IU Human Chorionic Gonadotropin Research Articles

Detrimental effects of electromagnetic radiation emitted from cell phone on embryo morphokinetics and blastocyst viability in mice.

Electromagnetic radiation (EMR) has deleterious effects on sperm motility and viability, as well as oocyte membrane and organelle structure. The aim was to assess the effects of cell phone radiation on preimplantation embryo morphokinetics and blastocyst viability in mice. For superovulation, 20 female mice were treated with intraperitoneal (IP) injections of 10 IU pregnant mare's serum gonadotropin (Folligon® PMSG), followed by 10 IU of human chorionic gonadotropin (hCG) after 48 h. The zygotes (n = 150) from the control group were incubated for 4 days. The experimental zygotes (n = 150) were exposed to a cell phone emitting EMR with a frequency range 900-1800 MHz for 30 min on day 1. Then, all embryos were cultured in the time-lapse system and annotated based on time points from the 2-cell stage (t2) to hatched blastocyst (tHDyz), as well as abnormal cleavage patterns. Blastocyst viability was assessed using Hoechst and propidium iodide staining. Significant increases (P < 0.05) were observed in the cleavage division time points of t2, t8, t10, and t12 of the experimental group compared with the controls. In terms of blastocyst formation parameters, a delay in embryo development was observed in the experimental group compared with the controls. Data analysis of the time intervals between the two groups showed a significant difference in the s3 time interval (P < 0.05). Also, the rates of fragmentation, reverse cleavage, vacuole formation, and embryo arrest were significantly higher in the experimental group (P < 0.05). Furthermore, the cell survival rate in the experimental group was lower than the control group (P < 0.05). Exposure to EMR has detrimental consequences for preimplantation embryo development in mice. These effects can manifest as defects in the cleavage stage and impaired blastocyst formation, leading to lower cell viability.

Ovarian function and pregnancy rates in dairy goats that received 300 IU of human chorionic gonadotropin (hCG) intravaginally at the time of artificial insemination

Two experiments were carried out to evaluate the effects of administering 300 IU hCG intravaginally (i.vag.) at the time of artificial insemination (AI) on ovulatory response, luteal function and pregnancy rates in cyclic Alpine (A) and Saanen (S) goats. All animals received two i.m. injections of 37.5 μg of d-cloprostenol (Prolise®; Tecnopec, São Paulo, SP, Brazil) at 11.5-day (Experiments 1 and 2) or 7.5-day intervals (Experiment 2). One day after the onset of estrus (day of AI=D0), the goats were randomly allocated to one of the two groups that received: 300 IU hCG (Vetecor®; Hertape-Calier, São Paulo, SP, Brazil) i.vag. (G-hCG; Experiment 1: n = 12 A; Experiment 2: n = 80 A + 63 S) or 0.3 mL or saline solution i.vag. (G-Control; Experiment 1: n = 12 A; Experiment 2: n = 82 A + 65 S). Blood samples for measurements of circulating progesterone concentrations were drawn on both days of d-cloprostenol injections and on D3, D7, D10, D13 and D21 (Experiment 1). Transrectal ovarian ultrasonography was done on D7, D10, D13, D17 and D21 (Experiment 1), and pregnancy detection was performed on D60 (Experiment 2). In Experiment 1, there were no differences (P > 0.05) in the mean diameter of ovulatory follicles, timing and number of ovulations, and incidence of early luteal regression between the two groups of goats. However, the total luteal area on D17 and D21 and luteal vascularization on D10 and D13 were higher (P < 0.05) in G-hCG compared with G-Control does. In Experiment 2, the pregnancy rate was higher (P < 0.05) in G-hCG compared with G-Control goats (80.4% compared with 67.3%, respectively). Human chorionic gonadotropin (300 IU) given i.vag. at the time of AI did not alter ovulatory response or progesterone secretory ability of resultant corpora lutea nor did it completely prevent premature luteal regression, but it increased the luteal area and luteal vascularization as well as the conception rate in dairy goats (the latter by 13.1%).

Reproduction, Early Development, and Larval Rearing of the Climbing Perch Anabas testudineus (Teleostei: Anabantidae) in Captivity

