bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of in vitro assays for the evaluation of cyclooxygenase inhibitors and predicting selectivity of nonsteroidal anti-inflammatory drugs in cats
Authors: Giraudel Jérôme M. , Toutain Pierre-Louis , Lees Peter
Published: December 2005

Objective—To develop and validate in cats suitable in vitro assays for screening and ranking nonsteroidal antiinflammatory drugs (NSAIDs) on the basis of their inhibitory potencies for cyclooxygenase (COX)-1 and COX-2.

Animals—10 cats.

Procedure—COX-1 and COX-2 activities in heparinized whole blood samples were induced with calcium ionophore and lipopolysaccharide, respectively. For the COX-2 assay, blood was pretreated with aspirin. The COX-1 and COX-2 assays were standardized, such that time courses of incubation with the test compounds and conditions of COX expression were as similar as possible in the 2 assays. Inhibition of thromboxane B2production, measured by use of a radioimmunoassay, was taken as a marker of COX-1 and COX-2 activities. These assays were used to test 10 to 12 concentrations of a COX-1 selective drug (SC-560) and of 2 NSAIDs currently used in feline practice, meloxicam and carprofen. Selectivities of these drugs were compared by use of classic 50% and 80% inhibitory concentration (ie, IC50and IC80) ratios but also with alternative indices that are more clinically relevant.

Results—These assay conditions provide a convenient and robust method for the determination of NSAID selectivity. The S(+) enantiomeric form of carprofen was found to be COX-2 selective in cats, but meloxicam was only slightly preferential for this isoenzyme.

Conclusions and Clinical Relevance—In vitro pharmacodynamic and in vivo pharmacokinetic data predict that the COX-2 selectivity of both drugs for cats will be limited when used at the recommended doses. This study provides new approaches to the selection of COX inhibitors for subsequent clinical testing. (Am J Vet Res2005;66:700–709)
Localization, morphology, and immunohistochemistry of spinal cord and dorsal root ganglion neurons that innervate the gastrocnemius and superficial digital flexor muscles in cattle
Authors: Chiocchetti Roberto , Grandis Annamaria , Bombardi Cristiano , Clavenzani Paolo , Spadari Alessandro , Gentile Arcangelo , Bortolami Ruggero
Published: December 2005

Objective—To determine the location, morphology,
and neurochemical code of spinal cord and dorsal root
ganglion neurons that innervate the gastrocnemius
muscle (GM) and superficial digital flexor muscle
(FDSM) in cattle.

Animals—5 healthy Friesian male calves.

Procedure—2 different types of neuronal retrograde
fluorescent tracers (fast blue and diamidino yellow)
were injected into the GM and FDSM, respectively.
The neurochemical code (substance P, calcitonin
gene-related peptide, galanin, and neuronal nuclear
protein) of labeled neurons was investigated by
immunohistochemistry.

Results—Neurons innervating the GM and FDSM
were located along the L6-S2 spinal cord segments
and ganglionic levels. A cranial-to-caudal topographic
distribution for each muscle was found, indicating that
the motor nuclei of the 2 muscles are organized by a
somatotopic pattern. The GM and FDSM motoneurons
were immunoreactive only for calcitonin generelated
peptide, whereas the afferent neurons were
immunoreactive for all of the neurochemical markers
considered.

Conclusions and Clinical Relevance—In our study,
location and the extent of neurons that supply the GM
and FDSM of cattle were characterized completely.
Because the GM and FDSM are involved in spastic
paresis of calves and it is thought that spastic paresis
results from an excessive activity of the neuromuscular
spindle reflex arc, findings in our study may be
useful for further electrophysiologic and clinical studies.
Knowledge of the neurochemical code of neurons
that supply the GM and FDSM in healthy calves could
be used to compare chemical alterations in the same
neuronal population of affected calves. (Am J Vet Res 2005;66:710–720)
Evaluation of a real-time quantitative polymerase chain reaction assay for detection and quantitation of virulent Rhodococcus equi
Authors: Harrington Jessica R. , Golding Michael C. , Martens Ronald J. , Halbert Natalie D. , Cohen Noah D.
Published: December 2005

Objective—To evaluate a real-time quantitative polymerase chain reaction (QPCR) assay in the detection and quantitation of virulentRhodococcus equi.

Sample Population—1 virulent, 2 intermediately virulent, and 2 avirulent strains ofR equiand 16 isolates of bacteria genetically related toR equi.

Procedure—The QPCR assay was evaluated for detection and quantitation of the virulence-associated gene (vapA) ofR equiin pure culture and in samples of tracheobronchial fluid, which were inoculated with known numbers of virulentR equi. Results were compared with those derived via quantitative microbial culture and standard polymerase chain reaction methods.

Results—The QPCR assay detected the vapAgene in pure culture ofR equiand in tracheobronchial fluid samples that contained as few as 20 CFUs of virulentR equi/mL and accurately quantitated virulentR equito 103CFUs/mL of fluid. The assay was highly specific for detection of thevapAgene of virulentR equiand was more sensitive than standard polymerase chain reaction for detection ofR equiin tracheobronchial fluid.

Conclusions and Clinical Relevance—The QPCR assay appears to be a rapid and reliable method for detecting and quantitating virulentR equi. The accuracy of the QPCR assay is comparable to that of quantitative microbial culture. The increased sensitivity of the QPCR method in detection of virulentR equishould facilitate rapid and accurate diagnosis ofR equipneumonia in foals. (Am J Vet Res2005;66:755–761)
Evaluation of a monoclonal antibody–based dot-blot ELISA for detection of Leptospira spp in bovine urine samples
Authors: Suwimonteerabutr Junpen , Chaicumpa Wanpen , Saengjaruk Patcharin , Tapchaisri Pramuan , Chongsa-nguan Manas , Kalambaheti Thareerat , Ramasoota Pongrama , Sakolvaree Yuwaporn , Virakul Prachin
Published: December 2005

Objective—To evaluate the efficacy of a novel monoclonal
antibody (MAb)–based dot-blot ELISA for
detection of Leptospira antigens in urine samples of
cattle.

