The Use of Injectable Estradiol in Transgender and Gender Diverse Adults: A Scoping Review of Dose and Serum Estradiol Levels.

OBJECTIVE: Feminizing gender-affirming hormone therapy is the mainstay of treatment for many transgender and gender diverse people. Injectable estradiol preparations are recommended by the World Professional Association for Transgender Health Standards of Care 8 and the Endocrine Society guidelines. Many patients prefer this route of administration, but few studies have rigorously assessed optimal dosing or route. METHODS: We performed a scoping review of the available data on estradiol levels achieved with various dosages of estradiol injections in transgender and gender diverse adults on feminizing gender-affirming hormone therapy. We also report on testosterone suppression, route (ie, subcutaneous vs intramuscular), and type of injectable estradiol ester as well as timing of blood draw relative to the most recent dose, where available. RESULTS: The data we reviewed suggest that the current guidelines, which recommend starting doses 2 to 10 mg weekly or 5 to 30 mg every 2 weeks of estradiol cypionate or valerate, are too high and likely lead to patients having supraphysiologic levels across much of their injection cycle. CONCLUSIONS: The optimal starting dose for injectable estradiol remains unclear and whether it should differ for cypionate and valerate. Based on the data available, we suggest that clinicians start injectable estradiol cypionate or valerate via subcutaneous or intramuscular injections at a dose ≤5 mg weekly and then titrate accordingly to keep levels within guideline-recommended range. Future studies should assess timing of injections and subsequent levels more precisely across the injection cycle and between esters.

Estradiol cypionate inhibits proliferation and promotes apoptosis of gastric cancer by regulating AKT ubiquitination.

Gastric cancer is a common gastrointestinal malignancy worldwide, with a high mortality rate and poor prognosis. Multidrug resistance remains a major obstacle to successful treatment for patients. Hence, it is of great significance to develop novel therapies to potentiate the anti-tumor effect. In this study, we have investigated the effect of estradiol cypionate (ECP) on gastric cancer in vitro and vivo. Our data show that ECP inhibited the proliferation, promoted apoptosis, and caused G1/S phase arrest of gastric cancer cells. The mechanism by which ECP promoted apoptosis of gastric cancer cells was related to the downregulation of AKT protein expression caused by the increased ubiquitination modification levels of AKT, which finally inhibited the over-activation of the PI3K-AKT-mTOR signaling pathway. In vivo tumorigenesis experiments showed that ECP significantly inhibited the growth of gastric cancer cells, showing promise for clinical application. The above findings indicate that ECP inhibited the growth of gastric cancer and induced apoptosis through the PI3K /Akt/mTOR pathway. In summary, the efficacy showed in our data suggests that ECP is a promising anti-tumor compound for gastric cancer.

Effects of estradiol cypionate dosage and body condition on reproductive performance of Nellore cattle synchronized for timed-artificial insemination.

This experiment was designed to evaluate the effects of increasing doses of estradiol cypionate (ECP) and different body condition score (BCS) on reproductive performance of Bos indicus beef females assigned to a timed-artificial insemination (TAI) management. In this experiment, 1683 Bos indicus Nellore cows were blocked by parity and assigned to receive 1) an intravaginal P4 device (1.9 g of P4) and 2.0 mg of estradiol benzoate on day -11, 12.5 mg (i.m.) of dinoprost tromethamine, 300 IU (i.m.) of equine chorionic gonadotrophin, 0.6 mg (i.m.) of estradiol cypionate and CIDR withdrawal on day -2, followed by TAI on day 0 (n = 849; 0.6ECP) or 2) the same synchronization protocol with 1.0 mg of ECP on day -2 (n = 834; 1.0ECP). In both treatments, estrus expression was measured between days -2 and 0. Body condition score (BCS) was evaluated on days -11, 31, and 71 of the experiment and the BCS variation (Δ) was also determined between these timepoints. Transrectal ultrasonography was performed on days 31, 71, and 111 for pregnancy rate determination. All binary data were analyzed using cow as the experimental unit with GLIMMIX, whereas continuous variables were analyzed with the MIXED procedure of SAS. No treatment effects were observed on estrus expression rate. Treatment × BCS interactions were observed for pregnancy rates in all time points (days 31, 71, and 111), as 1.0ECP cows with a LOW BCS also had a greater P/AI than cows assigned to 0.6ECP. In summary, increasing the dose of ECP benefited the reproductive performance of Nellore beef cows with a reduced BCS (≤2.75), whereas no benefits were seen when the BCS was considered adequate (>2.75).

