Pub Date : 2023-11-01DOI: 10.1016/j.xfss.2023.09.005
Daniel Nassau M.D. , Nicholas A. Deebel M.D. , Eliyahu Kresch M.D. , Davis Temple B.S. , Shathiyah Kulandavelu Ph.D. , Himanshu Arora Ph.D. , Ranjith Ramasamy M.D.
Objective
To study compensatory changes in testicular growth and the hormonal axis after unilateral orchiectomy in a neonatal, prepubertal, and pubertal/adult murine model. This is the first study to use a neonatal mouse survival surgery model.
Design
A laboratory-based study examining a control, neonatal, prepubertal, and pubertal/adult mouse model.
Setting
University-based basic science research laboratory.
Animals
Control, neonatal (2–4 days of life), prepubertal (12–21 days of life), and pubertal/adult (42–44 days of life) C57BL/6 mouse models.
Intervention
Unilateral orchiectomy in the neonatal, prepubertal, and pubertal/adult mouse models at their respective ages.
Main Outcome Measures
Body and testis weight and testicular length in the long axis were measured in a blinded fashion. In a similar way, testosterone, luteinizing hormone (LH), and follicle-stimulating hormone were assessed.
Results
Testes from neonatal and prepubertal mice weighed more (110.5, 12.2 and 103.0, 7.2 mg, respectively) than the control mice (91, 11.9 mg). There was no difference between the postpubertal group and the control group. The degree of compensatory hypertrophy was greater in the neonatal group but not in the prepubertal group when compared with the postpubertal group. Differences in follicle-stimulating hormone and testosterone were not statistically significant between the experimental and control arms. LH was significantly elevated in all experimental groups compared with the control.
Conclusions
This is the first study to assess testicular compensatory hypertrophy using a neonatal mouse survival surgery model. Testicular hypertrophy occurs when unilateral loss occurs before puberty, but not in adulthood in mice. Earlier testis loss may contribute to a greater degree of growth. Functionally, the unilateral testis can maintain eugonadal testosterone levels, but higher levels of LH are required after hemicastration to sustain eugonadal testosterone levels.
{"title":"Age-dependent effect on contralateral testicular compensation after testicular loss","authors":"Daniel Nassau M.D. , Nicholas A. Deebel M.D. , Eliyahu Kresch M.D. , Davis Temple B.S. , Shathiyah Kulandavelu Ph.D. , Himanshu Arora Ph.D. , Ranjith Ramasamy M.D.","doi":"10.1016/j.xfss.2023.09.005","DOIUrl":"10.1016/j.xfss.2023.09.005","url":null,"abstract":"<div><h3>Objective</h3><p>To study compensatory changes in testicular growth and the hormonal axis after unilateral orchiectomy in a neonatal, prepubertal, and pubertal/adult murine model. This is the first study to use a neonatal mouse survival surgery model.</p></div><div><h3>Design</h3><p>A laboratory-based study examining a control, neonatal, prepubertal, and pubertal/adult mouse model.</p></div><div><h3>Setting</h3><p>University-based basic science research laboratory.</p></div><div><h3>Animals</h3><p>Control, neonatal (2–4 days of life), prepubertal (12–21 days of life), and pubertal/adult (42–44 days of life) C57BL/6 mouse models.</p></div><div><h3>Intervention</h3><p>Unilateral orchiectomy in the neonatal, prepubertal, and pubertal/adult mouse models at their respective ages.</p></div><div><h3>Main Outcome Measures</h3><p><span>Body and testis weight and testicular length in the long axis were measured in a blinded fashion. In a similar way, testosterone, </span>luteinizing hormone (LH), and follicle-stimulating hormone were assessed.</p></div><div><h3>Results</h3><p>Testes from neonatal and prepubertal mice weighed more (110.5, 12.2 and 103.0, 7.2 mg, respectively) than the control mice (91, 11.9 mg). There was no difference between the postpubertal group and the control group. The degree of compensatory hypertrophy was greater in the neonatal group but not in the prepubertal group when compared with the postpubertal group. Differences in follicle-stimulating hormone and testosterone were not statistically significant between the experimental and control arms. LH was significantly elevated in all experimental groups compared with the control.</p></div><div><h3>Conclusions</h3><p>This is the first study to assess testicular compensatory hypertrophy using a neonatal mouse survival surgery model. Testicular hypertrophy occurs when unilateral loss occurs before puberty, but not in adulthood in mice. Earlier testis loss may contribute to a greater degree of growth. Functionally, the unilateral testis can maintain eugonadal testosterone levels, but higher levels of LH are required after hemicastration to sustain eugonadal testosterone levels.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 4","pages":"Pages 311-316"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41160588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To determine whether cyclic strain affects fibroid cell cytoskeletal organization, proliferation, and collagen synthesis differently than myometrial cells.
Design
A basic science study using primary cultures of patient-matched myometrial and fibroid cells.
Setting
Academic laboratory.
Patient(s)
Premenopausal women undergoing myomectomy or hysterectomy for the treatment of symptomatic uterine fibroids.