Sample Population—Blood and urine samples of 45
test cattle from 5 farms in Chonburi province and 20
control cattle from 2 farms in Khon Kaen province in
Thailand.

Procedure—Blood and urine samples were assayed
(microscopic agglutination test and urine antigen test)
for Leptospira infection by use of an MAb–based dotblot
ELISA, and results for the ELISA were compared
with those for dark-field microscopy (DFM), microbial
culture, and a polymerase chain reaction (PCR) assay.

Results—All urine samples with positive results for
DFM, microbial culture, PCR assay, or > 1 of these
tests also had positive results when tested by use of
the MAb–based dot-blot ELISA, except for 1 sample
that had positive results only for the PCR assay.
Detection limits of the dot-blot ELISA were 103 leptospires/
mL of urine and 9.3 ng of Leptospira
homogenate. Comparison revealed that the diagnostic
sensitivity, specificity, efficacy (accuracy), positive
predictive value, and negative predictive value for the
ELISA were in agreement with results for DFM
(100%, 72.72%, 80%, 57.14%, and 100%, respectively),
microbial culture (100%, 61.54%, 66.62%,
28.57%, and 100%, respectively), and PCR assay
(95.45%, 100%, 91.77%, 100%, and 95.83%, respectively).

Conclusions and Clinical Relevance—The
MAb–based dot-blot ELISA is suitable as a tool for
detecting leptospires in urine samples of cattle. (Am
J Vet Res 2005;66:762–766)
Pharmacokinetics after intravenous, subcutaneous, and oral administration of enrofloxacin to alpacas
Authors: Gandolf A. Rae , Papich Mark G. , Bringardner Amy B. , Atkinson Mark W.
Published: December 2005

Objective—To determine plasma concentrations of enrofloxacin and the active metabolite ciprofloxacin after PO, SC, and IV administration of enrofloxacin to alpacas.

Animals—6 adult female alpacas.

Procedure—A crossover design was used for administration of 3 single-dose treatments of enrofloxacin to alpacas, which was followed by an observational 14-day multiple-dose regimen. Single-dose treatments consisted of IV and SC administration of injectable enrofloxacin (5 mg/kg) and PO administration of enrofloxacin tablets (10 mg/kg) dissolved in grain to form a slurry. Plasma enrofloxacin concentrations were measured by use of high-performance liquid chromatography. The multiple-dose regimen consisted of feeding a mixture of crushed and moistened enrofloxacin tablets mixed with grain. Behavior, appetite, and fecal quality were monitored throughout the 14-day treatment regimen and for 71 additional days following treatment.

Results—Mean half-life following IV, SC, and PO administration was 11.2, 8.7, and 16.1 hours, respectively. For SC and PO administration, mean total systemic availability was 90.18% and 29.31%, respectively; mean maximum plasma concentration was 3.79 and 1.81 µg/mL, respectively; and area under the curve (AUC) was 50.05 and 33.97 (µg × h)/mL, respectively. The SC or PO administration of a single dose of enrofloxacin yielded a ratio for AUC to minimum inhibitory concentration > 100 for many grampositive and gram-negative bacterial pathogens common to camelids.

Conclusions and Clinical Relevance—The administration of enrofloxacin (5 mg/kg, SC, or 10 mg/kg, PO) may be appropriate for antimicrobial treatment of alpacas. (Am J Vet Res2005;66:767–771)
Evaluation of oral administration of cortisol as a model for prenatal stress in pregnant sows
Authors: Kranendonk Godelieve , Hopster Hans , Eerdenburg Frank van , Reenen Kees van , Fillerup Maaike , Groot Johanna de , Korte Mechiel , Taverne Marcel
Published: December 2005

Objective—To design a treatment that increases
plasma corticosteroid concentrations to mimic prenatal
stress in pregnant sows.

Animals—24 pregnant sows.

Procedure—Sows were assigned to 1 of 4 treatment
groups; treatment consisted of twice-daily oral administration
of a placebo or 20, 60, or 180 mg of hydrocortisone
acetate (HCA)/sow from 7 to 11 weeks of
gestation. Blood and saliva samples for determination
of cortisol concentrations were obtained hourly on
treatment days 3 and 25 and twice weekly for the
remainder of the treatment period. The WBC, neutrophil,
and lymphocyte counts and concentrations of
interleukin (IL)-2 and IL-4 were determined on 4 days
during treatment. Litter characteristics were recorded.

Results—Plasma and salivary cortisol concentrations
were significantly increased in sows that received 60
or 180 mg of HCA (0.30 to 0.37 mg/kg and 0.95 to
1.15 mg/kg, respectively), compared with control
sows. Except for the second day of treatment, the
number of WBCs and the IL-2:IL-4 ratio did not differ
among treatment groups. The neutrophil-to-lymphocyte
ratio was significantly higher in sows that
received 180 mg of HCA. Gestation duration was significantly
shorter in sows that received 180 mg of
HCA.

Conclusions and Clinical Relevance—Oral administration
of 60 mg of HCA is suitable to increase plasma
and salivary cortisol concentrations in pregnant sows
in a controlled manner to concentrations comparable
to concentrations detected after psychologic stressors.
This model seems to be suitable to study the
effects of increased maternal corticosteroid concentrations
on young pig behavior, physiologic variables,
and development. (Am J Vet Res 2005;66:780–790)
Climatic and aeroallergen risk factors for chronic obstructive pulmonary disease in horses
Authors: Ward Michael P. , Couëtil Laurent L.
Published: December 2005

Objective—To estimate the association between climate and airborne pollen and fungal factors and chronic obstructive pulmonary disease (COPD) in horses.

