Pub Date : 2022-11-01DOI: 10.1016/j.xfss.2022.05.001
Lauren Reschke M.D., Sadia Afrin Ph.D., Malak El Sabah M.D., Natasha Charewycz B.S., Mariko Miyashita-Ishiwata M.D., Ph.D., Mostafa A. Borahay M.D., Ph.D., M.B.A.
Objective
To investigate the molecular effects of leptin on uterine leiomyoma cells.
Design
Experimental study using in vitro culture of immortalized human leiomyoma (HuLM) cells.
Setting
Academic university center.
Patient(s)
Women with uterine fibroids who underwent a hysterectomy or myomectomy.
Intervention(s)
Administration of human recombinant leptin to the media of cultured HuLM cells separately or in combination with pharmacologic Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) or mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) inhibitors.
Main Outcome Measure(s)
We examined HuLM tissues and cells for the expression of the leptin receptor, termed OB-R. Cellular proliferation was measured at 6, 24, and 48 hours using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay. Protein expression levels of proliferating cell nuclear antigen, collagen 1, phosphorylated STAT3/total STAT3, and phosphorylated ERK1/2 and total ERK1/2 were quantified using immunoblotting. Pharmacologic inhibitors were employed to further assess the role of the JAK2/STAT3 and MAPK/ERK pathways in the proliferative response.
Result(s)
The presence of OB-R was confirmed in clinical leiomyoma and myometrial tissue obtained from 3 separate human subjects using immunofluorescence staining, and the expression of OB-R in HuLM cells was identified using immunoblotting. There was no significant difference in the expression of the leptin receptor in the myometrium compared with that in the leiomyoma tissue. Leptin stimulated cell proliferation and extracellular matrix (ECM) deposition at 24 hours after treatment. Pretreatment with a JAK2/STAT3 inhibitor attenuated ECM deposition, and pretreatment with a MAPK/ERK inhibitor significantly decreased leptin’s stimulatory effect on cell proliferation and ECM deposition.
Conclusion(s)
Leptin induces a proliferative response and ECM deposition in HuLM cells. These findings suggest that leptin, acting through the JAK2/STAT3 and MAPK/ERK pathways, is involved in the development of uterine leiomyomas, which may partly explain their increased incidence in obese women.
{"title":"Leptin induces leiomyoma cell proliferation and extracellular matrix deposition via JAK2/STAT3 and MAPK/ERK pathways","authors":"Lauren Reschke M.D., Sadia Afrin Ph.D., Malak El Sabah M.D., Natasha Charewycz B.S., Mariko Miyashita-Ishiwata M.D., Ph.D., Mostafa A. Borahay M.D., Ph.D., M.B.A.","doi":"10.1016/j.xfss.2022.05.001","DOIUrl":"10.1016/j.xfss.2022.05.001","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the molecular effects of leptin on uterine leiomyoma cells.</p></div><div><h3>Design</h3><p>Experimental study using in vitro culture of immortalized human leiomyoma (HuLM) cells.</p></div><div><h3>Setting</h3><p>Academic university center.</p></div><div><h3>Patient(s)</h3><p><span>Women with uterine fibroids who underwent a hysterectomy or </span>myomectomy.</p></div><div><h3>Intervention(s)</h3><p>Administration of human recombinant leptin<span> to the media of cultured HuLM cells separately or in combination with pharmacologic Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) or mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) inhibitors.</span></p></div><div><h3>Main Outcome Measure(s)</h3><p><span>We examined HuLM tissues and cells for the expression of the leptin receptor<span>, termed OB-R. Cellular proliferation was measured at 6, 24, and 48 hours using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2</span></span><em>H</em><span><span><span>-tetrazolium bromide assay. Protein expression<span> levels of proliferating cell nuclear antigen, </span></span>collagen 1, phosphorylated STAT3/total STAT3, and phosphorylated ERK1/2 and total ERK1/2 were quantified using </span>immunoblotting. Pharmacologic inhibitors were employed to further assess the role of the JAK2/STAT3 and MAPK/ERK pathways in the proliferative response.</span></p></div><div><h3>Result(s)</h3><p><span>The presence of OB-R was confirmed in clinical leiomyoma and myometrial tissue obtained from 3 separate human subjects using immunofluorescence staining, and the expression of OB-R in HuLM cells was identified using immunoblotting. There was no significant difference in the expression of the leptin receptor in the </span>myometrium<span> compared with that in the leiomyoma tissue. Leptin stimulated cell proliferation and extracellular matrix<span> (ECM) deposition at 24 hours after treatment. Pretreatment with a JAK2/STAT3 inhibitor attenuated ECM deposition, and pretreatment with a MAPK/ERK inhibitor significantly decreased leptin’s stimulatory effect on cell proliferation and ECM deposition.</span></span></p></div><div><h3>Conclusion(s)</h3><p>Leptin induces a proliferative response and ECM deposition in HuLM cells. These findings suggest that leptin, acting through the JAK2/STAT3 and MAPK/ERK pathways, is involved in the development of uterine leiomyomas, which may partly explain their increased incidence in obese women.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48076045","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 : 2022-11-01DOI: 10.1016/j.xfss.2022.10.001
William H. Catherino M.D., Ph.D.
{"title":"From the Editor-in-Chief","authors":"William H. Catherino M.D., Ph.D.","doi":"10.1016/j.xfss.2022.10.001","DOIUrl":"10.1016/j.xfss.2022.10.001","url":null,"abstract":"","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33513080","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 evaluate the phosphorylation of estrogen receptor α at serine-118 (phospho-ERα S118) in the endometrium, ovarian endometrioma, and deep infiltrating endometriosis (DIE).
Design
Experimental study.
Setting
University-affiliated hospital and academic research laboratory.
Patient(s)
Twenty-five patients underwent a hysterectomy, 18 patients underwent surgical removal of ovarian endometrioma, and 6 patients underwent DIE.
