Prostaglandin F2 Alpha Analogue Research Articles

Temporada corta de reprodución controlada com ovejas Santa Inês criadas em clima tropical húmedo

The objective of this work was to evaluate the adoption of management techniques that could allow for a shorter controlled breeding season. In three consecutive mating seasons (MS1: September/2021, n=45; MS2: June/2022, n=42 and MS3: March/2023, n=43), lactating Santa Ines ewes, multiparous, aged between two and four years, mean live weight of 62.20 ± 4.60 kg and mean body condition score of 3.4 ± 0.4 (scale 1-5), two vasectomized males and two breeding rams with prior andrological evaluation, one Santa Ines and one Dorper. A pre-mating period was implemented with biostimulation using vasectomized males for 21 days. After this period, on day 0 (D0), an IM injection of 120.5µg of cloprostenol, a prostaglandin F2alpha analogue (PGF2α), was performed and estrus was observed until D3; ewes with identified estrus were mated by the breeding rams, as per controlled breeding. Ewes that did not show estrus were submitted to a second IM injection of PGF2α seven days after the first dose (D7), then repeating the management of estrus observation and controlled mating. Ewes were allowed three attempts at conception. Pregnancy diagnosis was performed by ultrasonography (D30). During biostimulation, 100% of the ewes showed estrus in the three mating seasons. There was no significant difference in performance between seasons (P>0.05). From the total of ewes in season (n= 130), 90.0% (n=117) became pregnant, 71.5% (n=93) in the first, 13.9% (n=18) in the second and 4.6% (n= 6) in the third mating. Prior biostimulation was efficient in inducing estrus to lactating Santa Ines ewes and the use of PGF2α was sufficient to synchronize estrus and generate a shorter controlled breeding season with up to three matings, presenting an efficient method to concentrate births and standardize allotments of lambs in a production system under humid tropical climate.

GnRH34 with or without estradiol cypionate in timed AI in Bos indicus beef cows

Two experiments were performed to evaluate the effects of GnRH treatment on the fertility of suckled Nelore beef cows treated with an estradiol/progesterone (E2/P4)-based protocol for timed artificial insemination (TAI). Experiment 1 focused on determining the effects of estradiol cypionate (EC) on ovulation in TAI cows treated with GnRH 34 h after removal of the intravaginal P4 device (IPD). Suckled cows (n = 26) were treated with 2 mg estradiol benzoate (EB) and IPD containing 1 g P4. After 8 days, IPDs were removed, and all cows were treated with 150 μg of d-cloprostenol (prostaglandin F2 alpha analog) and 300 IU of equine chorionic gonadotropin (eCG), then separated into two treatment groups consisting of cows who received 1) saline 0.9% i.m. (GnRH34 group) or 2) 0.6 mg i.m. of EC (EC-GnRH34 group). On day 9 (05:00 p.m.), all cows were given GnRH (10.5 μg of buserelin acetate) i.m. No differences were observed between the groups (P > 0.05) in the time of ovulation after IPD removal or in the proportion of cows ovulating. Experiment 2 focused on determining the effects of GnRH34 along with or in the absence of EC on day 8 on pregnancy per AI (P/AI) in postpartum beef cows. Cows (n = 981) were treated similarly to those in Experiment 1, but an additional group, the EC-GnRH48 group, was included, in which cows received EC on day 8 whereas those that did not show estrus received GnRH at TAI. Thus, in this experiment, groups consisted of GnRH34 (n = 322), EC-GnRH34 (n = 335), and EC-GnRH48 (n = 324). A higher rate of estrus expression was observed in cows treated with EC following IPD removal (EC-GnRH34: 69%, EC-GnRH48: 64.8%) than in cows in the GnRH34 group (45.6%). No difference in P/AI was observed between the treatment groups (P = 0.45), but P/AI in cows in the EC-GnRH34 group (64.2%) tended to be greater (P = 0.1) than in cows in the GnRH34 group (58%). In summary, although ovulation synchrony did not differ among the groups, P/AI in cows treated with EC and GnRH 34 h after IPD removal tended to be greater than in cows treated solely with GnRH; this was most likely due to a shorter proestrus/estrus period, considering the lower proportion of cows that displayed estrus in the GnRH34 group. Finally, given that P/AI did not differ between the EC-GnRH34 and EC-GnRH48 groups, our results suggest that, for cows not displaying estrus, administration of EC at the time of IPD removal followed by treatment with GnRH 48 h afterward represents the most cost-efficient TAI strategy for South American Zebu-based beef operations.