Sample Population—Data from 1,444 horses with a diagnosis of COPD.

Procedure—The Veterinary Medical Database was used to identify records of horses admitted to veterinary teaching hospitals in the United States and Canada between 1990 and 1999. Rainfall, mean minimum and maximum temperature, and maximum monthly pollen and fungal spore (mold) counts recorded at the city closest to where the hospital is located were identified for each month data were reported to the Veterinary Medical Database. Associations between climatic and aeroallergen data and monthly prevalence of COPD were estimated by use of crosscorrelation and logistic regression models.

Results—Significant positive correlations were found between prevalence of COPD and rainfall 3 months previously, minimum temperature 1 and 2 months previously, total pollen counts measured 3 months previously, and total mold counts measured during the same month and 1 month previously.

Conclusions and Clinical Relevance—Outdoor aeroallergens and climatic factors may contribute to the occurrence of COPD in horses. (Am J Vet Res2005;66:818–824)
Use of serologic evaluation for antibodies against bovine viral diarrhea virus for detection of persistently infected calves in beef herds
Authors: Waldner Cheryl L. , Campbell John R.
Published: December 2005

Objective—To determine whether use of serologic
evaluation of a sentinel sample of calves or cows for
antibodies against bovine viral diarrhea virus (BVDV)
would accurately predict whether an animal persistently
infected with BVDV could be detected in beef
herds.

Sample Population—27 cow-calf herds in which the
status of persistently infected calves was not known and
11 herds known to have persistently infected calves.

Procedure—Detection of persistently infected calves
was determined through immunohistochemical testing
of tissue obtained at necropsy of all calves that
died during calving season and skin (ear notch) specimens
obtained from all young stock in the fall of
2002. Serum samples were collected from 30 springborn
calves and 10 mature cows.

Results—Optimum serologic test performance at
time of weaning was detected when 10 calves were
evaluated. At least 3 of 10 randomly selected calves
were likely to have a titer > 1:1,000 against BVDV
type I or II in 53% of herds in which a persistently
infected calf was detected during that year (sensitivity,
53%). However, at least 3 of 10 randomly selected
calves were also likely to have a titer > 1:1,000 in
20% of herds that did not have a persistently infected
calf detected during that year (specificity, 80%).

Conclusions and Clinical Relevance—Despite the
use of a number of various cutoff values and sample
sizes, serologic evaluation of a small number of calves
or cows could not be used to accurately predict the
presence of persistently infected cattle in a herd. (Am
J Vet Res 2005;66:825–834)
Correlation of results of pulmonary computed tomography and pathologic findings in mice with Pasteurella-induced pneumonia
Authors: Armbrust Laura J. , Mosier Derek A. , Nelson Ethany L. , Hart Marcy L. , Chapes Stephen K.
Published: December 2005

Objective—To determine correlation between results
of computed tomography (CT) versus pathologic
examination for determining the volume percentage
of affected lung in mice experimentally infected with
Pasteurella pneumotropica.

Animals—30 adult mice.

Procedure—After helical CT scans on day 0, mice
were inoculated intranasally with P pneumotropica.
Repeat CT scans were performed on days 1, 2, 3, 4,
6, 8, 10, and 13. Regions of interest (affected areas)
were manually drawn on the CT images, and percentage
volume of normal lung was calculated by use
of 3 methods: first-day volume, largest volume, and
last-day volume. Three mice were euthanatized for
pathologic evaluation after each scan day. The lungs
were examined with a dissection microscope, and
lesion scores were assigned on the basis of percentage
volume of pneumonia. Correlation coefficients
comparing results of the 3 CT methods with results
of gross examination were calculated.

Results—Lung abnormalities were detected via dissection
microscopy by postinfection day 2 and via CT
by days 2 or 3. Correlation coefficients for the 3 CT
methods of analysis, compared with pathologic findings,
were 0.7 via first-day lung volume, 0.8 via
largest lung volume, and 0.8 via last-day lung volume.

Conclusions and Clinical Relevance—Results of CT
correlated well with results of dissection microscopy
for estimating percentage volume of lung affected by
pneumonia in mice experimentally infected with
P pneumotropica. This method may be useful for longitudinal
studies of pneumonia in mice. (Am J Vet Res
2005;66:835–838)
Effect of daily administration of pyrantel tartrate in preventing infection in horses experimentally challenged with Sarcocystis neurona
Authors: Rossano Mary G. , Schott II Harold C. , Kaneene John B. , Murphy Alice J. , Kruttlin Elizabeth A. , Hines Melissa T. , Sellon Debra C. , Patterson Jon S. , Elsheikha Hany M. , Dubey J. P. , Mansfield Linda S.
Published: December 2005

Objective—To determine whether daily administration of pyrantel tartrate can prevent infection in horses experimentally challenged withSarcocystis neurona.

Animals—24 mixed-breed specific-pathogen-free weanling horses, 10 adult horses, 1 opossum, and 6 mice.

Procedure—Sarcocystis neurona-naïve weanling horses were randomly allocated to 2 groups. Group A received pyrantel tartrate at the labeled dose, and group B received a nonmedicated pellet. Both groups were orally inoculated with 100 sporocysts/d for 28 days, 500 sporocysts/d for 28 days, and 1,000 sporocysts/ d for 56 days. Blood samples were collected weekly, and CSF was collected monthly. Ten seronegative adult horses were monitored as untreated, uninfected control animals. All serum and CSF samples were tested by use of western blot tests to detect antibodies againstS neurona. At the end of the study, the number of seropositive and CSF-positive horses in groups A and B were compared by use of the Fisher exact test. Time to seroconversion on the basis of treatment groups and sex of horses was compared in 2 univariable Cox proportional hazards models.

