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.     

Role of PGF2α in luteolysis based on inhibition of PGF2α synthesis in the mare

The effects of inhibition of PGF2α synthesis on luteolysis in mares and on the incidence of prolonged luteal activity were studied in controls and in a group treated with flunixin meglumine (FM), a PGF2α inhibitor (n = 6/group). The FM was given every 8 hours (1.0 mg/kg) on each of Days 14.0 to 16.7. Concentration (pg/mL) of PGF2α metabolite averaged over 8 hours of hourly blood sampling at the beginning of each day, was lower in the FM group than in the controls on Day 14 after ovulation (6.7 ± 1.3 vs. 13.8 ± 2.9, P < 0.05), Day 15 (15.0 ± 3.9 vs. 35.2 ± 10.4, P < 0.10), and Day 16 (21.9 ± 5.7 vs. 54.7 ± 11.4, P < 0.03). Concentration (ng/mL) of progesterone (P4) was greater in the FM group than in the controls on Day 14 (10.1 ± 0.9 vs. 7.7 ± 0.9, P < 0.08), Day 15 (9.2 ± 1.0 vs. 4.3 ± 1.0, P < 0.008), and Day 16 (5.6 ± 1.6 vs. 1.2 ± 0.4, P < 0.02). The interval from ovulation to the beginning of a decrease in P4 and to the end of luteolysis (P4 < 1 ng/mL) was each delayed (P < 0.03) by ∼1 day in the FM group. Intervals involving the luteal phase were long (statistical outliers, P < 0.05) in two mares in the FM group, indicating prolonged luteal activity. Results supported the hypotheses that (1) inhibition of PGF2α synthesis interferes with luteolysis in mares and (2) inhibition of PGF2α at the expected time of luteolysis may lead to prolonged luteal activity.     

Ovarian and PGF2α responses to stimulation of endogenous PRL pulses during the estrous cycle in mares

The effects of a PRL-stimulating substance (sulpiride) on PRL and PGF2α secretion and on luteal and ovarian follicular dynamics were studied during the estrous cycle in mares. A control group (n = 9) and a sulpiride group (Sp; n = 10) were used. Sulpiride (25 mg) was given every 8 h from Day 13 postovulation to the next ovulation. Repeated sulpiride treatment did not appear to maintain PRL concentrations at 12-h intervals beyond Day 14. Therefore, the hypothesis that a long-term increase in PRL altered luteal and follicular end points was not testable. Hourly samples were collected from the hour of a treatment (Hour 0) to Hour 8 on Day 14. Concentrations of PRL increased to maximum at Hour 4 in the Sp group. The PRL pulses were more prominent (P < 0.008) in the sulpiride group (peak, 19.4 ± 1.9 ng/mL; mean ± SEM) than in the controls (11.5 ± 1.8 ng/mL). Concentrations of a metabolite of PGF2α (PGFM), number, and characteristics of PGFM pulses, and concentrations of progesterone during Hours 0 to 8 were not affected by the increased PRL. A novel observation was that the peak of a PRL pulse occurred at the same hour or 1 h later than the peak of a PGFM pulse in 8 of 8 PGFM pulses in the controls and in 6 of 10 pulses in the Sp group (P < 0.04), indicating that sulpiride interfered with the synchrony between PGFM and PRL pulses. The hypothesis that sulpiride treatment during the equine estrous cycle increases concentrations of PRL and the prominence of PRL pulses was supported.     

Temporal relationships of a pulse of prolactin (PRL) to a pulse of a metabolite of PGF2α in mares

