Equine chorionic gonadotropin (eCG) treatment in heifers: Double ovulation, twinning rate, and pregnancy losses in twin pregnancies.

This study aimed to evaluate the effects of different doses of equine chorionic gonadotropin (eCG; 200 and 300 IU) administered at the end of a fixed-time artificial insemination (FTAI) treatment protocol on ovulation, pregnancy, and twin rates in Bos taurus beef heifers. In addition, pregnancy losses in heifers with singleton and twin pregnancies were determined. A total of 2382 Angus heifers treated with a 6-day estradiol/progesterone-based protocol for FTAI (J-Synch protocol) were randomly allocated to two experimental groups to receive 200 or 300 IU of eCG administered intramuscularly at the time of intravaginal progesterone device removal; FTAI was performed from 60 to 72 h after device removal. The pregnancy rate did not differ (P = 0.89) between the 200 and 300 IU eCG groups. The number of corpus luteum induced by both eCG doses was determined by ultrasonographic examination 14 days after insemination and those treated with 300 IU of eCG had a greater double ovulation rate (P < 0.05). In addition, 300 IU eCG treated heifers had a higher twinning rate on day 30 of gestation (P < 0.05) and parturition (P < 0.05). Pregnancy losses from 30 days of gestation to calving did not differ between heifers treated with 200 and 300 IU of eCG (P = 0.70). However, regardless of the experimental group, heifers bearing twins had greater pregnancy losses than heifers with singletons (P < 0.05). In conclusion, reducing the dose of eCG from 300 to 200 IU under FTAI treatment protocol decreases double ovulation and twinning rates, maintaining a similar pregnancy rate in heifers. Nulliparous cows carrying two fetuses suffer greater pregnancy losses than cows with singletons.

Time of ovulation and pregnancy outcomes obtained with the prostaglandin-based protocol Synchrovine for FTAI in sheep.

The objective of the present study was to determine the ovarian response induced with the prostaglandin-based protocol Synchrovine (two doses of PGF2α given 7 d apart), as well as the fertility after FTAI. In Experiment 1, 15 females received the Synchrovine protocol using two different PGF2α analogues (Delprostenate vs. D-Cloprostenol). No differences in estrus response, time of ovulation and follicular dynamics were found between both groups (P < 0.05). The ovulation after Synchrovine was synchronized in a similar mean interval (68.8 ± 7.1 h) than when the females received a single dose of PGF2α (70.2 ± 20.7 h; P=NS), but the dispersion between the first and the last ovulation was reduced with this protocol (range 60-84 h vs. 24-96 h, respectively; P < 0.05). In experiment 2, 318 ewes were treated with the Synchrovine protocol and cervical FTAI was performed using different sperm cell concentrations. Pregnancy rate was higher using 200 × 106 and 100 × 106 sperm cells (38.2%, 39/102; and 34.9%, 38/109, respectively) than using 50 × 106 (23.4%, 25/107, P < 0.05). In Experiment 3, 444 ewes received the Synchrovine protocol and were assigned to receive 300 IU of eCG or not at the moment of the second dose of PGF2α, and cervical FTAI was performed 42 h or 48 h after the second dose of PGF2α. No effect was found related to the eCG administration nor the time of insemination. In Experiment 4, 342 received cervical or intrauterine insemination after treatment with the Synchrovine protocol, resulting in greater pregnancy rate for intrauterine insemination than cervical insemination (52.5%, 90/171 vs. 31%, 53/171, P < 0.05). These experiments demonstrate that the Synchrovine protocol effectively induces luteolysis, estrus and ovulation in most of the treated females, and ovulation is synchronized into a narrow window of 24 h. Pregnancy rate obtained with cervical FTAI is around 30-45%, with similar results using 100 × 106 or 200 × 106 sperm cells, the eCG administration seems not to be necessary, the type of PGF2α analogue does not appear relevant, and fertility is improved with intrauterine semen deposition.

Furoxan derivatives as cytotoxic agents: preliminary in vivo antitumoral activity studies.