Induced spawning, early development, and larval rearing of the climbing perch Anabas testudineus were examined under captive conditions. Five female climbing perch [125.33 ± 3.27 mm SL; 70.67 ± 5.59 g BW] were paired with apparently mature males (108.50 ± 3.97 mm SL; 39.27 ± 4.70 g BW) and induced to spawn with 5,000 IU human chorionic gonadotropin (hCG) + 0.5 mL Ovaprim kg BW-1. All hCG + Ovaprim-injected fish spawned 9 – 11 h after hormone administration but saline-injected fish (control group) failed to spawn. Mean egg production per female, fertilization and hatching rates, and larval production per female were 7,667 ± 1,313, 98.11 ± 1.63, 61.01 ± 19.92, and 4,435 ± 1,041 %, respectively. Fertilized eggs (1.52 – 1.96 mm in dia) were almost spherical, non-adhesive, transparent, and buoyant due to a large, single oil globule (515 ± 33 μm in dia). First cleavage appeared 30 min post-fertilization (MPF) and egg development lasted for 20 – 24 h post-fertilization (HPF) (29°C). Newly hatched climbing perch larvae [1.73 ± 0.08 mm in total length (TL)] with yolk volume (YV) and oil globule volume (OGV) of 0.545 ± 0.104 and 0.072 ± 0.013 mm3, respectively, had no pigmented eyes or functional mouth parts and digestive tract, and immobile with the yolk sac pointed upward. Eyes became pigmented and both mouth and anus opened in 1 d old larvae (2.09 ± 0.04 mm TL) in preparation for exogenous feeding. Except for body depth (BD) and pre-anal length (PAL), a general increase in TL (1.73 – 3.72 mm), eye diameter (ED) (0.164 – 0.378 mm), head length (HL) (0.205 – 0.923 mm), and mouth gape (MG) size (0.308 – 0.552 mm) was observed, coinciding with yolk resorption in 7 d post-hatched larvae. Climbing perch larvae were reared at 4 stocking densities (25, 50, 75, 100 larvae L-1) and fed solely on live food (Brachionus rotundiformis from 2 – 12 d and Artemia nauplii from 13 – 40 d) or co-fed live food and microparticulate diet (MPD) (Artemia nauplii from 2 – 15 d and Artemia + MPD from 16 – 40 d). Mean survival rates at lower densities (25 – 50 larvae L-1) were significantly higher than those reared at 75 and 100 larvae L-1 (P &lt; 0.05). Furthermore, climbing perch larvae co-fed Artemia nauplii and MPD showed better survival (28.67 – 81.00%) than those fed solely on live food (24.88 – 64.00%). However, at the end of the 40 d trial period, growth parameters did not vary significantly among densities (P &gt; 0.05), except for the condition factor in larvae co-fed Artemia nauplii and MPD. Taken together, results demonstrate the effectiveness of hCG + Ovaprim for captive breeding and observations on the early development of climbing perch, and the feasibility of rearing the larvae at 25 – 50 L-1 on a combination of Artemia nauplii and MPD under laboratory conditions.

Ovine endometrial estrogen receptor expression is altered following PG-600 administration.

Dysregulation of the estrogen receptor 1 (ESR1) expression during the establishment of pregnancy could contribute to reduce fertility reported in ewes treated with PG-600. The objective of this study was to evaluate the effect of treatment with PG-600 on ESR1 expression in the ovine endometrium during early diestrus. Polypay ewes (n = 24) that had exhibited an oestrous cycle of normal duration (16-18 days) were treated with intravaginal progesterone-releasing device (CIDR) for 9 days. Cloprostenol was administrated two days before CIDR withdrawal. On the day of CIDR withdrawal (day 0), ewes received a 5mL intramuscular injection of PG-600 (400 IU equine chorionic gonadotropin and 200 IU human chorionic gonadotropin) or saline. Blood samples were collected before treatment and on the day of tissue collection to determine serum estradiol-17β and progesterone concentrations using radioimmunoassay. On days 4 and 7, six ewes from each treatment group were anaesthetised and a laparotomy was performed to obtain intercaruncular endometrial samples. Endometrial samples were collected ipsilateral to the ovary bearing the highest number of corpora lutea. An estradiol exchange assay was used to determine ESR1 concentration. Estradiol concentrations did not differ by time or treatment, but progesterone concentrations were significantly higher in the PG-600-treated group on day 7 despite a similar number of corpora lutea. Endometrial ESR1 concentration was significantly reduced on day 7 in the PG-600 group. In conclusion, although progesterone was higher than controls on day 7, ESR1 did not differ from controls suggesting that treatment with PG600 is unlikely to impair reproductive potential.

The investigation of cholinergic receptor muscarinic 1 activity in the rat ovary with induced ovarian hyperstimulation.

We look at the immunoreactivity of cholinergic receptor muscarinic 1 (CHRM1) in the ovarian tissues of rats with ovarian hyperstimulation syndrome (OHSS) considering the possibility that the muscarinic activity may contribute to the pathophysiology of OHSS. In this study, 14 immature female Wistar Albino rats were divided into two groups at random. The rats were 22 days old. Rats in the control group (n=7) were 22 days old, while those in the OHSS group (n=7) received 10 IU follicle-stimulating hormone subcutaneously over the course of four days and 30 IU human chorionic gonadotropin (hCG) on the fifth day to induce ovarian hyperstimulation. All the rats were sacrificed after all the groups' ovaries and blood samples were collected at the conclusion of the experiment. The left ovarian tissues were kept in aluminum foil at -80 °C, while the right ovarian tissues were kept in 10% formalin. Tissue vascular endothelial growth factor (VEGF), interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor (TNF)-α and malondialdehyde (MDA) levels were measured by The Enzyme Linked Immunosorbent Assay technique in the ovarian tissues. CHRM1 immunoreactivity was scored immunohistochemically. Ovarian weight, tissue IL-10, TNF-α, VEGF and MDA levels, and CHRM1 immunoreactivity were significantly increased in the OHSS group. Increased levels of CHRM1 activity may play a role in the pathophysiology of OHSS. With further studies, the effect of luteinizing hormone and hCG on the ovarian and hypothalamic cholinergic system can be further investigated, and useful information can be obtained in determining OHSS prevention strategies.

