Transdermal 17-β estradiol or placebo patches were distributed to the subjects in a closed non-transparent envelope, and subjects were instructed to place a patch on the skin on the lower part of the abdominal region at the start of the intervention. The subjects were instructed to renew the patch twice a week. The ET patches were releasing 100 μg 17-β-estradiol per 24 h (Vivelle dot, Novartis Healthcare, Copenhagen, Denmark). The placebo patches (Matas blister patch, size small, Allerød, Denmark) and the ET patches both had a neutral, unlabeled appearance. Participants in the ET group were offered 10 days of treatment with progesterone (Provera, Pfizer, United States, 10 mg per day) after the intervention.
Provera
Manufactured by Pfizer
Sourced in United States, Italy
Provera is a laboratory diagnostic instrument designed for automated immunoassay testing. It is capable of performing various types of immunoassays, including enzymatic, chemiluminescent, and fluorescent. The core function of Provera is to provide accurate and reliable results for the detection and quantification of analytes in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cetrotide, by Merck Group (2 mentions)
Progynova, by Bayer (1 mentions)
Decapeptyl, by Ferring (1 mentions)
Menopur, by Ferring (1 mentions)
Pregnyl, by Merck Group (1 mentions)
Puregon, by Merck Group (1 mentions)
Cetrorelix, by Merck Group (1 mentions)
Gonal f, by Merck Group (1 mentions)
Pergoveris, by Merck Group (1 mentions)
Ganirelix, by Organon (1 mentions)
Insulin like growth factor 1 igf 1, by Thermo Fisher Scientific (1 mentions)
Insulin transferrin selenium mix, by Merck Group (1 mentions)
Deoxyribonuclease dnase 1, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Mccoy s 5a medium, by Thermo Fisher Scientific (1 mentions)
Percoll, by Merck Group (1 mentions)
Medium 199, by Thermo Fisher Scientific (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Collagenase type 1, by Worthington (1 mentions)
Estradiol valerate, by Bayer (1 mentions)
9 protocols using provera
1
Transdermal Estrogen Therapy Trial
2
Estradiol Valerate Dosing Protocol for Puberty Induction
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The drug administration method was applied in the same manner for all patients. First, estradiol valerate (Progynova®, Bayer Schering Pharma AG, Germany) 2 mg was administered for 6 months. Thereafter, the dose of estradiol valerate was increased by 2 mg every 2 months and up to a maximum dose of 12 mg/day. If the change in height was <1 cm, the estradiol dose was maintained. If no change in height was observed for 6 months (three consecutive outpatient visits), patients underwent radiographic assessment to evaluate fusion of the epiphyseal plate. When bone maturation was completed, the dose of estradiol valerate was decreased by 2 mg every 2 months. And eventually, patients stopped taking estradiol valerate. To protect endometrium, progesterone withdrawal bleeding was induced. Six months after initiating estradiol valerate treatment at 4 mg or if breakthrough bleeding occurred in the first 6 months of treatment, medroxyprogesterone acetate (Provera®, Pfizer, USA) 10 mg/day was administered from day 1 to 12 of each month.
3
Diverse COH Protocols for Infertility
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The COH protocol utilized in this study involved a standard GnRH agonist long protocol using leuprolide acetate (Lupron®; Takeda Chemical Industries, Tokyo, Japan), a standard GnRH antagonist protocol using cetrorelix acetate (Cetrotide®; Merck-Serono, Darmstadt, Germany), or progestin-primed ovarian stimulation (PPOS) with medroxyprogesterone acetate (Provera®; Pfizer, Milan, Italy). The choice of protocol was based on the physicians’ preference and aligned with our previous report [29 (link)]. Follitropin alfa (Gonal-f®; Merck-Sereno, Darmstadt, Germany) and/or hMG (Menopur®; Ferring, Pymble, NSW, Australia) were administered for COH starting from cycle day 3 until more than two dominant follicles reached a threshold diameter (≥17 mm). To trigger final oocyte maturation, recombinant human chorionic gonadotropin (hCG) (Ovidrel®; Merck-Sereno, Darmstadt, Germany) and/or triptorelin (Decapeptyl®; Ferring, Kiel, Germany) were injected. Ovum pick-up was performed approximately 36 h after the trigger injection.
