Exogenous Hormone Stimulation Research Articles

Dynamic changes in gene expression of growing nonhuman primate antral follicles.

The growth of the ovarian antral follicle is a complex process that is difficult to study, especially in human and nonhuman primates. Understanding the antral stage of development is key to new approaches to regulating reproduction. This study analyzed cohorts of three sizes of developing antral follicles obtained from adult rhesus macaque females using RNA sequencing of oocytes and cumulus and granulosa cells. The overall objective of this study was to identify key developmental changes in gene expression in oocytes, granulosa, and cumulus cells, as nonhuman primate antral stage follicles transition through progressively larger sizes in the absence of exogenous hormonal stimulation. Only a relatively small number of genes displayed altered mRNA expression levels in any of the three cell types during this period. Most of the identified differentially expressed genes (DEGs) decreased in the granulosa cells or increased in the cumulus cells. Although the number of DEGs observed was small, these DEGs indicate predicted effects on distinct upstream regulators in the cumulus and granulosa cells. This study is particularly important because it shows for the first time the gene expression changes during antral follicle growth in a medically relevant model.NEW & NOTEWORTHY Changes in gene expression in oocytes, granulosa, and cumulus cells were determined in nonhuman primate antral stage ovarian follicles transitioning through progressively larger sizes without exogenous hormonal stimulation. Only a small number of genes displayed altered mRNA expression levels in any of the three cell types. Most of the differentially expressed genes (DEGs) decreased in granulosa cells or increased in cumulus cells. These results identified upstream regulators of antral follicle development.

Osteoregenerative Potential of 3D-Printed Poly ε-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells.

The repair of orthopedic and maxillofacial defects in modern medicine currently relies heavily on the use of autograft, allograft, void fillers, or other structural material composites. This study examines the in vitro osteo regenerative potential of polycaprolactone (PCL) tissue scaffolding, fabricated via a three-dimensional (3D) additive manufacturing technology, i.e., a pneumatic micro extrusion (PME) process. The objectives of this study were: (i) To examine the innate osteoinductive and osteoconductive potential of 3D-printed PCL tissue scaffolding and (ii) To perform a direct in vitro comparison of 3D-printed PCL scaffolding with allograft Allowash® cancellous bone cubes with regards to cell-scaffold interactions and biocompatibility with three primary human bone marrow (hBM) stem cell lines. This study specifically examined cell survival, cell integration, intra-scaffold cell proliferation, and differentiation of progenitor cells to investigate the potential of 3D-printed PCL scaffolds as an alternative to allograft bone material for the repair of orthopedic injuries. We found that mechanically robust PCL bone scaffolds can be fabricated via the PME process and the resulting material did not elicit detectable cytotoxicity. When the widely used osteogenic model SAOS-2 was cultured in PCL extract medium, no detectable effect was observed on cell viability or proliferation with multiple test groups showing viability ranges of 92.2% to 100% relative to a control group with a standard deviation of ±10%. In addition, we found that the honeycomb infill pattern of the 3D-printed PCL scaffold allowed for superior mesenchymal stem-cell integration, proliferation, and biomass increase. When healthy and active primary hBM cell lines, having documented in vitro growth rates with doubling times of 23.9, 24.67, and 30.94 h, were cultured directly into 3D-printed PCL scaffolds, impressive biomass increase values were observed. It was found that the PCL scaffolding material allowed for biomass increase values of 17.17%, 17.14%, and 18.18%, compared to values of 4.29% for allograph material cultured under identical parameters. It was also found that the honeycomb scaffold infill pattern was superior to the cubic and rectangular matrix structures, and provided a superior microenvironment for osteogenic and hematopoietic progenitor cell activity and auto-differentiation of primary hBM stem cells. Histological and immunohistochemical studies performed in this work confirmed the regenerative potential of PCL matrices in the orthopedic setting by displaying the integration, self-organization, and auto-differentiation of hBM progenitor cells within the matrix. Differentiation products including mineralization, self-organizing "proto-osteon" structures, and in vitro erythropoiesis were observed in conjunction with the documented expression of expected bone marrow differentiative markers including CD-99 (>70%), CD-71 (>60%), and CD-61 (>5%). All of the studies were conducted without the addition of any exogenous chemical or hormonal stimulation and exclusively utilized the abiotic and inert material polycaprolactone; setting this work apart from the vast majority of contemporary investigations into synthetic bone scaffold fabrication In summary, this study demonstrates the unique clinical potential of 3D-printed PCL scaffolds for stem cell expansion and incorporation into advanced microstructures created via PME manufacturing to generate a physiologically inert temporary bony defect graft with significant autograft features for enhanced end-stage healing.

Genome-wide assessment of DNA methylation alterations induced by superovulation, sexual immaturity and in vitro follicle growth in mouse blastocysts