Results—After 134 days of sporocyst inoculation, no significant differences were found between groups A and B for results of western blot tests of serum or CSF. There were no significant differences in number of days to seroconversion on the basis of treatment groups or sex of horses. The control horses remained seronegative.

Conclusions and Clinical Relevance—Daily administration of pyrantel tartrate at the current labeled dose does not preventS neuronainfection in horses. (Am J Vet Res2005;66:846–852)
Risk factors associated with herd-level exposure of cattle in Nebraska, North Dakota, and South Dakota to bluetongue virus
Authors: Green Alice L. , Dargatz David A. , Schmidtmann Edward T. , Herrero Marco V. , Seitzinger Ann H. , Ostlund Eileen N. , Wagner Bruce A. , Moser Kathryn M. , Wineland Nora E. , Walton Thomas E.
Published: December 2005

Objective—To evaluate herd-level risk factors for
seropositive status of cattle to 1 or more bluetongue
viruses.

Animals—110 herds of cattle in Nebraska, North
Dakota, and South Dakota.

Procedure—Blood samples were collected before
and after the vector season. Samples were tested
for antibodies against bluetongue virus by use of a
commercially available competitive ELISA. Factors
evaluated included descriptors of geographic location
and management practices. Trapping of insect
vectors was conducted to evaluate vector status on
a subset of 57 operations. A multivariable logistic
regression model was constructed to evaluate associations.

Results—For the full data set, altitude and latitude
were associated with risk of having seropositive cattle
(an increase in altitude was associated with an
increase in risk, and a more northerly location was
associated with a decrease in risk of a premise having
seropositive cattle). Import of cattle from selected
states was associated with an increase in risk of having
seropositive cattle. From the subset of herds with
data on vector trapping, altitude and latitude were
associated with risk of having seropositive cattle, similar
to that for the full model. However, commingling
with cattle from other herds was associated with a
decrease in risk of seropositivity.

Conclusions and Clinical Relevance—Findings
reported here may be useful in generating additional
hypotheses regarding the ecologic characteristics of
bluetongue viruses and other vector-borne diseases
of livestock. Sentinel surveillance programs are useful
for documenting regionalization zones for diseases,
which can be beneficial when securing international
markets for animals and animal products. (Am J Vet
Res 2005;66:853–860)
Effect of desmopressin on von Willebrand factor multimers in Doberman Pinschers with type 1 von Willebrand disease
Authors: Callan Mary Beth , Giger Urs , Catalfamo James L.
Published: December 2005

Objective—To assess the effect of desmopressin
(DDAVP) administration in Doberman Pinschers with
type 1 von Willebrand disease (vWD) on plasma von
Willebrand factor (vWF) multimers through determination
of vWF collagen binding activity (vWF:CBA; a
functional vWF assay dependent on the presence of
high–molecular-weight [HMW] multimers), comparison
of vWF antigen concentration (vWF:Ag) to
vWF:CBA, and vWF multimer size distribution.

Animals—16 Doberman Pinschers with type 1 vWD
and 5 clinically normal control dogs.

Procedure—Plasma vWF:Ag and vWF:CBA assays
and vWF multimer analysis were performed before
and 1 hour after administration of DDAVP (1 µg/kg,
SC).

Results—Following DDAVP administration, dogs with
type 1 vWD had an increase in mean baseline values
of plasma vWF:Ag and vWF:CBA from 10% to 17%
for both variables. The mean vWF Ag:CBA ratio at
baseline (0.95) was similar after DDAVP administration
(0.97), indicating concordant increases in plasma
vWF concentration and activity. In control dogs, mean
plasma vWF:Ag and vWF:CBA increased from baseline
values of 64% to 113% and 58% to 114%,
respectively, and the vWF Ag:CBA ratios were
unchanged (1.1 vs 1.0) after DDAVP administration.
Plasma vWF multimer analysis revealed proportional
increases in band intensity for all multimer sizes following
DDAVP administration, in comparison to baseline
for the control dogs and Doberman Pinschers
with vWD, consistent with vWF Ag:CBA ratios of
approximately 1.

Conclusions and Clinical Relevance—Beneficial
effects of DDAVP on primary hemostasis in
Doberman Pinschers with type 1 vWD cannot be
explained by preferential increases in HMW vWF multimers.
(Am J Vet Res 2005;66:861–867)
Seroprevalence of Toxoplasma gondii antibodies in clinically ill cats in the United States
Authors: Vollaire Melissa R. , Radecki Steven V. , Lappin Michael R.
Published: December 2005

Objective—To determine regional seroprevalence estimates ofToxoplasma gondii-specific IgM and IgG in clinically ill cats throughout the United States.

Sample Population—Sera from 12,628 clinically ill, client-owned cats.

Procedure—Toxoplasma gondii-specific IgM and IgG antibodies were detected by use of ELISAs. Sera from clinically ill cats previously submitted forT gondiiantibody testing were sequentially selected from our serum bank and the sample submission paperwork reviewed. The country was divided into 12 geographic regions. Overall prevalence as well as prevalence for each region, age group, season, sex (male vs female), and breed (domestic shorthair vs other) was calculated. Data were analyzed by logistic regression analysis.

Results—Overall, 31.6% of the cats were seropositive forT gondii-specific IgM, IgG, or both. Percentage of cats seropositive forT gondiiantibodies ranged from 16.1% (southwestern United States) to 43.5% (northeastern United States). As age increased, odds of positiveT gondiiantibody assay results (IgM alone, IgG alone, and any combination of IgM or IgG) increased. Males were more likely than females to be seropositive forT gondiiantibodies (IgG alone and any combination of IgM or IgG). Domestic shorthair cats were more likely than other breeds to be seropositive forT gondiiantibodies (IgM alone, IgG alone, and any combination of IgM or IgG).

