Expression Profiles of Genes Related to Development and Progression of Endometriosis and Their Association with Paraben and Benzophenone Exposure.

Increasing evidence has been published over recent years on the implication of endocrine-disrupting chemicals (EDCs), including parabens and benzophenones in the pathogenesis and pathophysiology of endometriosis. However, to the best of our knowledge, no study has been published on the ways in which exposure to EDCs might affect cell-signaling pathways related to endometriosis. We aimed to describe the endometriotic tissue expression profile of a panel of 23 genes related to crucial cell-signaling pathways for the development and progression of endometriosis (cell adhesion, invasion/migration, inflammation, angiogenesis, and cell proliferation/hormone stimulation) and explore its relationship with the exposure of patients to parabens (PBs) and benzophenones (BPs). This cross-sectional study included a subsample of 33 women with endometriosis from the EndEA study, measuring their endometriotic tissue expressions of 23 genes, while urinary concentrations of methyl-, ethyl-, propyl-, butyl-paraben, benzophenone-1, benzophenone-3, and 4-hydroxybenzophenone were determined in 22 women. Spearman's correlations test and linear and logistic regression analyses were performed. The expression of 52.2% of studied genes was observed in >75% of endometriotic tissue samples and the expression of 17.4% (n = 4) of them in 50-75%. Exposure to certain PB and BP congeners was positively associated with the expression of key genes for the development and proliferation of endometriosis. Genes related to the development and progression of endometriosis were expressed in most endometriotic tissue samples studied, suggesting that exposure of women to PBs and BPs may be associated with the altered expression profile of genes related to cellular pathways involved in the development of endometriosis.

[Medical treatments for male infertility]. / Traitements médicaux de l'homme infertile.

BACKGROUND: Treatments to stimulate spermatogenesis and antioxidant food supplements are often offered to infertile patients either before sperm extraction surgery to improve results, or as part of medically assisted reproduction or spontaneous fertility to increase the likelihood of a live birth. METHODS: A bibliographic search limited to English-language literature on men published before 5/2023 was carried out, including clinical trials, literature reviews and meta-analyses on spermatogenesis-stimulating molecules and antioxidant treatments. RESULTS: Several medical treatments seem capable of improving male fertility: they act mainly by stimulating spermatogenesis through hormones, or by reducing the effects of oxidative stress. With regard to oligoasthenozoospermia, the literature shows that certain hormonal treatments stimulating spermatogenesis are useful. In the case of non-obstructive azoospermia, the value of treatment depends on the patient's FSH and testosterone levels. AOX supplementation appears to improve certain spermogram parameters and have an impact on pregnancy and live birth rates. CONCLUSION: This review should help urologists gain a better understanding of the various medical treatments and enable them to define an appropriate therapeutic strategy, tailored to the patient and the couple, in order to obtain the best results.

Safety of fertility preservation in breast cancer patients in a register-based matched cohort study.

PURPOSE: To determine if women with breast cancer that undergo fertility preservation (FP), with or without hormonal stimulation, present with an increased risk of breast cancer recurrence. METHODS: A matched cohort study on women with breast cancer attempting to ensure FP in Stockholm from 1999 to 2013 [exposed women (n = 188), age-matched unexposed controls (n = 378)] was designed using the Stockholm regional data from the Swedish National Breast Cancer Quality Register. Breast cancer relapse rates [incidence rate ratio (IRR)] and 95% confidence interval (CI) were estimated using Cox regression and adjusted for potential confounding factors. Completeness of the registry at the time of the study was close to 99%. RESULTS: Most women attempted FP by hormonal stimulation treatment (n = 148, 79%) with the objective of freezing their eggs or embryos. A smaller group elected FP methods without hormone stimulation (n = 40, 21%). Women who received hormone stimulation did not present with a higher relapse rate than unexposed control women in a model adjusted for age and calendar period of diagnosis (IRR 0.59, 95% CI 0.34-1.04). The results remained virtually unchanged after adjustment for tumor size, estrogen receptor status, affected lymph nodes, and chemotherapy treatment (IRR 0.66, 95% CI 0.37-1.17). CONCLUSION: Evidence was not found that fertility preservation, with or without hormonal stimulation, was associated with an increased risk of breast cancer recurrence. The high coverage rate of this population-based study supports the safe practice of fertility preservation in young women with breast cancer.