Intervention(s)
Application of uniaxial strain patterns mimicking periovulation, menses, or dysmenorrhea using the Flexcell tension system or static control. Secondarily, inhibition of G protein-coupled estrogen receptor-1 and phosphatidylinositol 3-kinase.
Main Outcome Measure(s)
Cell alignment, cell number, and collagen content.
Result(s)
Menses-strained cells demonstrated the most variation in cell alignment, cell proliferation, and procollagen content between myometrial and fibroid cells. Procollagen content decreased in myometrial cells with increasing strain amplitude and decreasing frequency. G protein-coupled estrogen receptor-1 inhibition decreases cellular alignment in the presence of strain.
Conclusion(s)
Mechanotransduction affecting cytoskeletal arrangement through the G protein-coupled estrogen receptor-1-phosphatidylinositol 3-kinase pathway is altered in fibroid cells. These results highlight the importance of incorporating mechanical stimulation into the in vitro study of fibroid pathology.
{"title":"Uterine fibroid cell cytoskeletal organization is affected by altered G protein-coupled estrogen receptor-1 and phosphatidylinositol 3-kinase signaling","authors":"Rachel Warwar M.D. , Andreja Moset Zupan B.S. , Carolyn Nietupski B.S. , Maricela Manzanares , Emily G. Hurley M.D. , Stacey C. Schutte Ph.D.","doi":"10.1016/j.xfss.2023.09.007","DOIUrl":"10.1016/j.xfss.2023.09.007","url":null,"abstract":"<div><h3>Objective</h3><p>To determine whether cyclic strain affects fibroid cell cytoskeletal organization, proliferation, and collagen synthesis differently than myometrial cells.</p></div><div><h3>Design</h3><p>A basic science study using primary cultures of patient-matched myometrial and fibroid cells.</p></div><div><h3>Setting</h3><p>Academic laboratory.</p></div><div><h3>Patient(s)</h3><p><span><span>Premenopausal women undergoing </span>myomectomy or </span>hysterectomy<span><span> for the treatment of symptomatic </span>uterine fibroids.</span></p></div><div><h3>Intervention(s)</h3><p>Application of uniaxial strain patterns mimicking periovulation, menses, or dysmenorrhea using the Flexcell tension system or static control. Secondarily, inhibition of G protein-coupled estrogen receptor-1 and phosphatidylinositol 3-kinase.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Cell alignment, cell number, and collagen content.</p></div><div><h3>Result(s)</h3><p>Menses-strained cells demonstrated the most variation in cell alignment, cell proliferation<span>, and procollagen content between myometrial and fibroid cells. Procollagen content decreased in myometrial cells with increasing strain amplitude and decreasing frequency. G protein-coupled estrogen receptor-1 inhibition decreases cellular alignment in the presence of strain.</span></p></div><div><h3>Conclusion(s)</h3><p><span>Mechanotransduction affecting cytoskeletal arrangement through the G protein-coupled estrogen receptor-1-phosphatidylinositol 3-kinase pathway is altered in fibroid cells. These results highlight the importance of incorporating </span>mechanical stimulation into the in vitro study of fibroid pathology.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 4","pages":"Pages 327-338"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41174145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-11-01DOI: 10.1016/j.xfss.2023.07.003
Erica D. Louden M.D. Ph.D. , Michael P. Dougherty M.D. , Lynn P. Chorich M.S. , Ali Eroglu Ph.D., D.V.M. , Lawrence C. Layman M.D.
Objective
To study if a pituitary or ovarian defect contributes to subfertility of the female Nsmf knockout (KO) mouse, an animal model of the hypogonadotropic hypogonadism gene NSMF.
Design
Analysis of hypothalamic, pituitary and ovarian gene expression at baseline, serum gonadotropin levels before and after gonadotropin-releasing hormone (GnRH) stimulation, ovarian response and implantation after superovulation, gonadotropin effects after ovariectomy, and ovarian NSMF protein expression.
Setting
University research laboratory.
Patients
None; mice were used.
Interventions
Gonadotropin-releasing hormone stimulation, superovulation, and ovariectomy in separate experiments.
Main Outcome Measures
Gene expression in the hypothalamus, pituitary, and ovary; ovarian response and implantation after superovulation; serum gonadotropins after GnRH stimulation and ovariectomy; Western blot to measure ovarian NSMF expression.
Results
We found increased hypothalamic Kiss1, Gnrh1, and Jak2 mRNA expression in female Nsmf KO vs. wild type (WT) mice. However, pituitary gonadotropin, and GnRH receptor gene expression was not affected, and serum gonadotropin levels were normal. Gonadotropins increased after ovariectomy for both groups. Baseline Kiss1, Fshr, Prkaca, Prkar1a, and Gdf9 ovarian mRNA expression was increased and Cyp19a1 expression was decreased in Nsmf KO mice, while superovulated Nsmf KO mice had reduced ovarian Kiss1r, Prkar1a, and Fshr mRNA expression, 50% less oocytes, and normal implantation. Western blot demonstrated NSMF protein expression in the ovary of WT mice.
