Objective—To determine the usefulness of canine
RBC with high concentrations of potassium, reduced
glutathione (GSH), and amino acid(ie, HK cells) for in
vitro cultivation of Babesia gibsoni.
Animals—RBC were obtained from 3 dogs that had
inherited HK cells and from 3 genetically unaffected
dogs that, therefore, had RBC with lower potassium
(LK) concentrations (ie, LK cells).
Procedures—First, B gibsoni were cultivated using
HK or LK cells in alpha-modification of Eagle medium,
consisting of Earle salts with glutamine and without
ribosides, deoxyribosides, and sodium bicarbonate
under a humidified atmosphere containing 5% CO2 at
37 C. Second, parasites were cultivated with LK- or
HK-cell lysates. Finally, HK cells were separated into 3
fractions (bottom, middle, top layers) by density gradient
centrifugation, and B gibsoni were cultivated
with each of the HK-cell fractions. In addition, the concentrations
of free amino acids and reduced glutathione
(GSH) in each HK-cell fraction were measured.
Results—B gibsoni preferentially multiplied in HK-cell
cultures rather than in LK-cell cultures. Furthermore,
the addition of HK-cell lysate to the culture medium
resulted in enhanced multiplication of the parasites.
Higher multiplication of the parasites was observed in
HK cells from the top layer, compared with HK cells
from the middle and bottom layers. The HK cells from
the top layer had higher concentrations of glutamate,
aspartate, and GSH, compared with HK cells from the
middle and bottom layer.
Conclusions—Canine HK cells are useful host cells
for in vitro cultivation of B gibsoni, and the high concentrations
of glutamate, aspartate, and GSH may
result in enhancement of multiplication of the parasites
in HK cells. (Am J Vet Res 2000;61:1520–1524)
Objective—To examine the expression and distribution of tight junction (TJ) and adherens junction (AJ) proteins in canine duodenal and colonic mucosa.
Sample—Mucosa obtained from 4 healthy Beagles.
Procedures—Biopsy specimens of the duodenum and colon were obtained via endoscopy from 4 healthy dogs. The expression patterns and subcelluar localization of claudin-1, -2, -3, -4, -5, -7, and -8; E-cadherin; and β-catenin in the duodenum and colon were analyzed by use of immunoblotting and immunofluorescence microscopy.
Results—In the duodenum, there was clear expression of claudin-3 and -5, E-cadherin, and β-catenin proteins and weak expression of claudin-7 protein. In contrast, there was clear expression of claudin-2 and -3, E-cadherin, and β-catenin proteins and weak expression of claudin-5 and -7 proteins in the colon, as determined by use of immunoblotting. As determined by the use of immunofluorescence microscopy, the duodenum and colon had staining for claudin-3 and -5, E-cadherin, and β-catenin in the most apical region and staining for claudin-7 in the basolateral region. Staining for claudin-2 was also observed in the colon.
Conclusions and Clinical Relevance—Information was provided about the expression patterns of TJ and AJ proteins in the duodenum and colon of clinically normal dogs. These results may provide valuable information for use in evaluating the importance of these TJ and AJ proteins in the pathogenesis of inflammatory bowel disease in dogs.
Objective—To evaluate left atrial phasic function in healthy dogs by means of 2-D speckle tracking echocardiography with time-left atrial area curve analysis and to assess repeatability and reproducibility of obtained measurements.
Animals—6 healthy Beagles.
Procedures—Each dog underwent echocardiography twice on different days (3 nonconsecutive examinations/d). Images were analyzed with offline software; area of the left atrium was automatically calculated in each frame throughout the cardiac cycle to derive time-left atrial area curves. Variables used to assess left atrial phasic function (total, passive, and active emptying area and emptying fractions and mean active and total emptying rates) were calculated. Agreement between variables measured via speckle tracking echocardiography and a manual tracing method was assessed with modified Bland-Altman analysis. Within-day and between-day coefficients of variation were determined.