Regulatory changes of local produced prostaglandins in corpus luteum after experimentally induced luteolysis in the cow.

The objective of the study was to evaluate the expression patterns of prostaglandin F2alpha (PGF), prostaglandin E2 (PGE), PGF receptor (FP), PGE receptors (EP2 and EP4), prostaglandin-endoperoxide synthase 2 (PTGS2) and prostaglandin synthases (PGFS and PGES) in corpora lutea (CL) during experimentally induced luteolysis in cow. The Fleckvieh cows in the mid-luteal phase (days 8-12, control group) were injected with cloprostenol (PGF analogue), and CL were collected by transvaginal ovariectomy before (days 8-12, control group) and at 0.5, 2, 4, 12, 24, 48 and 64h after PGF application (n=5 per group). The mRNA expression was determined by RT-qPCR, the hormone concentrations by enzyme immunoassay and localization by immunohistochemistry. PTGS2gene expression increased significantly 2h after PGF application, followed by continuous and significant downregulation afterwards. The PGF tissue concentration increased significantly just after PGF injection and again during structural luteolysis (after 12h), whereas PGE concentration significantly decreased during structural luteolysis. The FP receptor mRNA decreased significantly at 2h and again at 12h after PGF. In contrast, EP4 receptor mRNA increased significantly just after the PGF application (0.5h). The immunostaining of PGES and PTGS2 on day 15-17shows numerous positive luteal cells, followed by lower activity afterwards on day 18 (luteolysis). In conclusion, the changes of examined prostaglandin family members in CL tissue after PGF application may be key components of the local mechanisms regulating the cascade of actions leading to functional and subsequent structural luteolysis in the bovine ovary.

Prostaglandin analog effects on cerebrospinal fluid reabsorption via nasal mucosa.

Cerebrospinal fluid (CSF) outflow has been demonstrated along nasal lymphatics via olfactory nerve projections; flow may be increased by stimulating lymphatic contractility using agents such as noradrenaline and the thromboxane A2 analog U46619. Lymphatics elsewhere in the body show increased contractility upon exposure to the prostaglandin F2alpha analog isoprostane-8-epi-prostaglandin. We investigated the ability of ophthalmic prostaglandin F2alpha analogs to increase CSF outflow when applied to the nasal mucosa by inhalation. Latanoprost (0.1, 0.5, or 1mg/ml), bimatoprost (0.3 or 3mg/ml), travoprost (0.04 or 0.4mg/ml), latanoprostene bunod (0.24 or 2.4mg/ml), tafluprost (0.25 or 2.5mg/ml), or control vehicle (10% DMSO) was administered to awake adult C57B/6 mice by nasal inhalation of 2μl droplets. Multiday dosing (daily for 3 days) of latanoprost also was evaluated. A total of 81 animals were studied including controls. General anesthesia was induced by injection, and fluorescent tracer (AlexaFluor647-labelled ovalbumin) was injected under stereotaxic guidance into the right lateral ventricle. Nasal turbinate tissue was harvested and homogenized after 1 hour for tracer detection by ELISA and fluorometric analysis. Inhalation of latanoprost 0.5mg/ml and 1mg/ml led to a 11.5-fold increase in tracer recovery from nasal turbinate tissues compared to controls (3312 pg/ml vs 288 pg/ml, p<0.001 for 0.5mg/ml; 3355 pg/ml vs 288 pg/ml, p<0.001 for 1mg/ml), while latanoprost 0.1 mg/ml enhanced recovery 6-fold (1713 pg/ml vs 288 pg/ml, p<0.01). Tafluprost 0.25mg/ml and bimatoprost 0.3mg/ml showed a modest (1.4x, p<0.05) effect, and the remaining agents showed no significant effect on tracer recovery. After 3 days of daily latanoprost treatment and several hours after the last dose, a persistently increased recovery of tracer was found. Prostaglandin F2alpha analogs delivered by nasal inhalation resulted in increased nasal recovery of a CSF fluorescent tracer, implying increased CSF outflow via the nasal lymphatics. The greatest effect, partially dose-dependent, was observed using latanoprost. Further studies are needed to determine the efficacy of these agents in reducing ICP in short and long-term applications.