Intervention(s)
Tissue samples were obtained from patients who underwent surgical procedures.
Main Outcome Measure(s)
Immunostaining for phospho-ERα S118, ERα, or phosphorylated p44/42 mitogen-activated protein kinase (phospho-p44/42 MAPK) was performed to evaluate the endometrium with or without endometriosis, ovarian endometrioma, and DIE. For in vitro analysis, endometrial epithelial cells (Ishikawa cells) were stimulated with estradiol (E2) or tumor necrosis factor alpha (TNFα), and the expression levels of phospho-ERα S118 and phospho-p44/42 MAPK were evaluated via Western blotting.
Result(s)
First, phospho-ERα S118 level was significantly higher in the glands and stroma of ovarian endometriosis samples than in those of endometrial and DIE samples. Second, colocalization of phospho-p44/42 MAPK and phospho-ERα S118 was observed in the glands of ovarian endometrioma. The proportions of cells strongly expressing phospho-p44/42 and phospho-ERα were 87% in phosphor-p44/42 MAPK-positive cells and 79% in phosphor-ERα-positive cells. Third, E2 stimulation significantly enhanced phospho-ERα S118 after 15 and 30 minutes in in vitro analysis using endometrial epithelial cells. Fourth, TNFα stimulation modestly but significantly enhanced phospho-ERα S118 after 15 and 30 minutes. Fifth, in Ishikawa cells, treatment with a p44/42 inhibitor (PD98059) significantly reduced phospho-ERα S118 by TNFα but not by E2.
Conclusion(s)
ERα-S118 phosphorylation was increased in ovarian endometriosis. Our findings may provide a new perspective for understanding the mechanism of increased ERα action in the pathophysiology of endometriosis.
{"title":"Elevated phosphorylation of estrogen receptor α at serine-118 in ovarian endometrioma","authors":"Hui Sun M.D. , Tetsuya Hirata M.D., Ph.D. , Kaori Koga M.D., Ph.D. , Tomoko Arakawa M.D. , Natsuki Nagashima M.D. , Kazuaki Neriishi M.D., Ph.D. , Mohammed Elsherbini M.D. , Eiko Maki M.D. , Gentaro Izumi M.D., Ph.D. , Miyuki Harada M.D., Ph.D. , Yasushi Hirota M.D., Ph.D. , Osamu Wada-Hiraike M.D., Ph.D. , Yutaka Osuga M.D., Ph.D.","doi":"10.1016/j.xfss.2022.04.004","DOIUrl":"10.1016/j.xfss.2022.04.004","url":null,"abstract":"<div><h3>Objective</h3><p><span>To evaluate the phosphorylation of estrogen receptor α<span> at serine-118 (phospho-ERα S118) in the endometrium, ovarian </span></span>endometrioma<span>, and deep infiltrating endometriosis (DIE).</span></p></div><div><h3>Design</h3><p>Experimental study.</p></div><div><h3>Setting</h3><p>University-affiliated hospital and academic research laboratory.</p></div><div><h3>Patient(s)</h3><p>Twenty-five patients underwent a hysterectomy, 18 patients underwent surgical removal of ovarian endometrioma, and 6 patients underwent DIE.</p></div><div><h3>Intervention(s)</h3><p>Tissue samples were obtained from patients who underwent surgical procedures.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Immunostaining<span> for phospho-ERα S118, ERα, or phosphorylated p44/42 mitogen-activated protein kinase (phospho-p44/42 MAPK) was performed to evaluate the endometrium with or without endometriosis, ovarian endometrioma, and DIE. For in vitro analysis, endometrial epithelial cells (Ishikawa cells) were stimulated with estradiol (E2) or tumor necrosis factor alpha<span> (TNFα), and the expression levels of phospho-ERα S118 and phospho-p44/42 MAPK were evaluated via Western blotting.</span></span></p></div><div><h3>Result(s)</h3><p>First, phospho-ERα S118 level was significantly higher in the glands and stroma<span> of ovarian endometriosis samples than in those of endometrial and DIE samples. Second, colocalization of phospho-p44/42 MAPK and phospho-ERα S118 was observed in the glands of ovarian endometrioma. The proportions of cells strongly expressing phospho-p44/42 and phospho-ERα were 87% in phosphor-p44/42 MAPK-positive cells and 79% in phosphor-ERα-positive cells. Third, E2 stimulation significantly enhanced phospho-ERα S118 after 15 and 30 minutes in in vitro analysis using endometrial epithelial cells. Fourth, TNFα stimulation modestly but significantly enhanced phospho-ERα S118 after 15 and 30 minutes. Fifth, in Ishikawa cells, treatment with a p44/42 inhibitor (PD98059) significantly reduced phospho-ERα S118 by TNFα but not by E2.</span></p></div><div><h3>Conclusion(s)</h3><p>ERα-S118 phosphorylation was increased in ovarian endometriosis. Our findings may provide a new perspective for understanding the mechanism of increased ERα action in the pathophysiology of endometriosis.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47231905","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 : 2022-11-01DOI: 10.1016/j.xfss.2022.09.002
Samantha B. Schon M.D., M.T.R. , Kun Yang M.D. , Ronald Schindler B.S. , Li Jiang M.S. , Lisa M. Neff M.D., M.S. , Randy J. Seeley Ph.D. , Erica E. Marsh M.D., M.Sci.
Objective
To compare the proteomic composition of follicular fluid from women with normal weight vs. women with obesity but without a history of polycystic ovary syndrome or known ovarian dysfunction undergoing in vitro fertilization.
Design
Cross-sectional.
Setting
Academic medical center.
Patient(s)
Eight women with normal weight and 8 women with obesity undergoing in vitro fertilization and without a history of polycystic ovary syndrome, ovulatory dysfunction, diminished ovarian reserve, or known endometriosis were included in the analysis.
Intervention(s)
Not applicable.
