Injection Of Follicle-stimulating Hormone Research Articles

Responsiveness of rabbits to superovulation treatment by a single injection of follicle‐stimulating hormone with aluminum hydroxide gel

Aluminum hydroxide gel (Al-gel), which is used as an adjuvant, can absorb macromolecules. We investigated the applicability of Al-gel to the sustained release of follicle-stimulating hormone (FSH) as a simplified method of superovulation (SOV) in rabbits. The responsiveness of rabbits to SOV by a single injection of FSH dissolved in Al-gel suspension (3.2 mg Al/ml) and in 10% (w/v) polyvinylpyrrolidone (PVP), and by multiple injections of FSH in saline was examined. The numbers of total and fertilized eggs recovered from rabbits treated with FSH in Al-gel (40.5 and 26.3, respectively) were similar to multiple injections (47.4 and 28.6, respectively) and were significantly greater (P < 0.05) than single injection of FSH with PVP (17.3 and 11.5, respectively). We also compared the plasma FSH levels of rabbits which were induced SOV by multiple or a single injection of Al-gel. Al-gel provided sustained release of FSH to the blood stream at a high enough dose for SOV. Moreover, the developmental competence of the pups of DNA-injected embryos from rabbits treated with a single injection of FSH mixed with Al-gel (18%) was similar to that of DNA-injected embryos, recovered from rabbits treated with FSH dissolved in saline (21%). Two transgenic pups were obtained from embryos recovered from rabbits by a single injection of FSH with Al-gel. These results indicate that a single injection of FSH with Al-gel is an effective method for SOV of rabbit and that this technique is applicable to research requiring large numbers of rabbit embryos such as the production of transgenic rabbits.

Novel follicular-phase gonadotropin-releasing hormone antagonist stimulation protocol for in vitro fertilization in the poor responder

Poor responders continue to be vexing in infertility therapy. By using GnRH antagonists before ovarian stimulation, we demonstrate an improvement in oocyte, embryo, and zygote yield in patients with a prior poor response.

The Effect of Reduced Dose and Number of Treatments of FSH on Superovulatory Response in CIDR-Treated Korean Native Cows

The objective of this study was to investigate the effect of dosage and number of days of follicle stimulating hormone (FSH) treatment on superovulatory response in controlled internal drug release (CIDR)-treated Korean native cows. Forty cows underwent two superovulatory treatments with a crossover design. Cows, at random stages of the estrous cycle, received a CIDR together with injections of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 days later. The cows were divided into 2 groups based on the dosage and number of days of treatment with porcine FSH; a total of 28 mg FSH was given in twice daily intramascular injections in decreasing doses over 4 days (5, 5, 4, 4, 3, 3, 2 and 2 mg; T1 group, n=20) or a total of 24 mg FSH was given in twice daily decreasing doses over 3 days (5, 5, 4, 4, 3 and 3 mg; T2 group, n=20). This was followed by the alternate treatment in the subsequent superovulation. The cows were treated identically in all other respects. PGF(2alpha) (25 mg and 15 mg) was given with the 5th and 6th injections of FSH, CIDR were withdrawn at the 6th FSH injection and the cows received 200 microg GnRH 36 h after CIDR withdrawal. The cows were artificially inseminated twice, at 48 and 60 h after CIDR withdrawal, using commercial semen from four Korean native bulls, and embryos were recovered 6 or 7 days after the 2nd insemination. The numbers of corpora lutea (CL; 7.9+/-1.0 vs. 8.3+/-1.1) and large follicles (1.2+/-0.2 vs. 1.3+/-0.3) present at the time embryo recovery, as detected by ultrasonography, did not differ between the T1 and T2 groups (P>0.05). Similarly, the numbers of total ova/embryos (6.2+/-0.9 vs. 6.4+/-1.1), transferable embryos (3.4+/-0.8 vs. 3.2+/-0.7), degenerate embryos (0.8+/-0.2 vs. 1.0+/-0.3) and unfertilized ova (2.1+/-0.5 vs. 2.2+/-0.5) did not differ between the groups (P>0.05). These data indicate that a reduced dose (24 vs. 28 mg) and number of treatments (6 vs. 8) of FSH for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.