Hourly blood samples were collected from 10 mares during 24 h of each of the preluteolytic, luteolytic, and postluteolytic periods. The autocorrelation function of the R program was used to detect pulse rhythmicity, and the intra-assay CV was used to locate and characterize pulses of prolactin (PRL) and a metabolite of prostaglandin F2α (PGFM). Rhythmicity of PRL and PGFM concentrations was detected in 67% and 89% of mares, respectively. Combined for the three periods (no difference among periods), the PRL pulses were 5.2 ± 0.4 h (mean ± SEM) at the base, 7.5 ± 1.5 h between nadirs of adjacent pulses, and 12.3 ± 1.5 h from peak to peak. The peaks of PRL pulses were greater (P < 0.05) during the luteolytic period (46 ± 14 ng/mL) and postluteolytic period (52 ± 15 ng/mL) than during the preluteolytic period (17 ± 3 ng/mL). Concentrations of PRL during hours of a PGFM pulse were different (P < 0.003) within the luteolytic period and postluteolytic period and were greatest at the PGFM peak; PRL concentrations during a PGFM pulse were not different during the preluteolytic period. The frequency of the peak of PRL and PGFM pulses occurring at the same hour (synchrony) was greater for the luteolytic period (65%, P < 0.01) and postluteolytic period (50%, P < 0.001) than for the preluteolytic period (17%). This is the first report in mares on characterization and rhythmicity of PRL pulses, synchrony between PRL and PGFM pulses, and greater PRL activity during the luteolytic and postluteolytic periods than during the preluteolytic period.     

Metabolites of PGF2α in Blood Plasma and Urine as Parameters of PGF2α Release in Cattle

The metabolism of PGF2α in cattle results initially in the formation of 15-keto-13,14-dihydro-PGF2α (15-ketodihydro-PGF2α) and later the 11-ketotetranor PGF metabolites. Both types of metabolites appear in the peripheral circulation and finally the 11-ketotetranor PGF metabolites are found in large quantities in the urine in a species-related pattern. Several approaches can be made to the quantitative analysis of PGF2α release during reproductive studies. First, assay of the 15-ketodihydro-PGF2α metabolite in the peripheral circulation; second, analysis of the longer-lived 11-ketotetranor PGF metabolites in the peripheral circulation; and finally analysis of the latter metabolites in the urine. The antibodies used in radioimmunoassays of both types of metabolites of PGF2α were found to be specific and the results agree well with those obtained earlier by mass spectrometric analysis. The assay of 11-ketotetranor PGF metabolites was used to study the excretion of urinary metabolites in the cow after i.v. infusion of PGF2α and also during the normal estrous cycle and early pregnancy. These studies suggest that 11-ketotetranor PGF metabolites in cow urine serve as a good parameter of PGF2α release, especially for long–term studies, but when a precise pattern of PGF2α release is required, measurement of 15-ketodihydro-PGF2α levels in frequently collected plasma samples is preferable.     

Concentrations of circulating hormones during the interval between pulses of a PGF2α metabolite in mares and heifers

The temporal relationship of several hormones to a metabolite of prostaglandin F2α (PGFM) was studied in mares and heifers from the beginning of the first PGFM pulse during luteolysis to the end of the second pulse. Mares (n=7) were selected with a 9-h interval between the peaks of the two pulses. In mares, estradiol-17β (estradiol) increased (P<0.05) within each PGFM pulse and plateaued for a mean of 6h between the pulses, resulting in a stepwise estradiol increase. Progesterone decreased linearly (P<0.0001) throughout the intra-pulse and inter-pulse intervals of PGFM. In heifers (n=6), inter-pulse intervals were variable, and therefore Hours 1-4 of the first pulse (Hour 0=PGFM peak) and Hours -4 to -1 of the second pulse were used to represent the mean 8-h interval between peaks of the two pulses. Estradiol increased (P<0.05) during the ascending portion of each PGFM pulse and then decreased (P<0.05) beginning at Hour -1 of the first PGFM pulse and Hour 0 of the second pulse. The 1-h delay during the second pulse was accompanied by an apparent increase in PRL. A transient decrease in estradiol occurred in individuals between PGFM pulses at a mean of 5h after the first PGFM peak, concomitant with a transient LH increase (P<0.05). Results indicated that estradiol plateaued in mares and fluctuated in heifers during the interval between PGFM pulses. Heifers also showed temporal relationships between estradiol and LH and apparently between estradiol and PRL.     