Pediatric Penile and Glans Anthropometry Nomograms: An Aid in Hypospadias Management.

OBJECTIVE: To establish pediatric penile and glans anthropometry nomograms. This may be used as a reference model for penile assessment while managing hypospadias. PATIENTS AND METHODS: Between October 2012 and September 2013, 263 boys of varying ages (0-16 years) were included in the study. Those with genetic, endocrine disorders, having genital anomaly, undescended testis, neonates, and infants with a nonretractile prepuce, with multiple congenital anomalies and refusal to take part in the study were excluded. Evaluated outcome variables were stretched penile length, glans circumference (GC) at coronal sulcus, glans diameter at coronal sulcus (Gdcl), mid glans diameter, and ventral glans length. Glans ratios were generated by dividing Gdcl by GC. Data were expressed as mean, median, and standard deviation. Correlation between age and variables was evaluated using nonparametric Spearman's rank correlation coefficient. RESULTS: The patients were divided in six age groups, namely 0-1 (n = 61), 1-3 (n = 37), 3-5 (n = 36), 5-7 (n = 36), 7-12 (n = 45), and >12 years (n = 48). Gdcl was the maximum transverse glans diameter and based on it small glans size varied widely from 8.9 to 35.04 mm for various age groups. Although glans anthropometry showed age-related changes, glans ratio remained relatively constant between 0.49 and 0.53 (mean: 0.5 ± 0.051, r = 0.29). All the variables except glans ratio showed a significant positive correlation with age (r = 0.954-0.98, P < 0.01). CONCLUSION: Penile anthropometry nomograms provide a reference model for hypospadias. This may aid in (a) objective preoperative assessment of glans size (b) patient selection for preoperative hormonal stimulation (c) provides a yardstick for postoperative cosmesis.

Effect of temperature and spawning agents on wild female asp (Leuciscus aspius) reproductive efficiency under controlled conditions.

Artificial reproduction is a bottleneck to produce stocking material for many species of freshwater fish. One of these species is the asp, Leuciscus aspius. Research in the field of artificial reproduction of this species is very scarce and often incomplete. There are no breeding protocols specifying optimal environmental conditions and hormonal stimulation for many species of rheophilic cyprinids, including asp. Since the number of natural asp populations is constantly decreasing, it is important to support natural stocks by restocking with high quality stocking material. For this reason, optimized protocols are needed to breed this species under controlled conditions to produce stocking material with high biodiversity and good health. Such an approach will make it possible to maintain the population of natural asp at a constant level. The aim of this study was to develop the protocol of asp artificial reproduction using optimized thermal conditions and appropriate hormonal stimulation. In experiment I, the influence of constant temperature (10.0, 12.0 and 14.0 °C) on the effectiveness of artificial reproduction of asp. In experiment II, the effectiveness of asp reproduction was checked after the application of spawning agents: Ovopel, Ovaprim or a combination of these two agents The obtained results indicate that for the final maturation of oocytes (FOM) and artificial reproduction of asp in controlled conditions, water temperatures of 10-12 °C are the most useful. Under these thermal conditions, the highest percentages of female's ovulation and embryo survival, as well as the percentage of hatching, were obtained. Hormone injections are necessary to perform final oocyte maturation (FOM) in female asp in captivity. All spawning agents used were especially useful for artificial reproduction of asp, however, the best values of the studied indices, such as ovulation rate and embryo survival, were obtained after the application of Ovaprim or the combination of Ovopel and Ovaprim in water temperature at a range of 10-12 °C. It was found that the pH of ovarian fluid may be a preliminary indicator of the biological quality of eggs in asps. The optimal pH value is 8.0-8.4. At pH below 7.4, no viable embryos were observed.