BackgroundIn their attempt to fulfill the wish of having children, women who suffer from fertility issues often undergo assisted reproductive technologies such as ovarian stimulation, which has been associated with adverse health outcomes and imprinting disorders in children. However, given the crucial role of exogenous hormone stimulation in improving human infertility treatments, a more comprehensive analysis of the potential impacts on DNA methylation in embryos following ovarian stimulation is needed. Here, we provide genome-wide DNA methylation profiles of blastocysts generated after superovulation of prepubertal or adult mice, compared with blastocysts derived from non-stimulated adult mice. Additionally, we assessed the impact of the in vitro growth and maturation of oocytes on methylation in blastocysts.ResultsNeither hormone stimulation nor sexual maturity had an impact on the low global methylation levels characteristic of the blastocyst stage or was associated with extensive DNA methylation alterations. However, we found hormone- and age-associated changes at specific positions but dispersed throughout the genome. In particular, we detected anomalous methylation at a limited number of CpG islands. Additionally, superovulation in adult mice was associated with alterations at the Sgce and Zfp777 imprinted genes. On the other hand, in vitro culture of follicles from the early pre-antral stage was associated with globally reduced methylation and increased variability at imprinted loci in blastocysts.ConclusionsOur results indicate a minimal effect of ovarian stimulation of adult and prepubertal mice on the DNA methylation landscape attained at the blastocyst stage, but potentially greater impacts of in vitro growth and maturation of oocytes. These findings have potential significance for the improvement of assisted reproductive techniques, in particular for those related to treatments in prepubertal females, which could be crucial for improving human fertility preservation strategies.

Granulosa Cell Dysfunction Is Associated With Diminished Ovarian Response in FMR1 Premutation Carriers

FMR1 premutation (PM) carriers are at increased risk of ovarian impairment resulting in diminished ovarian response (DOR) to exogenous follicle-stimulating hormone (FSH) stimulation. Expanded CGG repeat transcript and RAN-associated protein (FMRpolyG) have been shown to accumulate in cellular aggregates and sequester proteins, thus impairing their function. Sam68 is a multifunctional RNA-binding protein highly expressed in the gonads involved in FSH receptor (FSHR) transcript maturation during FSH-dependent follicular development. The present study examined a possible pathophysiological explanation for DOR to exogenous FSH stimulation in FMR1 PM carriers. We used both a human granulosa cell (GC) line model and human GCs from FMR1 PM carriers to evaluate whether Sam68 is sequestered with expanded CGG repeat transcript. We show that Sam68 is sequestered in GCs, most likely by interaction with the expanded CGG repeat transcript. The sequestration may lead to reduced levels of free Sam68 available for FHSR precursor transcript processing, causing dysregulation of FSHR transcript maturation, and a consequent decrease in FSHR protein levels. Sam68 sequestration may underlie the diminished ovarian response to FSH stimulation in FMR1 PM carriers.

Understanding the Ovarian Interrelationship with Low Antral Follicle Counts (AFC) in the In Vivo Bos indicus Cow Model: Unilateral and Bilateral Main AFC as Possible Biomarkers of Ovarian Response to Hormonal Synchronisation.

Simple SummaryThe number of growing antral follicles visualised by ultrasound generates an antral follicle count (AFC), which can predict the number of female gametes remaining in an animal’s ovary. However, important information on ovarian biology with regard to the ovarian interrelationship with the main number of antral follicle populations (mAFC) is scarce; therefore, it is important to uncover additional information on ovarian biology that can be helpful in predicting the ovarian response after hormonal stimulation in mono-ovulatory animal species. Thus, mono-ovulatory Bos indicus beef cows with low numbers of AFC were induced using a hormonal programme in an effort to increase the understanding of the interrelationship with unilateral mAFC (main number of antral follicle populations appearing on only one side of the ovary). Based on the odds ratio, our results emphasise that the likelihood of an ovarian response was higher in cows with unilateral mAFC relationships on the day of exogenous hormonal stimulation.The antral follicle count (AFC) is a test in which the number of oocyte-containing follicles that are developing in both ovaries are visually counted. The count of these follicles strongly relates to the population of the growing follicle reserve on the ovaries. However, the importance of the main number of antral follicle populations (mAFC) in mono-ovulatory animal species has yet to be completely elucidated. Moreover, the investigation of the ovarian interrelationship with unilateral mAFC (main number of antral follicle populations appearing on only one side of the ovary) and bilateral mAFC (main number of antral follicle populations appearing in equivalent numbers on both sides of the ovary) and how understanding this interrelationship can offer possible indicators of ovarian response to hormonal induction have not yet been investigated in mono-ovulatory Bos indicus beef cows. The aim of this study is to investigate the different ovarian interrelationships of mAFC (unilateral and bilateral mAFC) at the time of exogenous hormonal stimulation on the total number of AFC (left and right ovaries) at the beginning of the hormonal protocol for ovarian stimulation and ovarian response at the completion of exogenous hormonal stimulation as well as their usefulness as possible biomarkers of successful hormonal stimulation in Bos indicus beef cattle. Beef cows (n = 104) with low total numbers of AFC (4.7 ± 2.4 follicles) were stimulated with a gonadotropin-releasing hormone-progesterone-prostaglandin F2α-based protocol. At the beginning of the hormonal protocol, ovarian ultrasound scans were performed to evaluate AFC from both ovaries of cows. Beef cows were divided into two groups, unilateral (n = 74) and bilateral mAFC (n = 30), according to the ovarian interrelationship. At the completion of the hormonal stimulation, ovarian ultrasound scans were performed to evaluate the dominant follicle (DF) and cows with DF > 8.5 mm in diameter emerging on their ovaries were defined as having experienced a response to hormonal stimuli. There was a difference of 19.1% between Bos indicus cows bearing unilateral mAFC that produced an increase in ovarian response (odds ratio = 2.717, p < 0.05) compared to the responsive rate of cows displaying bilateral mAFC (82.4% vs. 63.3%). In unilateral mAFC, cows bearing mAFC ipsilateral to the ovary of dominant follicle (DF) had a higher responsive rate than cows bearing mAFC contralateral to the DF ovary (50.0% vs. 32.4%, p < 0.05). In mAFC ipsilateral to the DF ovary, pregnancy rates were greatest in cows bearing mAFC and DF on the right ovary compared with cows bearing mAFC and DF on the left ovary (25.0% vs. 9.1%, p < 0.05). In primiparous and multiparous cows, unilateral mAFC occurs with a greater (p < 0.05) frequency than bilateral mAFC (69.0% and 72.0% vs. 31.0% and 28.0%, respectively). In unilateral mAFC, primiparous cows bearing mAFC ipsilateral to the DF ovary had a greater responsive rate than primiparous cows bearing mAFC contralateral to the DF ovary (55.0% vs. 20.0%, p < 0.05). In mAFC ipsilateral to the DF ovary, responsive and pregnancy rates were greatest (p < 0.05) in multiparous cows bearing mAFC and DF on the right ovary compared with multiparous cows bearing mAFC and DF on the left ovary (58.1% and 22.6% vs. 25.8% and 3.2%, respectively). Furthermore, there was a positive correlation between the mean diameter of AFC at the time of the exogenous hormonal trigger and the mean diameter of DF at the completion of hormonal synchronisation (p < 0.05). Our findings emphasise that the ovarian interrelationship with unilateral mAFC at the time of the hormonal trigger might be a promising biomarker for predicting success in ovarian response to hormonal stimulation of mono-ovulatory Bos indicus beef cows with low AFCs.

