Estradiol Cypionate Research Articles

Reproductive outcomes of lactating dairy cows submitted to first timed artificial insemination protocols with different strategies to induce final ovulation.

The study evaluated strategies for induction of ovulation at the end of timed AI (TAI) protocols initiated after a novel presynchronization strategy. A total of 909 lactating dairy cows from 6 dairy herds initiated a presynchronization protocol on d -15 with an intravaginal progesterone (P4) implant and 7 d later (d -8) were treated with 1.0 mg of estradiol cypionate (EC) and 0.530 mg of cloprostenol sodium (a synthetic analog of prostaglandin F2α; PGF), simultaneously with the P4 implant removal. On d 0, the protocol for synchronization of ovulation was initiated with 16.8 µg of buserelin acetate (a synthetic analog of GnRH) and insertion of a 2.0 g P4 implant, followed by a treatment with 0.530 mg of PGF on d 6, and a second PGF on d 7, concomitant with the P4 implant withdrawal. In Group EC, cows received 1.0 mg of EC on d 7 as an ovulation inducer. In G group, cows received 8.4 µg of GnRH 56 h after the first PGF (16 h before TAI). In Group EC/G, cows received both EC and GnRH. The TAI was performed on d 9 (48 h after P4 withdrawal) in all experimental treatments. There were no differences among treatments on pregnancy per AI (P/AI) on d 31 (40.4%; 367/909). There was a tendency for greater pregnancy loss in G group (19.8%) than in EC (12.2%) and EC/G (10.1%) groups. When the 2 groups that received EC were combined, the pregnancy loss was lower than when cows received only GnRH (11.2% [21/188] vs. 19.8% [17/86]; P = 0.05). There was an interaction between treatment and expression of estrus on P/AI, in which estrus positively affected P/AI in EC (53.3% vs. 20.0%) and EC/G (53.1% vs. 36.7%), but not in G group (41.5% vs. 38.7%). Overall, 81% of the cows had a corpus luteum (CL) on d 0 and 91% at PGF treatment of the breeding protocols. Fertility was greater in cows with CL at the time of the first PGF treatment than in cows without CL (45.9% vs. 17.7%). In summary, the strategies to induce final ovulation resulted in similar fertility despite the different physiological responses, such as expression of estrus, but further research is needed to definitively confirm the relationship between lack of EC and increased pregnancy loss.

Estrogens impair hypophagia and hypothalamic cell activation induced by vasoactive intestinal peptide, but not by pituitary adenylate cyclase-activating polypeptide

The neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) act in arcuate (ARC) and paraventricular (PVN) hypothalamic nuclei, reducing food intake and changing plasma parameters. Estrogens (E) also regulate energy homeostasis, and loss of ovarian function leads to hyperphagia and body weight gain. This study aimed to evaluate the effects of estradiol (E) in a postmenopausal rat model, ovariectomy (OVX), on PAC1 and VPAC2 receptors in the PVN and ARC, as well as on food intake, plasma parameters, and PVN and ARC cell activation in response to intracerebroventricular microinjection of VIP and PACAP. For this, the rats underwent intracerebroventricular and OVX surgeries, being treated daily with subcutaneous injections of 0.2 mL of corn oil or 10 μg/0.2 mL of estradiol cypionate, comprising the OVX+O and OVX+E groups, respectively. OVX+E showed reduced VPAC2 mRNA expression in the PVN. PACAP reduced food intake in both groups, and VIP-induced hypophagia was not observed in OVX+E. VIP increased plasma glucose in both groups, and PACAP increased plasma glucose only in OVX+O. VIP decreased free fatty acids in OVX+E. Furthermore, PACAP increased ARC cell activation in both groups, and in the PVN only in OVX+O. Cell activation induced by VIP in ARC and PVN was blocked by estradiol. Therefore, estrogens disrupted the hypophagia induced by VIP, but not by PACAP, and these differences seem to be, at least in part, due to an impairment of the activation of the ARC-PVN pathway.

Reproductive efficiency in Bos indicus and Bos indicus crossed: impact of timed artificial insemination (TAI) using a progesterone and estradiol benzoate-based protocol

ABSTRACT Data from 15,342 bovine females subjected to timed artificial insemination (TAI) were analysed with the objective of assessing the pregnancy rate (PR) and its association with breed, number of uses of intravaginal implant, body condition score (BCS), animal categories, number of AI, and estrus expression before TAI. The protocol involved administering intravaginal progesterone + estradiol benzoate (day zero), progesterone removal + prostaglandin F2α+estradiol cypionate + equine chorionic gonadotropin (day 8) and TAI + gonadotropin-releasing hormone (day 10) in animals without estrus. Crossbred cows demonstrated higher PR. Non-suckling cows, those with 1st – and 2nd-use implants, and a single-dose implant demonstrated higher PR between categories; the BCS (>2.75–3.5) resulted in better overall PR. There were no differences in the general PR between number of AI. 2nd-use implants resulted in a better overall PR. Primiparous cows resulted better PR with 3rd-use implants. Animals in estrus before TAI demonstrated a higher PR. In conclusion, this study observed no significant differences in general PR among number of AI. Second-use implants were effective. Crossed and non-suckling cows, and 1st – and 2nd-use and single-dose implants exhibited higher fertility. Estrus before TAI favoured PR.