Suppression of Follicle Wave Emergence in Cyclic Ewes by Supraphysiologic Concentrations of Estradiol-17beta and Induction with a Physiologic Dose of Exogenous Ovine Follicle-Stimulating Hormone1

Follicle waves are preceded by follicle-stimulating hormone (FSH) peaks in ewes. The purpose of the present study was to see whether estradiol implant treatment would block FSH peaks to create a model in which the effect of the timing and mode of FSH peaks could be studied by ovine FSH (oFSH) injection. In Experiment 1, 10 ewes received estradiol-17beta implants on Day 4 after ovulation (Day 0, day of ovulation); five ewes received large implants, and five ewes received small implants. Five control ewes received empty implants. In Experiment 2, 12 ewes received large implants on Day 4. On Day 9, six ewes received oFSH twice, 8 h apart (0.5 microg/kg; s.c.). Implants were left in place for 10 days in both experiments. In both studies, ovarian ultrasonography and blood sampling was done daily. In Experiment 1, estradiol concentrations were significantly higher in ewes with large implants (10.4 +/- 0.7 pg/ml) compared with controls (3.9 +/- 0.7 pg/ml) and ewes with small implants (5.4 +/- 0.7 pg/ml; P < 0.001). A significant reduction was found in mean FSH peak concentration (31%; P < 0.05) and FSH peak amplitude (45%; P < 0.05) in ewes with large implants compared with controls. Mean and basal FSH concentrations were unaffected by the large implants. The large implants halted follicle-wave emergence between Day 0 and 8 after implant insertion. The small follicle pool (2-3 mm in diameter) was unaffected by the large implants. When oFSH was injected into ewes with large implants, a follicle wave emerged 1.5 +/- 0.5 days after injection; however, in ewes given saline alone, a follicle wave emerged 4.8 +/- 0.8 days after injection (P < 0.01). We concluded that truncation of FSH peaks by estradiol implants prevented follicle-wave emergence, but injection of physiologic concentrations of oFSH reinitiated follicle-wave emergence.

Using Follicle Stimulating Hormone (FSH) for Superovulation in Buffalo

Studies of superovulation have been carried out in buffaloes and the progress has been achieved but the number of embryo collected was still small. Two studies were conducted to evaluate the use Follicle Stimulating Hormone (FSH) for superovulation in buffalo. First study, fifteen buffaloes were randomly divided into 3 groups of five buffaloes of each group and superovulated using three type of FSH (FSH-P; FSC-China and Folltropin). Hormones were given twice a day for 4 days in decreasing doses methods. First injection initiated on day 10 of estrus cycle, followed by prostaglandin on day 12 than artificially inseminated (AI) two days later. Recovery of embryo was conducted un-surgically method by flushing uterus horn on day 6 after AI. Second study, 10 buffaloes were randomly divided into 2 groups of each five buffaloes First group was superovulated with FSH followed methodology in the first study (control group) and second group were given a prime FSH on day 1 of estrus cycle before superovulation (treatment group). After 3 months the superovulation was repeated in the reverse condition where the control become treatment and the treatment become a control group. The type of FSH did not significantly affect the superovulatory response. The mean total corpus luteum (TCL), total number of embryo (TNE) and total number of viable embryo (TVE) were 6.8, 3.3 and 2.2, 6.2, 3.0 and 2.1 and 7.2, 3.6 and 2.4 for FSH-P, FSH China and Folltropin respectively. A single injection of FSH significantly increased (P<0.05) the number of embryo collected. The mean TCL, TNE and TVE were 6.9, 2.8 and 2.1 and 8.2, 3.3 and 2.5 for control and treatments respectively. The peak progesterone level was significantly higher (P<0.05) in treatment group (6.8 ng/ml) than those in control (5.6 ng/ml). Repeated the superovulation after 3 months did not significantly affect the number of embryo collected. In conclusion, superovulation in buffaloes can be performed using a types of FSH and prime injection of FSH on day 1 of estrus cycle increase the number of embryo recovered. The superovulation can be conducted after 3 months. Key Words : FSH, Superovulation, Embryo

Ovum Pick‐up in Cycling and Lactating Postpartum Swamp Buffaloes (Bubalis bubalis)

The objective of this study was to evaluate the efficiency of Ovum Pick Up (OPU) in cycling (n = 5) and lactating, postpartum, swamp buffaloes (n = 6) with and without gonadotropin stimulation. The OPU was performed every two weeks in all groups of animals, for a total of six sessions. Thirty collections were performed in five cycling buffaloes and 36 collections in six lactating postpartum buffaloes. Buffaloes that received hormonal stimulation were given a total of 400 mg, follicle stimulating hormone (FSH), administered twice daily over 3 days in decreasing doses, together with 100 microg of GnRH, 24 h after the last FSH injection. Following a resting period of 1 month, the two groups of buffaloes, were subjected to the same OPU regimen, but without any hormonal treatment for an additional six OPU sessions. The number of aspirated follicles recorded from the hormonal stimulated, cycling animals and lactating, postpartum buffaloes was not significantly different, 7.2 +/- 3.7 and 9.0 +/- 3.2, respectively (p > 0.05). Recovered oocytes collected from the two groups of hormonally stimulated animals were also not statistically different: 3.7 +/- 2.7 in the cycling and 5.9 +/- 3.5 in the lactating postpartum group (p > 0.05). In the two groups of buffaloes not receiving hormonal stimulation, the number of aspirated follicles was not significantly different: 2.1 +/- 1.4 and 1.4 +/- 0.7 in cycling and lactating postpartum buffaloes respectively (p > 0.05). Recovered oocytes in the non-treated groups were also similar: 1.4 +/- 1.3 vs 0.7 +/- 0.8 in cycling and lactating buffaloes (p > 0.05). Among stimulated buffaloes, most aspirated follicles were small in size (< or =5 mm), whereas they were mostly medium and large sizes in the non-treated buffaloes. The oocyte recovery rate in both the groups, cycling and lactating postpartum, were 51.6% and 69.5% in stimulated groups and 55.0% and 53.1% in non-stimulated groups (p > 0.05). The majority of recovered oocytes were single- and multi-layered, and the number was greater in the cycling than in the lactating, postpartum buffaloes. The number and quality of recovered oocytes was similar in all groups of buffaloes whether they were received or did not receive hormonal stimulation. Moreover no difference was found in multi- and single-layered oocytes between cycling and lactating, postpartum buffaloes. In conclusion, OPU can be performed successfully in swamp buffalo in different reproductive status and FSH administration was shown to increase the number of aspirated oocytes in both cycling and lactating, postpartum buffaloes.