Effects of estradiol cypionate dose as an ovulatory stimulus on reproductive performance of lactating dairy cows during the summer season.

This experiment was designed to evaluate the effects of increasing doses of estradiol cypionate (ECP) on reproductive function of lactating dairy cows during the summer. In Exp. 1, 643 lactating Holstein cows were blocked by parity and assigned to receive 1) an intravaginal P4 device (1.9 g of P4) and 2.0 mg of estradiol benzoate on day -11, 25 mg (i.m.) of dinoprost tromethamine on day -4, 1.0 mg (i.m.) of estradiol cypionate and CIDR withdrawal on day -2, followed by TAI on day 0 (n = 326; ECP-1) or 2) the same synchronization protocol with 2.0 mg of ECP on day -2 (n = 317; ECP-2). In both treatments, cows were TAI on day 0 of the protocol, and cow rectal temperature was measured on days -2, 0, and 7. In Exp. 2, 608 lactating crossbred Holstein × Gir dairy cows were blocked by parity and enrolled to the same treatments as in Exp. 1, but on day 7, cows received one viable embryo into the uterine horn. In Exp. 1, a greater percentage of ECP-2 cows were detected on estrus (81.3 vs. 91.1%, respectively). A treatment × body condition score (BCS) interaction was observed on day 60 pregnancy per AI (P/AI), as ECP-2 cows with a BCS <2.75 had a greater P/AI vs. ECP-1, but an opposite result was observed in cows with a BCS ≥2.75. Regardless of treatment, there were effects of mean rectal temperature and heat stress events on P/AI. Treatment affected the diameter of the ovulatory follicle at TAI (ECP-1 = 15.3 mm vs. ECP-2 = 14.8 mm) and a greater percentage of ECP-1 cows had larger follicles (≥16.5 mm), but ECP-2 resulted in a greater incidence of early ovulatory cows (ovulating before day 0). Therefore, follicle diameter at TAI affected P/AI on day 60 in cows receiving ECP-2 and tended to affect ECP-1 cows. A treatment effect was observed on time to estrus following ECP treatments and a reduced proportion of ECP-2 cows showed estrus at TAI. Regardless of treatment, cows detected on estrus 48 h after ECP administration had a greater P/AI on day 60 vs. cows detected on estrus 24 h. In Exp. 2, a greater percentage of ECP-2 cows were detected on estrus (68.3 vs. 81.4%, respectively). In summary, a greater dose of ECP increased the percentage of animals expressing estrus, but it did not benefit the reproductive function of lactating dairy cows during the summer, regardless if animals were assigned to a TAI or timed-embryo transfer (TET) protocol.

Estrus Synchronization of Replacement Gilts Using Estradiol Cipionate and PGF2α and Its Effects on Reproductive Outcomes.

In this study, we evaluated the effectiveness of using estrogen-induced prolonged luteal function followed by prostaglandin F2 alpha (PGF2α) treatment to synchronize estrus in gilts. On day12 of the estrus cycle (D0 = first day of standing estrus), 52 gilts were assigned at random to two experimental groups: non-treated gilts (CON, n = 22), serving as controls, and prolonged luteal function group (CYP, n = 30), receiving a single treatment with 10 mg of estradiol cypionate intramuscularly Starting on day 12, blood samples were collected for estradiol and progesterone assays. Estrus detection started on day 17. Gilts from the CON group were inseminated at the onset of natural estrus. On day 28 CYP gilts were treated with PGF2α to induce luteolysis and inseminated at the onset of estrus. Gilts were slaughtered 5 d after the last insemination. A single treatment with estradiol cypionate prolonged luteal function in 90% of treated gilts. The duration of the estrous cycle was longer (p < 0.0001) for CYP gilts compared to CON gilts. CYP gilts showed synchronized estrus 3.96 ± 0.19 d after induction of luteolysis. The conception rate was similar (p = 0.10) for CON and CYP gilts. No difference was observed in the embryo recovery rate (p = 0.18) and total number of embryos per female (p = 0.06). The percentage of unfertilized oocytes, fragmented embryos and viable embryos was similar among females from CON and CYP groups (p > 0.05). The treatment of gilts with a single application of 10 mg of estradiol cypionate on day 12 of the estrous cycle was effective in prolonging luteal function and treatment with PGF2α resulted in synchronized estrus. Additionally, the synchronization protocol had no deleterious effect on fertility and embryonic development.