Comparison of dual-trigger and human chorionic gonadotropin-only trigger among polycystic ovary syndrome couples who underwent controlled ovarian stimulation and intrauterine insemination: A retrospective cohort study.

The dual-trigger regime, consisting of gonadotrophin releasing hormone agonist and human chorionic gonadotropin (HCG), has been shown to offer advantage over the HCG-only trigger regime. However, little is known about the influence of dual-trigger or HCG-only trigger regime on the reproductive outcome of polycystic ovary syndrome (PCOS) couples undergoing controlled ovarian stimulation (COS) and intrauterine insemination (IUI). A total of 404 cycles of COS and IUI treatments from couples with PCOS were enrolled, and divided, according to the regime of trigger, into dual-trigger group (n = 109, 0.1-0.2 mg gonadotrophin releasing hormone agonist plus 6000 IU HCG) and HCG-only group (n = 295, 10,000 IU HCG or 250 μg recombinant HCG). Baseline characteristics of the 2 groups were comparable (all P > .05). In dual-trigger group, live birth rate, clinical pregnancy rate and β -HCG positive rate were all higher as compared to the HCG-only group (20.18% vs 18.98%, 25.69% vs 23.39% and 28.44% vs 25.08% respectively), despite the differences failed to achieve statistical significances (all P > .05). Moreover, early miscarriage rate and multiple pregnancy rate of the dual-trigger group were lower than those of the HCG-only group (17.86% vs 18.84% and 3.57% vs 7.25% respectively), although no statistical significances were found (all P > .05). Additionally, logistic regression analysis revealed that age contributed significantly to the live birth of couples with PCOS ( P = .043, OR = 0.900). Dual-trigger regime for oocyte maturation seems to associate with beneficial improvements in reproductive outcomes of PCOS couples undergoing COS and IUI. Instead of HCG-only trigger, dual-trigger regime might be an alternative option in COS and IUI cycles for couples with PCOS.

Effect of progesterone infused controlled internal drug releasing (CIDR) device and timing of gonadotropin stimulation using P.G. 600 on reproductive success in ewes bred out of season.

Objectives of this study were to determine effects of exogenous progesterone (via controlled internal drug releasing devices; CIDR) in combination with exogenous gonadotropins (PMSG/hCG) use either at CIDR removal or 1 d before CIDR removal to induce estrus and cyclicity and subsequently enhance the proportion of ewes lambing, lambing rate, prolificacy, and days to lambing in ewes bred out of season. Multiparous ewes (n = 414) were randomly assigned to one of four treatments: untreated (U, n = 122), 7 d CIDR (C, n = 97), 7 d CIDR plus P.G. 600 (240 IU pregnant mare serum gonadotropin [PMSG] and 120 IU human chorionic gonadotropin [hCG]) at CIDR removal (CPG0, n = 97), and 7 d CIDR plus P.G. 600 (240 IU PMSG and 120 IU hCG) 1 d before CIDR removal (CPG-1, n = 98). Rams (n = 15) were joined with ewes immediately after CIDR removal and remained with ewes for a 21 d breeding period. Lambing data were summarized for the first 10 days of the lambing season and overall. Categorical data were analyzed using the GLIMMIX procedure of SAS whereas non-categorical data were analyzed using the mixed procedure. Proportion of ewes lambing in the first 10 d was greater (P < 0.05) for CPG0 and CPG-1 ewes compared with C ewes, which was greater (P < 0.0001) compared with U ewes. Overall proportion of ewes lambing was greater (P ≤ 0.0001) in all treatments utilizing CIDR compared with U ewes, but no differences (P ≥ 0.84) due to P.G. 600 were detected compared with C. Lambing rate in the first 10 d was greater (P < 0.05) for CPG-1 than C, with CPGO being intermediate, and all CIDR-treated ewes being greater than U (P < 0.0001). Overall lambing rate increased (P ≤ 0.0001) in all treatments utilizing CIDR compared with U ewes, but no differences (P ≥ 0.76) due to P.G. 600 were detected compared with C. Prolificacy was similar among all treatments both for the first 10 d of the lambing season (P = 0.86) and overall (P = 0.80). Day of lambing in the lambing season was reduced (P ≤ 0.03) for CPG0 and CPG-1 compared with CIDR-treated ewes, which was reduced (P < 0.0001) compared with U ewes (days 10.6, 9.0, 13.4, and 24.4 ± 0.9 for CPG0, CPG-1, CON, and U, respectively). Though timing of P.G. 600 did not influence results, the combination of CIDR and P.G. 600 enhanced the proportion of lambs born earlier in the lambing season, and incorporating a CIDR with or without P.G. 600 enhanced the overall proportion of ewes lambing and lambing rate in out-of-season breeding scenarios.

Simultaneously administration of cabergoline and PMSG reduces the duration of estrus induction in anestrous bitches.