Effect of uterine luminal perfusion of PGF2α and PGE2 on uterine vasomotor response and PGF2α output in cyclic cattle

Six cyclic Holstein dairy cows were anesthetized on days 12–14 post-oestrus. Reproductive tract was exposed by midventral incision, and the ovarian (utero-ovarian) vein and facial artery cannulated. Oviduct was ligated, and a catheter (affluent) introduced into the tip of the uterine horn. The uterine horn was ligated above the uterine body, a second catheter (effluent) introduced into the uterine lumen, and an electromagnetic blood flow transducer placed around the uterine artery. On the day following surgery, the uterine horn was infused constantly for 9 h with PGF_2α dissolved in PBS (0.7 ml/min, 177 ng/ml). During periods 1 and 3 (first 3 h and last 3 h, respectively) only PGF_2α was perfused; during period 2 (between 3 h and 6 h) 101tgμg/ml of PGE₂ were added to the perfusate together with PGF_2α. Uterine venous and peripheral blood samples were collected simultaneously every 15 min, and uterine blood flow recorded continuously. Least-square means for PGF measured in uterine venous drainage for periods 1, 2 and 3 were 315 ± 26, 557 ± 24 and 511 ± 26 pg/ml, respectively (P < 0.05). Uterine blood flow values were 52 ± 5, 67 ± 4 and 61 ± 4 ml/min for periods 1, 2 and 3 (P < 0.08), respectively.Results do not support the hypothesis that the antiluteolytic effect of PGE₂ is associated with a suppression of uterine PGF_2α release into the circulation. Greater release of PGF to the circulation in period 2 (addition of PGE₂) is probably the result of the vasodilatory effect of PGE₂ on uterine endometrial vasculature.     

Physiologic and nonphysiologic effects of exogenous prostaglandin F2α on reproductive hormones in mares

Responses to intravenous treatment of mares with prostaglandin F2α (PGF) 8 d after ovulation were studied in three groups (n = 4/group): control (no treatment), bolus (single treatment with 2.5 mg PGF), and infusion (0.1 mg PGF during 2 h). Infusion resulted in a 13,14-dihydro-15-keto-PGF2α (PGFM) concentration (559 ± 44 pg/mL) that was not different from the mean concentration for the major portion of a natural PGFM pulse associated with luteolysis (569 ± 45 pg/mL; n = 5). Progesterone in the bolus group increased (P < 0.03) between 0 (17.8 ± 3.5 ng/mL) and 2 min (25.3 ± 4.8 ng/mL), peaked at 10 min (28.5 ± 4.6 ng/mL), and then decreased. In the infusion group, progesterone decreased (P < 0.05) during 1 min (17.2 ± 1.3 ng/mL) to 15 min (13.5 ± 1.5 ng/mL) after the beginning of infusion and decreased (P < 0.05) similarly to the bolus group during 2 to 12 h; concentrations were lower (P < 0.05) at each hour than in controls. Interval between ovulations was shorter (P < 0.05) in the bolus (19.3 ± 2.0 d) and infusion (18.8 ± 2.1 d) groups than in controls (24.3 ± 1.3 d). Concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and cortisol increased (P < 0.05) within 10 min in the bolus group but did not change in the infusion group. Results supported the hypothesis that increases in progesterone, FSH, LH, and cortisol after a bolus luteolytic PGF treatment are nonphysiologic. Past conclusions on the nature of the luteolytic mechanism are problematic if based on responses to treatment with a single luteolytic bolus of PGF.     

Antiluteogenic effects of serial prostaglandin F2α administration in cycling mares