Treatment with estradiol cypionate at progesterone withdrawal reduces handling without compromising the pregnancy rate to timed-AI in buffalo.

The aim of this study was to determine if treatment with estradiol cypionate (EC) at the time of P4 withdrawal induced ovulation in a synchronization/timed-AI (TAI) protocol in buffalo. In Experiment 1, 56 buffaloes received an intravaginal P4 device (1.0 g) plus estradiol benzoate (EB, 2.0 mg im) on Day 0 (D0). On Day 9, the P4 device was removed and buffaloes were given PGF2α (0.53 mg im sodium cloprostenol) plus eCG (400 IU im). Buffaloes were then randomly allocated to one of two groups: Group GEC (n = 29), treated with EC (1.0 mg im) at P4 device removal; Group GEB (n = 27), treated with EB (1.0 mg im) 24 h after P4 device removal. Ovarian ultrasound was undertaken on: D0, to ascertain general ovarian status; D9 to D11 (every 24 h), to measure diameter of the largest follicle (LF) and follicular growth rate; D11 to D13 (every 12 h for 72 h), to determine the time of ovulation and ovulation rate. Following P4 device removal, Groups GEC and GEB had a similar follicular growth rate (0.9 ± 0.1 and 1.1 ± 0.1 mm/day, respectively; P = 0.15) and similar LF diameter on D11 (11.4 ± 0.6 and 12.5 ± 0.5 mm; P = 0.12). Groups GEC and GEB also had a similar diameter of the ovulatory follicle (13.0 ± 0.5 and 13.4 ± 0.6 mm; P = 0.52), interval from P4 device removal to ovulation (68.2 ± 2.8 and 71.1 ± 1.4 h; P = 0.41) and ovulation rate (62.1% and 70.4%; P = 0.44). In Experiment 2, 199 buffaloes were assigned to the two treatments in Experiment 1 (GEC, n = 100; GEB, n = 99). All animals underwent TAI 56 h after P4 device removal and pregnancy diagnosis was preformed on D41. The pregnancy rate was similar for Groups GEC and GEB (50.0 and 45.5%, respectively; P = 0.45). The findings indicate that treatment with EC at the time of P4 withdrawal induces ovulation and achieves the same pregnancy rate to TAI as treatment with EB 24 h after P4 removal. The use of EC requires one less handling which is highly important in facilitating practical adoption of TAI in assisted breeding and genetic improvement in buffalo.

Amphotericin B-loaded Eudragit RL100 nanoparticles coated with hyaluronic acid for the treatment of vulvovaginal candidiasis.

The treatment of vulvovaginal candidiasis (VVC) is based on oral and vaginal formulations which show limited effectiveness. In this study, amphotericin B-loaded Eudragit RL100 nanoparticles coated with hyaluronic acid (AMP EUD nanoparticles/HA) were developed to overcome the drawbacks of the conventional formulations. AMP EUD nanoparticles/HA were synthesized by nanoprecipitation, formulated by statistical experimental design, and characterized. AMP release from EUD nanoparticles/HA and its antifungal activity in a murine model of VVC were evaluated. Nanoparticles showed 147.6 ±â€¯16.7 nm of diameter, 0.301 ±â€¯0.09 of polydispersity index, - 29.9 ±â€¯3.76 mV of zeta potential, and 87.27 % of encapsulation efficiency. They released about 81 % of AMP in 96 h; and provided the elimination of 100 % of the vaginal fungal burden in 24 h. It was suggested that the AMP EUD nanoparticles/HA penetrated into the vaginal epithelium via CD44 receptors. These AMP EUD nanoparticles/HA represent a non-conventional vaginal formulation to improve the treatment of VVC.