This study designed a protocol that would combine pregnant mare serum gonadotrophin (PMSG) and cabergoline (CAB) to induce estrus in bitches. Twenty clinically healthy adult and anestrous female dogs were randomly assigned into four groups. The first group was treated with 5.00 μg kg-1 CAB until the onset of proestrus or for 25 days. The second group was treated with 20.00 IU kg-1 PMSG for 5 days and 500 IU human chorionic gonadotropin (hCG) on the 5th day. The third group was treated with 5.00 μg kg-1 CAB for 10 days in combination with 20.00 IU kg-1 PMSG for 5 days and 500 IU hCG on the 10th day. The control group received 1.00 mL of normal saline. Ovarian changes were evaluated ultrasonographically, and the estrus cycle phase was examined by vaginal cytology. Respectively, three, three and four bitches showed clinical signs of proestrus in each treatment group. The intervals between treatment and proestrus for each group were 30.00 ± 3.05, 7.67 ± 1.20 and 13.00 ± 1.20 days, respectively. Two weeks after estrus, the progesterone mean was 14.51 ± 6.24, 19.96 ± 17.16 and 19.12 ± 9.26 ng mL-1 for each group, respectively. In ultrasonography examination, the largest follicle was identified at 15.66 ± 1.33, 11.66 ± 2.40 and 8.75 ± 2.17 days after the onset of proestrus and the largest follicle's size was measured 6.50 ± 0.55, 4.83 ± 1.64 and 7.07 ± 1.49 mm for each group, respectively. Although the combined use of CAB and PMSG reduced the duration of treatment, alteration of the duration or PMSG dosage can be helpful to improve the results.

Exogenous L-Glutathione Improves Vitrification Outcomes in Murine Preimplantation Embryos.

Vitrification is an important tool to store surplus embryos in assisted reproductive technology (ART). However, vitrification increases oxidative damage and results in decreased viability. Studies have reported that L-glutathione (GSH) supplementation improves the preimplantation development of murine embryos. Glutathione constitutes the major non-protein sulphydryl compound in mammalian cells, which confers protection against oxidative damage. However, the effect of GSH supplementation on embryonic vitrification outcomes has yet to be reported. This study aims to determine whether GSH supplementation in culture media improves in vitro culture and vitrification outcomes, as observed through embryo morphology and preimplantation development. Female BALB/c mice aged 6-8 weeks were superovulated through an intraperitoneal injection of 10 IU of pregnant mare serum gonadotrophin (PMSG), followed by 10 IU of human chorionic gonadotrophin (hCG) 48 h later. The mated mice were euthanized by cervical dislocation 48 h after hCG to harvest embryos. Two-cell embryos were randomly assigned to be cultured in either Group 1 (GSH-free medium), Group 2 (GSH-free medium with vitrification), Group 3 (0.01 mM GSH-supplemented medium), or Group 4 (0.01 mM GSH-supplemented medium with vitrification). Non-vitrified (Groups 1 and 3) and vitrified (Groups 2 and 4) embryos were observed for morphological quality and preimplantation development at 24, 48, 72, and 96 h. In the non-vitrified groups, there were significant increases in the number of Grade-1 blastocysts in GSH cultures (p &lt; 0.05). Similarly, in the vitrified groups, GSH supplementation was also seen to significantly increase blastocyst formation. Exogenous GSH supplementation resulted in a significant increase in intracellular GSH, a release of cytochrome c from mitochondria, and a parallel decrease in intracellular reactive oxygen species (ROS) levels in vitrified eight-cell embryos (p &lt; 0.05). GSH supplementation was shown to upregulate Bcl2 expression and downregulate Bax expression in the vitrified preimplantation embryo group. The action of exogenous GSH was concomitant with an increase in the relative abundance of Gpx1 and Sod1. In conclusion, our study demonstrated the novel use and practical applicability of GSH supplementation for improving embryonic cryotolerance via a decrease in ROS levels and the inhibition of apoptotic events by improvement in oxidative status.

The use of vinylcyclohexene diepoxide to create diminish ovarian reserve model on rats

Background: Infertility remains a significant issue in the world of health. Now Assisted Reproductive Technology (ART) is widely used to help couples with infertility. In ART diminish ovarian reserve patients have a very low success rate of pregnancy. To help patients with Diminished Ovarian Reserve (DOR), research is conducted using an animal model. vinylcychlohexene diepoxide (VCD) has ovotoxic effects. This study looked at the use of VCD in inducing targeted DOR conditions in animals (rats), create a model that could be used for future animal studies in infertility. Methods: Adult female Rattus norvegicus were used in this study. All were given VCD injections at 80 mg/kg intraperitoneally for 15 days. An examination of Follicle Stimulating Hormone (FSH) levels will be carried out on days 0,3,5,7, and 15. To mimic IVF process, all rats got an injection of 10 IU of Pregnant Mare Serum Gonadotropin (PMSG) and 10 IU human Chorionic Gonadotropin (hCG) to confirm DOR. Ovarectomy was performed, dominant follicles were taken, denudation was carried out, and the oocytes were cultured for 12 hours. Then, oocytes were assessed through a microscope. Results: Starting from day 5 post VCD induction, there was a significant increase in the FSH level in the group of rats that were induced with the administered VCD dose compared to the control group that experienced normal FSH fluctuations. The FSH concentration reached &gt;50%. It was found that the number of degenerated oocytes had an average of 1.11. Oocytes that were in Germinal-Vesicle (GV) stage had an average of 1.89, while in the Metaphase I (M1) stage the average is 2.882. Lastly, the number of oocytes in the Metaphase (MII) stage which is fully matured had an average of 1.117. Conclusions: VCD can become an important tool for future studies that needs an animal model with DOR.