A single dose of PGF2α does not consistently induce luteolysis in the equine CL until at least 5 days after ovulation, leading to the erroneous assumption that the early CL is refractory to the luteolytic effects of PGF2α. We hypothesized that serial administration of PGF2α in early diestrus would induce a return to estrus similar to mares treated with a single injection in mid-diestrus, and fertility of the induced estrus would not differ. The objectives of the study were to evaluate the effects of the 2 approaches as reflected by: (1) concentrations of plasma progesterone; (2) interovulatory and treatment-to-ovulation intervals; (3) the proportion of mares pregnant after artificial insemination. The study consisted of a balanced crossover design in which 10 reproductively normal Quarter Horse Mares were exposed to 2 treatments on 2 consecutive reproductive cycles. At detected ovulation (Day 0), mares were randomly allotted to 1 of 2 treatment groups: I, mid-diestrus treatment, administration of a single 10-mg dose of dinoprost tromethamine (PGF2α) im on Day 10; II, early diestrus treatment, administration of 10-mg PGF2α im twice daily on Days 0, 1, and 2 and once daily on Days 3 and 4. Mares in estrus and with a follicle 35 mm or greater in diameter were artificially inseminated with at least 2 billion motile sperm from a fertile stallion. Pregnancy was defined as detection of a growing embryonic vesicle on 2 consecutive examinations approximately 14 days after ovulation. Serial plasma samples were collected throughout the study period, and concentration of plasma progesterone was determined by RIA. A mixed-model ANOVA for repeated measures was used to analyze hormonal data. Interovulatory and treatment-to-ovulation intervals were compared by a paired t test and fertility by a McNemar chi-square analysis. All mares in group I underwent luteolysis after PGF2α administration denoted by mean (±SD) concentration of plasma progesterone of 0.25 ± 0.21 ng/mL detected 2 days after treatment. In group II, mean concentration of plasma progesterone remained below 1.0 ng/mL during treatment and until the onset of the next estrus. The mean interovulatory interval in group I was 18.5 ± 2.0 days compared with 13.1 ± 3.7 days in group II (P < 0.01). Treatment-to-ovulation intervals were 8.5 ± 2.0 days and 13.1 ± 3.7 days for groups I and II, respectively (P < 0.05). In both groups, 9 of 10 mares were pregnant (P = 1.0). Serial PGF2α administration beginning at ovulation consistently prevented luteal function in 10 of 10 mares in the present study without adversely affecting pregnancy rate of post-treatment cycles.     

Combination of the ram effect with PGF2α estrous synchronization treatments in ewes during the breeding season

The role of the ram effect on the reproductive performance of ewes that have initiated estrous cycles following lambing in combination with synchronization of estrus using PGF(2α) was examined. A total of 1264 Corriedale × Merino ewes in the breeding season (March-April) were allocated to one of three treatments. The control group (PG2) of ewes (n=415) were in permanent direct contact with vasectomized rams throughout the experiment from 60 d prior to the administration of the first luteolytic dose of PGF(2α) which was followed by a second dose 13 d later (Day 0 of the experiment). Ewes assigned to the other two treatments remained isolated from rams until Day 0. In the second treatment, ewes (PG2RE; n=445) were administered PGF(2α) in the same manner and were joined with vasectomized rams at Day 0. Ewes allocated to the third treatment (PGRE; n=404) did not receive the second dose of PGF(2α) but were introduced to vasectomized rams on Day 0. Sexual receptivity, as indicated by tail-head marking, was recorded until d 11. More PG2RE ewes (407/445; 92%) were observed in estrus by Day 11 than occurred for PG2 ewes (353/415; 85%; P=0.003). The accumulated frequency of PG2RE ewes in estrus was greater than for PG2 ewes for each period from Day 3 (P<0.001) to Day 11 (P<0.01). The onset of estrus was earlier in PG2RE ewes (2.98±0.07 d) than for PG2 ewes (3.31±0.07 d; P<0.0001). In contrast, the total frequency of PGRE ewes observed in estrus by Day 11 (356/404; 88%) was similar to that observed for PG2 ewes. However, the trajectory of the accumulated frequency of the incidence of estrus was less for the PGRE ewes initially, particularly during the period of Days 3-6 of observation (P<0.0001). Consequently, onset of estrus was earlier in PG2 ewes (3.31±0.07 d) than for PGRE ewes (5.30±0.11 d; P<0.0001). It was concluded that the introduction of vasectomized rams simultaneously with the second administration of PGF(2α) advanced the onset of estrus and increased the number of ewes that responded. The introduction of rams 13 d after a single dose of PGF(2α) did not substitute for the second administration of PGF(2α).     

