) social cues evoked differential increases in USVs across sex, with males showing a much more robust increase in USV production, both with respect to the frequency and complexity of USV production. Exposure to an estrous female in particular caused an extraordinary increase in USVs in male subjects. These data suggest that USVs may be a useful measure of social motivation in this species, including how social behaviors can be impacted by drugs of abuse.