Ovarian lavage / artificial insemination method as an alternative approach for controlled reproduction of the northern pike (Esox lucius).

This study aimed to compare the effectiveness of the ovarian lavage / artificial insemination method with the traditional hormonal administration and fertilization methods over the artificial reproduction of the northern pike (Esox lucius). For this purpose, groups of five females were treated as follows: intraperitoneal injection of saline (C1); ovarian lavage with saline (C2); intraperitoneal injection of carp pituitary extract (CPE, T1); ovarian lavage with CPE (T2); intraperitoneal injection of CPE and ovarian lavage with semen after 72 h (T3); ovarian lavage with CPE and ovarian lavage with semen after 72 h (T4). According to the results, no fish ovulated in the control groups (C1 and C2). There were no significant differences (n.s.) among experimental treatments (P > 0.05, n.s.) in the reproductive parameters, such as latency time, ovulation rate, stripped egg amount, and pseudo-gonadosomatic index. The lowest fertilization rate (54.8%) was observed in the T4 treatment and significantly differed from the T1 and T2 treatments (P < 0.05). Moreover, the highest survival at swim-up stage was measured in the T4 treatment with a significant difference compared to the T1 group (P < 0.05). The survival at the eyed-egg stage, hatching rate, and malformations were similar (P > 0.05, n.s.) in all applied strategies. The results demonstrated that the ovarian lavage / artificial insemination method could be applied to control northern pike reproduction, like the traditional fertilization method. Consequently, this novel technique can be suggested as an alternative strategy to facilitate the hatchery operations in the controlled reproduction of this species.

Artificial reproduction of wild and pond-cultured common tench (Tinca tinca) under controlled conditions.

The common tench is an important thermophilic freshwater fish used for aquaculture diversification. This species is farmed in ponds as well as caught in the wild in many European and China. Artificial reproduction under controlled conditions is one of the most important part of modern aquaculture. In this study, two experiments were conducted in which the influence of hormonal agents (Ovopel and Ovaprim - experiment I) and the origin of the spawning stock (wild and pond-cultured - experiment II) on the success of reproduction in terms of ovulation rate (%), latency time (h), pseudo-gonadosomatic index (PGSI, %), hatching rate (%), abnormal larvae rate (%), and spawning efficiency coefficient (Se) were examined. While the wild fish could not reproduce in the absence of hormonal treatment with 0 % ovulation rate. In all tested hormonal treatment allowed to obtain high-quality eggs with hatching rates of over 83 %. Significant differences in latency time were observed among groups from different origins, such as differences in ovulation rate (79 % vs. 90 %, for pond-cultured and wild fish, respectively), PGSI (8.2 vs. 8.9), and hatching rate (63.8 % vs. 84.3 %), which showed significantly better quality in fish from the wild than those from pond culture. This means that when spawners of common tench of various origins are available, it is better to choose spawners from natural waters for artificial reproduction.

In vitro matured oocytes have a higher developmental potential than in vivo matured oocytes after hormonal ovarian stimulation in Callithrix jacchus.