Effect the administration of human chorionic gonadotrophin (hCG) hormone following superovulation treatment in buffalo

A numbers of studies had been reported to evaluate the effect of hormone for superovulation treatment in buffaloes, however the numbers of embryo recovered is still very small. One of the limiting factors which affect the numbers of embryo recovered is preovulatery LH concentration. The experiment was carried out to study the effect of hCG following superovulation treatments in buffaloes as an effort to increase the embryo recovered. Ten (10) buffaloes in three different genotypes (Riverine, swamp and its crosses) were superovulated using follicle stimulating hormone (FSH). A total of 12 ml FSH (Folltropin) was injected intramuscularry twice a day with 12 hours interval in a decreasing doses for 4 days (2.5, 2.5; 2.0, 2.0; 1.0, 1.0 and 0.5, 0.5 ml). Prostaglandin was injected intramuscularry 2 days after the first injection of FSH followed by administration of 500 IU hCG two days later. At the same days of administration of hCG, artificial insemination (AI) using frozen semen was conducted and repeated again after 12 and 24 hours. Blood collecting was conducted 3 times a week for obtaining the progesteron level of plasma blood. Embryo was collected by non-surgically technique on day 6 of estrus cycle by flushing each horn of uterus with 500 ml Dubelco's Phosphat Buffer Saline (DBPS). Parameter recorded were diameter of ovary (DO), total corpus luteum (TCL), number of embryo collected (NE), percentage of recovery rate (%RR) and peak progesteron level (PP). Administration of hCG significantly increase the response of buffalo to superovulation treatment. The mean of DO (cm), TCL, NE and RR (%) was 4.0, 6.3, 2.1 and 37.2 and 4.5, 7.5, 3.9 and 48.1 for controll and 500 IU hCG respectively. The mean NE was significantly higher (P<0.05) in hCG than those control. The peak progesterone concentration was highly significant higher (P<0.01) in hCG (8.9 ng/ml) than those control (6.8 ng/ml). Both left and right ovary gave a similar response to superovulation treatments. In conclusion the admission of hCG following superovulation treatments using folltrophin increase the number of embryos recovered. Key Words: hCG, Superovulation, Buffalo, Embryo

A comparison of 3-day and daily follicle-stimulating hormone injections on stimulation days 1–6 in women undergoing controlled ovarian hyperstimulation

Objective To evaluate the efficacy of ovarian hyperstimulation by intermittent doses of 450 IU of recombinant human (h)FSH compared with daily 150-IU doses of recombinant hFSH. Design A pilot, open, randomized, parallel-group study. Setting Center for Reproductive Medicine, Düsseldorf, Germany. Patient(s) Infertile women with indication for IVF/intracytoplasmic sperm injection after at least 1 year of unprotected intercourse. Intervention(s) Recombinant hFSH was administered daily or intermittently (3-day intervals) from days 1 to 6 of stimulation and thereafter by daily injection. Main outcome measure(s) Number of preovulatory follicles, retrieved oocytes, two-pronuclei (2PN) zygotes, implantation, and pregnancy rates. Result(s) The number of oocytes in the daily-dose group was significantly greater. There were no significant differences in mean values for number of follicles ≥11 mm (except for day 7) and ≥14 mm, 2PN zygotes, and number of transferred embryos. Implantation and pregnancy rates per cycle were in favor of the intermittent 450-IU dose regimen; implantation rates were 17.0% and 9.8% in the 3-day-dose and daily-dose groups, respectively, and biochemical and clinical pregnancy rates were 33.3% and 15.7% and 25.5% and 13.7%, respectively. Conclusion(s) Intermittent administration of recombinant hFSH significantly reduces the total number of recombinant hFSH injections, compared with a conventional FSH regimen, without detrimental effects on implantation rate or pregnancy rate.