ANALYSIS OF PARAMETERS OF MORPHOFUNCTIONAL STATE OF PIG OOCYTE LIPIDOME IN CASE OF IN VITRO CULTIVATION

Identification of effective markers of oocyte maturation based on morphological and functional characteristics of intracytoplasmic lipid droplets will allow to increaseinformative value of the criteria for assessing the quality of donor gametes for improvement of the efficiency of using cellular reproductive technologies in animal husbandry, biomedicine, and veterinary medicine. The aim of the study is to analyze the parameters of morphofunctional state of the lipidome (the morphology of lipid droplets and the intensity of fluorescence of the Nile red/lipid droplets complex - Nile red IF) in Susscrofadomesticus oocytes before and after in vitro cultivation. Oocytes were matured using Sage Media Cleavage (CooperSurgical, USA) with 5% Serum Protein Substitute serum (CooperSurgical, USA), 10 IU human chorionic gonadotropin (Sigma, USA). As a result of ranking the population of donor oocytes according to morphology of lipid droplets (granules and clusters) and Nile red IF (low, medium, high), it was found that, the proportion of oocytes with an average Nile red IF and lipid droplets (LD) in the form of granules was 63% before cultivation, which was significantly higher than the proportion of gametes with an average IF of Nile red and LD of a mixed form of 41% (P&lt;0.05). After cultivation, the level of oocytes with medium Nile red IF and LD in the form of granules was 45%, which exceeded the level of gametes with the same Nile red IF and mixed LD of 23% (P&lt;0.05). The proportion of in vitro matured oocytes with high Nile red IF and mixed LD morphology (granules and clusters) was 33%, 14% were characterized by the above parameters before cultivation (P&lt;0.05). Comparative morphological analysis of the relation between the parameters of morphofunctional activity (morphology and IF Nile red) of lipidome in Susscrofadomesticus oocytes was carried out for the first time. Features of functional activity of lipidome before and after cultivation were revealed. The usage of a comprehensive analysis of parameters of functional activity of lipidomeforassessment of the quality of the oocyte is substantiated. Pig oocytes which morphology of lipid droplets is represented by a granular form are promising for cultivation, while Nile red IF is assessed as low. With regard to the analysis of the obtained results, it is recommended to use the proposed comprehensive assessment of lipidome based on morphology and Nile red IF to assess the initial population of donor oocytes in cellular reproductive technologies.

171 Sex-Sorted Sperm Used for Fixed-Time Artificial Insemination and in Vitro Fertilization in Wood Bison

Abstract Wood bison (Bison bison athabascae) are a threatened species native to Canada. In a conservation setting such as zoos, females are preferred over males and sex-sorted sperm may help skew the proportion of females born. The objective of this study was to determine the efficacy of sex-sorted sperm in fixed-time artificial insemination (FTAI, Experiment 1) and in vitro fertilization (IVF, Experiment 2). In Experiment 1, females in bison herds at the University of Saskatchewan and Toronto Zoo were synchronized with 3.3 mg estradiol-17ß + 50 mg progesterone and given a progesterone-releasing intravaginal device for 8 days. At device removal, bison were given 500 µg of cloprostenol, and 72 h later were given 2500 IU human chorionic gonadotropin (hCG) and inseminated with either sex-sorted or conventional sperm. A second insemination occurred 24 h after hCG. Pregnancy diagnosis and fetal sexing was done by transrectal ultrasonography at 30- and 70-days post-AI, respectively. In Experiment 2, cumulus oocyte complexes collected transvaginally from live bison were fertilized in vitro with either sex-sorted or conventional sperm. Cleavage and embryo rates were determined at 3- and 7-days post fertilization, respectively. Proportional data were compared by chi-square. In Experiment 1, there was no effect of herd (P=0.17) or sperm (P=0.68) on pregnancy per AI (P/AI) at 30 days (total=25/51 [49%]). The proportion of female calves was greater in the sex-sorted than conventional group (9/10 [90%] vs 3/11 [27%]; P&amp;lt; 0.01). In Experiment 2, the cleavage (122/222 [55%] vs 62/94 [66%];P=0.07) and embryo rate (18/222 [8%] vs 14/94 [15%]; P=0.07) tended to be lower in the sex-sorted than the conventional group. In summary, sex-sorted sperm increased the proportion of female bison calves born following FTAI. Future efforts will include improving embryo production rates following IVF with sex-sorted sperm. Funding provided by Toronto Zoo, Mitacs, and STGenetics.

Hormonal profile in early luteal phase after triggering ovulation with gonadotropin-releasing hormone agonist in high-responder patients.