Results—Mean ± SD total, passive, and active emptying fractions of the left atrium were 49.8 ± 3.5%, 277 ± 4.0%, and 30.5 ± 4.3%, respectively. Mean ± SD total and active emptying rates were 16.0 ± 2.5 cm2/s and 25.1 ± 4.9 cm2/s, respectively. Within-day and between-day coefficients of variation were < 20% (range, 0.41% to 16.4%) for all variables except mean active emptying rate (between-day coefficient of variation, 29.2%). Agreement between variables measured via speckle tracking echocardiography and the manual tracing method was good, and differences between methods were nonsignificant.
Conclusions and Clinical Relevance—Evaluation of left atrial phasic function via speckle tracking echocardiography was feasible; repeatability and reproducibility of measurements were adequate in healthy dogs. Studies are needed to determine clinical applicability in canine patients.
Objective—To determine the expression of tight junction and adherens junction proteins in duodenal mucosa samples of dogs with inflammatory bowel disease (IBD).
Animals—12 dogs with IBD and 6 healthy control Beagles.
Procedures—Duodenal mucosa biopsy samples were endoscopically obtained from dogs with IBD and healthy control Beagles. The expression of claudin-1, -2, -3, -4, -5, -7, and -8; E-cadherin; and β-catenin in the duodenal mucosa samples was determined by means of immunoblotting. The subcellular localization of E-cadherin in the duodenal mucosa samples was determined with immunofluorescence microscopy.
Results—The expression of each claudin and β-catenin was not significantly different between control dogs and dogs with IBD. However, expression of E-cadherin was significantly lower in duodenal mucosa samples of dogs with IBD than it was in samples obtained from healthy control dogs. Results of immunofluorescence microscopy indicated decreased intensity of E-cadherin labeling in the tips of villi in duodenal mucosa samples obtained from 6 dogs with IBD, compared with staining intensity for other dogs.
Conclusions and Clinical Relevance—Results of this study indicated expression of claudin-1, -2, -3, -4, -5, -7, and -8 and β-catenin was not significantly different between duodenal mucosa samples obtained from control dogs and those obtained from dogs with IBD. However, E-cadherin expression was significantly lower in the villus epithelium in duodenal mucosa samples obtained from dogs with IBD versus samples obtained from control dogs, which suggested that decreased expression of that protein has a role in the pathogenesis of IBD in dogs.
OBJECTIVE To assess the use of contrast-enhanced ultrasonography (CEUS) of the hepatic vein for the detection of hemodynamic changes associated with experimentally induced portal hypertension in dogs.
ANIMALS 6 healthy Beagles.
PROCEDURES A prospective study was conducted. A catheter was surgically placed in the portal vein of each dog. Hypertension was induced by intraportal injection of microspheres (10 to 15 mg/kg) at 5-day intervals via the catheter. Microsphere injections were continued until multiple acquired portosystemic shunts were created. Portal vein pressure (PVP) was measured through the catheter. Contrast-enhanced ultrasonography was performed before and after establishment of hypertension. Time-intensity curves were generated from the region of interest in the hepatic vein. Perfusion variables measured for statistical analysis were hepatic vein arrival time, time to peak, time to peak phase (TTPP), and washout ratio. The correlation between CEUS variables and PVP was assessed by use of simple regression analysis.
RESULTS Time to peak and TTPP were significantly less after induction of portal hypertension. Simple regression analysis revealed a significant negative correlation between TTPP and PVP.
CONCLUSIONS AND CLINICAL RELEVANCE CEUS was useful for detecting hemodynamic changes associated with experimentally induced portal hypertension in dogs, which was characterized by a rapid increase in the intensity of the hepatic vein. Furthermore, TTPP, a time-dependent variable, provided useful complementary information for predicting portal hypertension.
IMPACT FOR HUMAN MEDICINE Because the method described here induced presinusoidal portal hypertension, these results can be applied to idiopathic portal hypertension in humans.