Main Outcome Measure(s)
Proteomic assessment using liquid chromatography–mass spectrometry analysis.
Result(s)
The mean age of women with normal weight was similar to that of women with obesity (32.9 vs. 32.6 years, not significant). The mean body mass index of women with normal weight was 21.2 kg/m2 compared with a body mass index of 37.1 kg/m2 in women with obesity. A total of 1,174 proteins were identified with ≥2 peptides present. Twenty-five proteins were found to be significantly altered in the follicular fluid from women with obesity. Of these 25 proteins, 19 were up-regulated and 6 were down-regulated. Notably, C-reactive protein was 11-fold higher in the follicular fluid from women with obesity than in the follicular fluid from women with normal weight.
Conclusion(s)
Obesity is associated with dysregulation at the level of the follicle, including alterations in proteins related to inflammation and metabolism. These include proteins with emerging roles in energy homeostasis and follicular regulation.
{"title":"Obesity-related alterations in protein expression in human follicular fluid from women undergoing in vitro fertilization","authors":"Samantha B. Schon M.D., M.T.R. , Kun Yang M.D. , Ronald Schindler B.S. , Li Jiang M.S. , Lisa M. Neff M.D., M.S. , Randy J. Seeley Ph.D. , Erica E. Marsh M.D., M.Sci.","doi":"10.1016/j.xfss.2022.09.002","DOIUrl":"10.1016/j.xfss.2022.09.002","url":null,"abstract":"<div><h3>Objective</h3><p><span>To compare the proteomic composition of follicular fluid from women with normal weight vs. women with obesity but without a history of </span>polycystic ovary syndrome<span> or known ovarian dysfunction undergoing in vitro fertilization.</span></p></div><div><h3>Design</h3><p>Cross-sectional.</p></div><div><h3>Setting</h3><p>Academic medical center.</p></div><div><h3>Patient(s)</h3><p><span>Eight women with normal weight and 8 women with obesity undergoing in vitro fertilization and without a history of polycystic ovary syndrome, ovulatory dysfunction, diminished ovarian reserve, or known </span>endometriosis were included in the analysis.</p></div><div><h3>Intervention(s)</h3><p>Not applicable.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Proteomic assessment using liquid chromatography–mass spectrometry analysis.</p></div><div><h3>Result(s)</h3><p><span>The mean age of women with normal weight was similar to that of women with obesity (32.9 vs. 32.6 years, not significant). The mean body mass index of women with normal weight was 21.2 kg/m</span><sup>2</sup> compared with a body mass index of 37.1 kg/m<sup>2</sup> in women with obesity. A total of 1,174 proteins were identified with ≥2 peptides present. Twenty-five proteins were found to be significantly altered in the follicular fluid from women with obesity. Of these 25 proteins, 19 were up-regulated and 6 were down-regulated. Notably, C-reactive protein was 11-fold higher in the follicular fluid from women with obesity than in the follicular fluid from women with normal weight.</p></div><div><h3>Conclusion(s)</h3><p>Obesity is associated with dysregulation at the level of the follicle, including alterations in proteins related to inflammation and metabolism. These include proteins with emerging roles in energy homeostasis and follicular regulation.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33464012","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 investigate testis-specific histone 2B (TSH2B) and its gene anomalies in infertile men.
Design
Case-control study.
Setting
Basic science laboratory.
Patient(s)
Fertile and infertile men.
Intervention(s)
Not applicable.
Main Outcome Measure(s)
The histone and protamine status of sperm was studied by aniline blue and chromomycin A3 staining, respectively. Testis-specific histone 2B, total H2B, and phosphorylated TSH2B (pTSH2B) were estimated by Western blot analysis. The frequency of genetic polymorphisms and rare variants in H2BC1 was studied by Sanger sequencing. Phosphosites on TSH2B in sperm were identified by reverse-phase high-performance liquid chromatography purification of TSH2B followed by mass spectrometric analysis.
Result(s)
Aniline blue and chromomycin A3 staining revealed significantly higher histone retention and low protamine in sperm of infertile men. Sperm TSH2B and total H2B levels were significantly lower in oligozoospermic and oligoasthenozoospermic men (in both groups). The TSH2B levels were comparable in asthenozoospermic men; however, the pTSH2B level was significantly low. The H2BC1 gene sequencing identified 6 variants, of which 2 are rare variants (rs368672899 and rs544942090) and 4 (rs4711096, rs4712959, rs4712960 and rs4712961) are single nucleotide polymorphisms. Minor allele frequency of 5′-untranslated region variant rs4711096 was significantly lower in infertile men (OR = 0.65). The rare nonsynonymous variant, rs368672899, p.Ser5Pro was seen in 1 oligoasthenoteratozoospermic individual. Interestingly, mass spectrometric analysis identified a site on TSH2B to bear a phosphate group in the sperm of fertile men.
Conclusion(s)
Our study reveals a defect in the replacement of somatic histones with testis-specific variants in infertile men. Chromatin compaction positively correlates with sperm motility, which is suggestive of its utility in diagnostic semen analysis of infertile individuals. Our observations with TSH2B and its cognate gene in sperm of infertile men indicate an essential role for TSH2B in meiosis and its phosphorylation in sperm motility, respectively.
