Detection of estrous behavior in buffalo heifers by radiotelemetry following PGF2α administration during the early or late luteal phase

This study examined the usefulness of radiotelemetry for estrous detection in buffalo heifers and the impact of prostaglandin F_2α (PGF_2α) administration during the early or late luteal phase on estrous behavior and ovulatory follicle variables. Induction of estrus with PGF_2α at a random stage of the estrous cycle was followed by the arbitrary division of heifers into groups receiving a second dose of PGF_2α during either the early (n = 33) or late (n = 17) luteal phase (6–9 or 11–14 days after estrus, respectively) for the induction of synchronized estrus. The electronic detection of synchronized estrus by radiotelemetry was confirmed using ultrasonography every 6 h until ovulation. Radiotelemetry was 90% efficient and 100% accurate for estrous detection. Intervals between the PGF_2α dose and the beginning of synchronized estrus (40.7 ± 10.9 vs. 56.7 ± 12.8 h) or ovulation (70.0 ± 11.3 vs. 85.6 ± 12.5 h) were shorter (P < 0.05) for heifers receiving PGF_2α during the early luteal phase. PGF_2α administration during the early or late luteal phase produced similar (P > 0.05) results for the duration of estrus, the intervals from the beginning or end of estrus to ovulation, the number and duration of mounts per estrus, the duration of mounts, the diameter of the ovulatory follicle and the luteal profile on day 5 after estrus. In conclusion, radiotelemetry was a suitable tool for the efficient and accurate detection of estrus in buffalo heifers. Treatment with PGF_2α during the early luteal phase had a shorter interval to synchronized estrus and ovulation; however, estrous behavior, ovulatory follicle dynamics and subsequent luteal activity were similar following PGF_2α administration during the early or late luteal phase.     

Effect of prostaglandins E2 and F2α on in vitro development and hatching of caprine blastocysts

Prostaglandin E₂ has been shown to increase the ovine embryo hatching rate, and PGF_2α to reduce the development of rabbit, bovine, and rat embryos. The objective was to determine the effects of PGE₂ and PGF_2α on development of caprine embryos. Estrus was synchronized in does (n = 25) with medroxyprogesterone acetate (MAP) intravaginal sponges for 12 days, and superovulated with 20 units of FSH. On day 6 following estrus, embryos were flushed (n = 128) and incubated individually per well in 25 μl droplets of TCM-199 and BSA (8 mg/ml) for 6 days at 38.5 °C in a 5% CO₂: air with one of the following treatments: (1) control (0.0002% EtOH), (2) PGE₂ (7 ng/ml), (3) PGF_2α (7 ng/ml), (4) low PGE₂:high PGF_2α (3.5 ng/ml:14 ng/ml), (5) balanced PGE₂:PGF_2α (7 ng/ml:7 ng/ml), or (6) high PGE₂:low PGF_2α (14 ng/ml:3.5 ng/ml). Treatment with PGE₂ alone reduced (P < 0.05) the hatching rate (1/15; 7%). The hatching rate of embryos treated with PGF_2α alone (9/18; 50%), low PGE₂:high PGF_2α (8/16; 50%), and balanced PGE₂:PGF_2α (11/16; 69%) were similar to control (6/18; 33%). In contrast, the hatching rate was non-significantly increased (13/18; 72%) with the high PGE₂:low PGF_2α treatment. None of the treatments affected development from the morula to blastocyst stage. From the current data, it can be concluded that PGE₂ alone reduced hatching rate, and PGF_2α alone had no effect on the development of caprine embryos. High concentrations of PGE₂ with PGF_2α improved the hatching rates. Thus, uterine concentrations of PGE₂ may need to reach a threshold level to improve embryo hatching, as previously reported, while increased uterine concentrations of PGF_2α during early pregnancy would not affect development of the embryo.     

Comparison of the patterns of antral follicular development between hormonally synchronized and natural estrous cycles of non-seasonal,polyestrous goats in the tropics

The effects of estrus synchronization with prostaglandin F_2α (PGF_2α) and Controlled Internal Drug Release Device (CIDR) on ensuing antral follicular development were documented and compared to natural estrous cycles of non-seasonal tropical goats. Two to six follicular waves were observed, with the three-follicular wave pattern being most frequently observed (58%), followed by four follicular waves (31.6%) per estrous cycle. There were no significant differences (p > 0.05) between the PGF_2α- or CIDR-synchronized and natural estrous cycles nor between the synchronized and subsequent non-synchronized cycles in terms of the time of ovulation, the duration of inter-ovulatory intervals, daily numbers of antral follicles ≥3 mm in diameter, and the number of follicular waves per cycle in the goats of the present study.     