Caprine somatic cell nuclear transfer using in vivo matured oocytes collected by laparoscopic follicular aspiration

ABSTRACT In vivo matured oocytes collected by laparoscopic follicular aspiration (LFA) from hormone treated female goats were used as recipient ooplasts for somatic cell nuclear transfer (SCNT). Japanese native (Shiba) goats were used as donor females and some donor females were used repeatedly (two or three times) at intervals of a few months. To induce synchronization of estrus, a sponge containing 0.5 g of progesterone was inserted into the vagina of each goat for 14 days. These animals were also treated with follicle stimulating hormone (FSH) in a series of 8 injections over 4 days. The first FSH injection was administered on the morning of day 9 of sponge insertion. On the morning of day 13, 50 µg of gonadotropin‐releasing hormone (GnRH) was injected into each animal. Twenty‐nine hours after GnRH injection, LFA was performed. After removal of cumulus cells, collected oocytes with the first polar body were selected and enucleated for nuclear transfer. Anterior pituitary cells isolated from an adult male Shiba goat were transfected with a DNA fragment containing the enhanced green flourescent protein gene and the puromycin resistance gene. A single donor cell was inserted into the perivitelline space of each enucleated oocyte and fusion was induced with one electric pulse of 20 V for 10 µs. The SCNT goat eggs were cultured in chemically defined medium at 38.5°C in 5% CO2, 5% O2, 90% N2 for 9 days. By LFA, 396 oocytes were collected from a total of 30 females. After removal of cumulus cells, 64% of them extruded the first polar body. The percentage of SCNT goat eggs produced using in vivo matured oocytes which developed to the blastocyst stage (20–21%) was significantly higher (P &lt; 0.05) than that produced with in vitro matured oocytes (3–8%).

Effects of dosage of FSH, vehicle and time of treatment on ovulation rate and prolificacy in ewes during the anestrous season

The effects of dosage of follicle stimulating hormone (FSH), vehicle and time of injection on ovulation rate and prolificacy were evaluated during the anestrous season. Ewes on four farms ( n=445) were treated with a CIDR-G ® device for 5 days and exposed to rams upon removal of the insert (CRRI, day 0). A 3×2×2 factorial arrangement tested effects of dosage of FSH (Folltropin ®; 0, 42 or 68 mg NIH-FSH-P1), vehicle (saline:propylene glycol 1:4, v/v [PGL], or 50% polyvinylpyrrolidone K 29–32 [PVP]) and time of injection (12 or 36 h before CRRI). Follicular growth was monitored by ultrasonography in four ewes per treatment at injection of FSH, at CRRI, and on days 1–3 post-CRRI. All ewes were examined by ultrasonography on days 10–14 for ovulation rate, and on days 26–31 and 46–51 for pregnancy and number of embryos. The largest follicle (F 1) increased in diameter (mm) between FSH injection (5.3±0.1) and day 1 (6.1±0.1; P<0.01). The F 1 was larger at CRRI ( P≤0.05) in ewes receiving 42 than 0 mg FSH, and 68 than 42 mg FSH. The F 2 increased in diameter ( P<0.05) from FSH injection (4.7±0.2) to CRRI (5.2±0.2) and to day 1 (5.8±0.2), but was not affected by treatment. Number of small follicles (≤4 mm) did not differ with time or treatment. Number of medium follicles (5 mm) declined ( P<0.05) between FSH (1.5±0.2) and days 1 (0.8±0.2), 2 (0.9±0.2), and 3 (0.5±0.2). Number of large follicles (≥6 mm) increased from FSH (0.6±0.3) to CRRI (1.4±0.3; P<0.05), and day 1 (2.3±0.3; P<0.05), then declined by day 3 (0.6±0.3; P<0.05). There were more large follicles at CRRI ( P<0.05) with 68 mg (2.1±0.3) or 42 mg (1.6±0.2) than 0 mg (0.5±0.4) FSH. Ovulation rate (mean 2.12±0.05) increased with FSH given 12 h, but not 36 h before CRRI (dosage × time, P<0.05). Estrous response, conception rate, percentage of ewes lambing or prolificacy did not differ. However, number of corpora lutea not represented by embryos increased with dosage of FSH ( P<0.01; 0.25±0.14, 0.55±0.09, 0.71±0.09 for ewes treated with 0, 42, and 68 mg FSH, respectively).

Patterns of follicular growth in superovulated sheep and influence on endocrine and ovarian response.