Use of Injectable Progesterone as a Pre-Synchronisation Strategy in a Timed Artificial Insemination Protocol Based on Gonadotropin-Releasing Hormone and Progesterone in Bos indicus Beef Cows in Anoestrous.

This study evaluated the effects of pre-synchronisation with injectable progesterone (P4) on the ovarian follicular dynamics of Bos taurus indicus cows in anoestrous treated with a timed artificial insemination (TAI) protocol. Multiparous Nelore females (n = 47) at 30-60 days postpartum were used in this study. 10 days before (D-10) the TAI protocol, antral follicle count (AFC; follicles ≥ 3 mm), ovarian condition and body condition score (BCS; 1-5) were assessed and were randomly allocated into two groups: Pre-sync (n = 25), which underwent pre-synchronisation with 150 mg of injectableP4 intramuscularly (i.m.), and control (n = 22), which received the same volume of NaCL 0.9%. On D0, the ovarian assessment was repeated, and TAI protocol was initiated in all animals, with the insertion of an intravaginal P4 device and administration of 10.5 μg of buserelin acetate (gonadotropin-releasing hormone-GnRH). On D7, the P4 device was removed, and 300 IU of equine chorionic gonadotropin, 150 μg of D-cloprostenol and 1 mg of estradiol cypionate were administered i.m. On the same day (D7), the presence of the corpus luteum (CL) was assessed, the dominant follicle was measured, and the tail was painted to evaluate estrous expression. On D9, the largest follicle was remeasured, and TAI was performed. Animals that were not detected in oestrous at the time of AI were administered 10.5 μg of GnRH i.m. Numerical data were analysed using the Mann-Whitney U test. Binary data were analysed using the Fisher's exact test (5%). BCS, both at the beginning of pre-synchronisation (p = 0.45) and TAI protocol initiation (p = 0.20), and AFC (p = 0.36) did not differ between control and Pre-sync groups. The diameter of the largest follicle was similar between the control and Pre-sync groups on D-10 (p = 0.32), D0 (p = 0.33), D7 (p = 0.29) and D9 (p = 0.22). On D7 of the protocol, the Pre-sync group had a higher percentage of CL visible on transrectal ultrasonography (84.0%; p = 0.02) than the control group (54.5%); however, the expression during oestrous did not differ between groups (p = 0.59). The pregnancy rate was similar (p = 0.64) between groups and was not influenced by the CL rate on D7 (p = 0.48), oestrous expression (p = 0.20) or their interaction (p > 0.1). Pre-synchronisation effectively increased the proportion of cows with CL on D7 without altering the diameter of the largest follicle, oestrous expression or pregnancy rate in anoestrous cows treated with a GnRH/P4-based TAI protocol.

8309 Injectable Estradiol Dosing Regimens; A Retrospective Review of Hormone Therapy for Gender-Diverse Adults Assigned Male at Birth

Abstract Disclosure: M. Kanin: None. M. Slack: None. R. Patel: None. K. Chen: None. N. Jackson: None. K. Williams: None. S. Grock: None. Introduction: Many gender incongruent individuals assigned male at birth (AMAB) seek hormone therapy to achieve physical and emotional changes. Hormone therapy typically includes estradiol, with or without an anti-androgen. However, limited data with relatively weak justifications support the currently recommended regimens. In particular, dosing schedules for injectable estradiol can vary considerably. UCSF guidelines recommend initiating estradiol valerate at 5 mg parenterally weekly or estradiol cypionate 2 mg weekly. Endocrine Society Guidelines recommend estradiol valerate or cypionate parenterally 5-30 mg every 2 weeks or 2-10 mg weekly. Our clinical observations led to the concern that current guideline-based dosing of injectable estradiol can result in supratherapeutic estradiol levels. Thus, we designed a retrospective cohort study to evaluate injectable estradiol dosing requirements for gender diverse individuals at our institution. Hypothesis: We hypothesize that low dose estradiol injections can lead to therapeutic estradiol levels (100-200 pg/mL) and suppressed testosterone (<50 ng/dL). Design: We performed a retrospective cohort study of gender diverse individuals AMAB aged 18-89 who were newly prescribed injectable estradiol by a single clinic from January 2017 to March 2023. Exclusion criteria included history of orchiectomy or estradiol use within the previous six months. Patients were identified using structured query language to extract data from our institution’s electronic health record system. A total 33 patients were identified, 29 of whom were eligible for inclusion. Data was collected for 15 months after estradiol initiation. Results: All individuals were prescribed estradiol valerate except for 1 who was prescribed estradiol cypionate. Estradiol dose ranged from 2.5 to 10 mg weekly and an average initial dose of 4.26 mg weekly. Mean initial estradiol level obtained while on treatment for at least 30 days was 349.8 pg/mL, which is supratherapeutic. Mean initial testosterone level was 25.3 ng/dL, which is suppressed. Mean estradiol dose at completion of data collection was 3.68 mg with a mean final estradiol level of 245.2 pg/mL. Testosterone levels reached less than 50 ng/dL for everyone in the sample with a mean final testosterone of 23.3ng/dL despite significant reductions in estradiol dose (change=-0.58 mg per week, p=0.001) Conclusion: Our study demonstrates lower dosages of estradiol can be used to achieve therapeutic estradiol levels in gender incongruent patients seeking estrogen therapy. Using lower dosages can avoid supratherapeutic estradiol levels and the associated risks. Further prospective studies are needed with the aim to reevaluate current guideline dosing recommendations for injectable estradiol. Presentation: 6/2/2024