Prostaglandin E1 (PGE1), but not prostaglandin E2 (PGE2), alters luteal and endometrial luteinizing hormone (LH) occupied and unoccupied LH receptors and mRNA for LH receptors in ovine luteal tissue to prevent luteolysis

Loss of luteal progesterone secretion at the end of the ovine estrous cycle is via uterine PGF₂α secretion. However, uterine PGF₂α secretion is not decreased during early pregnancy in ewes. Instead, the embryo imparts a resistance to PGF₂α. Prostaglandins E (PGE; PGE₁ + PGE₂) are increased in endometrium and uterine venous blood during early pregnancy in ewes to prevent luteolysis. Chronic intrauterine infusion of PGE₁ or PGE₂ prevents spontaneous or IUD, estradiol-17β, or PGF₂α-induced premature luteolysis in nonbred ewes. The objective was to determine whether chronic intrauterine infusion of PGE₁ or PGE₂ affected mRNA for LH receptors, occupied and unoccupied receptors for LH in luteal and caruncular endometrium, and luteal function. Ewes received Vehicle, PGE₁, or PGE₂ every 4 h from days 10 to 16 of the estrous cycle via a cathether installed in the uterine lumen ipsilateral to the luteal-containing ovary.Jugular venous blood was collected daily for analysis of progesterone and uterine venous blood was collected on day-16 for analysis of PGF₂α and PGE. Corpora lutea and caruncular endometrium were collected from day-10 preluteolytic control ewes and day-16 ewes treated with Vehicle, PGE₁ or PGE₂ for analysis of the mRNA for LH receptors and occupied and unoccupied receptors for LH. Luteal weights on day-16 in ewes treated with PGE₁ or PGE₂ and day-10 control ewes were similar (P ≥ 0.05), but were greater (P ≤ 0.05) than in day-16 Vehicle-treated ewes. Progesterone profiles on days 10–16 differed (P ≤ 0.05) among treatment groups: PGE₁ > PGE₂ > Vehicle-treated ewes. Concentrations of PGF₂α and PGE in uterine venous plasma on day-16 were similar (P ≥ 0.05) in the three treatment groups. Luteal mRNA for LH receptors and unoccupied and occupied LH receptors were similar (P ≥ 0.05) in day-10 control ewes and day-16 ewes treated with PGE₂ and were lower (P ≤ 0.05) in day-16 Vehicle-treated ewes. PGE₂ prevented loss (P ≤ 0.05) of day-16 luteal mRNA for LH receptors and occupied and unoccupied LH receptors. Luteal and caruncular tissue mRNA for LH receptors and occupied and unoccupied LH receptors were greater (P ≤ 0.05) on day-16 of PGE₁-treated ewes than any treatment group. mRNA for LH receptors and occupied and unoccupied receptors for LH in caruncules were greater (P ≤ 0.05) in day-16 Vehicle or PGE₂-treated ewes than in day-10 control ewes. It is concluded that PGE₁ and PGE₂ share some common mechanisms to prevent luteolysis; however, only PGE₁ increased luteal and endometrial mRNA for LH receptors and occupied and unoccupied LH receptors. PGE₂ prevents a decrease in luteal mRNA for LH receptors and occupied and unoccupied receptors for LH without altering endometrial mRNA for LH receptors or occupied and unoccupied receptors for LH.     

Synchronization of estrus with short- and long-term progestagen treatments and the use of GnRH prior to short-term progestagen treatment in ewes