Furoxan derivatives with in vitro cytotoxic activity were investigated as antitumoral agents in vivo. The compounds were tested in murine models of both CCRFS-180 II sarcoma and mammary adenocarcinoma. Two of the furoxan derivatives considered here, 3-formyl-4-phenyl-1,2,5-oxadiazole N2-oxide and 3-carbonitrile-4-phenyl-1,2,5-oxadiazole N2-oxide, present in vivo antitumoral activity. They were able to produce more than 90% of tumoral necrosis under the experimental protocol of administration and posology employed. NO-releasing capacity of furoxans may explain the anti-neoplastic activity of these compounds.

Stabilization of blood homocysteine in an epidemiological setting.

The major problem in the determination of homocysteine (Hcy), which is thought to be a risk factor in colorectal cancer, is the rise in its concentration if blood is not centrifuged immediately after collection. We assess the interference of 3-deazaadenosine (which inhibits conversion of S-adenosylhomocysteine into Hcy within the erythrocyte), using the fluorescence polarization immunoassay (FPIA) assay, the stabilizing effect of 3-deazaadenosine and the impact of temperature on Hcy stabilization. To assess interference of 3-deazaadenosine, 12 blood samples were extracted; two aliquots were obtained from each and one of them was added 3-deazaadenosine (50 micromol/l). To assess the stabilizing value of 3-deazaadenosine, as well as the effect of temperature, two blood samples were extracted from 24 volunteers. One of the tubes was immediately placed on ice and centrifuged (reference concentration). To the second tube was immediately added 3-deazaadenosine (50 micromol/l), producing six aliquots, three of which were kept at room temperature (25 degrees C) for 1, 4 and 6 hours, the other three kept at 37 degrees C. The mean values (standard deviation) obtained for methodological interference were: 7.32 (3.58) micromol/l without stabilizer, and 7.11 (3.61) micromol/l with stabilizer. There were no statistically significant differences (P = 0.104) and intraclass correlation coefficient was 0.989, suggesting no methodological interference. We did not find any significant differences regarding our reference value in the samples kept at room temperature during the interval studied. A high Pearson correlation coefficient was obtained. Nevertheless, in those samples kept at 37 degrees C, a slight increase was observed in the 4-hour period (P = 0.009). The addition of 3-deazaadenosine may avoid problems in the critical pre-analytical phase in the Hcy measurement. There is no interference with the FPIA assay, nor any dilution effect, and new reference values are not necessary.

Ovulatory response and luteal function after eCG administration at the end of a progesterone and estradiol' based treatment in postpartum anestrous beef cattle.

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) administration associated to fixed-time AI (FTAI) on follicular dynamics, ovulation, corpus luteum (CL) development and serum progesterone concentrations. Multiparous suckled Hereford cows (n=46) in anestrus with 60-75 days postpartum were used. Females received an intravaginal device containing 0.5g of progesterone during 8 days and 2mg of estradiol benzoate i.m. at device insertion. At device removal 500µg of cloprostenol and 0.5mg of estradiol cypionate were administered i.m., and FTAI was performed 52-56h later. Cows were divided into two experimental groups to receive 400IU of eCG i.m. at device removal (n=23), while control group did not receive eCG (n=23). Daily ovarian ultrasonography (7.5MHz transducer) and progesterone concentrations determined by RIA were assayed from device removal until 30 or 14 days after FTAI, respectively. Treatment with eCG increased ovulation rate [65.2% (15/23) vs. 30.4% (7/23); P=0.018], ovulatory follicle diameter (14.5±0.4 vs. 13.1±0.7mm, mean±SEM; P=0.081), CL area from 6 to 14 days after FTAI (344.3±25.1 vs. 274.2±23.9mm(2); P=0.045) and mean serum progesterone concentrations from FTAI to 14 days later (3.0±0.2 vs. 1.8±0.2ng/ml; P=0.001), in comparison with control cows. In conclusion, the addition of eCG to a progesterone and estradiol' based treatment for FTAI improves ovulation rate and luteal function in anestrous cows. These findings have implications in order to increase pregnancy rates in FTAI treatments in Bos taurus beef cattle.