Conclusions and Clinical Relevance—Toxoplasma gondii-specific antibodies are common in serum samples of clinically ill cats from all regions of the United States. Seroprevalence increases as cats age and is higher in male and domestic shorthair cats, compared with females and other breeds. (Am J Vet Res2005; 66:874–877)
Comparison of thermal dilution and electrical impedance dilution methods for measurement of cardiac output in standing and exercising horses
Authors: Wilkins Pamela A. , Boston Raymond C. , Gleed Robin D. , Dobson Alan
Published: December 2005

Objective—To compare cardiac output measured in
the pulmonary artery and a carotid artery by use of
thermal and electrical impedance dilution.

Animals—7 fit, clinically normal Standardbreds
between 2 and 5 years of age.

Procedure—Transient changes in electrical impedance
and temperature of blood were induced by bolus
injections of ice-cold saline hypertonic (6% and 9%
NaCl) solutions. Cardiac output was calculated by
applying Stewart-Hamilton principles to the indicator
dilution transients. Measurements were made during
sequential exercise episodes on a level treadmill over
approximately an 8-fold range of cardiac output values.

Results—We detected agreement between cardiac
output determined by use of electrical impedance
dilution at the pulmonary artery and carotid artery.
Cardiac output from thermal dilution measured at the
carotid artery exceeded that measured at the pulmonary
artery. Cardiac output from the thermal dilution
technique exceeded cardiac output from the electrical
impedance dilution technique at both locations.

Conclusions and Clinical Relevance—The electrical
impedance indicator is conserved on first transit;
therefore cardiac output measured by electrical
impedance dilution at the carotid artery is reliable over
a large range of values. Thermal dilution provides a
larger estimate of cardiac output, compared with the
electrical impedance dilution technique, probably
because of a loss of indicator. The transpulmonary
electrical impedance dilution technique may have
potential for clinical application, particularly in animals
in which catheterization of the pulmonary artery is not
appropriate or blood loss must be minimized. (Am J
Vet Res 2005;66:878–884)
Investigation of the effect of pamidronate disodium on the in vitro viability of osteosarcoma cells from dogs
Authors: Ashton Jenna A. , Farese James P. , Milner Rowan J. , Lee-Ambrose Linda M. , Gilder James M. van
Published: December 2005

Objective—To determine the effect of pamidronate disodium on the in vitro viability of osteosarcoma cells and non-neoplastic cells from dogs.

Sample Population—3 osteosarcoma and 1 fibroblast cell lines derived from dogs.

Procedure—Cell counts and cell viability assays were performed in cultures of osteosarcoma cells (POS, HMPOS, and COS31 cell lines) and fibroblasts after 24, 48, and 72 hours of incubation with pamidronate at concentrations of 0.001 to 1,000µM or with no drug (control treatment). Percentage viability was determined in cell samples for each concentration of pamidronate and each incubation time. A DNA fragmentation analysis was performed to assess bisphosphonate- induced apoptosis.

Results—Osteosarcoma cell viability decreased significantly in a concentration- and time-dependent manner at pamidronate concentrations ranging from 100 to 1,000µM, most consistently after 48 and 72 hours' exposure. In treated osteosarcoma cells, the lowest percentage cell viability was 34% (detected after 72 hours' exposure to 1,000µM pamidronate). Conversely, 72 hours' exposure to 1,000µM pamidronate did not significantly reduce fibroblast viability (the lowest percentage viability was 76%). After 72 hours of exposure, pamidronate did not cause DNA fragmentation in POS or HMPOS cells.

Conclusions and Clinical Relevance—Results indicate that pamidronate may have the potential to inhibit osteosarcoma growth in dogs, possibly through a nonapoptotic mechanism. The clinical relevance of these in vitro findings remains to be determined, but administration of pamidronate may potentially be indicated as an adjuvant treatment in chemotherapeutic protocols used in dogs. (Am J Vet Res2005;66: 885–891)
Clinical and endocrine responses to treatment with deslorelin acetate implants in ferrets with adrenocortical disease
Authors: Wagner Robert A. , Piché Claude A. , Jöchle Wolfgang , Oliver Jack W.
Published: December 2005

Objective—To evaluate the clinical and endocrine responses of ferrets with adrenocortical disease (ACD) to treatment with a slow-release implant of deslorelin acetate.

Animals—15 ferrets with ACD.

Procedure—Ferrets were treated SC with a single slow-release, 3-mg implant of deslorelin acetate. Plasma estradiol, androstenedione, and 17-hydroxyprogesterone concentrations were measured before and after treatment and at relapse of clinical signs; at that time, the adrenal glands were grossly or ultrasonographically measured and affected glands that were surgically removed were examined histologically.

Results—Compared with findings before deslorelin treatment, vulvar swelling, pruritus, sexual behaviors, and aggression were significantly decreased or eliminated within 14 days of implantation; hair regrowth was evident 4 to 6 weeks after treatment. Within 1 month of treatment, plasma hormone concentrations significantly decreased and remained decreased until clinical relapse. Mean time to recurrence of clinical signs was 13.7 ± 3.5 months (range, 8.5 to 20.5 months). In 5 ferrets, large palpable tumors developed within 2 months of clinical relapse; 3 of these ferrets were euthanatized because of adrenal gland tumor metastasis to the liver or tumor necrosis.