8309 Injectable Estradiol Dosing Regimens; A Retrospective Review of Hormone Therapy for Gender-Diverse Adults Assigned Male at Birth

Abstract Disclosure: M. Kanin: None. M. Slack: None. R. Patel: None. K. Chen: None. N. Jackson: None. K. Williams: None. S. Grock: None. Introduction: Many gender incongruent individuals assigned male at birth (AMAB) seek hormone therapy to achieve physical and emotional changes. Hormone therapy typically includes estradiol, with or without an anti-androgen. However, limited data with relatively weak justifications support the currently recommended regimens. In particular, dosing schedules for injectable estradiol can vary considerably. UCSF guidelines recommend initiating estradiol valerate at 5 mg parenterally weekly or estradiol cypionate 2 mg weekly. Endocrine Society Guidelines recommend estradiol valerate or cypionate parenterally 5-30 mg every 2 weeks or 2-10 mg weekly. Our clinical observations led to the concern that current guideline-based dosing of injectable estradiol can result in supratherapeutic estradiol levels. Thus, we designed a retrospective cohort study to evaluate injectable estradiol dosing requirements for gender diverse individuals at our institution. Hypothesis: We hypothesize that low dose estradiol injections can lead to therapeutic estradiol levels (100-200 pg/mL) and suppressed testosterone (<50 ng/dL). Design: We performed a retrospective cohort study of gender diverse individuals AMAB aged 18-89 who were newly prescribed injectable estradiol by a single clinic from January 2017 to March 2023. Exclusion criteria included history of orchiectomy or estradiol use within the previous six months. Patients were identified using structured query language to extract data from our institution’s electronic health record system. A total 33 patients were identified, 29 of whom were eligible for inclusion. Data was collected for 15 months after estradiol initiation. Results: All individuals were prescribed estradiol valerate except for 1 who was prescribed estradiol cypionate. Estradiol dose ranged from 2.5 to 10 mg weekly and an average initial dose of 4.26 mg weekly. Mean initial estradiol level obtained while on treatment for at least 30 days was 349.8 pg/mL, which is supratherapeutic. Mean initial testosterone level was 25.3 ng/dL, which is suppressed. Mean estradiol dose at completion of data collection was 3.68 mg with a mean final estradiol level of 245.2 pg/mL. Testosterone levels reached less than 50 ng/dL for everyone in the sample with a mean final testosterone of 23.3ng/dL despite significant reductions in estradiol dose (change=-0.58 mg per week, p=0.001) Conclusion: Our study demonstrates lower dosages of estradiol can be used to achieve therapeutic estradiol levels in gender incongruent patients seeking estrogen therapy. Using lower dosages can avoid supratherapeutic estradiol levels and the associated risks. Further prospective studies are needed with the aim to reevaluate current guideline dosing recommendations for injectable estradiol. Presentation: 6/2/2024

Exploring the inhibitory potential of hormone replacement therapy drugs on glutathione transferase P1-1

Aim: Glutathione transferase P1-1 (GST P1-1) plays a crucial role in the human phase II detoxication system and is implicated in drug resistance in certain cancer cells. Analogs of drugs used in Hormone Replacement Therapy (HRT) have been reported to exhibit inhibition of various GSTs. This study aims to investigate the inhibitory potential of several HRT drugs on GST P1-1 for the possible use of counteracting drug resistance and for other therapeutic purposes. Materials and Methods: GST P1-1 was expressed and then purified in a single step using Nickel Sepharose affinity chromatography. Synthetic drugs that are used in HRT were screened for inhibition of GST P1-1 using 1-chloro-2,4-dinitrobenzene as the substrate. The IC50 value of the most potent compound was calculated and binding location and formation of bonds to GST P1-1 were identified through docking analysis. Results: Screening the inhibitory effect of eight synthetic estrogenic drugs reveals that estradiol valerate is the most potent inhibitor, showing 72 ± 4% inhibition of GST P1-1. This effect is followed by estradiol cypionate 53 ± 5%, mestranol 39 ± 4%, and estradiol propionate 35 ± 2%. The most potent compound estradiol valerate has an IC50 value of 30 ± 2 μM. According to docking analysis, it binds to the H-site of the enzyme where the residues Phe9, Arg14, Val36, Trp39, Ile105, Tyr109, Pro203, Asn207, and Gln210 were within 5 Å proximity of the ligand. Estradiol valerate forms Pi-alkyl interactions with Phe9 and Val36, as well as an alkyl interaction with Ile104. Conclusion: Estradiol valerate is a modest inhibitor for hGST P1-1, however it fits in the area of H-site of the enzyme and forms bonds with critical key residues. Understanding its binding site on the enzyme is critical for designing other inhibitors targeting GSTs or possibility for the potential use as a substrate with other GSTs.