{"title":"Proteomic and genetic dissection of testis-specific histone 2B in infertile men reveals its contribution to meiosis and sperm motility","authors":"Aniket Patankar M.Sc. , Digumarthi V.S. Sudhakar Ph.D. , Rahul Gajbhiye M.B.B.S., Ph.D. , Suchitra Surve M.D. , Kumarasamy Thangaraj Ph.D. , Priyanka Parte Ph.D.","doi":"10.1016/j.xfss.2022.07.003","DOIUrl":"10.1016/j.xfss.2022.07.003","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate testis-specific histone 2B (TSH2B) and its gene anomalies in infertile men.</p></div><div><h3>Design</h3><p>Case-control study.</p></div><div><h3>Setting</h3><p>Basic science laboratory.</p></div><div><h3>Patient(s)</h3><p>Fertile and infertile men.</p></div><div><h3>Intervention(s)</h3><p>Not applicable.</p></div><div><h3>Main Outcome Measure(s)</h3><p><span><span>The histone and protamine<span> status of sperm was studied by aniline blue and chromomycin A3 staining, respectively. Testis-specific </span></span>histone 2B<span>, total H2B, and phosphorylated TSH2B (pTSH2B) were estimated by Western blot analysis<span>. The frequency of genetic polymorphisms and rare variants in </span></span></span><span><em>H2BC1</em></span><span> was studied by Sanger sequencing. Phosphosites on TSH2B in sperm were identified by reverse-phase high-performance liquid chromatography purification of TSH2B followed by mass spectrometric analysis.</span></p></div><div><h3>Result(s)</h3><p>Aniline blue and chromomycin A3 staining revealed significantly higher histone retention and low protamine in sperm of infertile men. Sperm TSH2B and total H2B levels were significantly lower in oligozoospermic and oligoasthenozoospermic men (in both groups). The TSH2B levels were comparable in asthenozoospermic men; however, the pTSH2B level was significantly low. The <em>H2BC1</em><span> gene sequencing identified 6 variants, of which 2 are rare variants (rs368672899 and rs544942090) and 4 (rs4711096, rs4712959, rs4712960 and rs4712961) are single nucleotide polymorphisms. Minor allele frequency of 5′-untranslated region variant rs4711096 was significantly lower in infertile men (OR = 0.65). The rare nonsynonymous variant, rs368672899, p.Ser5Pro was seen in 1 oligoasthenoteratozoospermic individual. Interestingly, mass spectrometric analysis identified a site on TSH2B to bear a phosphate group in the sperm of fertile men.</span></p></div><div><h3>Conclusion(s)</h3><p><span>Our study reveals a defect in the replacement of somatic histones with testis-specific variants in infertile men. Chromatin compaction positively correlates with sperm motility, which is suggestive of its utility in diagnostic </span>semen analysis of infertile individuals. Our observations with TSH2B and its cognate gene in sperm of infertile men indicate an essential role for TSH2B in meiosis and its phosphorylation in sperm motility, respectively.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40607009","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 : 2022-11-01DOI: 10.1016/j.xfss.2022.06.002
Emma Giuliani M.D. , Samantha B. Schon M.D. , Kun Yang M.D. , Gregory W. Burns Ph.D. , Lisa M. Neff M.D. , Henriette A. Remmer Ph.D. , Jose M. Teixeira Ph.D. , Erica E. Marsh M.D., M.SCI.
Objective
Despite obesity’s significant impact on reproduction, its influence on the physiology of the human endometrium is largely understudied. We hypothesized that endometrial proteomic differences exist between obese (OW; body mass index [BMI] ≥30 kg/m2) and normal-weight women (NWW; BMI, 18.5–24.9 kg/m2).
Design
Clinical cross-sectional study.
Setting
Academic Medical Center.
Patient(s)
Healthy, normally-cycling, 18 to 40-year-old women (n = 6 OW and n = 6 NWW).
Main Outcome Measure(s)
Participants underwent screening and midfollicular phase visits. Demographic and anthropometric characteristics, blood samples, ultrasounds, and follicular phase endometrial biopsies were collected. Proteomic analyses of endometrial samples (liquid chromatography-mass spectrometry) were performed. Proteins with ≥2-fold difference and a false discovery rate of <0.1 were considered statistically significant (Benjamini-Hochberg adjustment).
Result(s)
Reproductive hormone levels did not differ between the two groups. Mean BMI, serum leptin concentration, and bioelectrical impedance analysis indices of adiposity were higher in OW than in NWW. Histological examination of the endometrial samples confirmed normal-appearing endometrium in both OW and NWW. A total of 2,930 proteins were detected across all samples, with an average number of proteins per sample of 2,059 ± 482 in NWW and 2,437 ± 187 in OW. A total of 17 proteins were differentially expressed in OW vs. NWW; 2 were more abundant, whereas 15 were underexpressed in OW, including the progesterone receptor.
Conclusion(s)
In this well-phenotyped population of healthy women, obesity was associated with significant endometrial proliferative phase proteomic differences affecting the hormonal and immunologic pathways. These could contribute to an increased risk of menstrual bleeding abnormalities and create an altered environment for future luteinization.