Stimulatory effect of α-synuclein on the tau-phosphorylation by GSK-3β

Hyperphosphorylation of tau protein (tau) causes neurodegenerative diseases such as Alzheimer's disease (AD). Recent studies of the physiological correlation between tau and α-synuclein (α-SN) have demonstrated that: (a) phosphorylated tau is also present in Lewy bodies, which are cytoplasmic inclusions formed by abnormal aggregation of α-SN; and (b) the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) increases the phosphorylation of tau as well as the protein level of α-SN in cultured neuronal cells, and also in mice. However, the molecular mechanism responsible for the α-SN-mediated hyperphosphorylation of tau remains to be elucidated. In this in vitro study, we found that: (a) α-SN directly stimulates the phosphorylation of tau by glycogen synthase kinase-3β (GSK-3β), (b) α-SN forms a heterotrimeric complex with tau and GSK-3β, and (c) the nonamyloid beta component (NAC) domain and an acidic region of α-SN are responsible for the stimulation of GSK-3β-mediated tau phosphorylation. Thus, it is concluded that α-SN functions as a connecting mediator for tau and GSK-3β, resulting in GSK-3β-mediated tau phosphorylation. Because the expression of α-SN is promoted by oxidative stress, the accumulation of α-SN induced by such stress may directly induce the hyperphosphorylation of tau by GSK-3β. Furthermore, we found that heat shock protein 70 (Hsp70) suppresses the α-SN-induced phosphorylation of tau by GSK-3β through its direct binding to α-SN, suggesting that Hsp70 acts as a physiological suppressor of α-SN-mediated tau hyperphosphorylation. These results suggest that the cellular level of Hsp70 may be a novel therapeutic target to counteract α-SN-mediated tau phosphorylation in the initial stage of neurodegenerative disease.     

Effects of estriol on PGF2α output by cultures of human endometrium and endometrial cells

We have shown that the rate of release of PGF_2α by monolayer cultures of epithelial cells from proliferative endometrium is markedly elevated by addition of estradiol to the medium. In cultures maintained in HAM F-10 medium containing charcoal stripped calf serum, estradiol (10⁻⁸M) increased the levels of PGF_2α several fold during the second and third days in culture. Similar responses were obtained with estradiol at 10⁻¹⁰M concentration.When this system was used to compare the effects of estradiol and estriol at equal concentrations (10⁻⁸M), similar elevation (10–16-fold) of PGF_2α levels were noted during 3 consecutive days in culture. When cultures of epithelial cells derived from secretory endometrium were used for these tests, estriol was as effective as than estradiol in elevating PGF_2α levels in the medium.When the effects of estradiol and estriol were compared using fragments of secretory endometrium in organ culture, the increases in PGF_2α levels noted in the medium were about equal (2- to 10-fold) for the two estrogens at the same concentration (10⁻⁹−10⁻⁸M).Exposure of the tissue to either estradiol or estriol for only 1 h resulted in increases in PGF_2α output for the following 3 days.These results clearly show that estriol is as effective as estradiol in stimulating PGF_2α output by human endometrial tissue.     

Inhibitory effect of novel somatostatin peptide analogues on human cancer cell growth based on the selective inhibition of DNA polymerase β

The present study was designed to investigate the anticancer activity of novel nine small peptides (compounds 1–9) derived from TT-232, a somatostatin structural analogue, by analyzing the inhibition of mammalian DNA polymerase (pol) and human cancer cell growth. Among the compounds tested, compounds 3 [tert-butyloxycarbonyl (Boc)-Tyr-Phe-1-naphthylamide], 4 (Boc-Tyr-Ile-1-naphthylamide), 5 (Boc-Tyr-Leu-1-naphthylamide) and 6 (Boc-Tyr-Val-1-naphthylamide) containing tyrosine (Tyr) but no carboxyl groups, selectively inhibited the activity of rat pol β, which is a DNA repair-related pol. Compounds 3–6 strongly inhibited the growth of human colon carcinoma HCT116 p53^+/+ cells. The influence of compounds 1–9 on HCT116 p53^?/? cell growth was similar to that observed for HCT116 p53^+/+ cells. These results suggest that the cancer cell growth suppression induced by these compounds might be related to their inhibition of pol. Compound 4 was the strongest inhibitor of pol β and cancer cell growth among the nine compounds tested. This compound specifically inhibited rat pol β activity, but had no effect on the other 10 mammalian pols investigated. Compound 4 combined with methyl methane sulfonate (MMS) treatment synergistically suppressed HCT116 p53^?/? cell growth compared with MMS alone. This compound also induced apoptosis in HCT116 cells with or without p53. From these results, the influence of compound 4, a specific pol β inhibitor, on the relationship between DNA repair and cancer cell growth is discussed.     