BACKGROUND: The common marmoset, Callithrix jacchus, is an invaluable model in biomedical research. Its use includes genetic engineering applications, which require manipulations of oocytes and production of embryos in vitro. To maximize the recovery of oocytes suitable for embryo production and to fulfil the requirements of the 3R principles to the highest degree possible, optimization of ovarian stimulation protocols is crucial. Here, we compared the efficacy of two hormonal ovarian stimulation approaches: 1) stimulation of follicular growth with hFSH followed by triggering of oocyte maturation with hCG (FSH + hCG) and 2) stimulation with hFSH only (FSH-priming). METHODS: In total, 14 female marmosets were used as oocyte donors in this study. Each animal underwent up to four surgical interventions, with the first three performed as ovum pick-up (OPU) procedures and the last one being an ovariohysterectomy (OvH). In total, 20 experiments were carried out with FSH + hCG stimulation and 18 with FSH-priming. Efficacy of each stimulation protocol was assessed through in vitro maturation (IVM), in vitro fertilization (IVF) and embryo production rates. RESULTS: Each study group consisted of two subgroups: the in vivo matured oocytes and the oocytes that underwent IVM. Surprisingly, in the absence of hCG triggering some of the oocytes recovered were at the MII stage, moreover, their number was not significantly lower compared to FSH + hCG stimulation (2.8 vs. 3.9, respectively (ns)). While the IVM and IVF rates did not differ between the two stimulation groups, the IVF rates of in vivo matured oocytes were significantly lower compared to in vitro matured ones in both FSH-priming and FSH + hCG groups. In total, 1.7 eight-cell embryos/experiment (OPU) and 2.1 eight-cell embryos/experiment (OvH) were obtained after FSH + hCG stimulation vs. 1.8 eight-cell embryos/experiment (OPU) and 5.0 eight-cell embryos/experiment (OvH) following FSH-priming. These numbers include embryos obtained from both in vivo and in vitro matured oocytes. CONCLUSION: A significantly lower developmental competence of the in vivo matured oocytes renders triggering of the in vivo maturation with hCG as a part of the currently used FSH-stimulation protocol unnecessary. In actual numbers, between 1 and 7 blastocysts were obtained following each FSH-priming. In the absence of further studies, FSH-priming appears superior to FSH + hCG stimulation in the common marmoset under current experimental settings.

In vitro mimicking of estrous cycle stages in porcine oviduct epithelium cells: estradiol and progesterone regulate differentiation, gene expression, and cellular function.

Throughout the estrous cycle the oviduct epithelium undergoes dramatic morphological and functional changes. To elucidate cyclic cellular events and associated regulation mechanisms of 17beta estradiol (E2) and progesterone (P4), we mimicked estrous cycle stages in vitro using a culture system of primary porcine oviduct epithelium cells (POEC). Cells were polarized in an air/liquid interface and then treated with E2 and P4 for physiological time periods: In experiment 1, high concentration of P4 with low concentration of E2 for 10 days resembled diestrus; in experiment 2, following the previous diestrus, sequential high E2 with low P4 for 2.5 days represented estrus. Histomorphometry and electron microscopy showed cyclic changes in cellular height, cell population, and cilia density under the influence of hormone stimulation. Transepithelial electrical resistance was high in simulated diestrus but reduced in estrus. Thus, E2 and P4 affect cellular polarity, transformation of ciliated and secretory cells, as well as electrical conductivity of oviduct epithelium. Simulation of diestrus led to significant decrease in expression of hormone receptors (PGR and ESR1) and other epithelial markers (MUC16, OVGP1, and HSP90B1), while sequential simulated estrus caused an increase in these markers. The hormonal regulation of some marker genes was clearly time-dependent. Furthermore, POEC showed increased sperm-binding capacity in simulated estrus. In this study, we also present a novel approach based on the AndroVision software, which can be routinely utilized as a parameter for ciliary activity, and for the first time, we showed fluid movement patterns along the epithelium lining in vitro.

Effect of preoperative hormonal stimulation on postoperative complication rates after proximal hypospadias repair: a systematic review.

PURPOSE: We conducted a systematic review and meta-analysis to summarize the effect of preoperative hormonal stimulation on complication rates following proximal hypospadias repair. MATERIALS AND METHODS: We comprehensively searched the published and unpublished literature between 1990 and 2010. Eligibility criteria were applied. Title, abstract and full text screening was carried out by 2 independent authors, and discrepancies were resolved by consensus. Heterogeneity between studies was tested using Cochran chi-square Q test and quantified by calculating I(2). Quality appraisal of included studies was performed. Meta-analysis was conducted when appropriate using a random effects model. RESULTS: Our search yielded 288 citations, of which 11 (622 patients) met inclusion criteria and were incorporated into the systematic review. Most series were retrospective observational studies of moderate or low methodological quality. Of the patients 45% underwent administration of preoperative hormonal stimulation, with intramuscular testosterone being the most commonly prescribed formulation. Four studies addressed postoperative complication rate stratified by preoperative hormonal stimulation use and were included in a meta-analysis. The odds ratio for a complication occurring with preoperative hormonal stimulation use was 1.67 (CI 0.96-2.91, p = 0.07, I(2) = 0%). No persistent side effects due to preoperative hormonal stimulation were reported. CONCLUSIONS: To our knowledge this is the only systematic review and meta-analysis thus far that has critically assessed the effect of preoperative hormonal stimulation on operative outcomes after hypospadias repair. The published literature is of low quality and lacks standardized reporting of important patient and surgical details. The effect of preoperative hormonal stimulation on operative outcomes after hypospadias repair remains unclear and requires further investigation.