The objective of the present study was to determine the efficacy of the synchronization of estrus using short- and long-term progestagen treatments in ewes at the onset of the breeding season, and to evaluate the effect of the exogenous GnRH administration immediately prior to short-term progestagen treatment on the reproductive performance. A total of 240 Tahirova cross-bred ewes, aged 18–24 months, and 40 rams, aged 2–4 years-old, were used in the trial. Ewes were divided equally into 3 groups (n = 80 per group). Intravaginal progestagen sponges containing FGA (30 mg) were inserted in the ewes for 7 d in the FGA1 (short-term) and GnRH treatment groups, and for 12 d in the FGA2 group (long-term). The ewes in the GnRH group received 10.5 μg busereline acetate i.m. at the time of sponge insertion. Tiaprost tromethamol (PGF_2α; 0.294 mg) and eCG (400 IU) were injected i.m. on the 6th day of progestagen treatment in the GnRH and FGA1 groups, and on the 11th day in the FGA2 group following sponge insertion. All ewes were hand-mated once at the detection of estrus. The estrous response, fertility rate, multiple birth rate and litter size recorded was 88.7, 87.3, 51.6% and 1.6 in the FGA1 group, 92.5, 71.6, 50.9% and 1.5 in the FGA2 group, and 96.2, 89.6, 71.0% and 1.8 in the GnRH group, respectively. No significant difference in estrous response between the groups was recorded, but the fertility rate in the FGA1 and GnRH groups was significantly (P < 0.05) higher than in the FGA2 group. The occurrence of multiple births and litter sizes were significantly (P < 0.05) higher in the GnRH group, compared to both the FGA1 and FGA2 groups, with the number of single lambs being significantly (P < 0.05) higher in the FGA1 (48.4%) and FGA2 (49.0%) groups than in the GnRH (29.0%) group. However, the differences recorded between any of the groups in terms of the number of twin and triplet lambs were insignificant. In conclusion, it can be said that estrous synchronization using the 12-d-FGA-eCG-PGF_2α regimen could be replaced with the 7-d-FGA-eCG-PGF_2α regime in sheep at the onset of the breeding season. However, the combination of GnRH with the latter regimen (7-d-GnRH-FGA-eCG-PGF_2α) increased the multiple birth rate and litter size in the ewes.     

Direct effect of PGF2α pulses on PRL pulses, based on inhibition of PRL or PGF2α secretion in heifers

The relationships between PRL and PGF_2α and their effect on luteolysis were studied. Heifers were treated with a dopamine-receptor agonist (bromocriptine; Bc) and a Cox-1 and -2 inhibitor (flunixin meglumine [FM]) to inhibit PRL and PGF_2α, respectively. The Bc was given (Hour 0) when ongoing luteolysis was indicated by a 12.5% reduction in CL area (cm²) from the area on Day 14 postovulation, and FM was given at Hours 0, 4, and 8. Blood samples were collected every 8-h beginning on Day 14 until Hour 48 and hourly for Hours 0 to 12. Three groups of heifers in ongoing luteolysis were used: control (n = 7), Bc (n = 7), and FM (n = 4). Treatment with Bc decreased (P < 0.003) the PRL concentrations averaged over Hours 1 to 12. During the greatest decrease in PRL (Hours 2-6), LH concentrations were increased. Progesterone concentrations averaged over hours were greater (P < 0.05) in the Bc group than in the controls. In the FM group, no PGFM pulses were detected, and PRL concentrations were reduced. Concentrations of PGFM were not reduced in the Bc group, despite the reduction in PRL. Results supported the hypothesis that a decrease (12.5%) in CL area (cm²) is more efficient in targeting ongoing luteolysis (63%) than using any day from Days 14 to ≥19 (efficiency/day, 10-24%). The hypothesis that PRL has a role in luteolysis was supported but was confounded by the known positive effect of LH on progesterone. The hypothesis was supported that the synchrony of PGFM and PRL pulses represents a positive effect of PGF_2α on PRL, rather than an effect of PRL on PGF_2α.     

The effects of confinement on periparturient behaviour and circulating prolactin,prostaglandin F2α and oxytocin in gilts with access to a variety of nest materials

In a 2×2 factorial experiment, the effects of gestation and farrowing housing on (1) periparturient behaviour and circulating prolactin, prostaglandin F_2α (PGF_2α) and oxytocin in gilts with access to peat, straw and branches, and (2) correlational relationships between the periparturient behaviour and hormones were studied. The treatments consisted of housing in stalls or pens from mating to day 110 of gestation followed by farrowing crates or pens until after parturition. Landrace×Yorkshire gilts were observed from video recordings (n=25) from 20 h prepartum and blood sampled via jugular catheters (n=16) from 24 h prepartum until 2 h after the birth of the first piglet.There was an interaction between gestation and farrowing housing affecting the start of nest-building (P=0.03). Gilts that experienced a change in type of housing accommodation commenced nest-building closer to parturition than gilts that were penned both during gestation and at farrowing (both P<0.05). There were no effects of the housing environment on the timing of termination of nest-building, behaviour during parturition, or the course of parturition. However, relative to base level, crated gilts sat more from 16 to 6 h prepartum, whereas this was the case for penned gilts only from 9 to 7 h prepartum. Crated gilts also tended to change posture more often (P=0.07) and lie more in sternal recumbency (P=0.095). This suggests that familiarity with the environment in combination with space to move about and/or availability of materials is important in the timing of nest-building. Confinement during farrowing did not appear to impair feed-back from the materials and the nest, although increased number of postural changes may reflect the motivation but inability to nest-build, or general discomfort in the crate.There was a development over time in postural and nest-building behaviours as well as in plasma concentrations of prolactin, PGF_2α (measured by the metabolite PGFM) and oxytocin, but there were only few effects of housing treatments on hormones or associations between behaviour and hormones. The results suggest that nest-building occurs independently of a prepartum rise in prolactin, but that oxytocin may be associated with the termination of nest-building as there was a negative correlational relationship with nosing (P<0.01) and arranging nest-building materials (P<0.001).Farrowing crate housing appeared to have fewer effects on periparturient behaviour and course of parturition than reported in previous studies where effects of confinement and provision of nest-building materials may have been confounded. Thus, provision of nest-building materials to crated sows may have beneficial effects on sow behaviour and welfare.     