{"title":"Obesity-induced follicular phase endometrial proteome dysregulation in a well-phenotyped population","authors":"Emma Giuliani M.D. , Samantha B. Schon M.D. , Kun Yang M.D. , Gregory W. Burns Ph.D. , Lisa M. Neff M.D. , Henriette A. Remmer Ph.D. , Jose M. Teixeira Ph.D. , Erica E. Marsh M.D., M.SCI.","doi":"10.1016/j.xfss.2022.06.002","DOIUrl":"10.1016/j.xfss.2022.06.002","url":null,"abstract":"<div><h3>Objective</h3><p><span><span>Despite obesity’s significant impact on reproduction, its influence on the physiology of the human endometrium<span> is largely understudied. We hypothesized that endometrial proteomic differences exist between obese (OW; </span></span>body mass index [BMI] ≥30 kg/m</span><sup>2</sup>) and normal-weight women (NWW; BMI, 18.5–24.9 kg/m<sup>2</sup>).</p></div><div><h3>Design</h3><p>Clinical cross-sectional study.</p></div><div><h3>Setting</h3><p>Academic Medical Center.</p></div><div><h3>Patient(s)</h3><p>Healthy, normally-cycling, 18 to 40-year-old women (n = 6 OW and n = 6 NWW).</p></div><div><h3>Main Outcome Measure(s)</h3><p><span>Participants underwent screening and midfollicular phase visits. Demographic and anthropometric characteristics, blood samples, ultrasounds, and </span>follicular phase<span> endometrial biopsies were collected. Proteomic analyses of endometrial samples (liquid chromatography-mass spectrometry) were performed. Proteins with ≥2-fold difference and a false discovery rate of <0.1 were considered statistically significant (Benjamini-Hochberg adjustment).</span></p></div><div><h3>Result(s)</h3><p><span>Reproductive hormone<span> levels did not differ between the two groups. Mean BMI, serum leptin concentration, and bioelectrical impedance analysis indices of adiposity were higher in OW than in NWW. Histological examination of the endometrial samples confirmed normal-appearing endometrium in both OW and NWW. A total of 2,930 proteins were detected across all samples, with an average number of proteins per sample of 2,059 ± 482 in NWW and 2,437 ± 187 in OW. A total of 17 proteins were differentially expressed in OW vs. NWW; 2 were more abundant, whereas 15 were underexpressed in OW, including the </span></span>progesterone receptor.</p></div><div><h3>Conclusion(s)</h3><p>In this well-phenotyped population of healthy women, obesity was associated with significant endometrial proliferative phase proteomic differences affecting the hormonal and immunologic pathways. These could contribute to an increased risk of menstrual bleeding abnormalities and create an altered environment for future luteinization.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9114480","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 : 2022-11-01DOI: 10.1016/j.xfss.2022.07.004
Shuang Tang Ph.D. , Nannan Yang M.Sc. , Mingxi Yu Ph.D. , Shuo Wang Ph.D. , Xiangdong Hu B.Sc. , Heliang Ni B.Sc. , Wenyang Cai Ph.D.
Objective
To establish an optimized autologous mitochondria transport technique for oocyte–aging rescue, which minimizes both the patient’s pains and the damage to oocytes.
Design
Experimental laboratory study.
Setting
Laboratory.
Animal(s)
Institute of Cancer Research mice.
Intervention(s)
The murine umbilical cord mesenchymal stem cells were isolated from the female pup and cryopreserved. After the female aged, its germinal vesicle (GV) oocytes were collected and treated to weaken the zona pellucida. Its autologous umbilical cord mesenchymal stem cells were induced into granulosa cells (iGCs). The zona-weakened GV oocytes were aggregated with iGCs into iGC-oocyte complexes. Then, these complexes were cultured in growth-differentiation factor 9-containing media for 3 days. Next, they were subjected to in vitro maturation and fertilization. Presumptive zygotes were cultured for 24 hours, and the cleaved 2-cell embryos were selected for embryo transfer.
Main Outcome Measure(s)
The oocyte quality was determined by examining mitochondrial ultrastructure using transmission electron microscopy, the adenosine triphosphate content using a luminometer, and intracellular reactive oxygen species levels by confocal microscopy. The spindle organization in mature oocytes was examined by confocal microscopy. The developmental potential of oocytes was evaluated by monitoring the in vitro embryo development and the birth rate after embryo transfer.
Result(s)
Mitochondria migrated from iGCs into the GV oocyte via transzonal filopodia. The maturation rate, quality, and developmental potential of these oocytes were substantially increased. Furthermore, the birth rate after embryo transfer has been improved.
Conclusion(s)
This approach used noninvasive procedures to collect mitochondria donor cells and optimized mitochondria transfer manipulations; thus, it may have potential in ameliorating oocyte–aging-related subfertility.
{"title":"Noninvasive autologous mitochondria transport improves the quality and developmental potential of oocytes from aged mice","authors":"Shuang Tang Ph.D. , Nannan Yang M.Sc. , Mingxi Yu Ph.D. , Shuo Wang Ph.D. , Xiangdong Hu B.Sc. , Heliang Ni B.Sc. , Wenyang Cai Ph.D.","doi":"10.1016/j.xfss.2022.07.004","DOIUrl":"10.1016/j.xfss.2022.07.004","url":null,"abstract":"<div><h3>Objective</h3><p>To establish an optimized autologous mitochondria transport technique for oocyte–aging rescue, which minimizes both the patient’s pains and the damage to oocytes.</p></div><div><h3>Design</h3><p>Experimental laboratory study.</p></div><div><h3>Setting</h3><p>Laboratory.</p></div><div><h3>Animal(s)</h3><p>Institute of Cancer Research mice.</p></div><div><h3>Intervention(s)</h3><p><span><span>The murine umbilical cord </span>mesenchymal stem cells were isolated from the female pup and cryopreserved. After the female aged, its </span>germinal vesicle<span> (GV) oocytes were collected and treated to weaken the zona pellucida<span><span>. Its autologous umbilical cord mesenchymal stem cells were induced into granulosa cells (iGCs). The zona-weakened GV oocytes were aggregated with iGCs into iGC-oocyte complexes. Then, these complexes were cultured in growth-differentiation factor 9-containing media for 3 days. Next, they were subjected to in vitro maturation and fertilization. Presumptive </span>zygotes<span> were cultured for 24 hours, and the cleaved 2-cell embryos were selected for embryo transfer.</span></span></span></p></div><div><h3>Main Outcome Measure(s)</h3><p><span>The oocyte quality was determined by examining mitochondrial ultrastructure<span> using transmission electron microscopy<span>, the adenosine triphosphate content using a luminometer, and intracellular reactive oxygen species levels by </span></span></span>confocal microscopy. The spindle organization in mature oocytes was examined by confocal microscopy. The developmental potential of oocytes was evaluated by monitoring the in vitro embryo development and the birth rate after embryo transfer.</p></div><div><h3>Result(s)</h3><p>Mitochondria migrated from iGCs into the GV oocyte via transzonal filopodia. The maturation rate, quality, and developmental potential of these oocytes were substantially increased. Furthermore, the birth rate after embryo transfer has been improved.</p></div><div><h3>Conclusion(s)</h3><p>This approach used noninvasive procedures to collect mitochondria donor cells and optimized mitochondria transfer manipulations; thus, it may have potential in ameliorating oocyte–aging-related subfertility.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40596944","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 the expression of enzymes in tryptophan (Trp) catabolism in fibroids and matched myometrium and determine the effects of race and mediator complex subunit 12 gene (MED12) mutation on their expression.