Effects of Two Types of Estrogen on the Follicular Wave for in Vivo Oocyte Collection in Brown Swiss Cows

The manipulation of follicular waves through hormonal treatments, such as estrogen administration, plays a crucial role in optimizing in-vivo oocyte collection for assisted reproductive technologies. The present study aimed to evaluate the effect of two specific types of estrogen on follicular wave dynamics and their impact on in-vivo oocyte collection in Brown Swiss cows. Fourteen cows, in their first lactation, weighing approximately 340 kg were randomly assigned to one of two treatments including T1 (estradiol cypionate) and T2 (estradiol benzoate). Both treatments were administered at 1.2 mg of estrogen, at day 0 of the experiment. All Brown Swiss cows were provided with a diet entirely consisting of alfalfa grazing. On day 7, follicular wave dynamics were assessed using a DP-50 vet ultrasound device equipped with a 7.5 MHz transducer for transvaginal follicular aspiration guidance. Follicle counts were categorized into three size ranges including 2-4 mm (small), 4-8 mm (medium), and greater than 8 mm (large). Additionally, the quantity and quality (viable oocytes) of the collected oocytes were evaluated by the Ovum Pick Up (OPU) team for oocyte viability on day 7. The study assessed the follicular dynamics (number of follicles) and efficiency of oocyte collection (viable oocytes) in cows treated with Estradiol Cypionate (T1) and Estradiol Benzoate (T2). The average number of small, medium-sized, and large follicles size were 6.048 ± 6.037, 3.16 ± 2.01, and 0.53 ± 0.67 respectively. The total number of follicles was 9.59 ± 3.56. The mean number of viable oocytes recovered was 3.024 ± 1.66, while the mean number of non-viable oocytes was 1.47 ± 1.01. The results indicated no significant differences between treatments in the size of small, medium, and large follicles, nor in the total number of follicles and viable oocytes recovered. However, a significant difference was observed in the number of non-viable oocytes recovered, with a higher mean in T2 (1.86) compared to T1 (1.09). The results indicated an adequate follicular response and viable oocyte recovery in both treatment groups (estradiol cypionate and estradiol benzoate). However, variations in oocyte viability were observed, with estradiol cypionate showing a slight advantage.

Optimisation of Early Pregnancy Diagnosis andResynchronisation in Beef Cows.

The objectives of this study were (1) to investigate the effects of the preovulatory follicle (POF) size on the accuracy of Doppler-based early pregnancy detection, and (2) to determine whether the removal of PGF2α (PGF) treatment during the resynchronisation protocol would affect fertility in beef cows. In Experiment 1, Nelore suckling cows (n = 224) were enrolled in an estradiol-progesterone-based timed artificial insemination (TAI) protocol. At TAI, cows were separated based on the range of POF diameters, as follows: ≤11.0 mm (n = 50), 11.1-12.9 mm (n = 64), 13.0-14.4 mm (n = 62) and ≥14.5 mm (n = 48). On day 22 after TAI, the corpus luteum (CL) blood flow (CLBF) of all cows was examined by colour Doppler ultrasonography to diagnose nonpregnant cows. The cows with the largest POF had the greatest positive predictive value (88.6%; 31 of 35) and diagnostic accuracy (91.7%; 44 of 48). In Experiment 2, Nelore cows (n = 233) were subjected to the same TAI protocol. Fourteen days after TAI, all cows were started on a resynchronisation protocol. Cows diagnosed as nonpregnant based on CLBF, on day 22, received 0.5 mg estradiol cypionate intramuscular (im) and were assigned to receive either 150 μg of PGF (PGF; n = 50) or 2 mL of saline (control; n = 47). Cows treated with PGF had a P/AI of 30.0% compared with a 48.9% P/AI in controls (p = 0.06). Our findings demonstrate that the POF size affects the accuracy of a CLBF-based early pregnancy diagnosis and that the removal of PGF treatment from the resynchronisation protocol tended to increase P/AI of the second TAI.

Investigation of potential ascorbate peroxidase inhibitors for anti-leishmaniasis therapy.

L eishmaniasis is a prevalent disease that impacts 98 countries and territories, mainly in Africa, Asia, and South America. It can cause substantial illness and death, particularly in its visceral manifestation that can be specifically targeted in the development of medications to combat leishmaniasis. This study has found natural compounds with possible inhibitory activity against APX using a reliable and accurate QSAR model. Despite the severe side effects of current treatments and the absence of an effective vaccination, these compounds show promise as a potential treatment for the disease. Nine hit compounds were found, and subsequent molecular docking was performed. Estradiol cypionate showed the lowest binding energy (- 10.5kcal/mol), thus showing the strongest binding, and also had the strongest binding affinity, with a ΔGTotal of - 26.31 ± 3.01kcal/mol, second only to the control molecule. Additionally, three hits viz. cloxacillin-sodium (- 16.57 ± 2.89kcal/mol), cinchonidine (- 16.04 ± 3.27kcal/mol), and quinine hydrochloride dihydrate (13.38 ± 1.06kcal/mol) also showed significant binding affinity. Multiple interactions between drugs and active site residues demonstrated a substantial binding affinity with the target protein. The identified compounds exhibited drug-like effects and were orally bioavailable based on their ADME-toxicology features. Overall, estradiol cypionate, cloxacillin sodium, cinchonidine, and quinine hydrochloride dihydrate all exhibited inhibitory effects on the APX enzyme of Leishmania donovani. These results suggest that further investigation is needed to explore the potential of developing novel anti-leishmaniasis drugs using these compounds.