4
Ovarian Antral Follicle Counting Protocol
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Transvaginal or transrectal sonographic examination was performed in the early follicular phase between days 2 and 4 of the menstrual cycle after spontaneous bleeding or withdrawal bleeding induced by medroxyprogesterone acetate Provera (Pfizer Inc., New York, NY, USA; 10 mg/day for 7 days) by using a Voluson S7 (GE Ultrasound Korea, Seongnam, Korea) equipped with a 7-MHz transvaginal transducer. For each ovary, the total number of visible antral follicles measuring 2 to 9 mm in diameter was counted using the continuous scanning method for both ovaries from the inner to outer margins in a longitudinal cross-section. The ovarian volume was calculated using the simplified formula for a prolate ellipsoid (0.5×length×width×thickness) [2 ]. The total ovarian volume was defined as the sum of both ovarian volumes, and the total antral follicle count was defined as the sum of both antral follicle counts.
5
Monza-Protocol for Ovarian Stimulation and Retrieval
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Cycle monitoring and oocyte retrieval were based on
the ‘Monza-protocol’ (7 (link)). At the start of the treatment, a
baseline ultrasound was performed on cycle day 2, 3 or
4. In patients with severe oligomenorrhoea (cycle length
>42 days) or amenorrhoea, a withdrawal bleeding was induced
with 7 days 10 mg progestagen (Provera®, Pfizer)
orally. The cycle was excluded if at baseline ultrasound
the endometrial thickness exceeded 4 mm or an ovarian
cyst larger than 12 mm was present.
Subsequently, ovarian priming was performed by the
administration of 150 IU rFSH s.c. (Puregon®, Merck
Sharp & Dohme) on cycle day 3 to 5. The second ultrasound
was scheduled at cycle day 6 to 8 and thereafter
at one- or two-day intervals until the identification of a
dominant follicle. A dominant follicle was defined as a
follicle that had grown to at least 8 mm (but not larger
than 12 mm) accompanied by a thickening of the endometrial
lining to 5 mm or more (14 (link)). Subsequently, 10.000
IU of hCG (Pregnyl®, Merck Sharp & Dohme) were administered
subcutaneously and the oocyte retrieval was
scheduled 38 hours later (15 (link)). A cycle was cancelled when
there was a follicle larger than 14 mm or an endometrium
less than 5 mm. Oocytes were retrieved by transvaginal
ultrasound-guided needle aspiration at a vacuum pressure
of 80-100 mmHg with a 16 gauge needle (Origio).
the ‘Monza-protocol’ (7 (link)). At the start of the treatment, a
baseline ultrasound was performed on cycle day 2, 3 or
4. In patients with severe oligomenorrhoea (cycle length
>42 days) or amenorrhoea, a withdrawal bleeding was induced
with 7 days 10 mg progestagen (Provera®, Pfizer)
orally. The cycle was excluded if at baseline ultrasound
the endometrial thickness exceeded 4 mm or an ovarian
cyst larger than 12 mm was present.
Subsequently, ovarian priming was performed by the
administration of 150 IU rFSH s.c. (Puregon®, Merck
Sharp & Dohme) on cycle day 3 to 5. The second ultrasound
was scheduled at cycle day 6 to 8 and thereafter
at one- or two-day intervals until the identification of a
dominant follicle. A dominant follicle was defined as a
follicle that had grown to at least 8 mm (but not larger
than 12 mm) accompanied by a thickening of the endometrial
lining to 5 mm or more (14 (link)). Subsequently, 10.000
IU of hCG (Pregnyl®, Merck Sharp & Dohme) were administered
subcutaneously and the oocyte retrieval was
scheduled 38 hours later (15 (link)). A cycle was cancelled when
there was a follicle larger than 14 mm or an endometrium
less than 5 mm. Oocytes were retrieved by transvaginal
ultrasound-guided needle aspiration at a vacuum pressure
of 80-100 mmHg with a 16 gauge needle (Origio).
6
Baboon Pharmacological Intervention Trial
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Immediately before lap-3, all baboons were randomized into one of the following four groups with the use of a sealed envelope system. The calendar dates of lap-3 are presented in Table 1.
Group 1, placebo (n ¼ 5; negative control): piece of banana orally once a day. Group 2, JNKI (n ¼ 5): JNKI AS602801 (bulk b/n 46517-04; molecular weight ¼ 457.55), 20 mg/kg once a day orally as a powder mixed with orange or pineapple juice administered 4 hours before feeding. Group 3, JNKI þ MPA (n ¼ 5): JNKI AS602801 (20 mg/kg once a day orally) þ MPA (Provera; Pfizer; 10 mg once a day orally) as a powder mixed with orange or pineapple juice administered 4 hours before feeding. Group 4, cetrorelix (n ¼ 5; positive control): GnRH antagonist cetrorelix acetate (Cetrotide; Merck Serono), 3 mg subcutaneously every 3 days.