Conclusions
Altered hypothalamic and ovarian gene expression was demonstrated in female Nsmf KO mice. It is possible that increased hypothalamic Gnrh1 and Kiss1 mRNA expression could compensate for reduced NSMF enabling a normal pituitary gonadotropin response. Impaired superovulation response, altered ovarian gene expression, and decreased number of oocytes indicate ovarian dysfunction, but a uterine factor cannot be excluded. These findings provide an anatomic basis for future mechanistic studies of subfertility in female Nsmf KO mice.
{"title":"Investigation of subfertility in the female Nsmf knockout mouse","authors":"Erica D. Louden M.D. Ph.D. , Michael P. Dougherty M.D. , Lynn P. Chorich M.S. , Ali Eroglu Ph.D., D.V.M. , Lawrence C. Layman M.D.","doi":"10.1016/j.xfss.2023.07.003","DOIUrl":"10.1016/j.xfss.2023.07.003","url":null,"abstract":"<div><h3>Objective</h3><p><span>To study if a pituitary or ovarian defect contributes to subfertility of the female </span><em>Nsmf</em><span> knockout (KO) mouse, an animal model of the hypogonadotropic hypogonadism gene </span><em>NSMF.</em></p></div><div><h3>Design</h3><p><span>Analysis of hypothalamic, pituitary and ovarian gene expression at baseline, serum gonadotropin levels before and after gonadotropin-releasing hormone (GnRH) stimulation, ovarian response and implantation after </span>superovulation<span><span>, gonadotropin effects after ovariectomy, and ovarian NSMF </span>protein expression.</span></p></div><div><h3>Setting</h3><p>University research laboratory.</p></div><div><h3>Patients</h3><p>None; mice were used.</p></div><div><h3>Interventions</h3><p>Gonadotropin-releasing hormone stimulation, superovulation, and ovariectomy in separate experiments.</p></div><div><h3>Main Outcome Measures</h3><p>Gene expression in the hypothalamus<span>, pituitary, and ovary; ovarian response and implantation after superovulation; serum gonadotropins after GnRH stimulation and ovariectomy; Western blot to measure ovarian NSMF expression.</span></p></div><div><h3>Results</h3><p>We found increased hypothalamic <em>Kiss1, Gnrh1</em>, and <em>Jak2</em> mRNA expression in female <em>Nsmf</em><span> KO vs. wild type (WT) mice. However, pituitary gonadotropin, and GnRH receptor gene expression was not affected, and serum gonadotropin levels were normal. Gonadotropins increased after ovariectomy for both groups. Baseline </span><em>Kiss1, Fshr, Prkaca, Prkar1a</em>, and <em>Gdf9</em> ovarian mRNA expression was increased and <em>Cyp19a1</em> expression was decreased in <em>Nsmf</em> KO mice, while superovulated <em>Nsmf</em> KO mice had reduced ovarian <em>Kiss1r, Prkar1a</em>, and <em>Fshr</em> mRNA expression, 50% less oocytes, and normal implantation. Western blot demonstrated NSMF protein expression in the ovary of WT mice.</p></div><div><h3>Conclusions</h3><p>Altered hypothalamic and ovarian gene expression was demonstrated in female <em>Nsmf</em> KO mice. It is possible that increased hypothalamic <em>Gnrh1</em> and <em>Kiss1</em><span> mRNA expression could compensate for reduced NSMF enabling a normal pituitary gonadotropin response. Impaired superovulation response, altered ovarian gene expression, and decreased number of oocytes indicate ovarian dysfunction, but a uterine factor cannot be excluded. These findings provide an anatomic basis for future mechanistic studies of subfertility in female </span><em>Nsmf</em> KO mice.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 4","pages":"Pages 286-293"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9943632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-11-01DOI: 10.1016/j.xfss.2023.09.003
Jesús Cadenas Ph.D. , Susanne Elisabeth Pors Ph.D. , Caroline Pulz Hansen M.Sc. , Sarah Maria Olufsen M.Sc. , Cristina Subiran M.Sc. , Jane Alrø Bøtkjær Ph.D. , Liv La Cour Poulsen M.D., Ph.D. , Jens Fedder M.D., Ph.D. , Margit Dueholm M.D., Ph.D. , Lotte Berdiin Colmorn M.D., Ph.D. , Stine Gry Kristensen Ph.D. , Linn Salto Mamsen Ph.D. , Claus Yding Andersen D.M.Sc.
Objective
To characterize the growth factor midkine (MDK) in the human ovary to determine whether MDK is produced locally within the ovary, examine whether different ovarian cell types are more likely to produce MDK, and determine whether there are any stage-specific variations during follicle growth. Previous studies have revealed that MDK potentially affects human follicle growth and oocyte maturation. Proteomic analyses in follicular fluid (FF) have identified MDK to functionally cluster together and follow a similar expression profile to that of well-known proteins involved in ovarian follicle development. Midkine has not yet been characterized in the human ovary.
Design
Descriptive study.
Setting
University Hospital.