Conclusions and Clinical Relevance—In ferrets with ACD, a slow-release deslorelin implant appears promising as a treatment to temporarily eliminate clinical signs and decrease plasma steroid hormone concentrations. Deslorelin may not decrease adrenal tumor growth in some treated ferrets. Deslorelin implants may be useful in the long-term management of hormone-induced sequelae in ferrets with ACD and in treatment of animals that are considered at surgical or anesthetic risk. (Am J Vet Res2005;66:910–914)
In vitro comparison of RNA preparation methods for detection of feline calicivirus in urine of cats by use of a reverse transcriptase-polymerase chain reaction assay
Authors: Scansen Brian A. , Kruger John M. , Wise Annabel G. , Venta Patrick J. , Bartlett Paul C. , Maes Roger K.
Published: December 2005

Objective—To compare 5 methods of preparation of
RNA from feline urine samples for use in a feline calicivirus
(FCV), p30 gene-based, real-time reverse-transcriptase
polymerase chain reaction (RT-PCR) assay.

Sample Population—Urine and blood samples from
6 specific-pathogen-free cats.

Procedures—Aliquots of each urine sample (unmodified,
centrifuged, or mixed with whole or hemolyzed
blood) were spiked with FCV and serially diluted in
urine. Serial dilutions of FCV in tissue culture medium
were used as positive controls. Viral RNA was prepared
via dilution and thermal inactivation (DT
method), polyethylene glycol precipitation (PEG
method), isolation with oligo(dT)25-coated magnetic
beads (dTMB method), or extraction by use of 2 silica
gel–based columns (RN or QA method). Lower detection
limits and mean RT-PCR threshold cycle (Ct) values
associated with each RNA preparation method
and sample type were compared.

Results—Because DT-prepared samples yielded negative
results via RT-PCR assay, this method was not evaluated.
Lower detection limits (TCID50/sample) for the
assay in urine were 1,950, 104, 11, and 7 for PEG-,
dTMB-, RN-, and QA-prepared samples, respectively. For
RN and QA preparations, Ct values were similar and significantly
lower than those for dTMB and PEG preparations.
Overall, urine modifications did not affect FCV RNA
detection in dTMB-, QA-, and RN-prepared samples.

Conclusions and Clinical Relevance—Of the methods
evaluated, the RN and QA methods of RNA
preparation were most appropriate for the FCV RTPCR
assay. An RT-PCR assay optimized for detection
of FCV in feline urine may aid investigations of FCVinduced
urinary tract diseases in cats. (Am J Vet Res
2005;66:915–920)
Comparison of antibody detection assays for the diagnosis of equine herpesvirus 1 and 4 infections in horses
Authors: Hartley Carol A. , Wilks Colin R. , Studdert Michael J. , Gilkerson James R.
Published: December 2005

Objective—To compare methods of detecting equine
herpesvirus type 1 (EHV1)- and EHV4-specific antibodies
in horse sera.

Sample Population—33 acute and convalescent
serum samples from experimentally or naturally
infected horses after confirmed EHV1 or EHV4 infection.

Procedure—For each sample, serum antibody titers
against EHV1 and EHV4 were determined by use of
virus neutralization (VN) and complement fixation (CF)
assays. The ELISA absorbance values for each serum
sample were determined against the EHV1 and EHV4
recombinant ELISA antigens. Values obtained for
acute and convalescent sera in each assay were compared.

Results—Following experimental infection of foals,
EHV1 or EHV4 antibodies that were specific for the
inoculating virus were detected only by use of the
ELISA. Results of VN and CF assays indicated that the
foals seroconverted to EHV1 and EHV4 following
infection with EHV4 only. After EHV1-induced abortion,
myeloencephalitis, or respiratory tract disease,
the VN and CF assay results revealed seroconversion
to EHV1 and EHV4, whereas results of the ELISA
revealed seroconversion to EHV1 only. Similarly, after
confirmed EHV4-induced respiratory tract disease,
increases in EHV4-specific antibodies were detected
only by use of the ELISA with no indication of an
increase in EHV1 antibodies. The CF and, to a lesser
degree, VN assays revealed that seroconversion to
EHV1 and EHV4 occurred between the time of
obtaining acute and convalescent serum samples.

Conclusions and Clinical Relevance—The EHV1/
EHV4 type-specific antibody ELISA clearly identifies
horses that have been infected with EHV1 or EHV4 by
use of acute and convalescent sera. Results of VN and
CF assays indicate that cross-reactive antibodies greatly
limit their use. (Am J Vet Res 2005;66:921–928)
Evaluation of glucose tolerance and intestinal luminal membrane glucose transporter function in horses with equine motor neuron disease
Authors: Benders Nikkie A. , Dyer Jane , Wijnberg Inge D. , Shirazi-Beechey Soraya P. , van der Kolk Johannes H.
Published: December 2005

Objective—To confirm whether the plasma glucose concentration curve obtained during oral glucose tolerance tests (OGTTs) in horses with equine motor neuron disease (EMND) is decreased, compared with that obtained in clinically normal horses, and determine whether that decrease is a result of defective glucose metabolism or intestinal glucose transport dysfunction.

Animals—8 horses with EMND and 44 matched control horses.

Procedure—Electromyography and OGTTs were performed in all 8 affected horses and 10 control horses. Intravenous GTTs (IVGTTs) were performed in 6 affected horses and another 11 control horses. The activity and levels of jejunal luminal membrane glucose transporter (Na+/glucose cotransporter isoform 1 [SGLT1]) were measured in 2 affected horses and 23 control horses.

Results—In horses with EMND, generalized neuropathy was detected via quantitative electromyography; the mean increase in plasma glucose concentration during the OGTT was significantly decreased, compared with the value in control horses. During the IVGTT, the mean increase in plasma glucose concentration was significantly lower than that of control horses. The activity and levels of SGLT1 in 2 affected horses were similar to those of control horses. Diagnosis of EMND was confirmed postmortem in all affected horses.