GnRH34: An alternative for increasing pregnancy in timed AI beef cows

Two experiments evaluated the effects of gonadotropin releasing hormone (GnRH) treatment on fertility of suckled Nelore beef cows treated with an estradiol/progesterone (P4)-based protocol for timed artificial insemination (TAI). Experiment 1 was designed to determine the effect of GnRH administered 34 h after P4 insert removal (GnRH34) on time of ovulation. Suckled cows (n = 34) were treated with 2 mg estradiol benzoate (EB) and an intravaginal insert containing 1.9 g of P4. Eight days later, P4 inserts were removed, and all cows received 150 μg of d-cloprostenol (prostaglandin F2 alpha analogue), 300 IU of eCG, and 1 mg of estradiol cypionate (ECP). On Day 9 (05:00 p.m.), cows were randomly distributed, according to the diameter of the pre-ovulatory follicle, in two treatments: 1) GnRH (n = 17) cows that received 10.5 μg of buserelin acetate, or 2) no further treatment (control, n = 17). Cows treated with GnRH 34 h after P4 insert removal ovulated earlier (P = 0.02) than control cows (66 ± 0.0 and 77.2 ± 4.3 h). Experiment 2 was designed to determine the effect of GnRH34 on the fertility of suckled beef cows. Nelore cows (n = 506) were treated as Experiment 1. On Day 8, cows were painted in the sacrocaudal region to identify cows that displayed estrus. On Day 9 (05:00 p.m.), cows were randomized to receive same treatment as Experiment 1, control (n = 252), or GnRH (n = 254). All cows were TAI 48 h after P4 insert removal. At TAI, estrus was evaluated, and deemed to have occurred in cows without a tail-head chalk mark (>75% paint loss). Cows treated with GnRH 34 h after P4 insert removal had greater (P = 0.01) pregnancy per AI (P/AI) than cows that only received ECP (63.0% and 50.4%). No difference (P = 0.5) was observed in the proportion of cows that displayed estrus between treatments. Furthermore, cows that displayed estrus had greater (P < 0.01) P/AI than cows that did not. Treatment with GnRH, given at 34 h after P4 insert removal, increased (P < 0.05) P/AI in cows that did not show estrus at TAI. In summary, treatment with GnRH 34 h after P4 insert removal was associated with earlier ovulation and resulted in greater P/AI in suckled Nelore cows treated with an estradiol/P4-based protocol for TAI.

Oestrus synchronization of West African Dwarf Does using prostaglandin-F2α analogue

Introduction: Successful livestock propagation through artificial insemination technique is achievable only when the different processes involved, including oestrus synchronization, are well executed. Methods: Dinoprost thromithamine (Lutalyse®) is an analogue of Prostaglandin F2-alpha (PGF2α) used extensively for oestrus synchronization in ruminants. Its efficacy as oestrus synchronizing agent was studied using six (6) healthy West African Dwarf (WAD) does aged between 2 and 4years and weighing 6-13 kg. The animals, randomly divided into three groups of 2 animals each, were kept in a standard goat pen and allowed to graze from morning to evening. They were fed with concentrates in the evening and supplied water ad libitum. The multiparous group (I) and nulliparous group (II) were administered two intramuscular injections of 5mg Lutalyse® at 11 days interval while the control group (III) was not given any treatment. The animals were closely monitored for signs of oestrus during and after the treatments. Oestrus detection was carried out by teasing the does twice daily (0800 and 1600 hours respectively) using one vasectomized buck per set of does. Results: The 2 does in group I responded to the treatment, while does in group II and III did not. Group I does were seen to come on heat between 48th and 96th hours after the second injection (100% response). The two does in Group I showed intense oestrus (90.00 %) while does in Group II presented weak oestrus. Significance: From this study, it can be concluded that 5mg of Dinoprost is effective as an agent of oestrus synchronization in multiparous WAD does.

Stanniocalcin-1 (STC-1), a downstream effector molecule in latanoprost signaling, acts independent of the FP receptor for intraocular pressure reduction.