Cortisol, progesterone, 17αOHprogesterone, and pregnenolone in foals born from mare's hormone-treated for experimentally induced ascending placentitis.

This study aimed to evaluate steroid hormones in foals born from mares treated for ascending placentitis with different combinations of trimethoprim-sulfamethoxazole (TMS), flunixin meglumine (FM), long-acting altrenogest (ALT) and estradiol cypionate (ECP) for ten consecutive days, starting two days after experimental induction of placentitis with Streptococcus zooepidemicus. Fourty-six pregnant mares and respective foals were assigned as healthy group (Control, n = 8) or treated groups as follows: TMS+FM (n = 8), TMS+FM+ALT (n = 8), TMS+FM+ALT+ECP (n = 6), TMS+FM+ECP (n = 6) and no treatment (NO TREAT n = 10). At delivery, foals were classified as high-risk or low-risk based on clinical and hematologic findings, and survival rates were recorded during the first week of life for comparisons across groups. Cortisol, progesterone, 17αOHprogesterone, and pregnenolone concentrations were determined via immunoassays in 31 of the 46 foals immediately after foaling (0 h), at 12, 24, 48 h, and seven days post-partum (168h). At birth, serum cortisol concentrations were higher in Control and TMS+FM+ECP foals than in remaining groups (p < 0.05). Foals in TMS+FM+ALT and TMS+FM groups had higher 17αOHprogesterone concentrations at 24 h and 48 h, respectively (p < 0.05). Pregnenolone concentrations were higher in TMS+FM than TMS+FM+ALT+ECP foals at 7 days (p < 0.05). High-risk and non-surviving foals had decreased concentrations of cortisol at parturition, but increased concentrations of progesterone from 0 h to 48 h. Pregnenolone and 17αOHprogesterone concentrations were increased and pregnenolone after 12 h in high-risk and non-surviving foals (p < 0.05). In conclusion, adding ECP to the treatment of experimentally-induced placentitis appears to improve foal viability and endocrine response. Cortisol and progestogen profiles were abnormal in high-risk and non-surviving foals, and those treated with ALT or TMS+FM only.

Quantification of estradiol cypionate in plasma by liquid chromatography coupled with tandem mass spectrometry: Application in a pharmacokinetic study in healthy female volunteers.

The combination of medroxyprogesterone acetate 25 mg + estradiol cypionate 5 mg is a highly effective, monthly injectable contraceptive. For the first time, this study presents the development and validation of a sensitive method for estradiol cypionate analysis in human plasma by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Aliquots (500 µL) of plasma were extracted with ethyl ether (100%) and derivatized with dansyl chloride. Its separation was performed on a Jones Chromatography Genesis C8 column and the quantification was performed with a mass spectrometer equipped with an electrospray interface operating in negative ion mode. The run time was 6 min and the calibration curve was linear over the range of 0.005-0.15 ng/mL. The method was applied to evaluate the pharmacokinetics of estradiol cypionate in plasma collected up to 1008 h (42 days) after a single intramuscular administration of 25 mg/mL medroxyprogesterone acetate +5 mg/mL estradiol cypionate to healthy female volunteers (n = 12). The estradiol cypionate maximum plasma concentration (Cmax) was 0.14 ± 0.08 ng/mL reached at 16.83 ± 21.07 h and the area under the plasma concentration versus time curve (AUC0-last) was 14.07 ± 6.32 ng.h/mL. Elimination half-life (t½), apparent volume of distribution (Vd/F), apparent clearance (CL/F) and mean residence time (MRT) were 89.65 ± 76.04 h, 28038 ± 9636 L, 49.02 ± 10.62 L/h and 576.05 ± 238.32 h, respectively, showing that the estradiol cypionate release from the administration site was prolonged and there was no drug accumulation.

Use of prostaglandin F2α as ovulatory stimulus for synchronizing dairy cattle.