Patients
The study included samples from 121 patients: 71 patients (aged 17–37 years) who underwent ovarian tissue cryopreservation provided granulosa cells (GC), cumulus cells, ovarian cortex, medulla tissue, and FF from small antral follicles (SAF); and 50 patients (aged 20–35 years) receiving in vitro fertilization treatment provided FF from preovulatory follicles before and after induction of final follicle maturation.
Interventions
None.
Main Outcome Measures
MDK relative gene expression was quantified using a real-time quantitative polymerase chain reaction in cumulus cells, GC, and medulla tissue. Additionally, immunostaining and western blotting assays were used to detect MDK protein in the ovarian cortex, which contains preantral follicles, SAF, and medulla tissue. Furthermore, enzyme-linked immunosorbent assay analyses were performed to measure the concentration of MDK in FF aspirated from SAF and preovulatory follicles both before and 36 hours after inducing the final maturation of follicles.
Results
Immunostaining and reverse transcription-quantitative polymerase chain reaction revealed a more prominent expression of MDK in GC compared with other ovarian cell types. Intrafollicular MDK concentration was significantly higher in SAF compared with preovulatory follicles. In addition, different molecular weight species of MDK were detected using western blotting in various ovarian sample types: GC and FF samples presented primarily one band of approximately 15 kDa and an additional band of approximately 13 kDa, although other bands with higher molecular weight (between 30 and 38 kDa) were detected in medulla tissue.
Conclusions
This is the first time that MDK has been immunolocalized in human ovarian cells at the protein level and that potentially different MDK variants have been detected in human FF, GC, and ovarian medulla tissue. Future studies are needed to sequence and identify the different potential MDK variants
{"title":"Midkine characterization in human ovaries: potential new variants in follicles","authors":"Jesús Cadenas Ph.D. , Susanne Elisabeth Pors Ph.D. , Caroline Pulz Hansen M.Sc. , Sarah Maria Olufsen M.Sc. , Cristina Subiran M.Sc. , Jane Alrø Bøtkjær Ph.D. , Liv La Cour Poulsen M.D., Ph.D. , Jens Fedder M.D., Ph.D. , Margit Dueholm M.D., Ph.D. , Lotte Berdiin Colmorn M.D., Ph.D. , Stine Gry Kristensen Ph.D. , Linn Salto Mamsen Ph.D. , Claus Yding Andersen D.M.Sc.","doi":"10.1016/j.xfss.2023.09.003","DOIUrl":"10.1016/j.xfss.2023.09.003","url":null,"abstract":"<div><h3>Objective</h3><p><span><span>To characterize the growth factor midkine (MDK) in the human ovary to determine whether MDK is produced locally within the ovary, examine whether different ovarian cell types are more likely to produce MDK, and determine whether there are any stage-specific variations during follicle growth. Previous studies have revealed that MDK potentially affects human follicle growth and </span>oocyte maturation<span>. Proteomic analyses in follicular fluid (FF) have identified MDK to functionally cluster together and follow a similar expression profile to that of well-known proteins involved in ovarian </span></span>follicle development. Midkine has not yet been characterized in the human ovary.</p></div><div><h3>Design</h3><p>Descriptive study.</p></div><div><h3>Setting</h3><p>University Hospital.</p></div><div><h3>Patients</h3><p>The study included samples from 121 patients: 71 patients (aged 17–37 years) who underwent ovarian tissue cryopreservation<span><span><span><span> provided granulosa cells (GC), </span>cumulus cells, ovarian cortex, medulla tissue, and FF from small </span>antral follicles (SAF); and 50 patients (aged 20–35 years) receiving in vitro fertilization </span>treatment<span> provided FF from preovulatory follicles before and after induction of final follicle maturation.</span></span></p></div><div><h3>Interventions</h3><p>None.</p></div><div><h3>Main Outcome Measures</h3><p>MDK relative gene expression was quantified using a real-time quantitative polymerase chain reaction in cumulus cells, GC, and medulla tissue. Additionally, immunostaining<span> and western blotting assays were used to detect MDK protein in the ovarian cortex, which contains preantral follicles, SAF, and medulla tissue. Furthermore, enzyme-linked immunosorbent assay analyses were performed to measure the concentration of MDK in FF aspirated from SAF and preovulatory follicles both before and 36 hours after inducing the final maturation of follicles.</span></p></div><div><h3>Results</h3><p>Immunostaining and reverse transcription-quantitative polymerase chain reaction revealed a more prominent expression of MDK in GC compared with other ovarian cell types. Intrafollicular MDK concentration was significantly higher in SAF compared with preovulatory follicles. In addition, different molecular weight species of MDK were detected using western blotting in various ovarian sample types: GC and FF samples presented primarily one band of approximately 15 kDa and an additional band of approximately 13 kDa, although other bands with higher molecular weight (between 30 and 38 kDa) were detected in medulla tissue.</p></div><div><h3>Conclusions</h3><p>This is the first time that MDK has been immunolocalized in human ovarian cells at the protein level and that potentially different MDK variants have been detected in human FF, GC, and ovarian medulla tissue. Future studies are needed to sequence and identify the different potential MDK variants","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 4","pages":"Pages 294-301"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41180627","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-11-01DOI: 10.1016/j.xfss.2023.06.003
Ayman Al-Hendy M.D., Ph.D. , Ying F. Zhou M.D. , Thomas Faustmann M.D. , Esther Groettrup-Wolfers M.D. , Kaisa Laapas M.Sc. , Susanne Parke M.D. , Christian Seitz M.D.