Effect of gonadotropin-releasing hormone and prostaglandin F2α on reproduction in postpartum dairy cows

Thirty dairy cows serving as the treated group (Group A) were injected intramuscularly with 100 mcg gonadotropin-releasing hormone (GnRH) at 10 to 16 days postpartum followed by 25 mg prostaglandin F₂α (PGF₂α) 14 days later. Twenty-nine herdmate dairy cows (Group B) serving as controls were treated in a similar manner using saline injections rather than GnRH or PGF₂α treatments. Only cows without obvious uterine infection were assigned to the experimental groups, and any uterine pathology that developed during the treatment interval was treated accordingly following the experimental period. Internal genitalia were evaluated via rectal palpation prior to each injection. Blood samples were collected for progesterone analysis before each injection and at 30 hours following the PGF₂α or the second saline injection. Experimental animals were artificially inseminated at the first detected postpartum estrus starting 35 to 40 days following calving. Results indicated evidence of enhanced cyclicity when Group A cows were compared with those in Group B. However, there were no significant differences between the two groups for interval to first observed estrus, interval to first serive, first serive pregnancy rate, services per pregnancy and days open. Furthermore, no difference in the incidence of follicular or luteal cysts, incidence of repeat breeders or number of reproductive culls was observed. From observations in this study, the GnRH and PGF₂α treatment scheudule might not be economically beneficial in lactating dairy cows as long as reproductive tract abnormalities are promptly diagnosed and subsequently treated by the attending practitioner.     

Structure-activity relationships of bergenin derivatives effect on α-glucosidase inhibition

The α-glucosidase inhibitory activities of bergenin derivatives were evaluated. Bergenin derivatives were synthesized from bergenin which is a characteristic compound of B. ligulata. A new bergenin derivative, 11-O-(3′,4′-dimethoxybenzoyl)-bergenin showed the highest potent inhibitory activity among those of bergenin derivatives. The presence of substituents at 3′,4′-position in bergenin derivatives altered the α-glucosidase inhibitory activity. 11-O-(3′,4′-dimethoxybenzoyl)-bergenin was noncompetitive inhibitor for α-glucosidase. The present study reveals that bergenin derivatives could be classified as a new group of α-glucosidase inhibitors.     

Effect of early postpartum PGF2α treatment on reproductive performance in dairy cows with calving and puerperal traits