Each baboon received her medication every day as specified above for a total of 60 days.
Group 1, placebo (n ¼ 5; negative control): piece of banana orally once a day. Group 2, JNKI (n ¼ 5): JNKI AS602801 (bulk b/n 46517-04; molecular weight ¼ 457.55), 20 mg/kg once a day orally as a powder mixed with orange or pineapple juice administered 4 hours before feeding. Group 3, JNKI þ MPA (n ¼ 5): JNKI AS602801 (20 mg/kg once a day orally) þ MPA (Provera; Pfizer; 10 mg once a day orally) as a powder mixed with orange or pineapple juice administered 4 hours before feeding. Group 4, cetrorelix (n ¼ 5; positive control): GnRH antagonist cetrorelix acetate (Cetrotide; Merck Serono), 3 mg subcutaneously every 3 days.
Each baboon received her medication every day as specified above for a total of 60 days.
7
Comparison of Gonadotropin Regimens for IVF
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The participants under 38 years of age received rFSH (Follitrope®: LG Chem, Jeollabuk-do, Korea, or Gonal F®: Merck-Serono, Geneva, Switzerland, or Puregon®; Organon Oss, Netherlands), while the other group used rFSH + hp-hMG (Menopur®: Ferring Pharmaceutical, Copenhagen, Denmark, or Follitrope® plus IVF-M®; LG, Jeollabuk-do, Korea), or rFSH + rLH (Gonal F® plus Pergoveris®: Merck-Serono, Geneva, Switzerland) for ovarian stimulation. Cetrorelix (Cetrotide®; Merck-Serono, Geneva, Switzerland), Ganirelix (Orgalutran: Organon®, Oss, Netherlands), progesterone (Utrogestan®; Besins Healthcare by OLIC, Ayutthaya, Thailand), or medroxyprogesterone acetate (Provera® Pfizer Italia S.r.l., Ascoli, Italy) were used for pituitary gonadotropin suppression. In each case, ovulation was induced with a single trigger (recombinant HCG; Ovidrel®; Organon Oss, Netherlands, or gonadotropin-releasing hormone agonist (GnRHa); Decapeptyl®: Ferring Pharmaceutical, Copenhagen, Denmark) or dual trigger (Ovidrel®; Organon Oss, Netherlands, plus Decapeptyl®: Ferring Pharmaceutical, Copenhagen, Denmark). The oocytes were retrieved transvaginally using ultrasound guidance. The data are presented in Table 1 .
8
Ovarian cell culture protocol
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Medium 199, McCoy’s 5A medium, and fetal bovine serum (FBS) were purchased from Gibco BRL (Grand Island, NY, USA). Percoll, insulin-transferrin-selenium mix, and deoxyribonuclease (DNase) I were obtained from Sigma-Aldrich (St. Louis, MO). Collagenase type 1 was purchased from Worthington (Lakewood, NJ), and insulin-like growth factor–I (IGF-I) was provided by PeproTech (Rocky Hill, NJ). Estradiol valerate was obtained from Bayer (Progynova; Bayer, Leverkusen, Germany), and progesterone was purchased from Pfizer (Provera; Pfizer Inc., NY, USA). Primary antibodies for PCNA and cyclin D1 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against FSH-R, LH-R, CYP17A1, and CYP19 were obtained from Abcam (Cambridge, UK). β-Actin was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Sex steroid (estrogen and progesterone) enzyme-linked immunosorbent assay (ELISA) kits were provided by Enzo, Life Sciences (Plymouth Meeting, PA, USA). The FSH and LH ELISA kits were purchased from Tsz Biosciences (San Francisco, CA, USA). The vendors for other chemicals and reagents are denoted in the corresponding sections of Methods.
9
Baseline Hormone Assays in Amenorrhoeic Women
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Baseline hormone assays were performed using blood samples collected on day 2 or 3 of the menstrual cycle. In amenorrhoeic women, blood samples were collected after withdrawal bleeding induced by oral progestin (10 mg of Provera for 10 days, Pfizer Korea, Seoul, Korea). Basal serum levels of AMH, FSH, LH, estradiol (E2), thyroid stimulating hormone, prolactin, total testosterone, and dehydroepiandrosterone sulfate (DHEA-S) were measured. Serum AMH concentrations were measured using a two-site enzyme immunoassay (AMH/MIS ELISA kit, Immunotech, Marseille, France), with a sensitivity of 0.14 ng/mL, and intra- and interassay coefficients of variation of 12.3% and 14.2%, respectively.
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