The major limitations associated with gonadotropin-releasing hormone agonist (GnRHa) triggering are inferior clinical outcomes in fresh embryo transfer cycles caused by luteal phase insufficiency following the GnRHa triggering. We included 153 high-risk patients in this study. In group I, the patients received gonadotropin-releasing hormone agonist (GnRHa) trigger + 1,500 IU human chorionic gonadotropin (hCG) support on the oocyte pick-up (OPU) day; in group II, the patients had a dual trigger (GnRHa + 1,500 IU hCG); and in group III (control), 10,000 IU hCG trigger was prescribed for the final oocyte maturation. The levels of LH, estradiol, and progesterone were evaluated in serum on the stimulation starting day, day 6 of stimulation, on the day of the trigger administration, OPU day, days 3 and 5 post-OPU, and day 14 post-ET, as well as in follicular fluid. Progesterone concentration was significantly lower in group I on OPU+5 compared to the hCG group (I vs. III, р = 0.0065). Progesterone levels were significantly lower in group II in serum on OPU+5 compared to groups I and III (I vs. II, р = 0.0068; II vs. III, р = 1.76 × 108). The progesterone levels were significantly higher in follicular fluid in group III compared to the study groups (I vs. III, р = 0.002; II vs. III, p = 0.009). However, no significant differences in clinical outcomes were found between the groups. Then, we divided all women into pregnant and non-pregnant groups and found that estradiol (p = 0.00009) and progesterone (p = 0.000036) on the day of the pregnancy test were significantly higher in the pregnant women group. Also, progesterone on OPU day was significantly higher in the non-pregnant group (p = 0.033). Two cases of moderate ovarian hyperstimulation syndrome (OHSS) late-onset occurred in group I (3.5%, 2/56), no case of moderate/severe OHSS late-onset in group II, and three cases of moderate late-onset in group III (5.7%, 3/53). The low-dose hCG supplementation improves the luteal phase insufficiency after GnRHa triggering, which is confirmed by the comparable pregnancy rates in fresh transfer cycles between the groups. However, low-dose hCG carries a similar risk of OHSS as the full dose of hCG in high-responder patients.

Triggering with 1,500 IU of human chorionic gonadotropin plus follicle-stimulating hormone compared to a standard human chorionic gonadotropin trigger dose for oocyte competence in in vitro fertilization cycles: a randomized, double-blinded, controlled noninferiority trial

To assess if triggering with 1,500 IU of human chorionic gonadotropin (hCG) with 450 IU of follicle-stimulating hormone (FSH) induces noninferior oocyte competence to a standard dose of hCG trigger used in invitro fertilization (IVF). The alternative trigger will be considered noninferior if it is at least 80% effective in promoting oocyte competence. Randomized, double-blinded, controlled noninferiority trial. Academic infertility practice. Women aged 18-41 undergoing IVF with antral follicle count ≥8, body mass index ≤30 kg/m2, and no history of ≥2 IVF cycles canceled for poor response were enrolled. Participants with a serum estradiol >5,000 pg/mL on the day of trigger were excluded because of high risk of ovarian hyperstimulation syndrome. Participants were randomized to receive an alternative trigger of 1,500 IU of hCG plus 450 IU of FSH or a standard trigger dose of hCG (5,000 or 10,000 IU) for final oocyte maturation. The primary outcome was total competent proportion, defined as the probability of 2 pronuclei from an oocyte retrieved. The alternative trigger will be considered noninferior to the standard trigger if a 1-sided 95% confidence interval (CI) of the relative risk (RR) is not <0.8. Secondary outcomes included oocyte recovery and maturity, intracytoplasmic sperm injection fertilization, embryo quality, pregnancy rates, as well as serum and follicular hormones. Secondary outcomes were compared using a 2-sided superiority test. Outcomes were analyzed by intention-to-treat and per-protocol. A total of 105 women undergoing IVF were randomized from May 2015 to June 2018. The probability of the primary outcome was 0.59 with the alternative trigger and 0.65 with the standard trigger, with a RR of 0.91 and a 1-sided 95% CI of 0.83. Noninferiority of the alternative trigger was demonstrated. Live birthrate from all fresh transfers in the alternative trigger group vs. standard trigger was 46.9 vs. 46.4% (RR, 1.01; 95% CI, 0.62-1.62), respectively. Live birthrate per randomized participant was 48.1% in the alternative trigger group vs. 62.7% with the standard trigger (RR, 0.73; 95% CI, 0.48-1.11). No participants had a failed retrieval. Triggering with 1,500 IU of hCG plus 450 IU of FSH promoted noninferior oocyte competence compared to a standard hCG trigger dose. NCT02310919.