Objectives of this study were to characterize patterns of follicular development in sheep superovulated with purified follicle stimulating hormone (FSH) (OVAGEN, ICP, Auckland, New Zealand) and to determine its influence on preovulatory events (onset of the oestrus behaviour and timing of the preovulatory luteinizing hormone surge) and ovarian response (ovulation rate and embryo yield). Number and size of all >/= 23 mm follicles from the first FSH injection to withdrawal of progestagen sponges was determined by transrectal ultrasonography just prior to every FSH injection in nine Manchega ewes superovulated with eight decreasing doses (ml) (1.5 x 3, 1.25 x 2 and 1 x 3) of OVAGEN injected twice daily from 60 h before to 24 h after the withdrawal of 40 mg fluorogestone acetate sponges. Oestrous detection and jugular blood sampling for LH radioimmunoassay were performed every 3 h from 14 to 53 h after sponge removal and ovulation rate and number of embryos were determined 4 days after progestagen withdrawal. Administration of OVAGEN induced a significant rise (p < 0.0005) in the number of follicles >/= 4 mm in size because of an increased growth in size of follicles from the first FSH injection to sponge removal, an increase in the number of newly detected follicles from 12 to 36 h of the first FSH dose (p < 0.005) and a decrease in regression rate from 24 h (p < 0.001). The number of follicles 2-3 mm in size at first FSH dose (10.4 +/- 1.5) was positively correlated with the number of >/= 4 mm follicles at 0 h (19.0 +/- 2.7, p < 0.01). A higher number of >/= 4 mm follicles at 0 h was related with an earlier appearance of oestrus (31.5 +/- 1.5 h, p = 0.08) and LH surge (45.0 +/- 2.3 h, p < 0.005), and a higher ovulation rate (18.2 +/- 3.8, p < 0.005). On the other hand, the rate of embryo recovery was decreased in ewes with earlier preovulatory LH peaks (p < 0.005), with a shorter interval between oestrus and LH peak (p < 0.05).

Premature follicle luteinization as a paracrine regulator of androstenedione (A4) in prenatally androgenized (PA) female rhesus monkeys receiving follicle-stimulating hormone (FSH) therapy for in vitro fertilization (IVF)

Objective: Early prenatal androgenization in female rhesus monkeys receiving FSH therapy for IVF induces premature follicle luteinization, reduces intrafollicular A4 and estradiol (E2) levels, and impairs blastocyst development (Dumesic et al. JCEM 2002;87:1111). Such premature follicle luteinization may reduce follicle fluid (FF) A4 levels by impairing paracrine regulation of theca A4 synthesis by granulosa cells, since granulosa cell-derived factors in rats enhance luteinizing hormone (LH)-responsive theca androgen production (Smyth et al. Endocrinology 1993;133:1532). The present study examines whether FSH-stimulated granulosa cells induce paracrine signalling of theca androgen production in female rhesus monkeys undergoing FSH therapy for IVF and, if so, whether such FSH-stimulated signalling is impaired by prenatal androgenization. Design: Prospective study. Materials/Methods: Five early-treated (E) and 4 late-treated (L) PA female (e.g., 15–35 days of testosterone (T) propionate exposure starting on gestational days 40–44 or 100–115, respectively) and 5 control (C) monkeys were studied. All females received recombinant (r) human (h) FSH injections (30 IU, IM twice daily) for 7–9 days (without human chorionic gonadotropin); oocyte retrieval was performed 1 day later. Serum FSH, LH, E2, progesterone (P4), A4, T and dihydrotestosterone (DHT) were measured basally, during rhFSH therapy and at retrieval. Sex steroids in FF also were analyzed. Results: Age, weight and serum FSH were similar in all female groups; serum LH bioactivity was unchanged during rhFSH therapy. Serum E2 were unaffected by female type and rose significantly during rhFSH therapy to reach maximal levels at retrieval. Serum P4 levels also were similar in all females and decreased on day 2 of rhFSH therapy before returning to basal values during rhFSH therapy and at retrieval. Serum A4, T and DHT levels were unaffected by female type. Serum A4 levels remained at basal values through day 4 of rhFSH therapy, and rose on day 6 of rhFSH therapy (P ≤0.025 vs day 4), reaching maximal levels at retrieval (P ≤0.01 vs day 6). Serum T and DHT also rose above basal values by day 6 of rhFSH therapy (P ≤0.005) and increased further at retrieval (P ≤0.01). The FF/serum P4 ratio was significantly higher in EPA females than in LPA or C females (P ≤0.025). Conversely, the FF/serum A4 ratio was significantly lower in EPA females than in LPA females or C females (P ≤0.01). FF/serum E2 and T ratios were similar in all female groups; FF/serum DHT levels were elevated in all PA females (P ≤0.025). While the number of oocytes retrieved and incidences of oocyte nuclear maturation and fertilization were similar among female types, 15.3% of zygotes developed to blastocysts in C females. No zygote developed to blastocyst in any PA female. Conclusions: Paracrine signalling of theca A4 production by granulosa cells occurs in female rhesus monkeys receiving rhFSH for IVF. Early prenatal androgenization impairs this phenomenon through premature follicle luteinization and reduces the amounts of aromatizable androgen and E2 in the follicle during oocyte developmental competence. Supported by: NIH grants RR14093/RR13635.