Placental antiangiogenic prolactin fragments are increased in human and rat maternal diabetes

Introduction/objectivesThe role of the placenta in diabetic mothers on fetal development and programming is unknown. Prolactin (PRL) produced by decidual endometrial cells may have an impact. Although full-length PRL is angiogenic, the processed form by bone morphogenetic protein-1 (BMP-1) and/or cathepsin D (CTSD) is antiangiogenic.The objectives were to investigate the involvement of decidual PRL and its antiangiogenic fragments in placentas from type-1 diabetic women (T1D) and from pregnant diabetic rats with lower offspring weights than controls.MethodsPRL, BMP-1, and CTSD gene expressions and PRL protein level were assessed in T1D placentas (n = 8) at delivery and compared to controls (n = 5). Wistar rats received, at day 7 of pregnancy, streptozotocin (STZ) (n = 5) or nicotinamide (NCT) plus STZ (n = 9) or vehicle (n = 9). Placental whole-genome gene expression and PRL western blots were performed at birth.ResultsIn human placentas, PRL (p < 0.05) and BMP-1 (p < 0.01) gene expressions were increased with a higher amount of cleaved PRL (p < 0.05) in T1D than controls. In rats, diabetes was more pronounced in STZ than in NCT–STZ group with intra-uterine growth restriction. Decidual prolactin-related protein (Dprp) (p < 0.01) and Bmp-1 (p < 0.001) genes were up-regulated in both diabetic groups, with an increased cleaved PRL amount in the STZ (p < 0.05) and NCT–STZ (p < 0.05) groups compared to controls. No difference in CTSD gene expression was observed in rats or women.ConclusionsAlterations in the levels of the PRL family are associated with maternal diabetes in both rats and T1D women suggesting that placental changes in these hormones impact on fetal development.     

Brain fixation for analysis of brain lipid-mediators of signal transduction and brain eicosanoids requires head-focused microwave irradiation: An historical perspective

Endothelin-1 (ET-1) has been reported to mediate prostaglandin (PG) F₂α (PGF₂α)-induced luteolysis. Prostaglandins E (PGE; PGE₁ + PGE₂) are associated with implantation, maternal recognition of pregnancy, and are antiluteolytic and luteotropic in vitro and in vivo. ET-1 increased PGE secretion by bovine luteal tissue in vitro from cows where estrus was not synchronized or when estrus was synchronized with lutalyse and did not affect luteal PGF₂α or progesterone secretion, which does not support the concept that ET-1 is luteolytic or mediates PGF₂α luteolysis. Therefore, the objective of this experiment was to determine whether ET-1 infused every 6 h from 2400 h on day 10–1800 h on day 18 of the ovine estrous cycle either into the interstitial tissue of the ovarian vascular pedicle (IP) or intrauterine (IU) adjacent to the luteal-containing ovary was luteolytic in ewes. Treatments were: Vehicle-IP; Vehicle-IU; ET-1-IP; or ET-1-IU. Weights of corpora lutea differed (P ≤ 0.05) among treatment groups. Weights of corpora lutea at 1800 h on day 18 were: VEH-IP—247 ± 38 mg; VEH-IU—195 ± 31 mg; ET-1-IP—626 ± 74 mg; and ET-1-IU—542 ± 69 mg. Luteal weights on day 18 in ET-1-IP or ET-1-IU-treated ewes did not differ (P ≥ 0.05), but were heavier (P ≤ 0.05) than in the Vehicle-IP or Vehicle-IU treatment groups which did not differ (P ≥ 0.05). Profiles of progesterone in jugular venous plasma of both control groups treated with Vehicle-IP or Vehicle-IU were lower (P ≤ 0.05) than in ewes treated with ET-1-IP or ET-1-IU, which did not differ (P ≥ 0.05) between ET-1-IP or ET-1-IU treatment groups. Treatment with ET-1-IP or ET-1-IU increased (P ≤ 0.05) the PGE:PGF₂α ratio when compared to the Vehicle-IP or Vehicle-IU treatment groups, which did not differ (P ≥ 0.05) between each other. In summary, ET-1 prevented the decrease in luteal weights and the decline in progesterone, but increased the PGE:PGF₂α ratio when compared to controls. Therefore, it is concluded that ET-1 is not luteolytic in ewes, but instead may be luteotropic or antiluteolytic by altering uterine secretion of the PGE:PGF₂α ratio, since PGE₁ or PGE₂ are luteotropic in vitro and in vivo, PGE₁ or PGE₂ prevent PGF₂α-induced luteolysis in vitro and in vivo, and PGE₁ and PGE₂ increase two-fold in ewe endometrium to prevent luteolysis during early pregnancy.     