The objective of this study was to evaluate the effects of early postpartum PGF two alpha treatment on reproductive performance in dairy cows with calving and puerperal traits. A total of 363 Holstein cows (128 primiparous and 235 multiparous) were selected based on the presence of at least one of calving and puerperal traits (dystocia, retained placenta, twin, abortion, and postpartum uterine infections) and were assigned to two groups (treatment and control) irrespective of presence or absence of luteal tissue. Cows in the treatment group were treated twice with 25 mg dinoprost 8 h apart on day 20 postpartum, and for the control group saline placebo was administered. As it was speculated that the timing of a second dose would mimic the release of endogenous PGF2α from the uterus, our hypothesis was that two doses of PGF2α 8 h apart may increase the duration of elevated plasma prostaglandin F2α metabolite concentration in these cows. Recorded reproductive variables included days to first estrus, days to first AI, first service conception rate, pregnancy by 150 days in milk, service per conception, open days, and the percentage of repeat breeder animals. The data were analyzed using SPSS (Version 15) (IBM North America, New York, NY, USA) and Minitab (Version 14) (Minitab, State College, PA, USA). Although early postpartum PGF2α treatment had no effect on days to first estrus (36.7 days vs. 34.9 days, P = 0.056) and days to first AI (70.5 days vs. 72.2 days, P = 0.537), it increased first service conception rate (47.1% vs. 27.6%, P < 0.001); and this was more remarkable in primiparous cows (64.7% vs. 25%, P < 0.001). PGF2α treatment reduced the mean service per conception (1.92 vs. 2.72, P < 0.001) and the mean open days (112 days vs. 144 days, P < 0.001), and increased pregnancy by 150 days in milk (DIM) (80% vs. 66%, P = 0.004). The prevalence of repeat breeder syndrome in cows with calving and puerperal traits was reduced by PGF2α treatment (10% vs. 29.8%, P < 0.001). In conclusion, treatment of cows with calving and puerperal traits twice with a luteolytic dose of PGF2α 8 h apart on Day 20 postpartum improved reproductive performance and reduced the prevalence of repeat breeder syndrome.     

Stimulatory effect of 17β-estradiol on osteogenic differentiation potential of rat adipose tissue-derived stem cells

Adipose tissue-derived stem cells (ADSCs) are considered as a potential cell source for regenerative medicine and tissue engineering. Although ADSCs have greater proliferation capacity than bone marrow stem cells (BMSCs), lower differentiation ability of these cells limits their utility in experimental and clinical studies. The purpose of this study was to investigate whether 17β-estradiol (E(2)) has a stimulatory effect on osteogenic differentiation potential of ADSCs in vitro. ADSCs were isolated from visceral adipose tissues of rats and treated with different concentrations of E(2) in osteogenic medium (OM) for 21 days. The differences in osteogenic differentiation potential of the cultures were assessed by von Kossa staining, measurement of alkaline phosphatase (ALP) activity and calcium levels. ADSCs cultured in OM supplemented with E(2) showed greater bone-like nodule formation and mineral deposition in comparing with the cells grown in OM. In addition, ALP activity and calcium levels also were significantly higher in the cultures exposed to E(2) than the cells treated only with OM (p < 0.005, n = 5). Our results suggest that E(2) may stimulate the osteogenic differentiation of ADSCs and therefore, can be used as an inducing agent to improve the efficiency of these cells in in vitro and in vivo studies.     

Effects of inhibition of prostaglandin F2α biosynthesis during preluteolysis and luteolysis in heifers

Flunixin meglumine (FM; 2.5 mg/kg) was given to heifers at three 8-h intervals, 16 d after ovulation (first treatment = Hour 0) to inhibit the synthesis of prostaglandin F_2α (PGF), based on plasma concentrations of a PGF metabolite (PGFM). Blood samples were collected at 8-h intervals from 15 to 18 d in a vehicle (control) and FM group (n = 16/group). Hourly samples were collected from Hours −2 to 28 in 10 heifers in each group. Heifers that were in preluteolysis or luteolysis at Hour 0 based on plasma progesterone (P4) concentrations at 8-h intervals were partitioned into subgroups. Concentration of PGFM was reduced (P < 0.05) by FM treatment in each subgroup. For the preluteolytic subgroup, the first decrease (P < 0.05) in P4 concentration after Hour 0 occurred at Hours 24 and 40 in the vehicle and FM groups, respectively. Plasma P4 concentrations 32 and 40 h after the beginning of luteolysis in the luteolytic subgroup were greater (P < 0.05) in the FM group. Concentration at the peak of a PGFM pulse in the FM group was greater (P < 0.05) in the luteolytic than in the preluteolytic subgroup. The peak of a PGFM pulse occurred more frequently (P < 0.001) at the same hour as the peak of an LH fluctuation than at the ending nadir of an LH fluctuation. In conclusion, a reduction in prominence of PGFM pulses during luteolysis delayed completion of luteolysis, and treatment with FM inhibited PGFM production more during preluteolysis than during luteolysis.