In vitro fertilization cycle medication administration: combining follicle-stimulating hormone with GnRH-agonist in a single injection.

Objective: In vitro fertilization (IVF) treatment routinely involves the administration of follicle-stimulating hormone (FSH) and a GnRH-agonist (Lupron; TAP Pharmaceuticals, Deerfield, IL, USA). These medications are usually given in two, separate subcutaneous injections. In an effort to simplify the medication regimen and reduce patient discomfort, we implemented a single, combined administration of FSH and Lupron (in one subcutaneous injection) for most IVF treatment cycles. We then compared the data to see if there were any differences in select outcome variables. Design: A retrospective review of all IVF cycles initiated between January 1, 1999 and April 30, 1999. The data obtained were compared using Chi-square analyses and Student’s t-test with GraphPad Prism software, San Diego, CA, USA. P < 0.05 was considered to be statistically significant. Materials/Methods: Five hundred twenty six cycles were performed using a single, combined injection of FSH and Lupron, and 180 cycles were performed using two, separate injections. Variables analyzed included patient age, peak estradiol measurement, number of treatment days, daily gonadotropin dose, number of follicles >14 mm on the day of hCG administration, number of embryos transferred, implantation rate, clinical pregnancy rate, and delivered pregnancy rate. Results: There were no statistically significant differences in combined v. separate injection groups for age (36.13 and 36.34 years, respectively, p = 0.57), peak estradiol (1502.98 and 1638.38 pg/ml, respectively, p = 0.10), daily gonadotropin dose (281.47 and 263.23 units, respectively, p = 0.08), number of follicles >14 mm on the day of hCG administration (6.56 and 6.29, respectively, p = 0.33), number of embryos transferred (2.81 and 2.97, respectively, p = 0.13), and implantation rate (15.60% and 14.31%, respectively, p = 0.50). There was a statistically significant difference in the number of treatment days (13.24 and 12.80, respectively, p = 0.02). This was not felt to be clinically significant. No statistically significant difference existed for the overall clinical pregnancy rate (27.95% and 28.33%, respectively, p = 0.92) or delivered pregnancy rate (24.52% and 22.22%, respectively, p = 0.53). Conclusions: Combined FSH and Lupron administration simplifies the IVF treatment regimen and reduces patient discomfort with no detrimental effect on treatment outcome.

Successful treatment of embryo implantation failure in patients with the Sjögren syndrome with low-dose prednisolone

Successful treatment of embryo implantation failure in patients with the Sjögren syndrome with low-dose prednisolone

Suppression by developing ovarian follicles of the low-dose endotoxin-induced glomerular inflammatory reaction in the pregnant rat

Objective: In the current study the role of developing ovarian follicles in the control of the endotoxin-induced pregnancy-specific inflammatory reaction was evaluated. Study Design: Follicular development was induced in pregnant rats (n = 20) by means of daily intraperitoneal injections of follicle-stimulating hormone from day 11 of pregnancy until the end of the experiment. Control pregnant rats (n = 20) received daily sodium chloride injections. All pregnant rats were infused for 1 hour with either 2 mL endotoxin solution (1.0 μg/kg body weight) or 2 mL sodium chloride solution on day 14 and killed 4 hours or 3 days later. At death, the left kidneys were snap-frozen and immunohistologically stained for the presence of polymorphonuclear leukocytes and monocytes. Results: The results show that in control pregnant rats endotoxin significantly increased glomerular polymorphonuclear leukocyte and monocyte numbers at both 4 hours and 3 days after endotoxin infusion. Induction of follicular development did not affect glomerular polymorphonuclear leukocyte number after endotoxin infusion but significantly decreased the number of monocytes in the glomeruli at both 4 hours and 3 days after endotoxin infusion. Conclusion: We conclude that follicles stimulated with follicle-stimulating hormone produce a follicular factor or factors that are able to prevent the endotoxin-induced influx of monocytes into the glomeruli of pregnant rats. It is suggested that these factors play a role in the control of inflammatory processes associated with reproduction, including the disease of pregnancy, preeclampsia. (Am J Obstet Gynecol 2000;183:89-93.)