Design
Experimental laboratory study.
Setting
Academic research laboratory.
Patient(s)
Women of reproductive age who underwent hysterectomy while on no hormonal medications before surgery.
Intervention(s)
Fibroids and matched myometrium were obtained from patients who underwent hysterectomy from different race or ethnic groups.
Main Outcome Measure(s)
The expression of enzymes in the Trp catabolic pathway, tryptophan transporters, and the cytochrome P450 1B1 gene (CYP1B1) in the fibroids and matched myometrium of women from different race and ethnic groups and in tumors bearing the MED12 mutation and tumors without the mutation was determined using quantitative reverse-transcription polymerase chain reaction. The levels of serotonin, kynurenic acid (KYNA), and nicotinamide adenine dinucleotide (NAD) were determined using enzyme-linked immunosorbent assay.
Result(s)
In fibroids, the expression of tryptophan hydroxylase 1 (TPH1), kynurenine amino transferase (KAT)2, large neutral amino acid transporter small subunit 2 (SLC7A8), and large neutral amino acid transporter small subunit 1 (SLC7A5) messenger RNA (mRNA) was high and that of kynureninase (KYNU) and tryptophanyl-tRNA ligase (WARS1) mRNA was low, with no changes in the expression of WARS2, kynurenine formamidase (AFMID), kynurenine 3-monooxygenase (KMO), KAT1, KAT3, and KAT4 compared with that in the matched myometrium (n = 81). The expression of CYP1B1 mRNA, a marker of the activation of the aryl hydrocarbon receptor, was higher in fibroids. Tumors bearing the MED12 mutation expressed higher levels of CYP1B1 and lower levels of WARS1, KAT1, KAT3, and KAT4 mRNAs compared with tumors without the MED12 mutation. Race or ethnicity affected the expression of KYNU, with tumors from African American and Hispanic patients expressing lower levels of KYNU mRNA compared with those from Caucasian patients. We also quantified the levels of serotonin, KYNA, and NAD, which are the end products of Trp catabolism. There were no significant differences in the levels of serotonin and KYNA, whereas the levels of NAD were lower in fibroids than in the paired myometrium. This reduction in the levels of NAD was independent of race or ethnicity.
{"title":"Further characterization of tryptophan metabolism and its dysregulation in fibroids","authors":"Tsai-Der Chuang Ph.D., Derek Quintanilla B.S., Drake Boos B.S., Omid Khorram M.D., Ph.D.","doi":"10.1016/j.xfss.2022.04.005","DOIUrl":"10.1016/j.xfss.2022.04.005","url":null,"abstract":"<div><h3>Objective</h3><p><span><span>To determine the expression of enzymes in tryptophan (Trp) catabolism in fibroids and matched </span>myometrium<span> and determine the effects of race and mediator complex subunit 12 gene (</span></span><em>MED12</em>) mutation on their expression.</p></div><div><h3>Design</h3><p>Experimental laboratory study.</p></div><div><h3>Setting</h3><p>Academic research laboratory.</p></div><div><h3>Patient(s)</h3><p>Women of reproductive age who underwent hysterectomy while on no hormonal medications before surgery.</p></div><div><h3>Intervention(s)</h3><p>Fibroids and matched myometrium were obtained from patients who underwent hysterectomy from different race or ethnic groups.</p></div><div><h3>Main Outcome Measure(s)</h3><p><span>The expression of enzymes in the Trp catabolic pathway, tryptophan transporters, and the cytochrome P450 1B1 gene (</span><em>CYP1B1</em>) in the fibroids and matched myometrium of women from different race and ethnic groups and in tumors bearing the <em>MED12</em><span> mutation and tumors without the mutation was determined using quantitative reverse-transcription polymerase chain reaction. The levels of serotonin, kynurenic acid<span> (KYNA), and nicotinamide adenine dinucleotide (NAD) were determined using enzyme-linked immunosorbent assay.</span></span></p></div><div><h3>Result(s)</h3><p><span>In fibroids, the expression of tryptophan hydroxylase 1 (</span><em>TPH1</em><span>), kynurenine amino transferase (</span><em>KAT</em>)<em>2</em><span>, large neutral amino acid transporter small subunit 2 (</span><em>SLC7A8</em>)<em>,</em> and large neutral amino acid transporter small subunit 1 (<em>SLC7A5</em><span>) messenger RNA (mRNA) was high and that of kynureninase (</span><em>KYNU</em>) and tryptophanyl-tRNA ligase (<em>WARS1</em>) mRNA was low, with no changes in the expression of <em>WARS2</em><span>, kynurenine formamidase (</span><em>AFMID</em>), kynurenine 3-monooxygenase (<em>KMO</em>), <em>KAT1</em>, <em>KAT3</em>, and <em>KAT4</em> compared with that in the matched myometrium (n = 81). The expression of <em>CYP1B1</em><span> mRNA, a marker of the activation of the aryl hydrocarbon receptor, was higher in fibroids. Tumors bearing the </span><em>MED12</em> mutation expressed higher levels of <em>CYP1B1</em> and lower levels of <em>WARS1, KAT1</em>, <em>KAT3</em>, and <em>KAT4</em> mRNAs compared with tumors without the <em>MED12</em> mutation. Race or ethnicity affected the expression of <em>KYNU</em>, with tumors from African American and Hispanic patients expressing lower levels of <em>KYNU</em> mRNA compared with those from Caucasian patients. We also quantified the levels of serotonin, KYNA, and NAD, which are the end products of Trp catabolism. There were no significant differences in the levels of serotonin and KYNA, whereas the levels of NAD were lower in fibroids than in the paired myometrium. This reduction in the levels of NAD was independent of race or ethnicity.</p>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45473171","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 : 2022-11-01DOI: 10.1016/j.xfss.2022.05.002
Nichole A. Garzia Ph.D. , Kara Cushing-Haugen M.S. , Yu-Han Chiu M.D., Sc.D. , Helena Sandoval-Insausti M.D., Ph.D. , Jorge E. Chavarro M.D., Sc.D. , Stacey A. Missmer Sc.D. , Holly R. Harris Sc.D.