Recovery and Characterization of Spermatozoa in a Neotropical, Terrestrial, Direct-Developing Riparian Frog (Craugastor evanesco) through Hormonal Stimulation.

The Vanishing Rainfrog (Craugastor evanesco) is an endemic and critically endangered frog species of Panama. It is suspected that 90% of the population has disappeared from the wild. Frogs were collected from the wild and brought to a Captive Breeding Program; however, accomplishing regular reproductive events for this species has been difficult. The objective of this study was to determine the effect of hormonal stimulation on the production and quality of C. evanesco spermatozoa, aiming to develop an efficient and safe sperm collection protocol as a tool to help reproduce this endangered species. Mature males received intra-peritoneal injections with one of six hormone treatments, including des-Gly10, D-Ala6, Pro-NHEt9-GnRH-A, Amphiplex or hCG. Urine samples were collected at 10 different time points post-injection. Quality assessments included sperm concentration, percentage motility, percentage forward progressive motility (FPM), osmolality, pH and morphology analysis. Our results indicate that the optimal treatment for the collection of highly concentrated sperm samples of C. evanesco is 4 µg/gbw GnRH, followed by Amphiplex and 2 µg/gbw GnRH as sub-optimal treatments and finally, 6 µg/gbw GnRH and 5 and 10 IU/gbw hCG as non-optimal treatments. GnRH-A at 4 µg/gbw and Amphiplex stimulated the production of samples with the highest sperm concentrations and quality, despite Amphiplex producing lower percentages of intact acrosome and tail. In contrast, hCG concentrations were not reliable inducers of sperm production, consistently showing lower concentrations, higher percentages of sperm abnormalities and more acidic spermic urine than that induced by Amphiplex and GnRH-A. Morphological assessments revealed that C. evanesco spermatozoa have a filiform shape with a large acrosome on the anterior part of an elongated head, a small midpiece and a long tail with two filaments joined together by an undulating membrane.

Non-invasive method of obtaining milk from mice using carbetocin, a synthetic analogue of oxytocin.

INTRODUCTION: The mouse is an important laboratory animal in pharmacological and toxicological research. However, there is a limited ability to analyse the penetration of tested substances in mouse breastmilk, which is technically difficult to obtain in sufficient quantities. The aim of the study is to verify the use of carbetocin for this purpose. METHODS: The effect of carbetocin (20 and 40 µg per animal) was tested in nursing ICR females that had milk collected for 15 min, started 1.5 min after administration. At a higher dose, carbetocin was also tested with a 20-min collection, started 7 min after application. Oxytocin (2 IU per animal) and saline were used as positive and negative controls, respectively. RESULTS: Milk yield using a lower dose of carbetocin was comparable to oxytocin. However, the duration of action of carbetocin was longer than that of oxytocin. A higher doses of carbetocin resulted in significantly higher milk volumes. DISCUSSION: The use of carbetocin has proven to be an effective non-invasive method to obtain up to 0.89 g of milk from one mouse in 20 min.

Comparison of different methods of wild vimba bream Vimba vimba (L.) spawning under controlled conditions.