Improving Milk Yield, Milk Quality, and Follicular Functionality Behavior in Dairy Cows from the Implementation of Microencapsulated Chili Pepper Supplements in Their Diets.

The present study evaluates the effect of including microencapsulated hot chili pepper (MHCP) in the diet of crossbred dairy cows on the volume and quality of milk and on ovarian morphofunctionality. Twenty-four crossbred females in their lactating period were used. The cows were divided into two experimental groups, a control (CT) and an MHCP -supplemented group (CP) given 1 g a day per animal of microencapsulated hot chili in concentrate for 42 days. Over seven weeks of daily milk production was measured, and sample milk was collected weekly for composition analysis. Animals were subject to an ovulation synchronization protocol on day 0 (D0), and an intravaginal progesterone (P4) implant, estradiol benzoate, and prostaglandin (PGF2α) were administered. On D8, the P4 implant was removed and PGF2α, equine chorionic gonadotropin, and estradiol cypionate were administered to the animals. The ovarian dynamics were evaluated in B mode and color Doppler. There were significant differences (p < 0.05) in the group X time interaction, the volume of milk produced, and the amount in kg/day of milk components. There was a higher percentage of vascularization in the preovulatory follicle in the CP group (p ≥ 0.10). The findings show that the inclusion of MHCP in the diet of dairy cows does influence their milk production and reproduction.

The use of a recombinant equine chorionic gonadotropin (reCG) in fixed-time AI programs in beef cattle

The aim of the current study was to evaluate the effect of a novel recombinant eCG (reCG) on pregnancy rates to AI (P/AI) in suckled beef cows of different breeds that were synchronized with an estradiol/progesterone (P4)-based protocol for fixed-time AI (TAI). In experiment 1, 1244 Bos taurus suckled cows were used. On Day 0 all cows received an intravaginal P4 device (600 mg P4) and 2 mg of estradiol benzoate. On Day 7, devices were removed, and all cows received 0.150 mg of D-cloprostenol plus 1 mg of estradiol cypionate and were randomly divided to receive 140 IU or 105 IU of reCG or no reCG treatment (controls) at that time. Cows were tail painted for estrus detection and those in estrus by 48 h after P4 device removal were inseminated; whereas those not showing estrus were also inseminated and received GnRH at the same time. In experiment 2, 818 Bos taurus x Bos indicus crossbred suckled cows received the same FTAI protocol used in Experiment 1. Cows were randomly divided at the time of P4 device removal into 4 groups to receive 140 IU, 105 IU or 84 IU of reCG or no reCG treatment. In experiment 3, 345 Bos indicus suckled cows were submitted to the same FTAI protocol as those in previous experiments and were randomly divided into three groups to receive 140 IU or 105 IU of reCG, or 300 IU of serum derived eCG (PMSG). In Experiment 1, estrus rate and P/AI was greater (P < 0.05) in cows treated with reCG (79.9 and 53.5 %, 76.9 and 52.3 % for the 105 UI and 140 UI reCG groups, respectively) than those in the control group (69.9 and 44.4 %, respectively). In Experiment 2, cows treated with reCG tended (P < 0.1) to achieve a greater P/AI than control cows (38.6 %, 37.1 %, 36.2 % and 28.2 % for those receiving 84 IU, 105 IU,140 IU of reCG, and those in the control group); but when P/AI of all cows treated with reCG was contrasted to that of control cows, the difference was significant (P < 0.01). In Experiment 3, P/AI in cows treated with 84 IU of reCG (54 %) did not differ from that of cows treated with serum derived eCG (59 %) but both were greater (P < 0.05) than cows treated with 105 UI of reCG (41 %). In conclusion, treatment with reCG improved fertility in suckled Bos taurus and Bos taurus x Bos indicus beef cows. In suckled Bos indicus cows, although treatment with reCG and serum derived eCG were comparable, the higher dosage of reCG was detrimental to their P/AI.

Bovine somatotropin increases the pregnancy rate in fixed-time artificial insemination in beef cattle.

This study evaluated the effect of bovine somatotropin (bST) on pregnancy rate (PR) and size of the dominant follicle (DF) on the day of intravaginal progesterone (P4) removal in protocols for fixed-time artificial insemination (FTAI). Bos indicus (Nellore) females (n = 392) were distributed into three groups. The control group (CG; n = 92) received an intravaginal P4 device + estradiol benzoate on day (d)0; prostaglandin F2α on d7 (first application); removal of P4 + estradiol cypionate (EC) + PGF2α (second application) + ultrasound (US) of the DF on d9; the FTAI was performed on d11; and pregnancy diagnosis (PD) was performed on d45. The bST group (bSTG; n = 142) underwent the same protocol as the CG, except that the animals received 125 mg of bST on d7. The equine chorionic gonadotropin (eCG) group (eCGG; n = 158) underwent the same protocol as the CG, except that the animals received 300 IU of eCG on d9. The PRs of the bSTG, eCGG, and CG were 48%, 48%, and 35%, respectively (p < .05); the bSTG and eCGG showed greater PRs, with follicles 6-7.9 mm (p < .05) and 8-8.9 mm in diameter, respectively. The bSTG exhibited a greater dimension of the DF on d9 of the protocol (p < .05). The eCGG had higher PRs with a body condition score (BCS) of 2.5, and the bSTG had a BCS of 3.0 (p < .05). It was concluded that bST increased PR, bST showed better performance in smaller DF and larger follicular diameter on d9 of the protocol, eCG acted better on animals with lower BCSs, and bST can be used in FTAI.