Contraceptive efficacy of a novel intrauterine device (IUD) in white-tailed deer

Overabundant white-tailed deer (Odocoileus virginianus) pose risks to property, health, and safety of human beings. Public concerns about lethal management can impair efforts to address these issues, particularly in urban settings. Several techniques developed for reducing reproductive output of deer have limited utility because they require repeated dosing to achieve permanent effect and face uncertain regulatory approval for use beyond experimentation. From 10 August 2006 through 30 December 2007, we evaluated the contraceptive efficacy of copper-containing intrauterine devices (IUDs) implanted trans-cervically in white-tailed deer at the E.S. George Reserve in Pinckney, Michigan. Intrauterine devices were implanted before (n = 9) and shortly after (n = 10) the breeding season. Post-breeding season IUD treatment was in conjunction with a 5 cm³ dose of 5 mg/ml prostaglandin F_2α (PGF_2α), delivered subcutaneously. Intrauterine devices reduced pregnancy rates when administered prior to breeding (P < 0.001) and prevented pregnancy for up to 2 years (the duration of the study). Two of 8 does that received IUDs prior to the breeding season and survived to the end of the study became pregnant (due to loss of the implant) during the second year while all (n = 16) does without implants conceived. Cervical changes associated with early pregnancy made trans-cervical implantation after the breeding season challenging, and resulted in improperly placed IUDs in 2 treated does. The apparent expulsion of IUDs by pregnant does that received the combined treatment after breeding suggests IUD treatment should be limited to the pre-breeding season. Intrauterine devices show potential as a tool for small-scale deer population management via non-steroidal reproductive inhibition.     

Feeding soybean meal increases the blood level of isoflavones and reduces the steroidogenic capacity in bovine corpora lutea,without affecting peripheral progesterone concentrations

Thirty-three Holstein-Friesian cows were followed from 14 days pre partum until the fourth ovulation post partum. Housing conditions and basic ration were identical for all animals. Concentrates were individually supplemented according to the daily milk production level, using two different types of protein rich concentrates: soybean meal and rapeseed meal. Soybean and rapeseed meal are known to be respectively high and low in isoflavones. Cows were randomly divided into three groups and blocked for parity. Group I (n = 11) was supplemented with soybean meal and acted as control group. Groups II (n = 11) and III (n = 11) were respectively supplemented with soybean and rapeseed meal and were subjected to a biopsy sampling of the corpus luteum at day 9 of the first three postpartal estrous cycles.Soybean meal supplementation to lactating dairy cows (1.72 kg on average) induced an increase in the blood concentration of equol, dihydrodaidzein, o-desmethylangolensin in both soy groups and resulted in a reduced area occupied by steroidogenic (P = 0.012) and endothelial cells (P = 0.0007) in the luteal biopsies. Blood concentrations of equol and glycitein were negatively correlated with the areas occupied by steroidogenic (r = −0.410 with P = 0.0002, respectively r = −0.351 with P = 0.008) and endothelial cells (r = −0.337 with P = 0.01, respectively r = −0.233 with P = 0.085) in the 3 first estrous cycles. The latter however did not affect the diestrous peripheral blood progesterone concentration.     