Objective
To examine the association between the intake of fruits and vegetables with high- vs. low-pesticide residue burden and diagnosis of laparoscopically confirmed endometriosis. The etiology of endometriosis is not well understood, but dietary factors may influence the risk. Pesticides may act as endocrine disruptors, and the intake of pesticide-contaminated food is a common exposure pathway.
Design
Prospective cohort study. Hazard ratios and 95% confidence intervals were calculated using multivariable Cox proportional hazard models to evaluate the intake of fruits and vegetables with high- and low-pesticide residues in relation to the diagnosis of laparoscopically confirmed endometriosis.
Setting
Not applicable.
Patient(s)
Premenopausal US women (N = 52,053) of the Nurses’ Health Study II, aged 34–53 years at study baseline (1999), were followed until 2013. The diet was assessed every 4 years using a validated food frequency questionnaire. A previously developed and validated pesticide residue burden score (PRBS), on the basis of the US Department of Agriculture Pesticide Data Program, was used to assign fruits and vegetables to pesticide residue groups (high/low).
Intervention(s)
Not applicable.
Main Outcome Measure(s)
Cases of laparoscopically confirmed endometriosis were identified from self-reports to validated questionnaires.
Result(s)
During 14 years of follow-up, 956 incidences of laparoscopically confirmed endometriosis were reported. No association was observed between the intake of high- or low-PRBS fruit and vegetable intake and endometriosis (hazard ratio for 5th vs. 1st quintile: high-PRBS intake = 0.94, 95% confidence interval = 0.73–1.23; low-PRBS intake = 1.07, 95% confidence interval = 0.82–1.40). No associations were observed for high- or low-PRBS fruit and vegetable intake by fertility status.
Conclusion(s)
No clear associations were observed between high- or low-PRBS fruit and vegetable intake and endometriosis risk among premenopausal women. To our knowledge, this is the first study to evaluate the association between dietary pesticide residue intake and endometriosis. Further research is needed, particularly to evaluate this association among a younger population of women (adolescence or early adulthood) and assess the dietary exposure to specific pesticides or chemical families.
{"title":"Pesticide residue intake from fruit and vegetable consumption and risk of laparoscopically confirmed endometriosis","authors":"Nichole A. Garzia Ph.D. , Kara Cushing-Haugen M.S. , Yu-Han Chiu M.D., Sc.D. , Helena Sandoval-Insausti M.D., Ph.D. , Jorge E. Chavarro M.D., Sc.D. , Stacey A. Missmer Sc.D. , Holly R. Harris Sc.D.","doi":"10.1016/j.xfss.2022.05.002","DOIUrl":"10.1016/j.xfss.2022.05.002","url":null,"abstract":"<div><h3>Objective</h3><p>To examine the association between the intake of fruits and vegetables with high- vs. low-pesticide residue burden and diagnosis of laparoscopically confirmed endometriosis. The etiology of endometriosis is not well understood, but dietary factors may influence the risk. Pesticides may act as endocrine disruptors, and the intake of pesticide-contaminated food is a common exposure pathway.</p></div><div><h3>Design</h3><p><span>Prospective cohort study. Hazard ratios and 95% confidence intervals were calculated using multivariable Cox </span>proportional hazard models to evaluate the intake of fruits and vegetables with high- and low-pesticide residues in relation to the diagnosis of laparoscopically confirmed endometriosis.</p></div><div><h3>Setting</h3><p>Not applicable.</p></div><div><h3>Patient(s)</h3><p><span>Premenopausal US women (N = 52,053) of the Nurses’ Health Study II, aged 34–53 years at study baseline (1999), were followed until 2013. The diet was assessed every 4 years using a validated food frequency questionnaire. A previously developed and validated </span>pesticide residue burden score (PRBS), on the basis of the US Department of Agriculture Pesticide Data Program, was used to assign fruits and vegetables to pesticide residue groups (high/low).</p></div><div><h3>Intervention(s)</h3><p>Not applicable.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Cases of laparoscopically confirmed endometriosis were identified from self-reports to validated questionnaires.</p></div><div><h3>Result(s)</h3><p>During 14 years of follow-up, 956 incidences of laparoscopically confirmed endometriosis were reported. No association was observed between the intake of high- or low-PRBS fruit and vegetable intake and endometriosis (hazard ratio for 5th vs. 1st quintile: high-PRBS intake = 0.94, 95% confidence interval = 0.73–1.23; low-PRBS intake = 1.07, 95% confidence interval = 0.82–1.40). No associations were observed for high- or low-PRBS fruit and vegetable intake by fertility status.</p></div><div><h3>Conclusion(s)</h3><p>No clear associations were observed between high- or low-PRBS fruit and vegetable intake and endometriosis risk among premenopausal women. To our knowledge, this is the first study to evaluate the association between dietary pesticide residue intake and endometriosis. Further research is needed, particularly to evaluate this association among a younger population of women (adolescence or early adulthood) and assess the dietary exposure to specific pesticides or chemical families.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46462254","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 : 2022-11-01DOI: 10.1016/j.xfss.2022.06.003
Anna Mueller, Josef Lehner Ph.D., Katharina Hancke M.D., Wolfgang Janni M.D., Karin Bundschu M.D., Ph.D.