The aim of this study was to evaluate if prostaglandin F2α (PGF) can be used to induce ovulation in a GnRH-progesterone based protocol. In Experiment 1 crossbred dairy cows (n=32) were synchronized with a progesterone-GnRH based protocol for seven days, where the luteolytic dose of 150µg PGF was given 24h prior progesterone device removal (CIDR). On Day 8 cows were separated into two groups to receive: 1) 2mL of Saline (Control Group, n=15) or 2) 150µg of PGF (PGF Group, n=17). Ovulation rate was higher in the PGF than Control group (100% vs 53.3%, P=0.001, Odds ratio=30.88). The percentage of cows that ovulated synchronously tended to be higher in the PGF than Control group (P=0.1, Odds ratio=9.6). Experiment 2 was performed in a cross-over (3×3) design. Crossbred dairy cows (n=25) received a CIDR for seven days and GnRH on Day 0. Seven days later 150µg of PGF was given and the progesterone device was removed, and 24h later cows were distributed into three groups to receive: 1) 2mL of Saline (Control Group, n=25), 2) 150µg of PGF (PGF Group, n=25) or 3) 1mg of ECP (ECP Group, n=23). Diameter of ovulatory follicle was larger in the PGF and Control than ECP Group (P=0.002, Effect size>4.0). Synchronized ovulation rate (between 72 and 96h after CIDR removal) tended to be higher in PGF group in Control group (P=0.1, Odds ratio=0.35). Results suggest that PGF is equally efficient to ECP to induce synchronized ovulation in dairy cows subjected to progesterone-GnRH based protocols.

A benign method for maintaining ovulatory estrogen levels in cycling rhesus macaques.

The goal of this study was to develop a relatively noninvasive technique for generating ovulatory estrogen levels in cycling females over extended periods of time. Eleven intact cycling rhesus macaques were given weekly injections of estradiol cypionate in an effort to obtain weekly levels which approximated ovulatory levels. A dose of 500 µg generated weekly estrogen values averaging 370 ± 18 pg/ml. This method is a satisfactory alternative to more traditional techniques. It involves no surgical procedures, it is relatively nonintrusive, it does not terminate a female's reproductive career, and it is not as time-consuming as daily injections. Female cycle state can be altered with a minimum of manipulation for studies involving the behavior of estrous females.

Effect of estradiol cypionate and amount of progesterone in the intravaginal device on synchronization of estrus, ovulation and on pregnancy rate in beef cows treated with FTAI based protocols.

Three experiments were conducted to evaluate the effect of estradiol cypionate (ECP) and amount of progesterone in the intravaginal device (PID) on synchronization of estrus and ovulation, follicular dynamics, luteal dynamics and function and on pregnancy rate in beef cows treated with fixed-time artificial insemination (FTAI) based protocols. In Experiment 1, we evaluated the synchronization of ovulation using 1mg of ECP at PID removal (day 8 after PID insertion) or 1mg of estradiol benzoate (EB) 24h later, in cows treated with 0.558 or 1g of progesterone (P4). The final subgroups were: 0.558g+ECP: n=10; 0.558g+EB: n=11; 1g+ECP: n=10; 1g+EB: n=10. Ovarian ultrasonic examinations were performed to detect the dominant follicle and ovulation. There was no effect of treatments on the diameter of dominant follicle at any time, and on the mean interval to estrus and to ovulation (P>0.05); however, ECP treated cows had scattered distribution of estrus (P<0.03) and ovulation (P<0.03). In Experiment 2, cows received the following treatments: 0.558gP4+ECP: n=52; 0.558gP4+EB: n=52; 1gP4+ECP: n=50; 1gP4+EB: n=52; and FTAI. Pregnancy rate did not differ (P>0.05) between progesterone content (0.558g: 52.9%, 55/104; 1g: 56.9%, 58/102) but differed between estradiol esters (P<0.05; ECP: 48.9%, 49/102; EB: 61.5%, 64/104). In Experiment 3, cows received: 0.558gP4+ECP: n=55; 0.558gP4+EB: n=53; 1gP4+ECP: n=54; 1gP4+EB: n=53; and FTAI. Pregnancy rate did not differ (P>0.05) between progesterone content (0.558g: 48.1%, 52/108; 1g: 53.3%, 57/107) and estradiol esters (ECP: 47.7%, 52/109; EB: 53.8%, 57/106). In conclusion, ECP administration at device removal and progesterone content of PID has no influence on the synchronization of estrus, follicular dynamics, luteal dynamics and function. However, ECP administration affected the synchronization of ovulation and pregnancy rate in non-suckling beef cows, but did not affected pregnancy rate in suckling beef cows. Future studies should evaluate the distribution of ovulations in suckling Bos taurus beef cows.