Conclusions and Clinical Relevance—Data suggest that the decreased plasma glucose curve obtained in horses with EMND during OGTTs (compared with control horses) is a result of overall enhanced glucose metabolism or abnormalities in the facilitated glucose transporters; definitive identification of the underlying mechanisms could aid in the development of appropriate treatments of EMND in horses. (Am J Vet Res2005;66:93–99)
Isolation and preliminary characterization of a novel Helicobacter species from swine
Authors: Krakowka Steven , Ringler Susan S. , Flores Joel , Kearns Robert J. , Eaton Kathyrn A. , Ellis John A.
Published: December 2005

Objective—To determine whether aHelicobactersp similar toHelicobacter pyloriin the stomachs of humans could be isolated from the stomachs of pigs.

Animals—4 young conventionally reared and 21 gnotobiotic pigs.

Procedure—Gastric mucosal homogenates (10% wt/vol) from 4 young conventionally reared pigs were cultured on Skirrow medium under microaerophilic conditions to assess the presence ofHelicobacterspp. Colonies with morphologic features compatible withHelicobacterorganisms were selected, tested for urease activity, and subpassaged on Skirrow medium. Isolates were examined via SDS-PAGE electrophoresis and reciprocal western blot analyses involving convalescent sera from monoinfected gnotobiotic pigs.

Results—Urease- and catalase-positive, gram-negative, microaerophilic, small, curved rod bacteria were isolated from the gastric mucosa of young healthy pigs. The first isolate (2662) was structurally and immunologically closely related toH pyloriisolated from humans. The second isolate (1268) displayed an SDS-PAGE profile dissimilar to that ofH pyloriand isolate 2662, yet it shared limited immunologic crossreactivity with these microbes.

Conclusions and Clinical Relevance—Findings of this study indicate that development of gastric mucosal ulcers and ulceration of the nonglandular pars esophagea in pigs may be associated with gastric colonization by swine-originHelicobacterspp, which are similar toH pyloriisolated from humans. (Am J Vet Res2005;66:938–944)
Use of quantitative two-dimensional color tissue Doppler imaging for assessment of left ventricular radial and longitudinal myocardial velocities in dogs
Authors: Chetboul Valerie , Sampedrano Carolina Carlos , Concordet Didier , Tissier Renaud , Lamour Thierry , Ginesta Jacques , Gouni Vassiliki , Nicolle Audrey P. , Pouchelon Jean-Louis , Lefebvre Hervé P.
Published: December 2005

Objective—To determine left ventricular free wall
(LVFW) radial and longitudinal myocardial contraction
velocities in healthy dogs via quantitative 2-dimensional
color tissue Doppler imaging (TDI).

Animals—100 dogs.

Procedure—TDI was used by a single trained observer
to measure radial and longitudinal myocardial
movement in the LVFW. Radial myocardial velocities
were recorded in segments in the endocardial and
epicardial layers of the LVFW, and longitudinal velocities
were recorded in segments at 3 levels (basal,
middle, apical) of the LVFW.

Results—LVFW velocities were higher in the endocardial
layers than in the epicardial layers. Left ventricular
free wall velocities were higher in the basal
segments than in the middle and apical segments.
Radial myocardial velocity gradients, defined as the
difference between endocardial and epicardial velocities,
were (mean ± SD) 2.5 ± 0.8 cm/s, 3.8 ± 1.5 cm/s,
and 2.3 ± 0.9 cm/s in systole, early diastole, and late
diastole, respectively. Longitudinal myocardial velocity
gradients, defined as the difference between basal
and apical velocities, were 5.9 ± 2.2 cm/s, 6.9 ± 2.5
cm/s, and 4.9 ± 1.7 cm/s in systole, early diastole, and
late diastole, respectively. A breed effect was detected
for several systolic and diastolic TDI variables. In all
segments, systolic velocities were independent of
fractional shortening.

Conclusions and Clinical Relevance—LVFW
myocardial velocities decreased from the endocardium
to the epicardium and from base to apex, thus
revealing intramyocardial radial and longitudinal velocity
gradients. These indices could enhance conventional
echocardiographic analysis of left ventricular
function in dogs. Breed-specific reference intervals
should be defined. (Am J Vet Res 2005;66:953–961)
Duplex ultrasonography of the common carotid artery and external jugular vein of cows
Authors: Braun Ueli , Föhn Josef
Published: December 2005

Objective—To determine blood flow patterns in the common carotid artery and external jugular vein in cows before and after sedation achieved by administration of xylazine hydrochloride.

Animals—30 clinically normal Swiss Braunvieh cows.

Procedure—A 5.0-MHz sector transducer was used to examine the common carotid artery and external jugular vein before and after cows were sedated by administration of xylazine. Several variables were calculated, including diameter of the blood vessels, blood flow velocity, and flow-time volume.

Results—The common carotid artery before sedation had a maximum systolic velocity of 89 ± 8.5 cm/s, maximum diastolic velocity of 36 ± 6.0 cm/s, mean velocity of 35 ± 5.4 cm/s, and flow-time volume of 28.2 ± 3.48 cm3/s. In all cows, sedation achieved by administration of xylazine resulted in a significant decrease in velocity of arterial blood flow and flowtime volume. The external jugular vein before sedation had a maximum velocity of 65 ± 8.3 cm/s, maximum velocity of retrograde venous blood flow of 20 ± 6.6 cm/s, and flow-time volume of 29.7 ± 5.42 cm3/s. These values decreased significantly after cows were sedated by administration of xylazine.