Does the HCG trigger dose used for IVF impact luteal progesterone concentrations? a randomized controlled trial

Research questionIs there an association between the ovulation trigger dose of human chorionic gonadotrophin (HCG) and endogenous progesterone production during the luteal phase? DesignThis randomized controlled four-arm study, at the Fertility Clinic, Odense University Hospital, Denmark, included women undergoing gonadotrophin-releasing hormone (GnRH) antagonist IVF treatment with ≤11 follicles ≥12 mm. Group 1–3 were triggered with 5000 IU, 6500 IU or 10,000 IU HCG, respectively, receiving 17α-hydroxyprogesterone caproate intramuscularly for luteal-phase support (LPS) to measure endogenous progesterone production. Group 4 received 6500 IU HCG trigger and vaginal progesterone. During the study, the 5000 IU and 10,000 IU HCG groups were switched from urinary to recombinant HCG, as urinary HCG was removed from market. Eight blood samples were drawn during the luteal phase. ResultsNinety-four participants completed the study. There was a significant positive association between the HCG trigger dose and the progesterone at 8 days (P < 0.001), 10 days (P < 0.001) and 14 days (P < 0.001) post-oocyte retrieval. Comparing the groups individually revealed a significant difference in progesterone concentration between low and high trigger doses at 4 days (P = 0.037) and 8 days (P = 0.007) post-oocyte retrieval and between all intervention groups at oocyte retrieval + 6 days: group 1 and 2 (P = 0.011), group 2 and 3 (P = 0.042) and group 1 and 3 (P < 0.001). Higher HCG trigger dose increased the progesterone from the individual follicle. ConclusionsIncreasing HCG trigger doses significantly increased endogenous progesterone concentration during the mid–late luteal phase.

Early Follicular Phase Human Chorionic Gonadotropin Addition May Improve the Outcomes of In Vitro Fertilization/Intracytoplasmic Sperm Injection in Patients With "Unpredictable" Poor Response to Gonadotropin-Releasing Hormone Antagonist Protocol.

PurposeTo compare the effects of early and mid-late follicular phase administration of 150 IU of human chorionic gonadotropin (hCG) on gonadotropin-releasing hormone (GnRH) antagonist protocol in “unpredictable” poor ovarian response (POR) women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment.MethodsA retrospective single-center cohort study was conducted on 67 patients with “unpredictable” POR in their first IVF/ICSI cycle receiving GnRH antagonist protocol. Patients were treated with a second IVF/ICSI cycle using the same GnRH antagonist protocol with the same starting dose of recombinant follicle-stimulating hormone (rFSH) as the first cycle; a daily dose of 150 IU of hCG was administrated on either stimulation day 1 (Group A, n = 35) or day 6 (Group B, n = 32). The number of oocytes retrieved, number of usable embryos, serum level of estradiol (E2) on day of hCG trigger, and clinical pregnant outcomes were studied.ResultsThe addition of 150 IU of hCG on either the first day or sixth day of stimulation increases the serum level of E2, luteinizing hormone (LH), and hCG on the day of hCG trigger. Only the use of 150 IU of hCG on the first stimulation day improved the number of oocytes retrieved, mature of oocytes, and usable embryos, but not the addition of hCG on stimulation day 6. Implantation rate, clinical pregnancy rate, and ongoing pregnancy rate showed an increasing trend in patients receiving 150 IU of hCG in the early phase compared with mid-late phase, even thought there was no statistically significant difference.ConclusionsOur study demonstrated that adding 150 IU of hCG in subsequent GnRH antagonist cycle in “unpredictable” poor responders is associated with the improvement of response to stimulation. Furthermore, early follicular phase addition of 150 IU of hCG significantly increased the number of oocytes retrieved and usable embryos than did the mid-late addition of the same dose.

Induction of oestrus by administering Inhibin antiserum along with equine chorionic gonadotropin in anoestrous bitches.

As dogs experience oestrus only once or twice a year, it is necessary to establish an effective method of oestrous induction for efficient breeding. In the present study, we evaluated inhibin antiserum (IAS) on oestrous induction in anoestrous females. Bitches were administered 0.5ml/kg IAS or a mixture of 50IU/kg equine chorionic gonadotropin (eCG) and 0.5ml/kg IAS and 500IU human chorionic gonadotropin (hCG) administered 7days after the mixture injection. As a control, bitches received 50IU/kg eCG, with 500IU hCG administered 7days after eCG injection. Blood-tinged vaginal discharge, vulvar swelling, plasma progesterone concentrations and ovarian follicular development were assessed from day 0 to day 14. IAS alone injection did not induce oestrus in bitches at the anoestrous stage. Conversely, vulvar swelling, blood-tinged vaginal discharge and an estimated luteinizing hormone (LH) surge appeared on days 3-7, days 3-6 and days 7-9 after the IAS+eCG mixture injection, respectively, in all five bitches at the anoestrous stage. The average number of developing and ovulated follicles in bitches administered IAS+eCG was 8.8 and 9.6 respectively. A single eCG injection followed by hCG induced oestrous signs, with an average of 8.3 developing follicles and 4.5 ovulated follicles. This study revealed that IAS alone did not induce oestrus, but when IAS was used in combination with eCG, it induced oestrus and promoted a considerable number of ovulations in anoestrous dogs.