Genome-Wide Identification of DNA Binding with One Finger (Dof) Gene Family in Tartary Buckwheat (Fagopyrum tataricum) and Analysis of Its Expression Pattern after Exogenous Hormone Stimulation.

Simple SummaryA number of studies have demonstrated that DNA binding with one finger (Dof) proteins are involved in multiple biological processes. In the present study, Dof genes or proteins in Tartary buckwheat (FtDofs) were systematically analysed, including their physical properties, phylogenetic relationships, structure, motif composition, cis-acting elements present in promoter regions, chromosomal distribution, gene duplication events, syntenic relationships, expression patterns in different tissues and different fruit developmental stages and responses to exogenous hormone stimulation. The results indicated that the expansion of FtDofs was mainly due to segmental duplication. The tissue-specific expression patterns of FtDofs and their positive responses to exogenous hormone stimulation suggest that they play important roles in the growth and development of Tartary buckwheat as well as in the adaptation to environmental changes. Collectively, this study lays a foundation for further exploration of the function of FtDof genes in Tartary buckwheat.DNA binding with one finger (Dof) proteins have been proven to be involved in multiple biological processes. However, genome-wide identification of the Dof gene family has not been reported for Tartary buckwheat (Fagopyrum tataricum). In this study, 35 FtDof proteins were identified, and they could be divided into nine phylogenetic subgroups. Proteins within the same subgroup had similar gene structure and motif composition. Moreover, abundant cis-acting elements were present in the promoter regions of FtDof genes. Segmental duplication was the primary driving force for the evolution of the FtDof gene family. Synteny analysis indicated that Tartary buckwheat was closer to dicotyledons, and more orthologous Dof genes existed among them. The expression pattern of FtDofs in different tissues and at different fruit developmental stages varied. Different tissues contained several genes that were specifically expressed. FtDof expression was mainly upregulated under methyl jasmonate treatment and downregulated under other hormone treatments. Taken together, FtDofs may play important roles in the growth and development of Tartary buckwheat and in response to abiotic and biotic stresses. Therefore, the genome-wide identification and expression pattern analysis of the Tartary buckwheat Dof gene family lays a foundation for further exploration of the functional characteristics of FtDofs in the future.

Functional differentiation of two lhx8 paralogs and possible regulatory role of lhx8a in Japanese flounder (Paralichthys olivaceus)

Lhx8, belonging to the LIM-Homebox family, is involved in the tooth, nervous system, and primordial follicles development in mammals. However, little is known about the regulatory roles of lhx8 in teleosts. In this study, two lhx8 duplicates were identified in Paralichthys olivaceus, termed Polhx8a and Polhx8b, respectively. Bioinformatic analysis showed that Polhx8a was more likely to be a teleost-specific paralog. According to expression analysis, Polhx8a transcripts were almost exclusively concentrated in the oocytes, while Polhx8b was weakly expressed in the spleen, gill, and some facial organs, indicating sub-functionalization of this gene pair during evolution. Furthermore, Polhx8a mRNA level elevated from perinucleolar oocyte (PNO) stage to vitellogenic oocyte (VO) stage transition and changed after exogenous hormone stimulation, proving that Polhx8a was involved in the oocyte development and could be regulated by sex hormones. Yeast two-hybrid, bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (co-IP) experiments captured the positive protein interactions between PoLhx8a and the other two oocyte-specific transcription factors: PoFigla and PoNobox. After knocking down lhx8a in embryos or adult ovaries in vivo, the expression of oocyte-associated genes was significantly down-regulated (P < 0.05). Our findings suggest the evolution and functional differentiation of lhx8 genes, and shed light on the potential role of lhx8a in protein interactions and gene regulation in teleosts.