Prostaglandin F2 alpha (PGF2α) analogues such as latanoprost are common first-line intraocular pressure (IOP) lowering medications. However, their clinical use is limited in some patient populations due to minimal or no IOP lowering response or side effects. In searching for a more targeted approach for IOP reduction, our lab recently identified Stanniocalcin-1 (STC-1) as a molecule that was required for latanoprost-mediated IOP reduction and also acted as a stand-alone IOP lowering agent. In order to determine whether latanoprost and STC-1 were equivalent and/or additive for IOP reduction, we treated C57BL/6J mice with one or a combination of these agents and measured IOP. Importance of the FP receptor for latanoprost- and STC-1-mediated IOP reduction was examined in C57BL/6J mice utilizing the pharmacologic FP receptor inhibitor AL-8810 as well as FP receptor knockout mice generated in our laboratory. Latanoprost-free acid (LFA) and STC-1 reduced IOP to a similar degree and were non-additive in C57BL/6J mice. As expected, the IOP lowering effects of LFA were abrogated by pharmacologic inhibition of the FP receptor with AL-8810 and in FP receptor knockout mice. In contrast, STC-1 maintained IOP-lowering effects in the presence of AL-8810 and also in FP receptor knockout mice. These results suggest that LFA and STC-1 show equivalent and non-additive IOP reduction in C57BL/6J mice and that unlike LFA, STC-1-mediated IOP reduction occurs independent of the FP receptor.

Statin-induced myopathic changes in primary human muscle cells and reversal by a prostaglandin F2 alpha analogue

Statin-related muscle side effects are a constant healthcare problem since patient compliance is dependent on side effects. Statins reduce plasma cholesterol levels and can prevent secondary cardiovascular diseases. Although statin-induced muscle damage has been studied, preventive or curative therapies are yet to be reported. We exposed primary human muscle cell populations (n = 22) to a lipophilic (simvastatin) and a hydrophilic (rosuvastatin) statin and analyzed their expressome. Data and pathway analyses included GOrilla, Reactome and DAVID. We measured mevalonate intracellularly and analyzed eicosanoid profiles secreted by human muscle cells. Functional assays included proliferation and differentiation quantification. More than 1800 transcripts and 900 proteins were differentially expressed after exposure to statins. Simvastatin had a stronger effect on the expressome than rosuvastatin, but both statins influenced cholesterol biosynthesis, fatty acid metabolism, eicosanoid synthesis, proliferation, and differentiation of human muscle cells. Cultured human muscle cells secreted ω-3 and ω-6 derived eicosanoids and prostaglandins. The ω-6 derived metabolites were found at higher levels secreted from simvastatin-treated primary human muscle cells. Eicosanoids rescued muscle cell differentiation. Our data suggest a new aspect on the role of skeletal muscle in cholesterol metabolism. For clinical practice, the addition of omega-n fatty acids might be suitable to prevent or treat statin-myopathy.

Efficacy of topical latanoprost in the treatment of eyelid vitiligo: A randomized, double-blind clinical trial study.

Numerous studies have demonstrated that the pigmentation of iris and around the eyelid is a common side effect of latanoprost, a prostaglandin F2alpha analogue used in the treatment of glaucoma. Hence, the authors decided to study the effectiveness of topical latanoprost on vitiligo patches around the eyelid. In this randomized, double-blind, clinical trial study, 31 patients with vitiligo vulgaris and focal vitiligo involving the eyelids were evaluated. Patients were randomly divided into two groups. First group received topical latanoprost gel twice daily for 12 weeks, whereas the second group received placebo with the same protocol. To evaluate severity of the disease the VIDA rating system was used. Serial photos of the patches were taken to compare and evaluate the repigmentation percentage of the patches. The patients in both groups had almost similar VIDA score (p > .05). First group showed improved pigmentation, whereas participants in the second group did not show any improvement in the pigmentation. The group treated with latanoprost showed significant reduction in the symptoms of the disease, whereas those treated with placebo did not show any alteration (p > .05). No significant complications were observed in either groups. Latanoprost proved effective in treating vitiligo disease involving eyelids.