This paper describes a study in which different types of the reproduction of wild vimba bream (Vimba vimba) under controlled conditions were compared. Reproduction was performed by using three methods: spontaneous (natural spawning- NS groups): without hormones but with nests, semi-artificial (S groups) with hormones and nests, and artificial (A groups) with hormones and manual stripping of gametes. In groups S and A, different types of hormonal stimulation were tested: common carp pituitary homogenate (CPH), human chorionic gonadotropin (hCG), and commercial pellets containing mammalian GnRHa and dopamine antagonist-metoclopramide (Ovopel). The number of spawning fish, i.e., spermiating males and ovulating females, were compared depending on the type of breeding and the stimulant used. The lowest number of finally matured fish: spermiating males (67 %) and ovulating females (0 %) were noted in the spontaneously reproduced group. The best method of reproducing wild vimba bream under controlled conditions was the semi-artificial approach with hormonal stimulation and spawning nests, where ovulation rates in hormonally treated groups were between 43 % and 71 %, vs 26-57 % in artificial spawning groups. No ovulation observe in control groups in S and A types of reproduction. The highest embryo survival to the eyed-egg-stage was noted in group A-hCG (average 88.2). A higher value of spawning effectiveness (Se) with the use of the same hormones was obtained for groups S (0.374-0.574) than for groups A (0.252-0.493). In both cases, the highest Se was recorded for Ovopel stimulated fish and the lowest in hCG stimulated groups.

The effect of two different experimental rearing temperatures on the quality and the large-scale cryopreservation of Eurasian perch (Perca fluviatilis) sperm.

Eurasian perch is an important fish species for European aquaculture diversification, but the quality of reproduction still remains one of the main limitations for further industry development. In particular, the optimal condition to obtain the best quality of sperm is poorly understood. The aim of our study was to measure the possible effects of two experimental rearing temperatures (6 °C and the conventionally used 12 °C) and of hormonal stimulation, on the motility parameters (pMOT, VCL, VSL, LIN, ALH, BCF), osmolality and fertilizing capacity of Eurasian perch sperm at the end of the reproductive cycle. A prior untested, large-scale (5 mL cryotube and Polystyrene box) cryopreservation method was implemented using fresh sperm obtained from the two above mentioned temperature groups. Males were injected with 100 µg body weight kg-1 sGnRHa. No significant difference was recorded between the two rearing temperatures and between the saline control and sGnRHa treated groups on the different features of sperm quality. A similar fertilization rate was monitored in all sGnRHa treated (6 °C: 69 ± 13%, 12 °C: 81 ± 11%) and saline control groups (6 °C: 79 ± 10%, 12 °C: 87 ± 4%). Correspondingly, no significant difference in hatching rate was observed in the sGnRHa injected (6 °C: 27 ± 9%, 12 °C: 40 ± 20%) and saline control (6 °C: 35 ± 18%, 12 °C: 36 ± 7%) males. However, a notable negative effect of freezing process on sperm movement was observed following thawing in both temperature groups. No significant difference in the motility parameters was measured between the two temperature groups following large-scale cryopreservation. Furthermore, a similar result was observed in the fertilizing capacity (6 °C: 79 ± 10%, 12 °C: 75 ± 8) of thawed sperm as well as in the hatching rate (6 °C: 52 ± 13%, 12 °C: 46 ± 19%). Our results indicate that fresh Eurasian perch sperm can tolerate a reduced rearing temperature following hormonal treatment. The adopted large-scale cryopreservation method could be used efficiently in the future for the fertilization of large amounts of Eurasian perch eggs following a precise standardization process.

Artificial Reproduction of Blue Bream (Ballerus ballerus L.) as a Conservative Method under Controlled Conditions.