Contribution to Natural Rubber Production without Exogenous Hormonal Stimulation

Rubber production and especially its increase absolutely depend on the activation of the latex-producing metabolism. Can the latex-producing metabolism activation mechanism developed by rubber tree lead to higher yield without exogenous hormonal stimulation? In order to verify this decisive hypothesis, several works, carried out for nine years in Southern Côte d’Ivoire, were conducted on some 15 clones (IRCA 18, IRCA 209, IRCA 111, IRCA 130, PB 235, PB 260, PB 280, PB 330, PB 310; GT 1, BPM 24, RRIC 100; PB 217 and PR 107) of the three metabolisms respectively, active, moderate and slow, with two main statistical designs. On a small scale, in a “one-tree plot” design, the same latex harvesting system, tapping in d4 with different hormonal stimulation regimes (0/y; 2/y; 4/y; 6/y; 8/y; 13/y; 18/y; 26/y, 39/y and 78/y), has been applied to all treatments. In a randomized complete block design, different tapping frequencies (d2, d3, d4, d5 and d6) and hormonal stimulation (0/y, 4/y, 8/y and 10/y) were applied. Whatever the stimulation regime, the average g.t-1.t-1 of the unstimulated control (56) of the active metabolism clones (PB 235, PB 310, IRCA 111 and IRCA 130) over 9 years was statistically the same order that of the stimulated patterns (54). The average g.t-1.t-1 of the control (60) was lower than that of the highest yielding stimulated patterns (67) of the clones (PB 330, PB 280, PB 260, IRCA 18 and IRCA 209). The average g.t-1.t-1of the unstimulated control (49) was significantly lower than that of the highest yielding stimulated treatment (57) of moderate metabolism clones (GT 1, BPM 24 and RRIC 100). The average g.t-1.t-1 of the unstimulated control (39) was significantly lower than that of the highest yielding stimulated treatment (70) of slow metabolism clones (PB 217 and PR 107). On a large scale and at tapping frequency (d2), the unstimulated latex harvesting system (d2 0/y) showed an average yield of (2341; 2266 and 1849 kg.ha-1.year-1 for the active, moderate and slow metabolisms, respectively) statistically comparable to those of the highest yielding tapping frequencies d3, d4, d5 and d6 all latex harvesting systems combined for the clones studied (2388; 2348 and 2256 kg.ha-1.year-1). These results show that it is possible to produce natural rubber without exogenous hormonal stimulation by judiciously playing on tapping intensity.

Evaluation of the usefulness of positron emission tomography with [18F]fluorodeoxylglucose performed to detect non-radioiodine avid recurrence and/or metastasis of differentiated thyroid cancer - a preliminary study.

About 30% of patients with disseminated differentiated thyroid cancer (DTC) may experience a loss of iodine uptake. It is associated with higher aggressiveness of the tumour and a reduced 10-year survival rate. The diagnosis of non-radioiodine avid DTC metastases remains a diagnostic challenge. A helpful technique for this diagnosis is positron emission tomography with 2-[¹⁸F]fluoro-2-deoxy-D-glucose (PET/CT with [¹⁸F]FDG). On the other hand, there are still discussions about the clinical value of using exogenous thyroid-stimulating hormone (TSH) stimulation before PET/CT with [¹⁸F]FDG. The aim of the study was the assessment of the usefulness of PET/CT with [¹⁸F]FDG under TSH suppression and stimulation of TSH performed in the detection of non-radioiodine avid DTC metastases, as well as determination of the thyroglobulin concentration under suppression and stimulation of TSH, which influences the result of PET/CT with [¹⁸F]FDG in patients with non-radioiodine avid DTC. Retrospective analysis of 37 PET/CT with [¹⁸F]FDG performed in patients with DTC diagnosed and treated at the Department of Endocrinology and Isotope Therapy of the Military Institute of Medicine from January 2018 to July 2020. Of these, PET/CT with [¹⁸F]FDG under exogenous rhTSH stimulation was performed in 22 patients and PET/CT with [¹⁸F]FDG under TSH suppression in 15 was performed. In all analyzed patients, the result of diagnostic whole-body scintigraphy (WBS) using 80 MBq ¹³¹I under rhTSH stimulation was negative, and the concentration of thyroglobulin after stimulation (sTg) was greater than 1.0 ng/mL. In the group of patients examined under TSH suppression, non-radioiodine avid in PET/CT with [¹⁸F]FDG were found in 6 out of 15 patients (40%) and in the group of patients examined under rhTSH stimulation in 10 out of 22 patients (45%). The differences between the groups were not statistically significant. The analysis of the receiver operating characteristic (ROC) curves allowed to determine the cut-off point for the positive result of PET/CT performed under TSH suppression with sTg concentration of 11.03 ng/mL. In the group of studies performed under rhTSH stimulation, the cut-off point for sTg was 6.3 ng/mL. There was no statistically significant difference between the baseline thyroglobulin (natTg) and sTg levels and the positive PET/CT result. The administration of rhTSH before the PET/CT examination also had no statistically significant effect on the maximum standard uptake value (SUVmax) of the dominant lesion identified in the PET/CT. 1) PET/CT with [¹⁸F]FDG is a useful tool for detection of non-radioiodine avid recurrence and/or metastases of DTC. 2) The concentration of natTg and sTg is highly correlated with a positive result of PET/CT with [¹⁸F]FDG. 3) The concentration of natTg is comparable with sTg in predicting a positive result of PET/CT with [¹⁸F]FDG. 4) The cut-off point for a positive result of PET/CT for natTg was 1.36 ng/mL and for sTg was 7.05 ng/mL.