Active immunization against inhibin-based peptides to increase ovulation rate in non-prolific Malpura ewes

The aim of this study was to explore the possibility of increasing the ovulation rate of Malpura, a non-prolific tropical breed of sheep by immunization against inhibin-based peptide immunogens. Ewes were divided into three groups (n = 5 each) and actively immunized against the synthetic peptides from the α_C [bIα(1–29)-Tyr³⁰] or α_N [bI-₄₃-Tyr¹⁵²(153–167)Cys¹⁶⁸] area of the bovine inhibin α-subunit conjugated to ovalbumin or against ovalbumin (control). Each ewe received a primary immunization of 400 μg immunogen and 3 boosters, each of 200 μg immunogen at 4-week intervals. Estrus was synchronized using a double PGF₂α injection schedule and laparoscopy was performed after each estrus to determine the ovulation response. Immunization against both the peptides did not affect the interval from PGF treatment to the onset of estrus, the duration of estrus and the number of large unovulated follicles. In contrast to the complete absence of multiple ovulations in the controls, all the ewes immunized against α_C or α_N peptides showed multiple ovulations (range 2–7) in all the three estrous cycles evaluated, except for one ewe immunized against the α_N peptide, which exhibited multiple ovulations in only 1 out of the 3 estrous cycles. Compared to that of the controls (1.0 ± 0.9, 1.0 ± 0.0 and 0.6 ± 0.2, respectively), the mean ovulation rate was higher (P < 0.01) in the ewes immunized against the αC (4.8 ± 1.02, 5.0 ± 1.05 and 5.0 ± 0.45, respectively) or against αN (4.5 ± 1.19, 2.5 ± 0.87 and 2.7 ± 0.75, respectively, P < 0.05) peptide in estrous cycles numbers 1, 2 and 3. These results show that active immunization against inhibin-based peptide immunogens is effective in increasing ovulation rate in Malpura, a non-prolific breed of sheep and that it may be an alternative to conventional superovulation regimes.     

The DNA–DNA spacing in gemini surfactants–DOPE–DNA complexes

Gemini surfactants from the homologous series of alkane-α,ω-diyl-bis(dodecyldimethylammonium bromide) (CnCS12, number of spacer carbons n = 2 – 12) and dioleoylphosphatidylethanolamine (DOPE) were used for cationic liposome (CL) preparation. CLs condense highly polymerized DNA creating complexes. Small-angle X-ray diffraction identified them as condensed lamellar phase L_α^C in the studied range of molar ratios CnGS12/DOPE in the temperature range 20 – 60 °C. The DNA–DNA distance (d_DNA) is studied in dependence to CnGS12 spacer length and membrane surface charge density. The high membrane surface charge densities (CnGS12/DOPE = 0.35 and 0.4 mol/mol) lead to the linear dependence of d_DNA vs. n correlating with the interfacial area of the CnGS12 molecule.     

Effect of progesterone administration on the release of uterine prostaglandin F2α and ovarian oxytocin in the goat

The effects of intramuscular progesterone administration (20 mg·day⁻¹) on plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM-pulmonary metabolite of prostaglandin F_2α) and oxytocin were examined in seventeen goats after either bilateral ovariectomy, hysterectomy or during days 12–16 of the estrous cycle. Daily mean values of PGFM in animals treated with progesterone after ovariectomy were significantly greater (P<0.001) than in their corresponding controls on the last two treatment days (10 and 11); concentrations of oxytocin, however, remained at or near the limits of assay sensitivity. In hysterectomized goats PGFM concentrations remained extremely low and oxytocin release appeared steady rather than pulsatile. In the intact animals, undergoing luteolysis, daily mean concentrations of both PGFM and oxytocin were significantly greater (P<0.01) in progesterone-treated goats than in their oil-treated controls; furthermore, in the progesterone-treated goats, increases in PGFM concentrations, observed after the peaks of progesterone, were either coincident with or prior to pulses of oxytocin. These results demonstrate that uterine PGF_2α stimulates the pulsatile release of oxytocin from the ovary during luteolysis in the goat.     

Effect of time interval between prostaglandin F2α and GnRH treatments on occurrence of short estrous cycles in cyclic dairy heifers and cows

Prostaglandin F_2α (PGF_2α) and GnRH treatments, when administered 24 h apart during early diestrus, cause short estrous cycles in some dairy cows and heifers [J. Taponen, M. Kulcsar, T. Katila, L. Katai, G. Huszenicza, H. Rodriguez-Martinez, Short estrous cycles and estrous signs after premature ovulations induced with cloprostenol and gonadotropin-releasing hormone in cyclic dairy cows, Theriogenology 2002; 58, 1291-1302]. We investigated the effect of a time interval between PGF_2α and GnRH administration on the appearance of short cycles. Estrus was induced in heifers with dexcloprostenol. A second luteolysis was induced similarly on day 7 after ovulation, and either 0 (T0) or 24 h (T24) later an injection of GnRH (0.1 mg of gonadorelin) was administered. We monitored ovarian activity with progesterone analyses from blood plasma samples and with ultrasonography. Fourteen cases (12 in T0 and 2 in T24) were excluded due to either incomplete luteolysis (2 cases) or unresponsiveness to GnRH (10 in T0 and 2 in T24). Short estrous cycles (7 to 8 d) were detected in 11/11 and 8/17 heifers in groups T0 and T24, respectively, with a significant difference in the incidence of short cycles (P < 0.01). In Experiment 2, estrus was induced in cows on day 8 (D8, n = 18), 9 (D9, n = 5), or 10 (D10, n = 3) with cloprostenol and gonadorelin administered simultaneously. Daily milk samples were collected for progesterone analysis until subsequent estrus was detected and ovarian ultrasound examinations were performed. Eight cases had to be excluded due to unresponsiveness to GnRH, leaving 18 cases eligible for the study. Short estrous cycles (7-12 d) were detected in 14/18 cows. In conclusion, shortening the time interval between PGF_2α and GnRH treatments increased the incidence of short estrous cycles and appeared to increase the proportion of females unresponsive to GnRH treatment.     