Effect of the Interval From Prostaglandin F2alpha Treatment to Ovulation on Reproductive Efficiency Rates in a Commercial Equine Embryo Transfer Program

In the present study, 2.228 cycles of 180 Polo Argentino donor mares from an embryo transfer program in Argentina were examined to evaluate the effects of: (1) Interval from Prostaglandin F2alpha analog treatment to ovulation (ITO) on embryo recovery rate (ERR); (2) ITO on number of embryos per flushing (EPF); (3) ITO on multiple ovulation (MO) rate; (4) ITO from donor mare on pregnancy rate (PR) in recipient mares. Mares were inseminated with fresh semen from 31 fertile stallions in the induced estrus. Embryo flushing was performed 7–8 days postovulation. Following embryo flushing, donor mares were treated with prostaglandin F2alpha analog (cloprostenol 250 μg). The ERR increased along with the ITO (P = .01), with the lowest ERR (30.7%) for mares with an ITO of <4 days, and the highest (78.3%) in mares with an ITO of 10 days. The ITO from the donor mare in which the embryo was recovered did not have a significant effect on PR: ITO <6, 6 to 10, and >10 days were 74.6, 81.4, and 77.3%, respectively. The number of EPF and MO rate increased gradually along with the ITO (P < .05). In conclusion, the results of this study indicated that the ITO is positively correlated with the embryo recovery and the multiple ovulation rate.

High Prevalence of Subclinical Endometritis in Problem Mares– Effect of Activation and Treatment on Fertility

In the present study, 2.228 cycles of 180 Polo Argentino donor mares from an embryo transfer program in Argentina were examined to evaluate the effects of: (1) Interval from Prostaglandin F2alpha analog treatment to ovulation (ITO) on embryo recovery rate (ERR); (2) ITO on number of embryos per flushing (EPF); (3) ITO on multiple ovulation (MO) rate; (4) ITO from donor mare on pregnancy rate (PR) in recipient mares. Mares were inseminated with fresh semen from 31 fertile stallions in the induced estrus. Embryo flushing was performed 7–8 days postovulation. Following embryo flushing, donor mares were treated with prostaglandin F2alpha analog (cloprostenol 250 μg). The ERR increased along with the ITO (P = .01), with the lowest ERR (30.7%) for mares with an ITO of <4 days, and the highest (78.3%) in mares with an ITO of 10 days. The ITO from the donor mare in which the embryo was recovered did not have a significant effect on PR: ITO <6, 6 to 10, and >10 days were 74.6, 81.4, and 77.3%, respectively. The number of EPF and MO rate increased gradually along with the ITO (P < .05). In conclusion, the results of this study indicated that the ITO is positively correlated with the embryo recovery and the multiple ovulation rate.

Response to estrus induction with abortion treatment in microminipigs on different days after insemination.

In microminipigs, estrus induction with abortion treatment, which is typically performed between 25 and 40 days after mating, is not always successful. Thus, the authors hypothesized that it may be more difficult to induce estrus by treating microminipigs approximately 40 days after mating. Accordingly, in this study, estrus induction was performed with abortion treatment in four microminipigs as follows: 0.3 mg of cloprostenol, a prostaglandin F2-alpha analog, was administered (day 0); after 24 h, 0.15 mg of cloprostenol and 250 IU of equine chorionic gonadotrophin were administered intramuscularly and simultaneously (day 1); after 96 h, 120 IU of human chorionic gonadotropin was injected intramuscularly (day 4). These treatments were compared at two different stages of pregnancy: early treatment (26.5 ± 0.7 days) and late treatment (38.3 ± 0.8 days). In the early treatment, all four microminipigs exhibited estrus on day 5, whereas in the late treatment, estrus was observed clearly in only two pigs on day 6 and slightly in 1 pig on day 10, whereas it was unclear in 1 pig. These results suggest that it is difficult to induce estrus with abortion treatment in microminipigs at approximately 40 days after mating.

Expression pattern of HIF1alpha and vasohibins during follicle maturation and corpus luteum function in the bovine ovary.