Effect of Exogenous Hormonal Stimulation on the Metabolic Partition between Plant Growth and Rubber Production of Hevea brasiliensis Clones According to the Class of Metabolic Activity in Cote d'ivoire

Aims: In order to achieve a good and sustainable harvest of Hevea brasiliensis rubber, the effect of exogenous hormonal stimulation on the metabolic partition growth-production of clones according to the class of metabolic activity was made.&#x0D; Place and Duration of Study: The study was conducted for 9 years in the San-Pedro region in south-west Ivory Coast, precisely in the experimental industrial plantations of the former HEVEGO, now Southwestern Agricultural Civil Society Société (SCASO).&#x0D; Methodology and Results: The rubber trees were planted at a density of 510 trees/ha in a completely randomised system. They were bled in S/2 d4 and stimulated to 2.5% Etephon at annual frequencies of 0, 1, 2, 4, 6, 8, 13, 18, 26, 39 and 78. The parameters measured were rubber production, trunk circumference, sucrose content, inorganic phosphorus, thiol groups and dry notch rate. The results showed that productivity increased with the intensity of stimulation (58.96 - 68.49 g.a-1.s-1 ; 39.83 - 66.69 g.a-1.s-1) over the intervals [0.6] and [0.26] stimulations respectively in clones with active and slow metabolism. The circumference of the trunk was marked by a less pronounced decrease in growth in slow metabolizing clones (0.52%) with good protection of the laticigene system (R-SH : 0.815 mmol.l-1) and low sensitivity to dry notching (1.6). Whatever the classes of metabolic activity of the clones, the agrophysiological parameters were strongly correlated with each other by a degree 2 polynomial function.&#x0D; Conclusion: This polynomial function seems to reflect the existence of a good state of equilibrium between laticigenic metabolism and vegetative growth according to the class of metabolic activity of the Hevea brasiliensis clones.

Evaluation of different temporal periods between hormone-induced ovulation attempts in the female Fowler's toad Anaxyrus fowleri.

Many amphibian species reinitiate the processes of preparing for reproduction (e.g. oogenesis) soon after breeding indicating hormone-induced ovulation could potentially be achieved out-of-season, which would lead to higher annual fecundity compared to mono-seasonal breeding. Such strategies would be beneficial to captive breeding programs for threatened species that are short-lived, have aging populations or need large numbers of offspring to meet reintroduction goals for species recovery. Unfortunately, little is known regarding how female anurans respond to multiple ovulation events within a year, which could lead to higher annual fecundity compared to mono-seasonal breeding. Thus, we evaluated the effect of temporal period between exogenous hormone stimulation events on egg production using the Fowler's toad Anaxyrus fowleri as a model species. Female toads (n = 21) were administered hormone therapy twice in 1year with toads randomly assigned to a treatment of either a 4-, 8- or 12-month recovery period between hormone stimulations. Ovulation was induced using two priming doses of human chorionic gonadotropin (100IU; hCG) 72h apart, followed by a resolving dose of hCG (500IU) plus gonadotropin releasing hormone analogue (GnRHa; 15μg) given 24h after the second priming injection. Measured response variables include the number of females ovulating after treatment, total number of eggs produced and percent fertilization, neurula and tadpole development. No significant treatment effects were observed for any response variable (P > 0.05). Findings from this study suggest that hormone therapy can be administered in a bufonid species every 4 or 8months without significantly affecting the number of ovulating females, egg production, fertilization, neurulation or tadpole development. By collecting gametes out-of-season or multiple times throughout the year, captive breeding programs could potentially increase tadpole production for reintroductions as well as extend the breeding window in captivity.

Profil hormon FSH, LH dan estradiol serta kadar glukosa darah sidat, Anguilla bicolor bicolor (Mc Clelland, 1844) yang dirangsang hormon HCG, MT, E2 dan anti dopamin

This study aims to analyze the hormone profile of FSH, LH and E2 to the eel after exogenous hormone stimulation and blood glucose levels. Six formulated treatment applied with combination of dopamine antagonize 10 mg.mL–1 (A), estradiol (E2) 3 mg.mL–1 + A (EA), metyltestosteron (MT) 3 mg.mL–1 + A (MTA), hCG 2 mg.mL–1 + EA (hEA) and hCG 2 mg.mL–1 + MTA (hMTA), with (F) physiologis 0.9% NaCl to control. Six group eel (200±15g) reared in a concrete tank with a capacity of 3,400 liters and filled with sea water of 35 mg L–1 as much as 2000 liters. Eels injected 1 mL.kg–1 hormone by intramusculary, were feed to apparent satiation daily for 10 weeks. The study used Completely Randomized Design with one treatment factor, namely hormonal factor and its combination. Fish blood that ware directly concentration to FSH, LH and E2, the enhanced significantly high in the blood plasma on treatment hMTA and hEA P&lt;0.05. Glucose concentration in the blood palsma is high enough in a row on a formula hMTA 67.33 mg.dL–1 and significantly different to P&lt;0.05. The result indicates that induction of exogenous hormone (hMTA) improve FSH, LH and E2. FSH and LH profiles show permutation patterns during the development of eel gonad, beginning with the increase of FSH in the early phases of gonadal development. LH profile moves in line with the increase in E2 during gonadal maturation process eels (A. bicolor bicolor), with blood glucose levels in the normal range. AbstrakThis study aims to analyze the hormone profile of FSH, LH and E2 to the eel after exogenous hormone stimulation and blood glucose levels. Six formulated treatment applied with combination of dopamine antagonize 10 mg.mL–1 (A), estradiol (E2) 3 mg.mL–1 + A (EA), metyltestosteron (MT) 3 mg.mL–1 + A (MTA), hCG 2 mg.mL–1 + EA (hEA) and hCG 2 mg.mL–1 + MTA (hMTA), with (F) physiologis 0.9% NaCl to control. Six group eel (200±15g) reared in a concrete tank with a capacity of 3,400 liters and filled with sea water of 35 mg L–1 as much as 2000 liters. Eels injected 1 mL.kg–1 hormone by intramusculary, were feed to apparent satiation daily for 10 weeks. The study used Completely Randomized Design with one treatment factor, namely hormonal factor and its combination. Fish blood that ware directly concentration to FSH, LH and E2, the enhanced significantly high in the blood plasma on treatment hMTA and hEA P&lt;0.05. Glucose concentration in the blood palsma is high enough in a row on a formula hMTA 67.33 mg.dL–1 and significantly different to P&lt;0.05. The result indicates that induction of exogenous hormone (hMTA) improve FSH, LH and E2. FSH and LH profiles show permutation patterns during the development of eel gonad, beginning with the increase of FSH in the early phases of gonadal development. LH profile moves in line with the increase in E2 during gonadal maturation process eels (A. bicolor bicolor), with blood glucose levels in the normal range.

