Relaxin peptide hormones are protective during the early stages of ischemic stroke in male rats.

The pregnancy hormone relaxin protects tissue from ischemic damage. The ability of relaxin-3, a relaxin paralog, to do so has not been explored. The cerebral expression levels of these peptides and their receptors make them logical targets for study in the ischemic brain. We assessed relaxin peptide-mediated protection, relative relaxin family peptide receptor (RXFP) involvement, and protective mechanisms. Sprague-Dawley rats receiving permanent (pMCAO) or transient middle cerebral artery occlusions (tMCAO) were treated with relaxin peptides, and brains were collected for infarct analysis. Activation of the endothelial nitric oxide synthase pathway was evaluated as a potential protective mechanism. Primary cortical rat astrocytes were exposed to oxygen glucose deprivation and treated with relaxin peptides, and viability was examined. Receptor involvement was explored using RXFP3 antagonist or agonist treatment and real-time PCR. Relaxin and relaxin-3 reduced infarct size after pMCAO. Both peptides activated endothelial nitric oxide synthase. Because relaxin-3 has not previously been associated with this pathway and displays promiscuous RXFP binding, we explored the receptor contribution. Expression of rxfp1 was greater than that of rxfp3 in rat brain, although peptide binding at either receptor resulted in similar overall protection after pMCAO. Only RXFP3 activation reduced infarct size after tMCAO. In astrocytes, rxfp3 gene expression was greater than that of rxfp1. Selective activation of RXFP3 maintained astrocyte viability after oxygen glucose deprivation. Relaxin peptides are protective during the early stages of ischemic stroke. Differential responses among treatments and models suggest that RXFP1 and RXFP3 initiate different protective mechanisms. This preliminary work is a pivotal first step in identifying the clinical implications of relaxin peptides in ischemic stroke.

Vaginal impedometry for detection of optimal breeding time in bitches.

OBJECTIVE: To compare the efficacy of canine vaginal impedometry in identifying the preovulatory luteinizing hormone (LH) peak to that of currently used methods (serum progesterone concentration measurement, vaginal cytologic evaluation, and vaginoscopy). DESIGN: Prospective study. ANIMALS: 12 sexually intact female dogs. PROCEDURES: 12 mature postpubertal Beagle (n = 3), Beagle-cross (2), and hound-cross (7) bitches ranging from 7.5 to 27.5 kg (16.5 to 60.6 lb) were enrolled in the study. After the onset of spontaneous proestrus, determined on the basis of appearance of serosanguineous vaginal discharge, serum progesterone assays, vaginoscopy, vaginal cytologic evaluation, and vaginal impedometry were performed daily until approximately 4 days after peak LH concentration (day 0) as measured by radioimmunoassay. Vaginal impedometry was compared against serum progesterone concentration measurement, vaginal cytologic evaluation, and vaginoscopy as a method for accurately identifying the LH peak and therefore the optimal breeding time. Ten of 12 bitches were bred with subsequent assessment of embryos. RESULTS: Vaginal impedometry accurately predicted the preovulatory LH peak in 5 of 11 bitches. One bitch was removed from the study because data were not collected. Of the remaining 11 bitches, 6 had their LH peak on the day serum progesterone concentration first exceeded 2 ng/mL. Crenulation scores reached 1 (mean, 1.3; 95% confidence interval, 0.8 to 1.7) on day 0 as expected; however, these scores were not significantly different from those on days -1 or 1. Vaginal epithelial cell populations did not change noticeably on day 0. Nine of the 10 bitches that were bred produced viable embryos. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that daily use of vaginal impedometry in bitches was unreliable as a method for monitoring periovulatory events. All techniques evaluated (ie vaginal impedometry, serum progesterone concentration assays, vaginoscopy and vaginal cytologic evaluation) frequently produced inaccurate results when used individually. Multiple methods should be used to identify optimal breeding time in dogs.

An evaluation of B-mode and color Doppler ultrasonography for detecting periovulatory events in the bitch.

When determining optimal breeding time in the bitch, specific periovulatory events must be identified. The main objectives were to relate ultrasonographic changes in ovarian blood flow, follicle/corpora lutea count and echotexture to periovulatory events, and to assess the efficacy of each for identifying these events. Twelve Beagle (N = 3), Beagle-cross (N = 2) and hound-cross (N = 7) bitches (body weight range, 7.5-27.5 kg) were examined daily from the onset of proestrus to approximately 4 days post-LH peak. Follicle and corpora lutea count and echotexture analyses were performed using B-mode ultrasound and ovarian blood flow analysis was performed using color Doppler ultrasound. Serum LH concentrations were analyzed by validated RIA. There was an increase (P < 0.05) in ovarian blood flow from the day of the preovulatory LH peak (605 pixels; confidence interval, 397-856), to 1 day after this peak (1092 pixels; confidence interval, 724-1535), enabling detection of the preovulatory LH peak. There were no significant changes in follicle/corpora lutea echotexture relative to days from the preovulatory LH peak. There were significant decreases in follicle/corpora lutea number between Days -1 and 3; Days -1 and 4; and Days 0 and 3, relative to the preovulatory LH peak. We concluded that color Doppler ultrasound performed once daily was more accurate in identifying the preovulatory LH peak than B-mode ultrasound and enabled prospective determination of ovulation.