The aim of this study was to characterize expression patterns of hypoxia-inducible factor-1alpha (HIF1A) and vasohibin family members (VASH1 and VASH2) during different stages of ovarian function in cow. Experiment 1: Antral follicle classification occurred by follicle size and estradiol-17beta (E2) concentration in the follicular fluid into 5 groups (<0.5, 0.5-5, 5-40, 40-180 and >180 E2 ng/ml). Experiment 2: Corpora lutea (CL) were assigned to the following stages: days 1-2, 3-4, 5-7, 8-12, 13-16 and >18 (after regression) of oestrous cycle and of pregnancy (months 1-2, 3-4, 6-7, >8). Experiment 3: Cows on days 8-12 were injected with a prostaglandin F2alpha (PGF) analogue and CL were collected before and 0.5, 2, 4, 12, 24, 48 and 64hr after PGF injection. Expression of mRNA was measured by qPCR, steroid hormone concentration by EIA and localization by immunohistochemistry. HIF1A mRNA expression in our study increases significantly in follicles during final maturation. The highest HIF1A mRNA expression was detected during the early luteal phase, followed by a significant decrease afterwards. In contrast, the mRNA of vasohibins in small follicle was high, followed by a continuous and significant downregulation in preovulatory follicles. The obtained results show a remarkable inverse expression and localization pattern of HIF1A and vasohibins during different stages of ovarian function in cow. These results lead to the assumption that the examined factors are involved in the local mechanisms regulating angiogenesis and that the interactions between proangiogenic (HIF1A) and antiangiogenic (vasohibins) factors impact all stages of bovine ovary function.

Efficacy of induction of luteolysis in superovulated cows is dependent on time of prostaglandin F2alpha analog treatment: effects on plasma progesterone and luteinizing hormone profiles

The objectives were to (1) evaluate the effectiveness of induction of luteolysis in superovulated (SOV) cows at two distinct time points after embryo flushing; and (2) compare the pattern of LH release after treatment with PGF in cows with single vs. multiple ovulations. In the first experiment, Holstein cows were SOV with 400 IU of FSH following standard procedures. Uterine flushing for embryo recovery was performed 7 days after artificial insemination (Day 0), and cows were randomly allocated into two groups to receive PGF (0.5-mg sodium cloprostenol, intramascular) either immediately after flushing (Day 7 group, N = 19) or 4 days later (Day 11 group, N = 20). Time of luteolysis was determined on the basis of plasma progesterone (P4) concentrations. There was no difference (P > 0.05) in plasma P4 before treatment between Day 7 and Day 11 groups. A decline in plasma P4 was observed 48 hours after PGF treatment in both the groups (P < 0.0001). In Day 11 cows, P4 continued to decrease thereafter, whereas Day 7 animals had no further reduction in plasma P4. Luteolysis (P4 < 1 ng/mL) occurred in all Day 11 cows. In the Day 7 group, however, luteolysis failure was observed for 11 of 19 cows (57.9%). In cows without luteolysis, plasma P4 increased after the initial PGF–induced decline. The second experiment compared luteolysis in (SOV, N = 6) vs. non-SOV (control, N = 8) cows. Both groups received a single PGF treatment on Day 11 after estrus, and luteolysis was monitored daily by ovarian ultrasonography and plasma P4 measurements. In addition, plasma LH was measured in blood samples taken every 20 minutes for 1 hour during five consecutive days after treatment. A similar percentage of reduction in P4 was observed in both groups 24 hours after treatment; however, SOV cows only reached plasma P4 values similar (P > 0.05) to controls 96 hours after treatment. There was no difference in initial LH values between SOV and controls (P > 0.05). The slower decrease in plasma P4 in the SOV group prevented an increase in LH for up to 96 hours after luteolysis induction, whereas LH values increased (P < 0.05) in controls 24 hours after treatment. In conclusion, (1) luteolysis may fail or be incomplete when PGF treatment is given on the day of uterine flushing (Day 7) in SOV cows; (2) induction of luteolysis 4 days later (Day 11) is effective, but the initial high-plasma P4 concentrations result in a slower slope of P4 decline to basal levels, and consequently, delayed increase in LH pulses.

Expression and localization of members of the thrombospondin family during final follicle maturation and corpus luteum formation and function in the bovine ovary.

The aim of this study was to characterize the expression patterns and localization of the thrombospondin family members (THBS1, THBS2) and their receptors (CD36 and CD47) in bovine ovaries. First, the antral follicles were classified into 5 groups based on the follicle size and estradiol-17beta (E2) concentration in the follicular fluid (< 0.5, 0.5–5, 5–40, 40–180 and >180 E2 ng/ml). Second, the corpus luteum (CL) was assigned to the following stages: days 1–2, 3–4, 5–7, 8–12, 13–16 and >18 of the estrous cycle and of pregnancy (month 1–2, 3–4, 6–7 and > 8). Third, the corpora lutea were collected by transvaginal ovariectomy before and 0.5, 2, 4, 12, 24, 48 and 64 h after inducing luteolysis by injecting a prostaglandin F2alpha analog. The mRNA expression of examined factors was measured by RT-qPCR, steroid hormone concentration by EIA, and localization by immunohistochemistry. The mRNA expression of THBS1, THBS2, CD36, and CD47 in the granulosa cells and theca interna was high in the small follicles and reduced in the preovulatory follicles. The mRNA expression of THBS1, THBS2, and CD47 in the CL during the estrous cycle was high, but decreased significantly during pregnancy. After induced luteolysis, thrombospondins increased significantly to reach the maximum level at 12 h for THBS1, 24 h for THBS2, and 48 h for CD36. The temporal expression and localization pattern of the thrombospondins and their specific receptors in the antral follicles and corpora lutea during the different physiological phases of the estrous cycle and induced luteolysis appear to be compatible with their inhibitory role in the control of ovarian angiogenesis.