Objective
To investigate the advantages of cryopreserved medulla-containing ovarian cortex grafts with those of commonly used sole cortex grafts for fertility preservation by analyzing tissue quality, neovascularization processes, and the number of vital follicles.
Design
Experimental setting of cryopreserved bovine ovarian cortex tissue grafts with or without medulla tissue.
Setting
Laboratory animal research at Ulm University, Ulm, Germany.
Animals
Bovine ovaries and fertilized chicken eggs.
Intervention(s)
Experimental setting of bovine ovarian tissue samples grafted on the chicken chorioallantoic membrane (CAM) after cryopreservation and thawing to examine histologic tissue integrity, apoptosis and proliferation immunohistochemically, blood vessel counts and determine the presence of neutral red-stained vital follicles.
Main Outcome Measure(s)
We used hematoxylin and eosin staining to visualize tissue structures, immunohistochemistry with anti-caspase 3 to detect apoptosis, anti-Ki67 to examine proliferation, blood vessel count on the chicken CAM to investigate neovascularization processes, and neutral red staining to evaluate vital follicles.
Result(s)
We demonstrated that in all analyzed tissue samples, after cryopreservation, thawing, and grafting on the chicken CAM, there was excellent tissue integrity and quality, as shown by extremely rare apoptosis processes analyzed using immunohistochemical caspase 3 staining (sole cortex, 0.54%; thin medulla-containing cortex, 0.43%; thick medulla-containing cortex, 0.13%; and sole medulla, 2.82%). Moreover, we detected increased neovascularization in the vicinity of medulla and medulla-containing grafts (small blood vessels: cortex 8.7, thin medulla-containing cortex 9.9, thick medulla-containing cortex 9.7, and medulla 9.8; very small blood vessels: cortex 7.0, thin medulla-containing cortex 13.0, thick medulla-containing cortex 12.0, and medulla 15.0), with higher Ki67-detected proliferation (cortex, 17.58%; thin medulla-containing cortex, 20.28%; thick medulla-containing cortex, 20.56%; and medulla, 29.9%). Additionally, we identified an increased number of vital follicles in medulla-containing cortex grafts compared with the number of vital follicles in sole cortex tissue (cortex, 256.1; thin medulla-containing cortex, 338.2; thick medulla-containing cortex, 346.6; and medulla, 8.1).
Conclusion(s)
In this experimental setting, bovine medulla-containing cortex tissue had excellent tissue structure and quality after cryopreservation and thawing and increased neovascularization and an augmented vital follicle count after grafting than the commonly used sole cortex tissue. Therefore, we suggest reconsidering the current cryopreservation and grafting processes in humans for fertility pr
{"title":"Fertility preservation: improved neovascularization and follicle viability in cryopreserved bovine ovarian cortex transplants by remaining medulla tissue","authors":"Anna Mueller, Josef Lehner Ph.D., Katharina Hancke M.D., Wolfgang Janni M.D., Karin Bundschu M.D., Ph.D.","doi":"10.1016/j.xfss.2022.06.003","DOIUrl":"10.1016/j.xfss.2022.06.003","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the advantages of cryopreserved medulla-containing ovarian cortex grafts with those of commonly used sole cortex grafts for fertility preservation by analyzing tissue quality, neovascularization processes, and the number of vital follicles.</p></div><div><h3>Design</h3><p>Experimental setting of cryopreserved bovine ovarian cortex tissue grafts with or without medulla tissue.</p></div><div><h3>Setting</h3><p>Laboratory animal research at Ulm University, Ulm, Germany.</p></div><div><h3>Animals</h3><p>Bovine ovaries and fertilized chicken eggs.</p></div><div><h3>Intervention(s)</h3><p>Experimental setting of bovine ovarian tissue samples grafted on the chicken chorioallantoic membrane (CAM) after cryopreservation and thawing to examine histologic tissue integrity, apoptosis and proliferation immunohistochemically, blood vessel counts and determine the presence of neutral red-stained vital follicles.</p></div><div><h3>Main Outcome Measure(s)</h3><p>We used hematoxylin<span><span> and eosin staining to visualize tissue structures, </span>immunohistochemistry with anti-caspase 3 to detect apoptosis, anti-Ki67 to examine proliferation, blood vessel count on the chicken CAM to investigate neovascularization processes, and neutral red staining to evaluate vital follicles.</span></p></div><div><h3>Result(s)</h3><p>We demonstrated that in all analyzed tissue samples, after cryopreservation, thawing, and grafting on the chicken CAM, there was excellent tissue integrity and quality, as shown by extremely rare apoptosis processes analyzed using immunohistochemical caspase 3 staining (sole cortex, 0.54%; thin medulla-containing cortex, 0.43%; thick medulla-containing cortex, 0.13%; and sole medulla, 2.82%). Moreover, we detected increased neovascularization in the vicinity of medulla and medulla-containing grafts (small blood vessels: cortex 8.7, thin medulla-containing cortex 9.9, thick medulla-containing cortex 9.7, and medulla 9.8; very small blood vessels: cortex 7.0, thin medulla-containing cortex 13.0, thick medulla-containing cortex 12.0, and medulla 15.0), with higher Ki67-detected proliferation (cortex, 17.58%; thin medulla-containing cortex, 20.28%; thick medulla-containing cortex, 20.56%; and medulla, 29.9%). Additionally, we identified an increased number of vital follicles in medulla-containing cortex grafts compared with the number of vital follicles in sole cortex tissue (cortex, 256.1; thin medulla-containing cortex, 338.2; thick medulla-containing cortex, 346.6; and medulla, 8.1).</p></div><div><h3>Conclusion(s)</h3><p>In this experimental setting, bovine medulla-containing cortex tissue had excellent tissue structure and quality after cryopreservation and thawing and increased neovascularization and an augmented vital follicle count after grafting than the commonly used sole cortex tissue. Therefore, we suggest reconsidering the current cryopreservation and grafting processes in humans for fertility pr","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40150921","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}
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