Hormonal Effects on Breast Density, Fibroglandular Tissue, and Background Parenchymal Enhancement.

Breast density, fibroglandular tissue, and background parenchymal enhancement (BPE) are recognized independent biomarkers for breast cancer risk. For this reason, reproducibility and consistency in objective assessment of these parameters at mammography (breast density) and at magnetic resonance imaging (fibroglandular tissue and BPE) are clinically relevant. However, breast density, fibroglandular tissue, and BPE are manifestations of dynamic physiologic processes and may change in response to both endogenous and exogenous hormonal stimulation. It is therefore important for the radiologist to recognize settings in which hormonal stimulation may alter the appearance of these biomarkers at imaging and to appreciate how such changes may affect risk assessment, cancer detection, and even prognosis. The purpose of this review article is therefore to review key features and means of evaluating breast density, fibroglandular tissue, and BPE at imaging; to detail how endogenous and exogenous hormonal stimuli may affect breast density, fibroglandular tissue, and BPE, potentially affecting radiologic interpretation; and, finally, to provide an update regarding current hormone treatment guidelines and indications that may result in imaging changes through hormone modulation. ©RSNA, 2018.

Hormonal stimulation in 4 to 7 months old Nelore (Bos taurus indicus) females improved ovarian follicular responses but not the in vitro embryo production

The inclusion of pre-pubertal bovine females in reproductive management could allow in vitro embryo production and reduce generation interval, thereby causing faster genetic gain of the herd. However, oocytes of pre-pubertal females have lower competence, blastocyst production, and pregnancy rates than those collected from pubertal animals. This study aimed to evaluate the effect of an induced hormonal stimulation on the serum concentrations of Anti-Mullerian hormone (AMH) and FSH, ovarian responses, ovum pick up (OPU), and in vitro produced embryos (IVP) from oocytes obtained from four-to seven-months old Nelore female cattle. In a crossover design, these females were randomly allocated into: 1) Treated Group (TG, n = 9): the animals were subjected to a hormonal protocol (implanted progesterone device, estradiol benzoate, LH, and FSH) from Day 0 (the start of the treatment) to Day 7 (OPU day), and 2) Control Group (CG, n = 9): the females did not receive any hormonal stimulation, but they had ablation of their largest follicles on Day 2 of experiment. Blood collection for serum FSH measurements was done on Days 5, 6, 7, and 8, and collection for serum AMH measurements was done on Days 5 and 8. As hypothesized, TG had higher serum FSH concentrations (p < 0.05) on Day 5 (1.16 ± 0.31 ng/mL), Day 6 (1.21 ± 0.45 ng/mL), and Day 7 (0.95 ± 0.26 ng/mL) than CG (0.56 ± 0.17 ng/mL on Day 5, 0.60 ± 0.25 ng/mL on Day 6, and 0.60 ± 0.14 ng/mL on Day 7). However, serum AMH concentrations were neither significantly different (p > 0.05) between CG and TG, nor between the collection days. Hormonal stimulation also increased (p < 0.05) total follicular population (20.0 ± 4.95 CG vs 26.66 ± 4.24 TG), ovarian diameter (13.08 ± 1.0 mm CG vs 14.81 ± 1.38 mm TG) and number of follicles ≥2.5 mm (6.88 ± 2.14 CG vs 11.55 ± 4.09 TG). In TG, grades I and II oocytes predominated, whereas, in CG grades III and IV oocytes were more abundant (p < 0.05). No significant increases (p > 0.05) in the cleavage (49.33% CG vs 51.42% TG), cleavage > 4 cells (9.33% CG vs 16.19% TG), and blastocysts rates (1.33% CG vs 8.57% TG) were seen in TG. This hormonal protocol increased serum FSH concentrations that possibly contributed to increases in the observed follicle, as well as improving oocyte quality. This exogenous hormonal stimulation increased available oocytes numbers for IVP, despite no increase in the in vitro embryo production efficiency.