Intraocular Pressure-Lowering Activity of NCX 470, a Novel Nitric Oxide-Donating Bimatoprost in Preclinical Models.

The prostaglandin F2alpha (PGF2α) analogue bimatoprost lowers intraocular pressure (IOP) by increasing uveoscleral outflow at doses shown to elicit redness of the eye. With the aim to enhance the IOP-lowering effect of bimatoprost we studied NCX 470 [(S,E)-1-((1R,2R,3S,5R)-2-((Z)-7-(ethylamino)-7-oxohept-2-enyl)-3,5-dihydroxycyclopentyl)-5-phenylpent-1-en-3-yl 6-(nitrooxy)hexanoate], a dual-acting compound combining bimatoprost with nitric oxide (NO) known to mainly act via relaxation of trabecular meshwork and Schlemm's canal. New Zealand white rabbits with transient hypertonic saline-induced IOP elevation (tOHT-rabbits), cynomolgus monkeys with laser-induced ocular hypertension (OHT-monkeys), and normotensive dogs (ONT-dogs) were used. The levels of NCX 470, bimatoprost, and bimatoprost acid were determined in aqueous humor (AH), cornea (CR), and iris/ciliary body (ICB) by liquid chromatography-mass spectrometry/mass (LC-MS/MS), while cGMP in AH and ICB was monitored using an enzyme immunoassay (EIA) kit in pigmented Dutch Belted rabbits. NCX 470 (0.14%, 30 μL) lowered IOP in tOHT-rabbits with an E(max) of -7.2 ± 2.8 mm Hg at 90 minutes. Bimatoprost at equimolar dose (0.1%, 30 μL) was noneffective in this model. NCX 470 (0.042%, 30 μL) was more effective than equimolar (0.03%, 30 μL) bimatoprost in ONT-dogs (IOP change, -5.4 ± 0.7 and -3.4 ± 0.7 mm Hg, respectively, P < 0.05) and in OHT-monkeys (IOP change, -7.7 ± 1.4 and -4.8 ± 1.7 mm Hg, respectively, P < 0.05) at 18 hours post dosing. NCX 470 (0.042%, 30 μL) or bimatoprost (0.03%, 30 μL) resulted in similar bimatoprost acid exposure in AH, CR, and ICB while cGMP was significantly increased in AH and ICB at 18 and 24 hours after NCX 470 dosing. NCX 470 lowers IOP more than equimolar bimatoprost in three animal models of glaucoma by activating PGF2α and NO/cGMP signaling pathways.

Hirsutism following the use of bimatoprost eyedrops for glaucoma

Prostaglandin F2 alpha (PGF2α) analogues including bimatoprost are often the first line drugs used in the treatment of glaucoma. We present a case of a 62-year-old female patient who was started on bimatoprost in both the eyes for primary open angle glaucoma. The intraocular pressures reached the target level but she developed hair growth over the chin and upper lip after six months of commencing of the treatment. The regional hypertrichosis did not reduce much after stopping the drug. Hirsutism is a potential side effect of prostaglandin analogues which has rarely been reported. Doctors and patients need to be aware of this noticeable and unwanted side effect. The effect seems to occur in patients already having few non-vellus hairs in these areas.