Objective
Vilaprisan is a highly potent selective progesterone receptor modulator shown to reduce heavy menstrual bleeding, induce amenorrhea, and diminish uterine fibroid volume in phase 2 studies. The objective of ASTEROID 3 was to demonstrate the superiority of vilaprisan compared with placebo in the treatment of heavy menstrual bleeding in women with uterine fibroids.
Women with ≥1 uterine fibroid of ≥3 cm and heavy menstrual bleeding of >80 mL/cycle.
Intervention(s)
Women were randomly assigned to 1 of 4 treatment arms, which were planned to comprise 2 treatment periods of 12 weeks, each with vilaprisan (2 mg/d) or placebo that were continuous or separated by a break of one bleed.
Main Outcome Measure(s)
Amenorrhea (primary end point; <2 mL in the last 28 days of treatment) and heavy menstrual bleeding response (key secondary end point; <80 mL/cycle and >50% reduction in bleeding from baseline) were measured with the alkaline hematin method. Change in volume of the 3 largest fibroids from baseline to end of treatment was assessed by ultrasound. Safety was monitored throughout the study.
Result(s)
Overall, 75 women completed the first 12 weeks of treatment. Statistically significant and clinically meaningful differences were observed between the vilaprisan- and placebo-treated groups in both the full analysis and per-protocol sets. In the per-protocol set (n = 36 and n = 12 for the vilaprisan and placebo groups, respectively), amenorrhea was observed more frequently in women treated with vilaprisan than in those who received placebo (83.3% vs. 0%, P<.0001), with a median time to onset of 3 days in the vilaprisan group. Similarly, more vilaprisan- than placebo-treated women achieved a response in heavy menstrual bleeding (91.7% vs. 25.0%, P<.0001). Serious adverse events were reported for 22 (27.8%) of 79 women and were evenly distributed among the 4 groups receiving vilaprisan and/or placebo. None of these events led to study discontinuation or were related to the liver, and no new safety findings were identified compared with the earlier phase 2 ASTEROID studies.
Conclusion(s)
Vilaprisan is efficacious and well tolerated over 12 weeks in the treatment of heavy menstrual bleeding associated with uterine fibroids. Further investigations of the long-term efficacy and safety of vilaprisan are warranted.
{"title":"Efficacy and safety of vilaprisan in women with uterine fibroids: data from the ASTEROID 3 randomized controlled trial","authors":"Ayman Al-Hendy M.D., Ph.D. , Ying F. Zhou M.D. , Thomas Faustmann M.D. , Esther Groettrup-Wolfers M.D. , Kaisa Laapas M.Sc. , Susanne Parke M.D. , Christian Seitz M.D.","doi":"10.1016/j.xfss.2023.06.003","DOIUrl":"10.1016/j.xfss.2023.06.003","url":null,"abstract":"<div><h3>Objective</h3><p>Vilaprisan is a highly potent selective progesterone receptor modulator<span><span> shown to reduce heavy menstrual bleeding, induce </span>amenorrhea<span>, and diminish uterine fibroid<span> volume in phase 2 studies. The objective of ASTEROID 3 was to demonstrate the superiority of vilaprisan compared with placebo in the treatment of heavy menstrual bleeding in women with uterine fibroids.</span></span></span></p></div><div><h3>Design</h3><p>Randomized, double-blind, placebo-controlled, multicenter phase 3 study.</p></div><div><h3>Setting</h3><p>Hospitals and medical centers.</p></div><div><h3>Patient(s)</h3><p>Women with ≥1 uterine fibroid of ≥3 cm and heavy menstrual bleeding of >80 mL/cycle.</p></div><div><h3>Intervention(s)</h3><p>Women were randomly assigned to 1 of 4 treatment arms, which were planned to comprise 2 treatment periods of 12 weeks, each with vilaprisan (2 mg/d) or placebo that were continuous or separated by a break of one bleed.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Amenorrhea (primary end point; <2 mL in the last 28 days of treatment) and heavy menstrual bleeding response (key secondary end point; <80 mL/cycle and >50% reduction in bleeding from baseline) were measured with the alkaline hematin method. Change in volume of the 3 largest fibroids from baseline to end of treatment was assessed by ultrasound. Safety was monitored throughout the study.</p></div><div><h3>Result(s)</h3><p>Overall, 75 women completed the first 12 weeks of treatment. Statistically significant and clinically meaningful differences were observed between the vilaprisan- and placebo-treated groups in both the full analysis and per-protocol sets. In the per-protocol set (n = 36 and n = 12 for the vilaprisan and placebo groups, respectively), amenorrhea was observed more frequently in women treated with vilaprisan than in those who received placebo (83.3% vs. 0%, <em>P</em><.0001), with a median time to onset of 3 days in the vilaprisan group. Similarly, more vilaprisan- than placebo-treated women achieved a response in heavy menstrual bleeding (91.7% vs. 25.0%, <em>P</em><.0001). Serious adverse events were reported for 22 (27.8%) of 79 women and were evenly distributed among the 4 groups receiving vilaprisan and/or placebo. None of these events led to study discontinuation or were related to the liver, and no new safety findings were identified compared with the earlier phase 2 ASTEROID studies.</p></div><div><h3>Conclusion(s)</h3><p>Vilaprisan is efficacious and well tolerated over 12 weeks in the treatment of heavy menstrual bleeding associated with uterine fibroids. Further investigations of the long-term efficacy and safety of vilaprisan are warranted.</p></div><div><h3>Clinical Trial Registration Number</h3><p>NCT03400943 (<span>ClinicalTrials.gov</span><svg><path></path></svg>).</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 4","pages":"Pages 317-326"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9940805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-11-01DOI: 10.1016/j.xfss.2023.09.001
Ryan H. Miller M.S. , Elizabeth A. DeVilbiss Ph.D. , Kristin R. Brogaard Ph.D. , Carter R. Norton , Chad A. Pollard B.S. , Benjamin R. Emery M.S. , Kenneth I. Aston Ph.D. , James M. Hotaling M.D., M.S. , Tim G. Jenkins Ph.D.