Effect of intrauterine infusion of recombinant bovine interferon αI1 on luteal phase duration and oxytocin-induced release of 13,14-dihydro-15-keto-prostaglandin F2α in postpartum beef cows

The objective of this study was to determine if oxytocin-induced release of prostaglandin F₂α (PGF_2α; measured by the stable metabolite, 13,14-dihydro-15-keto-prostaglandin F₂α (PGFM)) was inhibited following intrauterine infusion of bovine interferon-α_I1 (rboIFNα_I1) into postpartum cows anticipated to have short estrous cycles following first ovulation postpartum. Cows expected to have short estrous cycles were assigned to receive twice daily intrauterine infusions of either placebo (SCP; n = 11) or 2 mg rboIFNα_I1 (SCIFN; n = 14) on Days 1–16 following hCG injection (2500 IU; day 0) on Days 30 or 31 postpartum. On Day 5 following hCG, each cow was injected with 100 IU oxytocin (i.v.) to induce the release of uterine PGF_2α (as measured by PGFM). Other treatment groups consisted of cows expected to have normal estrous cycle lengths following pretreatment with a 9 day norgestomet implant on Days 21 or 22 postpartum followed by hCG injection to induce ovulation. Cows expected to have normal estrous cycle lengths received twice daily intrauterine infusions of either placebo from Days 1 to 16 of the cycle and 100 IU oxytocin (i.v.) on Day 5 (NCPE; n = 11) or twice daily infusions of placebo (NCPL; n = 7) or rboIFNα_I1 (NCIFN; n = 10), from Day 13 post-hCG injection until luteolysis. Oxytocin was injected (100 IU; i.v.) into cows in the NCPL and NCIFN groups on Day 16. The calculated areas under the curve (arbitrary PGFM units) were: 164 ± 18 units, 96 ± 16 units, 93 ± 18 units, 137 ± 27 units and 53 ± 20 units for SCP, SCIFN, NCPE, NCPL and NCIFN, respectively (SCIFN < SCP; NCIFN < NCPL; P < 0.015). Mean luteal phase length was calculated as the number of days from injection of hCG until progesterone declined to below 0.5 ng ml⁻¹ and was: 6.7 ± 1.0 days, 10.5 ± 0.9 days, 12.0 ± 1.0 days, 18.0 ± 1.3 days and 20.7 ± 1.1 days for SCP, SCIFN, NCPE, NCPL and NCIFN, respectively (SCP < SCIFN = NCPE < NCPL = NCIFN; P < 0.01). In summary, luteal phase lengths were increased and oxytocin-induced release of PGFM was reduced by rboIFNα_I1 infusion in cows anticipated to have short luteal phases.     