The blue bream Ballerus ballerus (L.) is one of two species of the Ballerus genus occurring in Europe. The biotechnology for its reproduction under controlled conditions needs to be developed to conserve its local populations. Therefore, a range of experiments were performed to obtain valuable gametes from blue bream following hormonal stimulation with carp pituitary homogenate (CPH), Ovopel and Ovaprim. CPH and Ovopel were injected twice and Ovaprim-once, under a pectoral fin. The spawners were inspected 12 h after the last injection, and the gametes were collected. Eggs were collected from spawning females and fertilized with sperm from three males. The smallest number of ovulating blue bream (four females) was obtained from individuals stimulated with Ovaprim. There were six to nine ovulating females in the other groups, accounting for 60-90% of the females in the respective groups. The latency period was the shortest in the spawners stimulated with Ovopel. The fish in this group ovulated 14 h after the hormonal injection. Spawning was synchronized and all females spawned simultaneously. The highest average weight of eggs per fish (75.4 g) was obtained from the females stimulated with Ovopel, whereas the individuals stimulated with Ovaprim provided the smallest average amount of eggs (49.5 g). The study showed that blue bream spawners are not very sensitive to reproduction-related handling under controlled conditions.

Constant darkness negatively affects the outcome of hormonally induced reproduction in cultured Eurasian perch females.

This study aimed to assess the effect of constant darkness applied to fish during controlled breeding on reproductive traits in domesticated females of Eurasian perch. Based on the assumption that keeping fish in constant darkness during the reproduction operation may reduce stress, suspected to be responsible for variable spawning effectiveness in this species. Two conditions were assessed (16 h light per day [group 16L] and constant darkness [group 0L], two tank replicates per condition). The reproductive protocol involved a 7-day-long adaptation period for group 0L where photoperiod was reduced by 2.3 h a day down to constant darkness. After the adaptation period, two hormone injections (salmon gonadoliberin analogue) were applied to both groups: priming (10 µg/kg) and resolving (25 µg/kg) with a 7-day interval between them. During the study, morphometric indices were recorded and blood, brain, and pituitary samples were collected to assess stress markers and determine hypothalamic-pituitary-gonadal axis functioning via measuring blood plasma hormones, as well as gonadoliberin and gonadotropins (luteinising hormone [LH] and follicle-stimulating hormone [FSH]) transcript abundance (n = 7 for each group at each sampling point). In addition, kinetics of the final oocyte maturation (FOM) process, ovulation rate, and egg quality of each group was monitored (n = 12 for each group). The results indicated that there were no differences in terms of morphometry, FOM kinetics, and most stress indices between groups throughout the experiment, except haematocrit, which increased immediately following the acclimation period in fish kept in darkness. Constant darkness negatively affected plasma levels of 17α,20ß-dihydroxy-4-pregnen-3-one (DHP) and LH transcript expression at the time of the second hormone injection. This indicated that exposure to constant darkness negatively affected priming of the hormonal dose applied, resulted in the disruption of ovulation, and reduced ovulation rates (50%) for group 0L, as compared to 16L (91%). The findings of this study clearly indicate that constant darkness may have significant deleterious effects on reproductive traits throughout out-of-season induced, hormonally supported, controlled reproduction. Therefore, we advise against the use of constant darkness when managing broodstock reproduction in domesticated Eurasian perch.

Effect of hormonal stimulation on captive broodstock maturation, induced breeding and spawning performance of striped snakehead, Channa striata (Bloch, 1793).

In the present study, there was assessment of effects of gonadotropin treatments on broodstock maturation, induced breeding, and spawning outcomes of striped snakehead in captivity. The striped snakehead (n = 128) were equally distributed in four concrete tanks (15 m2) and hormone implants (500 IU human chorionic gonadotropin (hCG)/kg body weight) were inserted intramuscularly and striped snakehead broodstock administered this treatment were confined in two tanks and striped snakehead of a non-implanted group were confined in two tanks. The hormone implanted striped snakehead had a greater (P < 0.05) gonadosomatic index (GSI) and oocyte diameter in comparison to non-implanted striped snakehead. In a subsequent experiment, hCG and carp pituitary homogenate (CPH) were evaluated for inducing breeding. Dosages of hCG used were, 2,000 (TH1), 3000 (TH2), and 4000 (TH3) IU hCG/kg body weight of females. Dosages of CPH were, 20 (TP1), 30 (TP2), and 40 (TP3) mg CPH/kg body weight of females. Males were administered 0.75 of the dosage administered to females. The values for reproductive variables were estimated. Fertilization (89.0 ±â€¯3.0 %) and hatching (92.0 ±â€¯1.0 %) rates were greater (P < 0.05) in the TH1 group of implanted striped snakehead. Relative fecundity (19,023 ±â€¯2211), as well as fertilization (96.2 ±â€¯2.4 %), and hatching (96.6 ±â€¯1.7 %) rates were greater in the TP2 group of the implanted striped snakehead. The results from the present study indicate broodstock treated with gonadotropins had greater spawning outcomes which might facilitate mass scale breeding and fertilized egg as well as juvenile production of striped snakehead in captivity.