Objective
To investigate the power of DNA methylation variability in sperm cells in assessing male fertility potential.
Design
Retrospective cohort.
Setting
Fertility care centers.
Patients
Male patients seeking infertility treatment and fertile male sperm donors.
Intervention
None.
Main Outcome Measures
Sperm DNA methylation data from 43 fertile sperm donors were analyzed and compared with the data from 1344 men seeking fertility assessment or treatment. Methylation at gene promoters with the least variable methylation in fertile patients was used to create 3 categories of promoter dysregulation in the infertility treatment cohort: poor, average, and excellent sperm quality.
Results
After controlling for female factors, there were significant differences in intrauterine insemination pregnancy and live birth outcomes between the poor and excellent groups across a cumulative average of 2–3 cycles: 19.4% vs. 51.7% (P=.008) and 19.4% vs. 44.8% (P=.03), respectively. Live birth outcomes from in vitro fertilization, primarily with intracytoplasmic sperm injection, were not found to be significantly different among any of the 3 groups.
Conclusion
Methylation variability in a panel of 1233 gene promoters could augment the predictive ability of semen analysis and be a reliable biomarker for assessing intrauterine insemination outcomes. In vitro fertilization with intracytoplasmic sperm injection appears to overcome high levels of epigenetic instability in sperm.
{"title":"Epigenetic determinants of reproductive potential augment the predictive ability of the semen analysis","authors":"Ryan H. Miller M.S. , Elizabeth A. DeVilbiss Ph.D. , Kristin R. Brogaard Ph.D. , Carter R. Norton , Chad A. Pollard B.S. , Benjamin R. Emery M.S. , Kenneth I. Aston Ph.D. , James M. Hotaling M.D., M.S. , Tim G. Jenkins Ph.D.","doi":"10.1016/j.xfss.2023.09.001","DOIUrl":"10.1016/j.xfss.2023.09.001","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the power of DNA methylation variability in sperm cells in assessing male fertility potential.</p></div><div><h3>Design</h3><p>Retrospective cohort.</p></div><div><h3>Setting</h3><p>Fertility care centers.</p></div><div><h3>Patients</h3><p>Male patients seeking infertility treatment and fertile male sperm donors.</p></div><div><h3>Intervention</h3><p>None.</p></div><div><h3>Main Outcome Measures</h3><p>Sperm DNA methylation data from 43 fertile sperm donors were analyzed and compared with the data from 1344 men seeking fertility assessment or treatment. Methylation at gene promoters with the least variable methylation in fertile patients was used to create 3 categories of promoter dysregulation in the infertility treatment cohort: poor, average, and excellent sperm quality.</p></div><div><h3>Results</h3><p>After controlling for female factors, there were significant differences in intrauterine insemination pregnancy and live birth outcomes between the poor and excellent groups across a cumulative average of 2–3 cycles: 19.4% vs. 51.7% (<em>P</em>=.008) and 19.4% vs. 44.8% (<em>P</em>=.03), respectively. Live birth outcomes from in vitro fertilization, primarily with intracytoplasmic sperm injection, were not found to be significantly different among any of the 3 groups.</p></div><div><h3>Conclusion</h3><p>Methylation variability in a panel of 1233 gene promoters could augment the predictive ability of semen analysis and be a reliable biomarker for assessing intrauterine insemination outcomes. In vitro fertilization with intracytoplasmic sperm injection appears to overcome high levels of epigenetic instability in sperm.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 4","pages":"Pages 279-285"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666335X23000496/pdfft?md5=43c638c46631220e714347868ce6e311&pid=1-s2.0-S2666335X23000496-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10316832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To develop a spatiotemporal model for de prediction of euploid and aneuploid embryos using time-lapse videos from 10–115 hours after insemination (hpi).