Comparison of antagonist mild and long agonist protocols in terms of follicular fluid total antioxidant capacity

ObjectiveA high dose of prolonged gonadotropins can yield higher numbers of oocytes and embryos. The high dose or prolonged regimens can be associated with ovarian hyperstimulation syndrome (OHSS), multiple gestations, emotional stress, economical burden and treatment dropout. In mild stimulation lower doses and shorter duration times of gonadotropin are used in contrast to the conventional long stimulation protocol in IVF. It has been proposed that supraphysiologic levels of hormones may adversely affect endometrium and oocyte/embryo. Also it has been proposed that oxidative stress (OS) may alter ovarian hormone dynamics and could be further affected by additional exogenous hormonal stimulation. Therefore our aim was to compare follicular fluid total antioxidant capacity (TAC) in antagonist mild and long agonist stimulations. Materials and MethodsForty patients received antagonist mild stimulation, starting on the 5th day of their cycle and forty patients received long agonist treatment. Seventy-five patients undergoing their first IVF cycle were included in the final analysis. Follicular fluid (FF) samples were analyzed for estradiol (E2), antimullerian hormone (AMH) and TAC. ResultsFF-Total antioxidant capacity (TAC) levels were higher in the long agonist group as opposed to the antagonist group [1.07 ± 0.04 mmol Trolox equivalent/L vs 1 ± 0.13 mmol Trolox equivalent/L] (Fig. 1). Pregnancy rates were not significantly different between the two treatments. The FF-TAC levels were not different among infertility etiologies (Fig. 3). FF-TAC levels did not have a direct correlation with pregnancy but a positive correlation with the total gonadotropin dose was observed. ConclusionPatients with good ovarian reserves and under the age of 35 effectively responded to mild stimulation treatment. Using lower amounts of gonadotropin, yielded less FF-TAC levels in patients who underwent antagonist mild protocol. In patients under the age of 35, antagonist mild stimulation is a patient friendly and effective procedure when undergoing their first IVF cycle.

Repetitive Maturation of Oocytes From Non-Stimulated Xenografted Ovarian Tissue From a Prepubertal Patient Indicating the Independence of Human Ovarian Tissue.

Introduction Modern anti-cancer strategies have distinctly increased survival rates; nevertheless, often accompanied by sterility. Currently, the only option for preserving fertility in prepubertal females is to cryopreserve ovarian tissue and re-transplant frozen-thawed tissue to restore fertility after treatment. Our aim was to report the occurrence of repetitive antral follicle formation and oocyte maturation in a prepubescent ovarian tissue xenograft without exogenous hormone stimulation. Material and Methods Frozen-thawed ovarian tissue from a 6-year-old patient suffering from nephroblastoma was xenotransplanted in oophorectomized severe combined immunodeficiency (SCID) mice to evaluate follicle development. Ergebnisse Repetitive follicle development to the antral stage occurred in the same xenograft of prepubertal ovarian tissue without exogenous hormone administration; 37 days after retrieving a maturing oocyte (this first retrieval has been previously published), another, completely mature oocyte was harvested from the xenograft. Subsequent histological evaluation of the grafted tissue showed primordial follicles, nearly all stages of developing follicles, as well as large atretic ones. Many clusters with dormant primordial follicles were also present. Conclusion Xenotransplanted prepubertal ovarian tissue has the potential for repetitive oocyte retrieval cycles without administering exogenous hormones. The results indicate that the human ovarian tissue might be able to synchronize the hypothalamus-hypophysis-axes of the mouse to the physiological human cycle; this should be investigated in future studies.

The usefulness of fluorine-18 fluorodeoxyglucose PET in the detection of recurrence in patients with differentiated thyroid cancer with elevated thyroglobulin and negative radioiodine whole-body scan.

PET/computed tomography (CT) using fluorine-18 fluorodeoxyglucose (F-FDG) has been used in the diagnosis of recurrence and metastases of differentiated thyroid cancer (DTC) in cases of negative whole-body scan (WBS) despite elevated concentrations of stimulated thyroglobulin (Tg). To assess the utility of PET/CT in the detection of recurrence among patients with DTC with increased Tg levels and negative results of WBS. PET/CT results were retrospectively analyzed in patients with DTC with increased Tg and negative results of WBS as well as negative cervical ultrasonography and chest radiography. PET-CT was performed 1-2 weeks after recent diagnostics under conditions of endogenous or exogenous thyroid-stimulating hormone stimulation. PET/CT was performed using a Discovery ST scanner 1 h after an intravenously F-FDG injection (activity 4-5 MBq/kg). To determine the cutoff value of Tg, receiver operating characteristic curves were analyzed. Sixty-nine patients with DTC (48 women, 21 men) aged 22-83 years (mean 50.9±17.5 years) were qualified. In 44 patients (63.8%), PET/CT indicated lesions of DTC. Thirty (43.5%) patients had F-FDG positive findings. In the remaining 14 patients (20.3%), lesions were found in CT only. Patients with a positive PET/CT scan had significantly higher Tg values than patients with a negative PET/CT (mean 143.8 vs. 26.5 ng/ml, P=0.03). The cutoff value of Tg concentration measured with the receiver operating characteristic analysis was 32.9 ng/ml. PET/CT is a useful tool in the detection of recurrence among thyroid cancer patients in cases of conflicting results of standard procedures, particularly for those with high Tg levels and negative WBS. The probability of obtaining a positive PET-CT result increases with the level of Tg.

Human fallopian tube epithelium co-culture with murine ovarian follicles reveals crosstalk in the reproductive cycle.

