Comparison of follitropin beta administered by a pen device with follitropin beta administered by a conventional syringe in patients undergoing IVF-ET / 대한생식의학회지

OBJECTIVE: To compare the effectiveness and convenience of a pen device for the self-administration of follitropin beta with a conventional syringe delivering follitropin beta solution in patients undergoing IVF-ET. METHODS: GnRH agonist long protocol was used for controlled ovarian stimulation (COS) in all subjects. A total of 100 patients were randomized into the pen device group or the conventional syringe group on the first day of COS. Local tolerance reactions were assessed within 5 minutes, at 1 hour and at 3 hours after each injection. On the day of hCG injection, patients were asked to rate their overall pain and convenience experienced with self-injection on a visual anlaogue scale (VAS). RESULTS: There were no differences in patients' characteristics between the two groups. The duration of COS was significantly shorter in the pen device group than in the conventional syringe group. Patients included in the pen device group needed a significantly smaller amount of follitropin beta. However, no differences between the two groups were found in IVF results and pregnancy outcome. The incidence of local pain within 5 minutes, at 1 hour and at 3 hours after the injection was significantly lower in the pen device group. VAS scores indicated that injections using the pen device were significantly less painful and more convenient. CONCLUSION: The pen device for self-administration of follitropin beta is less painful, safer and more convenient for the patients, and can be more effective because of the shorter duration and smaller dose of follitropin beta when compared with the conventional syringe.

Comparison of follitropin beta administered by a pen device with follitropin beta administered by a conventional syringe in patients undergoing IVF-ET / 대한생식의학회지

OBJECTIVE: To compare the effectiveness and convenience of a pen device for the self-administration of follitropin beta with a conventional syringe delivering follitropin beta solution in patients undergoing IVF-ET. METHODS: GnRH agonist long protocol was used for controlled ovarian stimulation (COS) in all subjects. A total of 100 patients were randomized into the pen device group or the conventional syringe group on the first day of COS. Local tolerance reactions were assessed within 5 minutes, at 1 hour and at 3 hours after each injection. On the day of hCG injection, patients were asked to rate their overall pain and convenience experienced with self-injection on a visual anlaogue scale (VAS). RESULTS: There were no differences in patients' characteristics between the two groups. The duration of COS was significantly shorter in the pen device group than in the conventional syringe group. Patients included in the pen device group needed a significantly smaller amount of follitropin beta. However, no differences between the two groups were found in IVF results and pregnancy outcome. The incidence of local pain within 5 minutes, at 1 hour and at 3 hours after the injection was significantly lower in the pen device group. VAS scores indicated that injections using the pen device were significantly less painful and more convenient. CONCLUSION: The pen device for self-administration of follitropin beta is less painful, safer and more convenient for the patients, and can be more effective because of the shorter duration and smaller dose of follitropin beta when compared with the conventional syringe.

A prospective and randomized trial comparing 225IU and 300 IU follitropin-α in a fixed-dose regimen for controlled ovarian stimulation / Estudo prospectivo e randomizado comparativo de 225 UI e 300 UI defolitropina-α em regime de dose fixa para estimulação ovariana controlada

Purpose: To compare the outcomes of 225 IU and 300 IU follitropin-α in a fi xed-dose regimen for controlled ovarian stimulation in women ≥35 years old. Material and methods: We studied 120 normo-ovulatory women ≥35 years old, undergoing IVF or ICSI cycles. After pituitary suppression, patients were randomly divided into two groups: G225 and G300. In G225 (n=60), ovarian stimulation was performed with a fixed daily dose of 225 IU of follitropin-α and in G300 (n=60), with a fixed daily dose of 300 IU, until hCG administration. The main outcomes were: the number of metaphase II oocytes retrieved, the percentage of MII oocytes, the cancellation rates, the number of days of stimulation and the fertilization rates. Data were analyzed statistically by the χ2 and Mann-Whitney tests, as p<0.05 was considered significant. Results: In G225, six cycles were cancelled (10%) and in G300, five cycles were cancelled(8.3%). The cancellation rates did not present statistical differences between groups (p>0.05). In G225, 301 oocytes were retrieved (5.02±1.32 per cycle); 261 were at MII stage. In G300, 338 o ocytes were collected (mean: 5.63±1.68 per cycle); 300 were at MII stage (p<0.05). The percentage of MII oocytes (86.7% in G225 versus 88.7% in G300), fertilization rate (69.7% in G225 versus 72.7% in G300), and the mean number of days of stimulation (9.7±0.6 in G225 versus 9.7±0.7 in G300) were not statistically different in both groups (p>0.05). Conclusions: We conclude that the dose of 225 IU r-FSH, rather than 300 IU, may be the dose of choice for ovarian stimulation in a fixed-dose regimen in this group of patients.

