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- Author or Editor: Robert O. Gilbert x
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Abstract
Objective—To determine the effects of 2 conjugated linoleic acid (CLA) isomers (cis-9, trans-11 and trans-10, cis-12) on synthesis of prostaglandin (PG) E2 and F2α and expression of prostaglandin H synthase-2 (PGHS-2) of adult and fetal bovine endometrial epithelial cells in vitro.
Sample—Primary cultures of endometrial epithelial cells obtained from 4 adult cows and 4 fetal bovine carcasses.
Procedures—Cells were exposed to 0, 50, 100, or 200μM cis-9, trans-11 or trans-10, cis-12 CLA isomers for 24 hours. Culture media collected before and after 6 hours of stimulation of cells with phorbol 12-myristate 13-acetate were assayed to detect PGE2 and PGF2α via ELISA. After stimulation, cells were collected for western blot analysis to quantify PGHS-2.
Results—Concentrations of PGF2α and PGE2 were significantly lower in culture media of adult and fetal endometrial epithelial cells exposed to any concentration of either CLA than they were in media of cells not exposed to CLAs. The trans-10, cis-12 CLA isomer seemed to decrease PG production more markedly than did the cis-9, trans-11 CLA isomer. Most concentrations of both CLAs significantly reduced culture media PGE2:PGF2α concentration ratios of cells. Exposure of cells to CLAs did not affect expression of PGHS-2 protein.
Conclusions and Clinical Relevance—Results of this study indicated CLAs significantly decreased PGF2α and PGE2 concentrations and PGE2:PGF2α concentration ratios for cultures of adult and fetal endometrial epithelial cells with no apparent effect on PGHS-2 expression. Similar effects in cows could have effects on maternal recognition of pregnancy and immune function.
Abstract
Objective—To evaluate the cytopathic effects of Tritrichomonas foetus and a purified cysteine protease (ie, CP30) of T foetus on cultured bovine uterine epithelial cells (BUECs) in vitro.
Sample Population—10 reproductive tracts were obtained from late-term bovine fetuses at a commercial abattoir.
Procedure—An in vitro culture system of BUECs was developed to study the cytopathic effects of T foetus and purified CP30 of T foetus on host cells. Cytotoxicity of T foetus or CP30 on exposed BUECs was determined. Fluorescence microscopy and flow cytometry analyses were used to detect apoptosis. A fluorometric assay was used to detect BUEC caspase 3 activation. The CP inhibitor E-64 and a caspase inhibitor were used to inhibit apoptosis.
Results—Cytopathic effects were observed in BUECs treated with parasites or CP30 and were concentration and time dependent. The BUECs underwent apoptosis in the presence of parasites or CP30. The specific CP inhibitor E-64 abolished the induction of apoptosis in BUECs by CP30. The caspase inhibitor reduced the amount of apoptosis in BUECs.
Conclusions and Clinical Relevance—T foetus and its CP30 induce apoptosis in cultured BUECs in vitro. Induction of apoptosis by CP30 is correlated with protease activity. Endometrial cell death as a result of a T foetus infection is likely to be more important in mediating infertility than a direct effect on the conceptus. Provoking an apoptotic reaction in the host may mitigate an inflammatory reaction or immune response and therefore favor survival of the parasite in a chronic infection. (Am J Vet Res 2005;66:1181–1186)
Abstract
OBJECTIVE To evaluate use of flunixin meglumine as a treatment to postpone ovulation in mares, mare fertility after flunixin meglumine treatment during estrous cycles, and effects of flunixin meglumine on function of the corpus luteum after ovulation.
ANIMALS 13 healthy mares.
PROCEDURES A single-blinded, placebo-controlled, crossover study was conducted. Flunixin meglumine (1.1 mg/kg, IV, q 24 h) or lactated Ringer solution (placebo treatment) was administered for 2 days to mares with a dominant follicle (≥ 35 mm in diameter) and behavioral signs of estrus. Mares then were bred by artificial insemination. Number of days to ovulation from initial detection of a follicle ≥ 30 mm in diameter, uterine edema score, and pregnancy were determined by ultrasonography; the examiner was unaware of the treatment of each mare. Serum progesterone concentrations were evaluated 5 and 12 days after ovulation by use of radioimmunoassay.