Development of fixed-time artificial insemination protocols for locally adapted Curraleiro Pé-Duro cows / Desenvolvimento de protocolos de inseminação artificial em tempo fixo em vacas da raça localmente adaptada Curraleiro Pé-Duro

Two experiments were conducted aiming to evaluate the effects of two ovulatory inducers (Exp.1) and equine chorionic gonadotropin (eCG; Exp.2) on follicular and luteal dynamics in a fixed-time AI (FTAI) protocol in locally adapted Curraleiro Pé-Duro cows. In Exp. 1 multiparous cows (n=12) received an intravaginal device containing 1g of progesterone (P4) for 8 days and 2mg of estradiol benzoate (EB) intramuscularly (IM) at device insertion (Day 0). At device removal (Day 8) 0.150mg of Sodium D-Cloprostenol was administered IM and the cows were randomly assigned to receive 1mg of EB (EB8) or 1mg of estradiol cypionate (EC8) IM, or to not receive any ovulatory inducer (Control). All the animals participated in all treatments (crossover). The interval from P4 removal to ovulation was shorter and less variable in the EB8 treatment group (P≤0.05). In Exp. 2 (crossover), multiparous cows (n=12) received the same hormonal treatment as the EB8 group in Exp.1. At device removal (Day 8) cows were randomly assigned to receive 300UI of eCG IM or to not receive eCG (Control). No difference was ascertained on follicular and luteal parameters in Exp. 2 (P>0.05). We concluded that EB can be used as the ovulatory inducer (Exp. 1) in a FTAI protocol in Curraleiro Pé-Duro cows. However, eCG (Exp. 2) was not able to stimulate follicular and luteal development. This result is probably due to the adaptive capacity of Curraleiro Pé-Duro cows that maintained a satisfactory body condition score even in dry and hot environments.(AU)

Foram realizados dois experimentos com o objetivo de avaliar o efeito de dois indutores da ovulação e da gonadotrofina coriônica equina (eCG) na dinâmica folicular e luteal, em um protocolo de inseminação artificial em tempo fixo (IATF) em vacas localmente adaptadas da raça Curraleiro Pé-Duro. No experimento 1, vacas pluríparas receberam um dispositivo intravaginal contendo 1g de progesterona (P4) durante oito dias e 2mg de benzoato de estradiol (BE) intramuscular (IM) no momento da inserção do dispositivo (dia zero). Na retirada do dispositivo (dia oito), as vacas receberam 0,150mg de D-cloprostenol sódico IM e foram separadas aleatoriamente para receber 1mg de BE IM (BE8) ou 1mg de cipionato de estradiol IM (CE8), ou nenhum indutor da ovulação (controle). Todos os animais participaram de todos os tratamentos (crossover). O intervalo entre a retirada da P4 e a ovulação foi menor e menos variável no tratamento BE8 (P≤0,05). O momento da ovulação foi mais precoce e mais concentrado nos animais do grupo BE 8. No experimento 2 (crossover), vacas pluríparas receberam o mesmo tratamento hormonal do grupo BE8 do experimento1. Na retirada do dispositivo (dia 8), as vacas foram separadas aleatoriamente para receberem 300UI de eCG IM, enquanto o controle não. Não houve diferença nos parâmetros foliculares e luteais avaliados no experimento 2 (P>0,05). Em conclusão, o BE pode ser utilizado como indutor da ovulação (experimento 1) em protocolos de IATF em vacas Curraleiras Pé-Duro. Entretanto, o eCG (experimento 2) não foi capaz de estimular o desenvolvimento folicular e luteal. Esse resultado é devido provavelmente à capacidade adaptativa das vacas Curraleiras Pé-Duro em manter uma condição corporal satisfatória mesmo em condições de clima seco e quente.(AU)