Design
Retrospective study.
Main Outcome Measures
The research used an end-to-end approach to develop an automated artificial intelligence system capable of extracting features from images and classifying them, considering spatiotemporal dependencies. A convolutional neural network extracted the most relevant features from each video frame. A bidirectional long short-term memory layer received this information and analyzed the temporal dependencies, obtaining a low-dimensional feature vector that characterized each video. A multilayer perceptron classified them into 2 groups, euploid and noneuploid.
Results
The model performance in accuracy fell between 0.6170 and 0.7308. A multi-input model with a gate recurrent unit module performed better than others; the precision (or positive predictive value) is 0.8205 for predicting euploidy. Sensitivity, specificity, F1-Score and accuracy are 0.6957, 0.7813, 0.7042, and 0.7308, respectively.
Conclusions
This article proposes an artificial intelligence solution for prioritizing euploid embryo transfer. We can highlight the identification of a noninvasive method for chromosomal status diagnosis using a deep learning approach that analyzes raw data provided by time-lapse incubators. This method demonstrated potential automation of the evaluation process, allowing spatial and temporal information to encode.
{"title":"Deep learning system for classification of ploidy status using time-lapse videos","authors":"Elena Paya M.Sc. , Cristian Pulgarín M.Sc. , Lorena Bori M.Sc. , Adrián Colomer Ph.D. , Valery Naranjo Ph.D. , Marcos Meseguer Ph.D.","doi":"10.1016/j.xfss.2023.06.002","DOIUrl":"10.1016/j.xfss.2023.06.002","url":null,"abstract":"<div><h3>Objective</h3><p>To develop a spatiotemporal model for de prediction of euploid and aneuploid<span> embryos using time-lapse videos from 10–115 hours after insemination (hpi).</span></p></div><div><h3>Design</h3><p>Retrospective study.</p></div><div><h3>Main Outcome Measures</h3><p>The research used an end-to-end approach to develop an automated artificial intelligence system capable of extracting features from images and classifying them, considering spatiotemporal dependencies. A convolutional neural network extracted the most relevant features from each video frame. A bidirectional long short-term memory layer received this information and analyzed the temporal dependencies, obtaining a low-dimensional feature vector that characterized each video. A multilayer perceptron classified them into 2 groups, euploid and noneuploid.</p></div><div><h3>Results</h3><p>The model performance in accuracy fell between 0.6170 and 0.7308. A multi-input model with a gate recurrent unit module performed better than others; the precision (or positive predictive value) is 0.8205 for predicting euploidy. Sensitivity, specificity, F1-Score and accuracy are 0.6957, 0.7813, 0.7042, and 0.7308, respectively.</p></div><div><h3>Conclusions</h3><p>This article proposes an artificial intelligence solution for prioritizing euploid embryo transfer. We can highlight the identification of a noninvasive method for chromosomal status diagnosis using a deep learning approach that analyzes raw data provided by time-lapse incubators. This method demonstrated potential automation of the evaluation process, allowing spatial and temporal information to encode.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 3","pages":"Pages 211-218"},"PeriodicalIF":0.0,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10393453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-08-01DOI: 10.1016/j.xfss.2023.05.003
Mohamed Ali Ph.D. , David Stone M.D. , Archana Laknaur Ph.D. , Qiwei Yang Ph.D. , Ayman Al-Hendy M.D., Ph.D.
Objective
To investigate the link between EZH2 and Wnt/β-catenin signaling and its role in uterine fibroids (UFs) pathogenesis and explore the potential effect of natural compound methyl jasmonate (MJ) against UFs.
Design
EZH2 overexpression or inhibition was achieved in human uterine leiomyoma (HuLM) cells using EZH2-expressing adenovirus or chemical EZH2 inhibitor (DZNep), respectively. The HuLM and normal uterine smooth muscle cells were treated with 0.1–3 mM of MJ, and several experiments were employed.
Setting
Laboratory study.
Patients(s)
None.
Intervention(s)
Methyl jasmonate.
Main Outcome Measure(s)
Protein expression of EZH2, β-catenin, and proliferating cell nuclear antigen (PCNA) was measured by Western blot as well as gene expression alterations of Wnt ligands (Wnt5A, Wnt5b, and Wnt9A), WISP1, CTNNB1, and its responsive gene PITX2 using quantitative real-time polymerase chain reaction. The protein and ribonucleic acid (RNA) levels of several markers were measured in MJ-treated or untreated HuLM cells, including EZH2 and β-catenin, extracellular matrix markers collagen type 1 (COL1A1) and fibronectin (FN), proliferation markers cyclin D1 (CCND1) and PCNA, tumor suppressor marker p21, and apoptotic markers (BAX, cytochrome c, and cleaved caspase 3).