Suppression by developing ovarian follicles of the low-dose endotoxin-induced glomerular inflammatory reaction in the pregnant rat

<h2>Abstract</h2> <b>Objective:</b> In the current study the role of developing ovarian follicles in the control of the endotoxin-induced pregnancy-specific inflammatory reaction was evaluated. <b>Study Design:</b> Follicular development was induced in pregnant rats (n = 20) by means of daily intraperitoneal injections of follicle-stimulating hormone from day 11 of pregnancy until the end of the experiment. Control pregnant rats (n = 20) received daily sodium chloride injections. All pregnant rats were infused for 1 hour with either 2 mL endotoxin solution (1.0 μg/kg body weight) or 2 mL sodium chloride solution on day 14 and killed 4 hours or 3 days later. At death, the left kidneys were snap-frozen and immunohistologically stained for the presence of polymorphonuclear leukocytes and monocytes. <b>Results:</b> The results show that in control pregnant rats endotoxin significantly increased glomerular polymorphonuclear leukocyte and monocyte numbers at both 4 hours and 3 days after endotoxin infusion. Induction of follicular development did not affect glomerular polymorphonuclear leukocyte number after endotoxin infusion but significantly decreased the number of monocytes in the glomeruli at both 4 hours and 3 days after endotoxin infusion. <b>Conclusion:</b> We conclude that follicles stimulated with follicle-stimulating hormone produce a follicular factor or factors that are able to prevent the endotoxin-induced influx of monocytes into the glomeruli of pregnant rats. It is suggested that these factors play a role in the control of inflammatory processes associated with reproduction, including the disease of pregnancy, preeclampsia. (Am J Obstet Gynecol 2000;183:89-93.)

Non-Surgical Transfer of Fresh or Frozen-Thawed Ovine Embryos by Laparoscopy.

The present study was conducted to examine survival of fresh and frozen-thawed ovine embryos after embryo transfer (ET) using laparoscopy or in vitro culture. Embryos were collected on Day 7.5 (Day 0 =sponge removal) from donor ewes superovulated by pre-treatment of progestogen-impregnated intravaginal sponges (FGA) for 12 days and a single injection of follicle stimulating hormone (FSH) and equine chorionic gonadotrophin (eCG), at 2 days and 1 day before sponge removal. The donor ewes showed estrus between 12 h and 30 h (the mean time: 15.6 h) after sponge removal. Embryo transfer of fresh or frozen-thawed embryos was carried out on Day 7.5 in 37 recipient ewes using laparoscopic technique. Frozen-thawed morulae or blastocysts were cultured to examine their viability in a modified synthetic oviduct fluid medium (SOFM) for 24 h. The lambing rates after ET with fresh and frozen-thawed embryos were 33.3% (6/18) and 26.3% (5/19), respectively. The developmental stage (morula or blastocyst) affected the survival rate of frozen-thawed embryos transferred (54.5% and 0%, respectively). The survival rate (development to blastocyst or hatched blastocyst) of frozen-thawed embryos after 24 h in culture was 40.9% (9/22). The present results indicate that ovine ET using laparoscopy is a useful technique. But, the lambing and survival rates of both fresh and frozen-thawed embryos were low, and remained to be improved.

A less acidic human follicle-stimulating hormone preparation induces tissue-type plasminogen activator enzyme activity earlier than a predominantly acidic analogue in phenobarbital-blocked pro-oestrous rats.

Follicle-stimulating hormone (FSH) exists in multiple molecular forms. In both experimental animals and in humans the production and secretion of less acidic, short-lived FSH glycoforms significantly increase during the peri-ovulatory period. To gain further insights on the physiological role of these FSH variants, we analysed the ability of two FSH compounds, recombinant FSH (rFSH) and purified FSH from urinary origin (uFSH), (less acidic and acidic pattern of FSH charge isoform distributors respectively) to induce ovarian tissue-type plasminogen activator (tPA) enzyme activity in vivo. FSH produced by Chinese hamster ovary cells and highly purified uFSH were injected at 15:00 h on the pro-oestrous day into phenobarbital-blocked rats and the ovaries were analysed for tPA enzyme activity and tPA mRNA concentrations at different times after FSH injection. Induction of tPA enzyme activity by uFSH and rFSH showed distinct dynamics depending on the particular preparation administered. In animals treated with uFSH, maximum tPA enzyme activity was detected at 20:00 h, and maximum tPA mRNA concentrations were detected at 17:00 h. tPA enzyme activity induction by rFSH was at the maximum at 17:00 h, and maximum tPA mRNA concentration was at 16:00 h (P< 0.05 for uFSH versus rFSH). All animals in the uFSH- and rFSH-treated groups and none in phenobarbital-blocked, saline-treated controls ovulated. No significant differences were present in the number of ova shed by rats treated with uFSH or rFSH and spontaneously ovulating rats (10.7+/-1.7, 10.0+/-2.6 and 11.3+/-1.6 respectively). These data indicate that the increased biological activity exhibited by less acidic FSH glycovariants at the target cell level may compensate for the drawback imposed by their relatively short plasma half-life.

A Simple Superovulation Method of A Single Injection of Follicle-Stimulating Hormone Combined with Equine Chorionic Gonadotropin for Superovulation of Suffolk Ewes During the Breeding Season: I. Effects of Different Treatments on the Endocrine Profiles.