The effect of short-term lupin (Lupinus angustifolius) feed supplementation on serum steroid hormones, insulin-like growth factor I, and ovarian follicular development and atresia in Merino ewes

The effects of short-term lupin supplementation on ovulation rate in ewes during oestrous season are well known. In this study, we tested the effects of an 8-day lupin supplementation on folliculogenesis during anoestrus applied over the hormonal treatment of ewes. Fourteen anoestrous Merino ewes were assigned into 2 groups (n = 7 for each) in which the control group was fed a basal diet and the experimental group was supplemented with lupin groats (500 g per head per day). Both groups were treated intramuscularly with 12.5 µg/head lecirelinum (luteinising hormone releasing hormone super analogue) on the first day of supplementation and 5 days later with 37.5 µg/head cloprostenolum D (prostaglandin F2 alpha analogue). Serum sexual steroid and insulin-like growth factor I (IGF-I) concentrations were assessed using radioimmunoassay and enzyme-linked immunosorbent assay methods, respectively. Folliculogenesis was evaluated using a light microscope. Lupin feeding did not affect progesterone or IGF-I, but it significantly decreased the concentration of oestradiol-17beta. Lupin significantly increased the number of follicles of 3-5 mm in diameter and the total rate of atresia. The follicles mainly collapsed and underwent late atresia. The size of healthy and collapsed follicles increased in the lupin-fed group. These results indicated that 8-day lupin supplementation affected follicular development and atresia through an IGF-I/oestradiol feedback system.

緑内障点眼薬の薬物相互作用の解明と点眼薬物療法の費用最小化分析の確立

Latanoprost, a prostaglandin F2alpha analog (PG), has been shown to be an effective ocular hypotensive agent for treating glaucoma. Carbonic anhydrase (CA) inhibitors are also used to reduce ocular hypertension by decreasing aqueous humor secretion, and are given in combination with PG.Timolol malate has been shown to be an effective ocular hypotensive agent when used alone or with CA inhibitor on glaucoma patients.However, the effects of latanoprost and timolol malate on CA have not been clarified. Therefore, we studied the effects of latanoprost free acid and timolol hemihydrate on human CA (hCA)-I and hCA-II using stopped flow method.Latanoprost free acid inhibited the hydration activity of hCA-I or hCA-II by a noncompetitive mechanism. Timolol hemihydrate activates the enzyme activities of hCA-I and hCA-II. In hCA-I and hCA-II, the enzyme kinetic results clearly showed that timolol hemihydrate increases the value of Vmax but does not influence the value of Km. The noncompetitive inhibition mechanism and the binding mode of latanoprost free acid indicate that the behavior of latanoprost free acid is similar to that of simple anions. Timolol hemihydrate has a heterocyclic moiety and secondary amino group, which are typical structures in efficient activators of carbonic anhydrase.Moreover, we compared the cost of lowering intraocular pressure (IOP) by 1 mmHg in the treatment of glaucoma. Latanoprost is less expensive for lowering IOP by 1 mmHg than other products. These data should be helpful in selecting topical ophthalmic solutions from the viewpoint of cost-minimization.

Latanoprost Stimulates Ocular Lymphatic Drainage: An In Vivo Nanotracer Study

Ocular lymphatics have been recently shown to contribute to aqueous humor outflow. It is not yet known whether lymphatic outflow can be stimulated by pharmacological agents. Here we determine whether latanoprost, a prostaglandin F2 alpha analog commonly used to lower IOP to treat glaucoma, increases lymphatic drainage from the eye. Lymphatic drainage in mice was assessed in vivo, in 11 latanoprost-treated and 11 control animals using hyperspectral imaging at multiple times following quantum dot (QD) injection into the eye. QD signal intensity was also measured in tissue sections using hyperspectral imaging. In the latanoprost-treated group, lymphatic drainage rate into the submandibular lymph node was increased compared with controls (1.23 ± 1.06 hours-1 vs. 0.30 ± 0.17 hours-1, mean ± SD, P < 0.02). Total QD signal intensity in the submandibular lymph node was greater in the latanoprost-treated group compared with controls (10.55 ± 1.12 vs. 9.48 ± 1.24, log scale, P < 0.05). This is the first evidence that latanoprost increases lymphatic drainage from the eye. The pharmacological manipulation of this newly identified lymphatic outflow pathway may be relevant to treatments aimed at lowering intraocular pressure in glaucoma. This is the first evidence that a prostaglandin drug widely prescribed for glaucoma, enhances lymphatic drainage from the eye. The pharmacological stimulation of this newly identified outflow pathway may be highly relevant to treatments aimed at lowering IOP to prevent blindness from glaucoma.