Result(s)
EZH2 overexpression significantly increased the gene expression of several Wnt ligands (PITX2, WISP1, WNT5A, WNT5B, and WNT9A), which increased nuclear translocation of β-catenin and PCNA and eventually HuLM cell proliferation. EZH2 inhibition blocked Wnt/β-catenin signaling activation where the aforementioned genes significantly decreased as well as PCNA, cyclin D1, and PITX2 protein expression compared with those in untreated HuLM. Methyl jasmonate showed a potent antiproliferative effect on HuLM cells in a dose- and time-dependent manner. Interestingly, the dose range (0.1–0.5 mM) showed a selective growth inhibitory effect on HuLM cells, not on normal uterine smooth muscle cells. Methyl jasmonate treatment at 0.5 mM for 24 hours significantly decreased both protein and RNA levels of EZH2, β-catenin, COL1A1, FN, CCND1, PCNA, WISP1, and PITX2 but increased the protein levels of p21, BAX, cytochrome, c and cleaved caspase 3 compared with untreated HuLM. Methyl jasmonate–treated cells exhibited down-regulation in the RNA expression of 36 genes, including CTNNB1, CCND1, Wnt5A, Wnt5B, and Wnt9A, and up-regulation in the expression of 34 genes, including Wnt antagonist genes WIF1, PRICKlE1, and DKK1 compared with control, confirming the quantitative real-time polymerase chain reaction results.
{"title":"EZH2 activates Wnt/β-catenin signaling in human uterine fibroids, which is inhibited by the natural compound methyl jasmonate","authors":"Mohamed Ali Ph.D. , David Stone M.D. , Archana Laknaur Ph.D. , Qiwei Yang Ph.D. , Ayman Al-Hendy M.D., Ph.D.","doi":"10.1016/j.xfss.2023.05.003","DOIUrl":"10.1016/j.xfss.2023.05.003","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the link between EZH2 and Wnt/β-catenin signaling and its role in uterine fibroids (UFs) pathogenesis and explore the potential effect of natural compound methyl jasmonate (MJ) against UFs.</p></div><div><h3>Design</h3><p>EZH2 overexpression or inhibition was achieved in human uterine leiomyoma (HuLM) cells using EZH2-expressing adenovirus or chemical EZH2 inhibitor (DZNep), respectively. The HuLM and normal uterine smooth muscle cells were treated with 0.1–3 mM of MJ, and several experiments were employed.</p></div><div><h3>Setting</h3><p>Laboratory study.</p></div><div><h3>Patients(s)</h3><p>None.</p></div><div><h3>Intervention(s)</h3><p>Methyl jasmonate.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Protein expression of EZH2, β-catenin, and proliferating cell nuclear antigen (PCNA) was measured by Western blot as well as gene expression alterations of Wnt ligands (<em>Wnt5A</em>, <em>Wnt5b</em>, and <em>Wnt9A</em>), <em>WISP1</em>, <em>CTNNB1</em>, and its responsive gene <em>PITX2</em> using quantitative real-time polymerase chain reaction. The protein and ribonucleic acid (RNA) levels of several markers were measured in MJ-treated or untreated HuLM cells, including EZH2 and β-catenin, extracellular matrix markers collagen type 1 (COL1A1) and fibronectin (FN), proliferation markers cyclin D1 (CCND1) and PCNA, tumor suppressor marker p21, and apoptotic markers (BAX, cytochrome c, and cleaved caspase 3).</p></div><div><h3>Result(s)</h3><p>EZH2 overexpression significantly increased the gene expression of several Wnt ligands (<em>PITX2</em>, <em>WISP1</em>, <em>WNT5A</em>, <em>WNT5B</em>, and <em>WNT9A</em>), which increased nuclear translocation of β-catenin and PCNA and eventually HuLM cell proliferation. EZH2 inhibition blocked Wnt/β-catenin signaling activation where the aforementioned genes significantly decreased as well as PCNA, cyclin D1, and PITX2 protein expression compared with those in untreated HuLM. Methyl jasmonate showed a potent antiproliferative effect on HuLM cells in a dose- and time-dependent manner. Interestingly, the dose range (0.1–0.5 mM) showed a selective growth inhibitory effect on HuLM cells, not on normal uterine smooth muscle cells. Methyl jasmonate treatment at 0.5 mM for 24 hours significantly decreased both protein and RNA levels of EZH2, β-catenin, COL1A1, FN, CCND1, PCNA, WISP1, and PITX2 but increased the protein levels of p21, BAX, cytochrome, c and cleaved caspase 3 compared with untreated HuLM. Methyl jasmonate–treated cells exhibited down-regulation in the RNA expression of 36 genes, including <em>CTNNB1</em>, <em>CCND1</em>, <em>Wnt5A</em>, <em>Wnt5B</em>, and <em>Wnt9A</em>, and up-regulation in the expression of 34 genes, including Wnt antagonist genes <em>WIF1</em>, <em>PRICKlE1</em>, and DKK1 compared with control, confirming the quantitative real-time polymerase chain reaction results.</p></div><div><h3>Conclusion(s)</h3><p>Our studies provide a novel lin","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 3","pages":"Pages 239-256"},"PeriodicalIF":0.0,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10527015/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10151011","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号