Two trials were conducted to investigate a simple superovulation method for Suffolk ewes during the breeding season. The method involves a single injection of porcine follicle-stimulating hormone (pFSH) combined with equine chorionic gonadotropin (eCG) and was evaluated by measuring the concentrations of plasma hormones (estradiol-17β(E2), progesterone (P4), luteinizing hormone (LH) and follicle-stimulating hormone (FSH)). The ewes were treated with a vaginal sponge containing synthetic steroid with progestative action (P sponge) for 12 days in both trials. In Trial 1 (conducted in 1995), 18 ewes received a single injection (F1) or multiple injections (F6) of a total of 20 mg pFSH, and in combination with an injection of 500 IU eCG (F1+P, F6+P). The injections were conducted on Day -2 (Day 0 was the day of P sponge removal). In trial 2 (conducted in 1996), 24 ewes were given single injections of pFSH and eCG at different times; (i) both hormones on Day -2 (F-2/P-2), (ii) pFSH on Day -2 and eCG on Day -1 (F-2/P-1), and (iii) both hormones on Day -1 (F-1/P-1). One hundred micrograms of an analogue of gonadotropin-releasing hormone (GnRH) was administered on Day 1 in Trial 1 and at the detection of estrus in Trial 2. Frozen-thawed semen was inseminated in utero using a laparoscope at 30 h and 38 h after P sponge removal in Trial 1 and at 18 hours after the detection of estrus in Trial 2. Blood samples were collected from the jugular vein. E2, P4, LH and FSH were assayed by second-antibody enzyme immunoassays. F1+P showed similar endocrine profiles to those of F6 and F6+P with a preovulatory E2 peak after P sponge removal, but F1 did not show a clear preovulatory E2 peak. The E2 concentration in F-2/P-1 rose biphasically; there was a rise just after injection of pFSH and then another rise after P sponge removal. Simultaneous injections of 20 mg pFSH and 500 IU eCG may reduce the E2 elevation. We concluded that a good ovarian response comparable to the multiple injection method can be achieved by the present simple superovulation method.

A Simple Superovulation Method of A Single Injection of Follicle-Stimulating Hormone Combined with Equine Chorionic Gonadotropin for Superovulation of Suffolk Ewes During the Breeding Season: II. The Ovarian Responses and the Embryo Qualities.

Two trials were conducted to investigate the effects of a simple superovulation method using a single injection of porcine follicle-stimulating hormone (pFSH) combined with equine chorionic gonadotropin (eCG) on ovarian responses and embryo qualities in Suffolk ewes during the breeding season. The ewes were treated with a vaginal sponge containing synthetic progestative steroid (P sponge) for 12 days in both trials. In Trial 1 (conducted in 1995), 18 ewes received a single injection (F1) or multiple injections (F6) of a total of 20 mg pFSH, and in combination with an injection of 500 IU eCG (F1+P, F6+P). The injections were conducted on Day -2 (Day 0, was the day of P sponge removal). In trial 2 (conducted in 1996), 24 ewes were given single injections of pFSH and eCG at different times: (i) both hormones on Day -2 (F-2/P-2), (ii) pFSH on Day -2 and eCG on Day -1 (F-2/P-1), and (iii) both hormones on Day -1 (F-1/P-1). One hundred micrograms of an analogue of gonadotropin-releasing hormone (GnRH) was administered on Day 1 in Trial 1 and at the detection of estrus in Trial 2. Frozen-thawed semen was inseminated in utero using a laparoscope at 30 h and 38 h after P sponge removal in Trial 1 and at 18 h after the detection of estrus in Trial 2. Embryos were recovered on Day 7 or Day 8. In Trial 1, the mean number of ovulations and the rate of normal embryos were 6.8, 16.7% for F1, 15.0, 36.0% for F1+P, 15.5, 27.0% for F6 and 20.3, 44.2% for F6+P. The F1+P group showed the results comparable to those of F6 and F6+P. In Trial 2, the mean number of ovulations were 17.8 for F-2/P-1, 13.1 for F-2/P-2 and 12.4 for F-1/P-1. The rate of normal embryos in F-2/P-1 (63.0%) was significantly higher than in F-2/P-2 (41.1%, P<0.05) and tended to be higher than in F-1/P-1 (43.6%, P=0.050). The rate of freezable embryos in F-2/P-1 (56.2%) tended to be higher than in F-2/P-2 (39.3%, P=0.057) and in F-1/P-1 (38.5%, P=0.076). It is concluded that the simple superovulation method of a single injection of 20 mg pFSH combined with 500 IU eCG was as effective as multiple injctions of pFSH, and in combined with eCG on superovulation of Suffolk ewes during the breeding season. Administration of pFSH on Day -2 and eCG on Day -1 was most efficient to induce superovulation in Suffolk ewes.