RESULTS Data were available for 45 estrus cycles of the 13 mares. Number of days to ovulation from initial detection of a follicle ≥ 30 mm was not significantly affected by administration of flunixin meglumine versus the placebo. Per-cycle pregnancy rate was not significantly different between flunixin meglumine (20/24 [83%] breedings) and the placebo (13/19 [68%] breedings). Flunixin meglumine did not significantly affect behavioral signs of estrus, uterine edema, or serum progesterone concentrations.
CONCLUSIONS AND CLINICAL RELEVANCE Findings did not support the use of flunixin meglumine to postpone ovulation in mares.
Abstract
Objective—To estimate the number of dogs required to find linkage to heritable traits of hip dysplasia in dogs from an experimental pedigree.
Animals—147 Labrador Retrievers, Greyhounds, and their crossbreed offspring.
Procedure—Labrador Retrievers with hip dysplasia were crossed with unaffected Greyhounds. Age at detection of femoral capital ossification, distraction index (DI), hip joint dorsolateral subluxation (DLS) score, and hip joint osteoarthritis (OA) were recorded. Power to find linkage of a single marker to a quantitative trait locus (QTL) controlling 100% of the variation in a dysplastic trait in the backcross dogs was determined.
Results—For the DI at the observed effect size, recombination fraction of 0.05, and heterozygosity of 0.75, 35 dogs in the backcross of the F1 to the Greyhound generation would yield linkage at a power of 0.8. For the DLS score, 35 dogs in the backcross to the Labrador Retriever generation would be required for linkage at the same power. For OSS, 45 dogs in the backcross to the founding Labrador Retrievers would yield linkage at the same power. Fewer dogs were projected to be necessary to find linkage to hip OA. Testing for linkage to the DLS at 4 loci simultaneously, each controlling 25% of the phenotypic variation, yielded an overall power of 0.7.
Conclusions and Clinical Significance—Based on this conservative single-marker estimate, this pedigree has the requisite power to find microsatellites linked to susceptibility loci for hip dysplasia and hip OA by breeding a reasonable number of backcross dogs. (Am J Vet Res 2003;222:418–424)
Abstract
Objective—To determine the genetic influence on expression of traits associated with canine hip dysplasia.
Animals—193 dogs from an experimental canine pedigree.
Procedure—An experimental canine pedigree was developed for linkage analysis of hip dysplasia by mating dysplastic Labrador Retrievers with nondysplastic Greyhounds. A statistical model was designed to test the effects of Labrador Retriever and Greyhound alleles on age at detection of femoral capital epiphyseal ossification, 8-month distraction index, and 8-month dorsolateral subluxation score.
Results—The additive effect was significant for age at detection of femoral capital epiphyseal ossification. Restricted maximum likelihood estimates (± SD) for this trait were 6.4 ± 1.95, 10.2 ± 2.0, 10.8 ± 3.1, 11.4 ± 2.1, and 13.6 ± 4.6 days of age for Greyhounds, Greyhound backcross dogs, F1 dogs, Labrador Retriever backcross dogs, and Labrador Retrievers, respectively. The additive effect was also significant for the distraction index. Estimates for this trait were 0.21 ± 0.07, 0.29 ± 0.15, 0.44 ± 0.12, 0.52 ± 0.18, and 0.6 ± 0.17 for the same groups, respectively. For the dorsolateral subluxation score, additive and dominance effects were significant. Estimates for this trait were 73.5 ± 4.1, 71.3 ± 6.5, 69.1 ± 6.0, 50.6 ± 12.9, and 48.4 ± 7.7%, respectively, for the same groups.
Conclusions—In this canine pedigree, traits associated with canine hip dysplasia are heritable. Phenotypic differences exist among founder dogs of each breed and their crosses. This pedigree should be useful for identification of quantitative trait loci underlying the dysplastic phenotype. (Am J Vet Res 2002;63: 1029–1035)