Objetivo: Comparar os resultados do uso de 250 UI e 300 UI de folitropina-α em regime de dose fixa em pacientes ≥35 anos de idade. Material e métodos: Foram estudadas 120 pacientes normo-ovulatórias submetidas a ciclos de fertilização in vitro ou injeção intracitoplasmática de espermatozoide. Apósa supressão hipofisária, as pacientes foram randomizadas nos grupos G225 e G300. No G225 (n=60), a estimulação ovariana foi realizada com 225 UI defolitropina-α e no G300 (n=60) com 300 UI, em regime de dose fixa até o dia do exame de hCG. Os resultados observados foram o número e porcentagem de oócitos em metáfase II coletados, taxa de cancelamento, número de dias de estímulo e taxa de fertilização. A análise estatística foi feita pelos testes χ2 e Mann-Whitney, considerando-se significante p<0,05. Resultados: No G225, houve seis ciclos cancelados (10%) e no G300 cinco (8,3%) (p>0.05). No G225, foram coletados 301 oócitos (5,02±1,32 por ciclo); 261 eram MII. No G300, foram coletados 338 oócitos (5,63±1,68 por ciclo), sendo 300 MII. A recuperação de oócitos MII (86,7% no G225 versus 88,7% no G300), as taxas de fertilização (69,7 versus 72,7%) e o número médio de dias de estimulação(9,7±0,6 versus 9,7±0,7) não foram estatisticamente diferentes entre os grupos (p>0,05). Conclusões: Concluímos que a dose de 225 UI de r-FSH podeser a dose de escolha para estimulação ovariana em regime de dose fixa nesse grupo de pacientes.

[Isolation and cloning of the beta subunit of human follicle stimulating hormone [hFSH beta] with its native gene's signal sequence in methylotroph yeast Pichia pastoris pPIC9 shuttle vector]

Follicle Stimulating Hormone [FSH] is one of the pituitary glycoproteines that it consists of two subunits; alpha and beta. The beta subunit is responsible for the biological activity of FSH. The aim of present study was isolation of the beta subunit coding sequence containing its signal sequence from human genome and then cloning of the isolated sequence in pPIC9 shuttle vector under the control of AOX1 promoter and ? factor signal sequence. the gene sequence of interest was isolated as a 2kb DNA fragment and cloned in pTZ57R vector resulting to pTV-2019 plasmid. The construct was used as template for modification of 5? region of gene upstream to ATG codon using PCR. Finally, amplicon was cloned in pPIC9 and the new construct named pPIC9F1. The sequence of FSH beta gene in pTV-2019 was confirmed by restriction analysis and DNA sequencing. In addition, restriction analysis and AOX1 primer-mediated PCR showed that pPIC9F1 has correct construction. The new construct, pPIC9F1, contains the coding sequence of FSH beta gene and its signal sequence [E2-IVS2-E3]. Therefore, this construct can be used for integration of FSH beta gene into yeast genome exactly downstream to AOX1 promoter. Under this condition, a fusion protein is produced that it contains two signal peptides, ? factor and FSH signal peptides. Yeast expression system is able to cleavage ? factor. It seems this is the first attempt for cloning of human FSH beta in yeast expression system

Comparison of two different fixed doses of follitropin-beta in controlled ovarian hyperstimulation: A prospective randomized, double blind clinical trial.

A prospective randomized, double blind, single centre study was conducted to compare the efficacy, efficiency and clinical side effects of daily fixed dose regimen of either 100 IU or 200 IU of recombinant follicle stimulating hormone(rFSH) Follitropin beta in down-regulated women undergoing controlled ovarian hyperstimulation(COH) for either conventional in vitro fertilization(IVF) or intracytoplasmic sperm injection(ICSI). A total of sixty women were randomly allocated according to the criteria for the treatment by either 100 IU(n = 30) or 200 IU (n = 30) of FSH. Although more cycle cancellations due to low response were observed in the 100 IU group (n = 9 vs n = 2), two cases of mild and moderate ovarian hyperstimulation syndrome were noted in the higher dose group. Subjects in the group treated with 200 IU appeared to yield more follicles > 17 mm (4.4 vs 3.3, p = 0.05) and more oocytes compared to the group treated with 100 IU (9.2 versus 6.0 oocytes, NS). The total dosage required to develop at least three follicles according to the protocol was significantly lower in the group treated with 100 IU (1203.33 versus 2106. 67, P < 0.0001). In conclusion, a fixed daily dose of 200 IU of rFSH Follitropin beta compared to a fixed daily dose of 100 IU is more effective in terms of follicles > 17 mm development and the number of oocytes retrieved along with a lower cancellation rate, but less efficient as indicated by a higher total rFSH dose needed

Continuous measurement of urine beta-FSH excretion in men with hypogonadism / 中华男科学杂志