Conclusions and Clinical Relevance—Doppler ultrasonography is particularly suitable for evaluation of blood flow patterns in the common carotid artery and external jugular vein of healthy cows. The results reported here provide a basis for use in examination of cows with cardiac and blood vessel disease. (Am J Vet Res2005;66:962–965)
Cardiopulmonary changes induced during one-lung ventilation in anesthetized dogs with a closed thoracic cavity
Authors: Riquelme Miriam , Monnet Eric , Kudnig Simon T. , Gaynor James S. , Wagner Ann E. , Corliss Denise , Salman M. D.
Published: December 2005

Objective—To evaluate the effects of one-lung ventilation
(OLV) on oxygen delivery (DO2) in anesthetized
dogs with a closed thoracic cavity.

Animals—7 clinically normal adult Walker Hound
dogs.

Procedure—Dogs were anesthetized. Catheters
were inserted in a dorsal pedal artery and the pulmonary
artery. Dogs were positioned in right lateral
recumbency. Data were collected at baseline (PaCO2
of 35 to 45 mm Hg), during two-lung ventilation, and
15 minutes after creating OLV. Hemodynamic and respiratory
variables were analyzed and calculations performed
to obtain DO2
, and values were compared
among the various time points by use of an ANOVA
for repeated measures.

Results—OLV induced a significant augmentation of
shunt fraction that resulted in a significant reduction
in PaO2, arterial oxygen saturation, and arterial oxygen
content. Cardiac index was not significantly changed.
The net result was that DO2 was not significantly
affected by OLV.

Conclusions and Clinical Relevance—Use of OLV in
healthy dogs does not induce significant changes in
DO2, which is the ultimate variable to use when evaluating
tissue oxygenation. One-lung ventilation can
be initiated safely in dogs before entering the thoracic
cavity during surgery. Additional studies are necessary
to evaluate OLV in clinically affected patients and
variations in age, body position, and type of anesthetic
protocol. (Am J Vet Res 2005;66:973–977)
Effects of aerosolized class C fly ash in weanling goats
Authors: Purdy Charles W. , Straus David C. , Ayers J. R.
Published: December 2005

Objective—To determine effects of repeated aerosol
exposures to fly ash dust on respiratory tracts of tent-confined
goats.

Animals—12 weanling Boer-Spanish crossbred
goats.

Procedure—Goats were randomly assigned to 2
groups: fly ash treatment group (principal goats,
n = 6) or control group (control goats, 6). Aerosolized fly
ash dust was provided during a 4-hour period for each
of 6 applications given over 3 months and one 2-hour
application prior to necropsy. Fly ash particle diameters
ranged from 0.1 to 130 µm and averaged 17.8 µm, with
1.5% of fly ash particles in the 0.1- to 5-µm-diameter
range. A mean ± SD of 748 ± 152 g/treatment was
delivered inside a tent containing principal goats; control
goats were placed inside a similar tent for 4-hour
treatments without dust. Following treatment, rectal
temperatures were taken at 0, 4, 6, 8, 24, and 72
hours; Hcts were recorded at 0, 24, and 72 hours.

Results—Rectal temperatures were significantly
increased at 4, 6, and 8 hours and decreased at 72
hours, compared with 0 hours. Mean ± SEM Hct values
were significantly increased for principal goats
(37.47 ± 0.39%), compared with control goats (36.17
± 0.42%). A significant increase in the mean area of
gross atelectatic lung lesions (1,410 mm2) was found
in principal goats (n = 6), compared with control goats
(440 mm2; 5).

Conclusions and Clinical Relevance—An increase
in atelectatic lung lesions was observed in principal
goats, compared with control goats; however, overall,
fly ash dust effects were nontoxic. ( Am J Vet Res 2005;66:991–995)
Cytokine gene transcription in feline nasal tissue with histologic evidence of inflammation
Authors: Johnson Lynelle R. , De Cock Hilde E. V. , Sykes Jane E. , Kass Philip H. , Maggs David J. , Leutenegger Christian M.
Published: December 2005

Objective—To correlate gene transcription of
cytokines and chemokines with histologic inflammation
in nasal biopsy specimens of cats.

Animals—25 study cats and 4 specific pathogen–free
cats.

Procedure—One nasal biopsy specimen from each
cat was submitted for routine histologic evaluation; a
second was submitted for evaluation by use of a
quantitative real-time polymerase chain reaction
analysis with a fluorogenic probe (ie, TaqMan) for
detection of cytokines and chemokines (interleukin
[IL]-4, IL-5, IL-6, IL-10, IL-12 p40, IL-16, IL-18, interferon
[IFN]-γ, tumor necrosis factor [TNF]-α, and the regulated
on activation normal T cell expressed and secreted
[RANTES] protein). Specimens were grouped histologically
by degree of inflammation (none, mild,
moderate, or severe). Linearized TaqMan signals for
each gene were compared among histologic groups.

Results—Nasal biopsy specimens from specific
pathogen–free cats were histologically normal, and
cytokine transcription was low in these samples. As
nasal inflammation in study cats worsened from
absent (n = 3) to mild (4) to moderate (8) or severe
(10), progressively and significantly increasing transcription
of IL-6, IL-10, IL-12 p40, IFN-γ, TNF-α, and the
RANTES protein was detected. Transcription of IL-4, IL-
5, IL-16, and IL-18 did not correlate with worsened histologic
inflammation.

Conclusions and Clinical Relevance—Transcription
of specific cytokines and chemokines in nasal tissue
of cats progressively increased with severity of histologic
evidence of inflammation, and IL-6, IL-10, IL-12
p40, IFN-γ, TNF-α, and the RANTES protein were
markers of inflammation. Our data suggest that the
nasal cavity of cats is biased toward a Th1 cytokine
profile that is augmented by inflammation. (Am J Vet
Res 2005;66:996–1001)