Determinants of platelet activation in hypertensives with microalbuminuria

Microalbuminuria is a predictor of adverse outcome in hypertension. We evaluated in vivo platelet activation, by urinary 11-dehydrothromboxane (TX)B₂ and plasma P-selectin, in hypertensives with or without microalbuminuria, and its possible association with oxidative stress, by urinary 8-iso-prostaglandin (PG)F_2α and endothelial dysfunction. Sixty essential hypertensive patients, with (n = 30) or without (n = 30) microalbuminuria, and 30 controls were studied. Endothelial function was assessed by nitric oxide products, intercellular adhesion molecule (ICAM)-1, and asymmetric dimethylarginine (ADMA) levels. Urinary 11-dehydro-TXB₂ excretion was higher in microalbuminuric (median 805 pg/mg creatinine) compared to nonmicroalbuminuric patients or controls (414 and 291 pg/mg, respectively; P < 0.0001). Plasma P-selectin was significantly higher in patients with microalbuminuria (median 136 ng/ml) as compared to those without microalbuminuria or controls (85 and 65 ng/ml; P < 0.0001). Urinary 8-iso-PGF_2α excretion was also enhanced in microalbuminuric (median 279 pg/mg creatinine) compared to nonmicroalbuminuric patients or controls (157 and 146 pg/mg, respectively; P < 0.0001). A significant impairment in endothelial function was found in microalbuminuric patients, with decreased nitric oxide and increased ICAM-1 and ADMA levels. Multivariate regression analysis showed that urinary 8-iso-PGF_2α excretion (beta = 0.49; P < 0.0001) and microalbuminuria (beta = 0.36; P < 0.001) were independently related to 11-dehydro-TXB₂ in hypertensives. Vitamin E supplementation (900 mg daily for 1 month) in 10 hypertensives with microalbuminuria was associated with normalization in median 11-dehydro-TXB₂ and 8-iso-PGF_2α. We conclude that lipid peroxidation is a major determinant of persistent platelet activation in hypertensive patients with microalbuminuria.     

Relation between leptin and estradiol levels in Egyptian lactating Arab mares during foaling heat

Sixteen Arab lactating mares belonging to Al-Zahraa Arab Horse Stud underwent two ultrasound examinations at 3 weeks interval starting from the day of demonstration of foaling heat. In addition, daily blood samples were collected from parturition until after exhibiting first postpartum estrus (day 11) with daily observation of estrous signs. Both leptin and estradiol hormones were assayed. Mean day of foaling heat was 8.9 ± 0.9 day. Most mares came in foaling heat during days 9 and 10 had high conception rate compared to those who came in estrus earlier or later. Estradiol levels were high after day of foaling then decrease after expression of foaling heat. But leptin levels increase from day 8 to day 10 compared to other days before and after the first ovulation. A significant positive correlation was found between estradiol and leptin (r = 0.58, p < 0.025). The positive correlation between leptin and estradiol led us to suggest that leptin hormone plays an important role in ovulation of the first postpartum estrus in mares.     

Synthesis and monoamine transporter affinity of 3α-arylmethoxy-3β-arylnortropanes

A series of 3-arylnortrop-2-enes and 3α-arylmethoxy-3β-arylnortropanes were synthesized and evaluated for binding affinity at monoamine transporters. The 3-(3,4-dichlorophenyl)nortrop-2-ene (6e) exhibited high affinity for the SERT (K_i = 0.3 nM). The 3α-arylmethoxy-3β-arylnortropanes were generally SERT selective with the 3α-(3.4-dichlorophenylmethoxy)-3βphenylnortrop-2-ene (7c) possessing subnanomolar potency (K_i = 0.061 nM). However, 3α-(3,4-dichlorophenylmethoxy)-3β-phenylnortrop-2-ene (7b) exhibited high affinity at all three transporters [(DAT K_i = 22 nM), (SERT K_i = 6 nM) and (NET K_i = 101 nM)].     

Opposite changes in cannabinoid CB1 and CB2 receptor expression in human gliomas

Gliomas are the most important group of malignant primary brain tumors and one of the most aggressive forms of cancer. During the last years, several studies have demonstrated that cannabinoids induce apoptosis of glioma cells and inhibit angiogenesis of gliomas in vivo. As the effects of cannabinoids rely on CB₁ and CB₂ receptors activation, the aim of the present study was to investigate both receptors protein expression in cellular membrane homogenates of human glial tumors using specific antibodies raised against these proteins. Additionally, we studied the functionality of the cannabinoid receptors in glioblastomas by using WIN 55,212-2 stimulated [³⁵S]GTPγS binding.Western blot analysis showed that CB₁ receptor immunoreactivity was significantly lower in glioblastoma multiforme (?43%, n = 10; p < 0.05) than in normal post-mortem brain tissue (n = 16). No significant differences were found for astrocytoma (n = 6) and meningioma (n = 8) samples. Conversely, CB₂ receptor immunoreactivity was significantly greater in membranes of glioblastoma multiforme (765%, n = 9; p < 0.05) and astrocytoma (471%, n = 4; p < 0.05) than in control brain tissue (n = 10). Finally, the maximal stimulation of [³⁵S]GTPγS binding by WIN 55,212-2 was significantly lower in glioblastomas (134 ± 4%) than in control membranes (183 ± 2%; p < 0.05). The basal [³⁵S]GTPγS binding and the EC₅₀ values were not significantly different between both groups.The present results demonstrate opposite changes in CB₁ and CB₂ receptor protein expression in human gliomas. These changes may be of interest for further research about the therapeutic effects of cannabinoids in glial tumors.