Optimizing ReBreed21 I: Evaluation of endocrine and ovarian dynamics in non-bred Bos indicus heifers

The present study evaluated follicular and endocrine dynamics during ReBreed21, a reproductive strategy that allows resynchronization of ovulation every 21 days in Bos indicus (Nelore) heifers. A synchronized estrous cycle was induced using a standard timed ovulation protocol (d −10: P4 implant inserted + 2 mg estradiol benzoate; d −2: P4 removed+ 0.5 mg cloprostenol + 0.6 mg estradiol cypionate + 200 IU equine chorionic gonadotropin (eCG); d0: 8.4 μg buserelin) without AI to ensure nonpregnancy in heifers. Day of GnRH was designated d0 of estrous cycle. On d12, heifers (n = 80) were randomized into three experimental groups: (1) ReBreed21 (n = 28) d12 P4 device inserted, d19 P4 device withdrawal plus 200 IU eCG, and d21 8.4 μg buserelin (GnRH); (2) ReBreed21+G (n = 26) same as ReBreed21 plus GnRH (16.8 μg) treatment on d12; and (3) Control (n = 26) no treatment. ReBreed21+G increased two-fold (62.9%; 18/26) percentage of heifers with synchronized follicular wave emergence compared to Control (34.6%; 9/26) whereas ReBreed21 (53.6%; 15/28) was intermediate. The ReBreeed21 groups (eCG on d19) increased (P < 0.01) follicular growth between d19 and d21 in ReBreed21 (2.3 ± 0.2 mm) and ReBreed21+G (3.4 ± 0.2 mm) compared with Control (1.2 ± 0.3 mm), resulting in greater (P < 0.01) follicle diameter on d21 for ReBreed21 (10.7 ± 0.4 mm) and ReBreed21+G (10.8 ± 0.4 mm) compared with Control (9.1 ± 0.5 mm). Structural luteolysis was similar among groups (P = 0.51), although the average day when P4 was <1 ng/mL was later (P < 0.01) for ReBreed21 (20.5 ± 0.2) and ReBreed21+G (20.7 ± 0.2) compared to Control (19.2 ± 0.4). Overall ovulation at the end of the estrous cycle was increased (P = 0.03) for ReBreed21 groups (83.3%; 45/54) compared with Control (57.7%; 15/26). Synchronized ovulation on day 22–23 was greater (P < 0.01) for ReBreed21 (78.6%; 22/28) and ReBreed21+G (76.9%; 20/26) compared with Control (30.8%; 8/26). Thus, the ReBreed21 resynchronization program produced acceptable endocrine and follicular dynamics, including synchronized ovulation at the end of the protocol in nonpregnant heifers providing good rationale for testing the fertility and practical implementation of this protocol under field conditions.

The effectiveness of low-dose of eCG in timed AI Nelore (Bos indicus) heifers.

This study evaluated the efficacy of the administration of different doses of equine chorionic gonadotropin (eCG; 0 IU, 200 IU, or 300 IU) at the time of the progesterone device removal in 2-year-old Nelore (Bos indicus) heifers synchronized for fixed-timed artificial insemination (FTAI). On day 0 (D0), a total of 398 heifers received 2 mg of oestradiol benzoate i.m., 0.53 mg of cloprostenol i.m., and an eight-day previously used (second use) intravaginal device containing 1 g of progesterone (P4). Eight days later (D8), simultaneous with the P4 device removal, 0.5 mg of oestradiol cypionate i.m. and 0.53 mg of cloprostenol i.m. were administered. At the same time, heifers were randomly assigned to receive one of the following treatments: G-0 IU (n = 141; no eCG treatment), G-200 IU (n = 132; treated with 200 IU of eCG), and G-300 IU (n = 125; treated with 300 IU of eCG). FTAI was performed 48 h after the P4 device removal (D10). Ultrasonographic evaluations were performed at D0, D10, and D17. Heifers were scanned to measure the size of the largest follicle (LF), the presence, number, and size of the corpus luteum (CL), and the ovulation rate. Subsequently, at D40, the heifers underwent scanning to determine the pregnancy rate and identify any twin pregnancies. Additionally, at D70, scans were performed to assess pregnancy loss (PG). Data were analysed by orthogonal contrasts [C1 (eCG effect): control x (200 IU + 300 IU) and C2 (eCG dose effect): 200 IU × 300 IU]. On D0, CL presence was similar between the groups [G-0 IU = 65.2% (92/141), G-200 IU = 55.3% (73/132), and G-300 IU = 63.2% (79/125); p = .16]. No interactions between the presence of CL on D0 and eCG treatment were found for any of the variables (p > .05). The diameter of the LF at FTAI (D10) was not influenced by eCG treatment (p = .22) or eCG dose (p = .18). However, treatment with eCG increased the diameter of the CL at D17 (G-0 IU = 15.7 ± 0.3 mmb , G-200 IU = 16.6 ± 0.2 mma , and G-300 IU = 16.6 ± 0.3 mma ; p = .001), regardless of the dose used (p = .94). The ovulation rate was higher in heifers treated with eCG [G-0 IU = 79.4%b (112/141), G-200 IU = 90.2%a (119/132), and G-300 IU = 93.6%a (117/125); p = .002], but there was no effect of eCG dose (p = .36). Pregnancy per AI (P/AI) on D40 [G-0 IU = 32.6%b (46/141), G-200 IU = 42.4%a (56/132), and G-300 IU = 42.4%a (53/125); P = 0.05] and D70 [G-0 IU = 29.1%b (41/141), G-200 IU = 40.9%a (54/132), and G-300 IU = 40.8%a (51/125); p = .02] were higher on heifers that received eCG; however, no dose effect was observed for P/AI on D40 (p = .89) nor D70 (p = .98). Pregnancy loss between D40 and D70 tended to reduce (p = .07) in eCG-treated heifers without dose effect (p = .91). Heifers with CL at D0 presented a greater follicle diameter (LF) on D10 (With CL = 11.2 ± 0.2 mm and Without CL = 10.2 ± 0.2 mm; p = .05), CL diameter on D17 (With CL = 15.8 ± 0.03 mm and Without CL = 11.8 ± 0.6 mm; p = .01), and ovulation rate [With CL = 95.5% (233/244) and Without CL = 74.7% (115/154); p = .01]. However, no difference in pregnancy rate at D40 (p = .52) and D70 (p = .84) was found. In conclusion, eCG treatment increases ovulation and pregnancy rates of heifers submitted to a FTAI protocol. Furthermore, eCG treatment increases the diameter of the CL after FTAI and reduces pregnancy losses. No dose effect was observed, suggesting Nelore (Bos indicus) heifers respond to 200 IU of eCG treatment for FTAI.