Laparoscopic Ovum Pick-Up Followed by In Vitro Embryo Production and Transfer in Assisted Breeding Programs for Ruminants.

The potential of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production (IVEP) as a tool for accelerated genetic programs in ruminants is reviewed in this article. In sheep and goats, the LOPU-IVEP platform offers the possibility of producing more offspring from elite females, as the procedure is minimally invasive and can be repeated more times and more frequently in the same animals compared with conventional surgical embryo recovery. On average, ~10 and ~14 viable oocytes are recovered by LOPU from sheep and goats, respectively, which results in 3-5 transferable embryos and >50% pregnancy rate after transfer. LOPU-IVEP has also been applied to prepubertal ruminants of 2-6 months of age, including bovine and buffalo calves. In dairy cattle, the technology has gained momentum in the past few years stemming from the development of genetic marker selection that has allowed predicting the production phenotype of dairy females from shortly after birth. In Holstein calves, we obtained an average of ~22 viable oocytes and ~20% transferable blastocyst rate, followed by >50% pregnancy rate after transfer, declaring the platform ready for commercial application. The present and future of this technology are discussed with a focus on improvements and research needed.

Influence of Environmental Temperature and Hormonal Stimulation on the In Vitro Sperm Maturation in Sterlet Acipenser ruthenus in Advance of the Spawning Season.

Sturgeon sperm maturation occurs outside the testes during the transit of testicular spermatozoa (TS) through the kidneys and the Wolffian ducts. A method of in vitro TS maturation in sterlet Acipenser ruthenus was used to investigate the effects of temperature and hormonal stimulation of spermiation on the ability of TS to complete this process. Spermatozoa motility parameters after in vitro maturation of testicular sperm, concentrations of sex steroid hormones and testis morphology were studied in three groups of sterlet: (1) after overwintering in ponds (OW), (2) adapted to spawning temperature (ST), and (3) adapted to spawning temperature with hormonal induction of spermiation (ST-HI). Blood plasma concentrations of testosterone, 11-ketotestosterone and 17,20ß-dihydroxy-pregnenolone increased significantly after hormonal induction of spermiation (group ST-HI). In all groups, TS were not motile. After in vitro sperm maturation, motility was up to 60% only in group ST-HI. The data suggest that the ability of TS to be matured in vitro was not related to the environmental temperature, while hormonal stimulation of spermiation during the spawning season was an absolute requirement for optimal in vitro maturation.

Hormonal Stimulation of Human Ovarian Xenografts in Mice: Studying Folliculogenesis, Activation, and Oocyte Maturation.

Ovarian tissue cryopreservation and banking provides a fertility preservation option for patients who cannot undergo oocyte retrieval; it is quickly becoming a critical component of assisted reproductive technology programs across the world. While the transplantation of cryopreserved ovarian tissue has resulted in over 130 live births, the field has ample room for technological improvements. Specifically, the functional timeline of grafted tissue and each patient's probability of achieving pregnancy is largely unpredictable due to patient-to-patient variability in ovarian reserve, lack of a reliable method for quantifying follicle numbers within tissue fragments, potential risk of reintroduction of cancer cells harbored in ovarian tissues, and an inability to control follicle activation rates. This review focuses on one of the most common physiological techniques used to study human ovarian tissue transplantation, xenotransplantation of human ovarian tissue to mice and endeavors to inform future studies by discussing the elements of the xenotransplantation model, challenges unique to the use of human ovarian tissue, and novel tissue engineering techniques currently under investigation.