<p><b>OBJECTIVES</b>To measure continuously the urine beta-FSH excretion in the patients with male hypogonadism, and to evaluate the significance of urine beta-FSH when used in the clinical practice and pathophysiological study on male hypogonadism.</p><p><b>METHODS</b>Four health male volunteers (aged 19, 22, 27 and 33 years), four patients with the hypogonadotropic hypogonadism (aged 17, 17, 19 and 24 years) and five patients with idiopathy hypogonadism (hypergonadotropic, aged 16, 16, 17, 20 and 22 years) were asked to collect their morning-first urine samples for 30 to 32 days. One normal men collected his urine samples for 63 days. The urine beta-FSH was assayed with the method of EIA, then corrected by creatinine (Cr) concentration.</p><p><b>RESULTS</b>The urine beta-FSH level of normal men was (1.16 +/- 0.20) micrograms/mg Cr, with the peak variation in their curves, peak level at 2.76 micrograms/mg Cr. The levels of urine beta-FSH of 4 patients with the hypogonadotropic hypogonadism were lower significantly than those of normal men [(0.58 +/- 0.31) (0.93 +/- 0.47) (0.47 +/- 0.33) and (0.60 +/- 0.40) micrograms/mg Cr], without fluctuation in their curves. beta-FSH levels of 5 patients with idiopathy hypogonadism were higher significantly [(3.02 +/- 0.93), (4.36 +/- 1.12), (4.79 +/- 0.78), (4.64 +/- 1.42) and (3.88 +/- 1.42) micrograms/mg Cr], with irregular fluctuation, the highest peak level at 6.83 micrograms/mg Cr. The second sexual characteristics of hypogonadal patients were poor and serum testosterone levels low.</p><p><b>CONCLUSIONS</b>The urine beta-FSH level raised with irregular fluctuation in patients with idiopathy hypogonadism, while lowed without any fluctuation in patients with the hypogonadism. These findings suggested that the urine beta-FSH excretion was useful for the clinically classified diagnoses and pathophysiological study on male hypogonadism, and for observing the treatment reaction of androgen replacement.</p>

Regulation of FSH Gene Expression and Release in Cultured Rat Anterior Pituitary Cells / 대한내분비학회지

BACKGROUND: FSH is a heterodimeric glycoprotein and is composed of alpha and beta subunits. alpha subunit is common to FSH and LH, while an unique beta subunit determines the biological specificity of each hormone. The synthesis of beta subunit is the primary rate-limiting step in the synthesis of each hormone. Although FSH plays a pivotal role in folliculogenesis and ovulation, very little studies have been performed on the regulation of FSH beta gene expression. Therefore, the present study attempted to examine the effect of GnRH or activin on the expression of FSH beta mRNA as well as FSH release and signaling pathway involved in their actions. METHODS: The primary cultures of rat anterior pituitary were used for this study. To determine FSH beta mRNA levels, northern blotting method was used. The concentration of FSH in the culture medium was evaluated by using a specific radioimmunoassay for rat FSH. RESULTS: PMA, an activator of PKC, increased FSH beta mRNA levels and FSH release, whereas forskolin, an activator of adenylate cyclase, showed no effect. The application of GnRH augmented FSH release, but not FSH beta mRNA levels. However, the administration of activin increased FSH beta mRNA levels as well as FSH release. Staurosporine, an inhibitor of PKC, suppressed activin-induced increment of FSH beta mRNA levels and FSH release. CONCLUSION: The present study demonstrated that activin rather than GnRH is a major regulator for FSH beta mRNA expression, and suggest that PKC-dependent pathway is also involved in the action of activin on the expression of FSH beta mRNA and FSH release.

The Effect of Ovariectomy on The Secretion of Follicle Stimulating Hormone ( FSH ) and mRNA Levels of FSHbeta subunit in Rat / 대한산부인과학회잡지

Follicle stmulating hormone ( FSH ) consist of alpha and beta subunits, which are encoded by se-parate genes. Pituitary release of FSH appears to be regulated by the hypothalamic GnRH and the gonadal steroid hormones. In addition, inhibin and follistatin produced by the gonad have been known to inhibit FSH secretion selectively. However, little is known about their regulation of the biosynthesis of FSH subunits at transcriptional and posttranscriptional levels. In the pre-sent study, we studied the time course of changes in alpha and FSH beta subunit mRNA concentrati-ons after castration and the effects of ovarian steroids of changes in alpha and FSH beta subunit mRNA concentrations after castration and the effects of ovarian steroids on alpha and FSH beta subunit mRNA in ovariectomized rats in order to determine Whether FSH subunit synthesis is modulated at the pretranslational levels, and whether synthesis and secretion are differently regulated. Results are as follows : 1. The time course of the rise in the steady state alpha subunit and FSH beta subunit mRNA levels were observed after ovariectomy, which paralleled the increases in serum and pituitary FSH concentrations. The time course experiments revealed differences in the patterns of alpha and FSH beta subunit mRNA responses, the rise in FSH beta subunit mRNA levels being more pro- minent than the rise in alpha subunit mRNA. 2. FSH beta mRNA levels were negatively regulated by the single injection of progesterone but not by estradiol, suggesting that FSH beta subunit mRNA seemed to be more sensitive to ne-gative feedback by progesterone than estradiol. Similar results were obtained by the continuous treatment of ovarian steroids for 1~4 days, but inhibition was more prominent with continuous treatment. It is, therefore, concluded that estradiol and progesterone inhibit the synthesis of FSH at the pretranslational level by modulating the steady state levels of alpha and FSH beta subunit mRNA, progesterone effect being more promiment than that of estradiol and alpha and FSH beta subunit are regulated in a different manner.