The Discovery of inhibitors of the SARS-CoV-2 S protein through computational drug repurposing

SARS-CoV-2 must bind its principal receptor, ACE2, on the target cell to initiate infection. This interaction is largely driven by the receptor binding domain (RBD) of the viral Spike (S) protein. Accordingly, antiviral compounds that can block RBD/ACE2 interactions can constitute promising antiviral agents. To identify such molecules, we performed a virtual screening of the Selleck FDA approved drugs and the Selleck database of Natural Products using a multistep computational procedure. An initial set of candidates was identified from an ensemble docking process using representative structures determined from the analysis of four 3 μ s molecular dynamics trajectories of the RBD/ACE2 complex. Two procedures were used to construct an initial set of candidates including a standard and a pharmacophore guided docking procedure. The initial set was subsequently subjected to a multistep sieving process to reduce the number of candidates to be tested experimentally, using increasingly demanding computational procedures, including the calculation of the binding free energy computed using the MMPBSA and MMGBSA methods. After the sieving process, a final list of 10 candidates was proposed, compounds which were subsequently purchased and tested ex-vivo. The results identified estradiol cypionate and telmisartan as inhibitors of SARS-CoV-2 entry into cells. Our findings demonstrate that the methodology presented here enables the discovery of inhibitors targeting viruses for which high-resolution structures are available.

Revisiting Injectable Estrogen Dosing Recommendations for Gender-Affirming Hormone Therapy.

Injectable estrogens are options for gender-affirming hormone therapy per guidelines, which suggest intramuscular dosages of 5-30 mg every 2 weeks or 2-10 mg weekly with estradiol cypionate or valerate interchangeably. Data among transgender and gender-diverse patients are limited due to local unavailability and concerns around laboratory assay variability and estradiol (E2) level fluctuation. We note a concerning trend where patients are prescribed high-dose injections based on the guidelines leading to serum E2 levels well above the range recommended in the same guidelines. Our review indicates that 5 mg weekly or lower should be prescribed when initiating injectable estrogens to avoid supraphysiologic E2 levels.

Comparison of gonadotropin-releasing hormone and estradiol benzoate plus gonadotropin-releasing hormone to initiate a progesterone-based timed artificial insemination resynchronization protocol in lactating dairy cows