Do interactions between human fallopian tube epithelium and murine follicles occur during an artificial reproductive cycle in a co-culture system in vitro? In a co-culture system, human fallopian tissues responded to the menstrual cycle mimetic by changes in morphology and levels of secreted factors, and increasing murine corpus luteum progesterone secretion. The entire fallopian tube epithelium, including ciliated and secretory cells, can be regulated in the reproductive cycle. Currently, there are no in vitro culture models that can monitor fallopian tissues in real time in response to factors produced by the ovary. In addition, there are no reports on the impact of fallopian tissue on ovarian function during the menstrual cycle. Human fallopian tissue (n=24) was obtained by routine hysterectomies from women (aged 26-50 years, mean age=43.6) who had not undergone exogenous hormonal treatment for at least 3 months prior to surgery. CD1 female mice were used for ovarian follicle isolation. The human fallopian epithelium layers were either co-cultured with five murine multilayer secondary follicles (150-180 μm follicles, encapsulated in one alginate gel bead) for 15 days or received stepwise steroid hormone additions for 13 days. The fallopian tissue morphology and cilia beating rate, as measured by an Andor Spinning Disk Confocal, were investigated. Oviduct-specific glycoprotein 1 (OVGP1), human insulin-like growth factor 1 (hIGF1), vascular endothelial growth factor A (VEGF-A) and interleukin 8 (IL8) as biological functional markers were measured either by ELISA or western blot to indicate dynamic changes in the fallopian epithelium during the reproductive cycle generated by mouse follicles or by stepwise steroid hormone induction. Three or four patients in each experiment were recruited for replicates. Data were presented as mean±SD and further analyzed using one-way ANOVA followed by Tukey's multiple comparisons test. The cultured fallopian tube epithelium responded to exogenous steroid hormone stimulation, as demonstrated by enhanced cilia beating rate (~25% increase, P=0.04) and an increase in OVGP1 secretion (P=0.02) in response to 1 nM estradiol (E2) treatment when compared with 0.1 nM E2. Conversely, 10 nM progesterone plus 1 nM E2 suppressed cilia beating rate by ~30% (P=0.008), while OVGP1 secretion was suppressed by 0.1 nM E2 plus 50 nM progesterone (P=0.002 versus 1 nM E2 alone). Human fallopian tube epithelium was co-cultured with murine secondary follicles to mimic the human menstrual cycle. OVGP1 and VEGF-A secretion from fallopian tissue was similar with stepwise hormone treatment and when cultured with murine follicles. However, the secretion patterns of hIGF1 and IL8 differed in the luteal phase when comparing steroid treatment with follicle co-culture. In co-culture, hIGF1 secretion was suppressed in the luteal versus follicular phase (P=0.005) but stepwise hormone treatment had no effect on hIGF1. In co-culture, IL8 secretion was also suppressed on luteal phase day 15 (P=0.013) versus follicular phase day 7, but IL8 secretion increased continuously under high E2/progesterone treatment (P=0.003 for D13 versus D3). In the co-culture system, the corpus luteum continuously produced progesterone in the presence of fallopian tube tissue until Day 18 while, without fallopian tissue, progesterone started to drop from Day 13. One limitation of this study is that murine follicles were used to mimic the human menstrual cycle. However, although secretion patterns of peptide hormones such as inhibins and activins differ in mice and humans, the co-culture system used here did reveal interactions between the tissues that govern reproductive function. In vitro co-culture models of fallopian reproductive tissues with ovarian follicles can provide an important tool for understanding fertility and for uncovering the mechanisms responsible for reduced fertility. In addition, the role of oviductal secretions and how they influence ovarian function, such as the production of progesterone during the menstrual cycle, can be uncovered using this model. None. This work was funded by grants from the NIH (UH3TR001207), the American Cancer Society (RSG-12-230-01-TBG) and NIH (R01EB014806). The authors declare no competing financial interest.

Spontaneous antral follicle formation and metaphase II oocyte from a non-stimulated prepubertal ovarian tissue xenotransplant.

BackgroundCurrent strategies in cancer treatment have markedly increased the rates of remission and survival for cancer patients, but are often associated with subsequent sterility. While there are various options available to an adult female depending on the patient’s particular situation, the only realistic option for preserving fertility in prepubertal females is to cryopreserve ovarian tissue. This is the first report of a morphologically mature oocyte collected from non-stimulated prepubertal ovarian tissue xenotransplants.MethodsOvarian tissue from a 6 year old patient suffering from nephroblastoma was removed and cryopreserved for fertility preservation. The frozen-thawed ovarian tissue fragments were xenotransplanted to bilaterally oophorectomized severe combined immunodeficiency (SCID) mice to assess follicle development.ResultsAntral follicle formation occurred post-xenotransplantation in a single ovarian fragment without exogenous hormone stimulation. A morphologically maturing oocyte was harvested from these follicles.ConclusionsPrepubertal human ovarian follicles and oocytes can be matured after xenotransplantation even without exogenous hormone stimulation. These results indicate that tissue collected from prepubertal patients can support fertility in cancer survivors.

Association of Genetic Polymorphisms of Anti-Müllerian Hormone (AMH) and Its Type II Receptor with Ovarian Hyperstimulation Syndrome

Objective To explore the association of genetic polymorphisms in the genes encoding the anti-Mullerian hormone (AMH) and its type II receptor (AMHRII) with ovarian hyperstimulation syndrome (OHSS). Methods Using polymerase chain reaction (PCR) and DNA sequencing techniques, the exons of AMH and AMHRII were analyzed in 27 OHSS patients (OHSS group) and 22 non-OHSS patients (control group) who were applied controlled ovarian hyperstimulation (COH). Single nucleotide polymorphisms (SNPs) were also analyzed. Results SNPs G>T at position 146 of AMH exon 1 and G>A at position 134 of AMH exon 2 showed significant differences between the OHSS group and control group (P T at position 303 of AMH exon 1 showed no significant difference between the OHSS group and control group (P>0.05). No SNP was detected from the AMHR II exons 1 to 11 in either groups. Conclusions Genetic polymorphisms in the AMH gene may be a cause of ovarian hypersensitivity to exogenous hormone stimulation and the development of OHSS.