PERKEMBANGAN EMBRIO PREIMPLANTASI SECARA IN VIVO SETELAH PEMBERIAN EKSTRAK ETHANOL AKAR ALANG-ALANG (IMPERATA CYLINDRICA L) PER ORAL

Cogon grass (Imperata cylindrica L) is known as a medicinal plant that is scattered almost worldwide. Despite its role that inhibits another plant’s growth, cogon grass possesses several benefits in health. This research has to identify the effect of short-term gavage ethanolic extract of cogon grass roots (CGG) to in-vivo mice preimplantation embryo development. A total of 60 female mice were divided into control and treatment groups, dosages at 90 and 115 mg/kg of body weight of CGG, orally gavage for 20 days. The superovulation of mice was done at the end of the CGG treatment by injecting 5 IU Pregnant Mare Serum Gonadotropin (PMSG) and after 48 hours, followed by 5 IU Human Chorionic Gonadotropin (hCG) injection and directly the mice were mated. The mating rate was checked by the appearance of the vaginal plug 12 hours after hCG injection. Mice were sacrificed, the oviducts and cornua of uteri were isolated to collect the oocytes and embryonic cells by flushing the oviducts and cornua uteri with Phosphate-buffered saline (PBS). The effects of CGG as an antifertility were evaluated by measuring the number of oocytes, fertilization, and in-vitro embryo development rates. The results showed significantly reduced about half of the mating rate in the 115 mg/kg BW group (p&lt;0.05) compared to control. However, the 90 mg/kg BW dose reduced 20% mating rate compared to control, and not significant (p&gt;0.05). In all treatment groups, only half oocytes fertilized. The cleavage and blastocyst rate in 115 mg/kg BW group were significantly lower compared to the control group (p&lt;0.05). In conclusion, oral gavage of cogon grass root ethanolic extract disrupts the mating process and development of in-vivo mice preimplantation embryo development.

Transient receptor potential melastatin 2 ion channel activity in ovarian hyperstimulation syndrome physiopathology

Background/aimOvarian hyperstimulation syndrome (OHSS) is a complication of ovarian stimulation with increased vascular endothelial growth factor (VEGF) and vascular permeability in the ovarian tissue. Transient receptor potential melastatin 2 (TRPM2) is known to be associated with angiogenesis and vascular permeability. In this experimental study, we aimed to investigate the activity of TRPM2 in the development of OHSS. Materials and methodsFourteen immature female rats were divided into two groups. Group 1 was the control group, and Group 2 was the OHSS group that was exposed to 10 IU of subcutaneous application of FSH for four days and 30 IU of human chorionic gonadotropin (hCG) on the 5th day. At the end of the experiment, the ovaries were removed. The right ovarian tissues were stored in 10% formol for histopathological and immunohistochemical examination. The left ovarian tissues were stored at –80 °C for biochemical examinations. VEGF, tumor necrosis factor-alpha (TNF‐α) and malondialdehyde (MDA) levels were measured in the ovarian tissue. Congestion, edema, apoptosis and TRPM2 immunoreactivity were evaluated.ResultsThere was a significant increase in ovarian weight in the OHSS group compared to the control group. There was a significant increase in congestion, edema, apoptosis and TRPM2 immunoreactivity in the OHSS group. A significant increase in tissue levels of VEGF, TNF‐α and MDA was also found in the OHSS group compared to the control group. Conclusion As a result of our experiment, it was found that increased TRPM2 immunoreactivity on hyperstimulated rat ovary may be the reason or result of edema and congestion. Further studies are needed to discuss our results.

The exogenous progesterone-free luteal phase: two pilot randomized controlled trials in IVF patients

Research questionIs the reproductive outcome similar after gonadotrophin-releasing hormone agonist (GnRHa) trigger followed by luteal human chorionic gonadotrophin (HCG) boluses compared with HCG trigger and a standard luteal phase support (LPS)? DesignTwo open-label pilot randomized controlled trials (RCT) with 250 patients from 2014 to 2019, with a primary outcome of ongoing pregnancy per embryo transfer. Patients with ≤13 follicles on the trigger day were randomized (RCT 1) to: Group A (n = 65): GnRHa trigger followed by a bolus of 1500 IU HCG s.c. on the oocyte retrieval day (ORD) and 1000 IU HCG s.c. 4 days later, and no vaginal LPS; or Group B (n = 65): 6500 IU HCG trigger, followed by a standard vaginal progesterone LPS. Patients with 14–25 follicles on the trigger day were randomized (RCT 2) to Group C (n = 60): GnRHa trigger followed by 1000 IU HCG s.c. on ORD and 500 IU HCG s.c. 4 days later, and no vaginal LPS; or Group D (n = 60): 6500 IU HCG trigger and a standard vaginal LPS. ResultsIn RCT 1, the ongoing pregnancy rate was 44% (22/50) in the GnRHa group versus 46% (25/54) in the HCG trigger group (RR 0.95, 95% CI 0.62–1.45). No ovarian hyperstimulation syndrome (OHSS) was seen in Groups A or B. In RCT 2, the ongoing pregnancy rate was 51% (25/49) in the GnRHa group versus 60% (31/52) in the HCG trigger group (RR 0.86, 95% CI 0.60–1.22). The OHSS rates were 3.3% and 6.7%, respectively. ConclusionsAlthough a larger-scale study is needed before standard clinical implementation, the present study supports that the exogenous progesterone-free LPS is efficacious, simple and patient-friendly.