The present study compared 2 strategies to initiate a progesterone (P4)-based timed artificial insemination (TAI) protocol for lactating dairy cows: only GnRH or estradiol benzoate (EB) plus GnRH (EB+GnRH). Lactating Holstein cows (n = 487; 184 primiparous and 303 multiparous) from 2 commercial dairy herds were used for their second or greater services postpartum. Each week, cows that were nonpregnant at the pregnancy diagnosis 32 d after a previous AI were randomly assigned to 1 of 2 experimental groups that differed only in the strategy to initiate (d 0) the TAI protocol. On d 0, every cow received a 2.0-g P4 implant; in the EB+GnRH group, cows were treated with 2.0 mg i.m. of EB and 16.8 µg i.m. of the GnRH analog buserelin acetate, whereas in the GnRH group, cows received only 16.8 µg i.m. of GnRH. On d 7 after the initial treatment, 0.530 mg i.m. of cloprostenol sodium (PGF) was administered in all cows, followed by a second dose on d 8, concomitant with 1.0 mg i.m. of estradiol cypionate and P4 implant withdrawal. The TAI was performed on d 10 (48 h after P4 device withdrawal) in both experimental groups. Only conventional Holstein semen was used throughout the study. The percentage of cows with corpus luteum (CL) on d 0 (73%) and overall ovulation rate after d 0 (54%) did not differ between groups. The CL regression between d 0 and the first PGF treatment was greater in the EB+GnRH group than the GnRH group (42% vs. 31%). Consequently, the proportion of cows with CL at PGF was greater when only GnRH was used on d 0 compared with EB+GnRH (86% vs. 82%), and the mean number of CL at PGF was greater (1.23 vs. 1.11). The expression of estrus near TAI was greater in GnRH group (84% vs. 77%), and cows showing estrus had greater (44% vs. 10%) pregnancy per AI (P/AI) on d 32 for both treatments. We found no effect of the presence of CL on d 0 or at PGF, nor of ovulation after d 0 or CL regression between d 0 and d 7 on fertility. However, fertility was critically impaired when cows did not have CL at both times, d 0 and at PGF treatment. We did not observe any interaction between treatment and other variables, and the P/AI was similar in cows receiving EB+GnRH or only GnRH on d 0 (37.8% vs. 36.6%). In summary, although there was no detectable difference in P/AI between treatments, this study demonstrated potential negative physiological outcomes caused by EB treatment on d 0 (greater incidence of luteolysis after d 0 and fewer cows with CL at PGF treatment). Overall, we found no benefit of adding EB at the initiation of a P4-based TAI protocol on fertility compared with using GnRH alone, despite differences in ovarian dynamics and expression of estrus.

Berat Badan Pada Akseptor yang Menggunakan Kontrasepsi Suntik di Puskesmas Singgani Kota Palu

The Family Planning (KB) program is carried out in order to regulate the number of births or space births. Injectable contraception is one of the contraceptive methods that is widely used in Indonesia because it works effectively, is practical to use, is relatively cheap, and is safe. One of the ways injectable contraception works is to cause changes in the menstrual cycle to increase body weight which is the most frequently complained about by injectable contraceptive acceptors. Injection methods are divided into two types, namely 1-month injectable contraceptives and 3-month injectable contraceptives. 1-month contraceptive injection is a type of birth control injection that contains a combination of the hormone Medroxyprogesterone Acetate (progestin hormone) and Estradiol cypionate (estrogen hormone), and 3-month contraceptive injection is an injectable contraceptive containing the Depo Medroxyprogesterone Acetate hormone p (progestin hormone) with a volume of 150 mg. The aim of this study was to analyze differences in body weight among injectable contraceptive acceptors. This type of research uses analytical observational research methods. This research sample was taken using the Accidental Sampling technique with a sample size of 42 respondents. Analysis uses the sign difference test. The sign test results showed that there was a relationship between the use of injectable birth control (P &lt; 0.05) and weight gain.

Effects of mineral and vitamin supplementation on pregnancy rates in Nelore cattle submitted to FTAI programs with different reactivity scores

A high-reactivity herd temperament reduces productivity and profitability in cattle farming. In Bos taurus indicus (Nellore) cattle, we hypothesized that a higher reactivity score is associated with a lower pregnancy rate, and that injectable supplementation with minerals (Cu, Zn, Mn, Se) and vitamins (A, E) would increase the pregnancy rate and reduce embryonic mortality in very reactive cows. Injectable supplement or control treatments were administered on days 25 and 10 before fixed-time artificial insemination (FTAI; Day 0 : day of FTAI). We selected 1722 cows 45 ± 7 days postpartum, with different behavioral reactivity levels, and a series of measures related to reproduction were measured. The cows were randomly grouped and received sc 1 mL/50 kg live weight of the supplement (n = 872) or placebo (n = 850), 25 and 10 days before the FTAI protocol (Day–25 and Day–10 respectively). On Day −10, devices containing 1 g progesterone were inserted into the vagina, and 2 mg estradiol benzoate was applied intramuscularly. On Day–2, the devices were removed and 530 µg of sodium d-cloprostenol, 300 IU of equine chorionic gonadotropin, and 1 mg of estradiol cypionate were applied. At 48 h after final injection, the cows were subjected to FTAI (Day 0). On Day - 25 (25 days before IATF), temperament was graded as 3 to 12 using composite reactivity score (RS), which evaluates movement and breathing while the animal is in the cattle chute, and their escape velocity, and was used to classify cows as calm (RS ≤ 4; n = 590), reactive (RS 5 to 8; n = 573), and very reactive (RS ≥ 9; n = 559). On Day 0, calm cows had a higher largest follicle diameter on the FTAI compared to reactive and very reactive (14.4 + 0.2 vs. 13.6 + 0.4 and 13.2 + 0.2 mm, respectively; P = 0.03). Interactions were observed, where supplemented cows compared to those no supplemented had higher pregnancy on Day 30 (P = 0.04; 60.00 vs. 54.10 % calm and 58.00 vs. 52.50 % very reactive) and Day 60 (P = 0.03; 58.70 vs. 47.90 % calm, 52.20 vs. 49.10 % reactive and 48.40 vs. 41.30 % very reactive) and reduced embryonic loss at 30 to 60 d (P = 0.04; 2.00 vs. 6.10 % calm, 4.20 vs. 8.40 % reactive and 9.60 vs. 11.20 % very reactive). Supplementation on Days–25 and–10 improved pregnancy rates and reduced embryonic in calm and very reactive cows.