F&S science最新文献_第3页

Embryos from vitrified vs. fresh oocytes in an oocyte donation program: a comparative morphokinetic analysis 卵母细胞捐献计划中来自玻璃化卵母细胞和新鲜卵母细胞的胚胎：形态动力学比较分析

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2024.03.002

Mary Karagianni M.Sc. , Maria Ioanna Papadopoulou M.Sc. , Chara Oraiopoulou M.Res. , Nikolaos Christoforidis M.D. , Achilleas Papatheodorou Ph.D. , Alexia Chatziparasidou M.Sc.

Objective

To compare the morphokinetic patterns of human embryos originating from vitrified oocytes (VITRI group) with those derived from freshly collected oocytes (CONTROL group) in oocyte donation cycles.

Design

This is a retrospective observational study.

Setting

Embryolab Fertility Clinic, Embryology Lab, Thessaloniki, Greece.

Patient(s)

The study included embryos from 421 vitrified oocytes from 58 oocyte donation cycles and 196 fresh oocytes from 23 oocyte donation cycles.

Intervention(s)

None.

Main Outcome Measure(s)

Key time parameters, dynamic events, fertilization rates, degeneration rates, cleavage rates, blastocyst rates, pregnancy rates, clinical pregnancy rates, implantation rates, and live birth rates were estimated.

Results

The mean survival rate of vitrified oocytes was 92.58% (±7.42%). Fertilization rates were significantly different between the 2 groups (VITRI group: 71.92% ± 20.29% and CONTROL group: 80.65% ± 15.22%) whereas the degeneration, cleavage, blastocyst, pregnancy, clinical pregnancy, ongoing pregnancy, implantation, and live birth rates were not significantly different between embryos derived from fresh or vitrified oocytes. Time-lapse analysis showed no significant difference in any key time parameter. However, when examining dynamic parameters, first cell cycle (CC1) (t2 − tPB2: from the second polar body extrusion (tPB2) up to 2 cells (t2)) showed a significant difference whereas CC1a (t2 − tPNf: from fading of the pronuclei (tPNf) up to 2 cells (t2)) was at the threshold of significance.

Conclusion(s)

CC1 in vitrified oocytes exhibited a comparatively slower progression in contrast to fresh oocytes. Conversely, CC1a in vitrified oocytes demonstrated faster progression compared with fresh oocytes. It is worth noting that these temporary deviations had minimal impact on the subsequent development. Despite the clinical outcomes showing a decrease in the vitrified group, none of them reached statistical significance. This lack of significance could be attributed to the limited sample size of the study.

目的比较卵母细胞捐献周期中来自玻璃化卵母细胞的人类胚胎（VITRI 组）与来自新鲜采集卵母细胞的人类胚胎（CONTROL 组）的形态发生模式。主要结果指标对关键时间参数、动态事件、受精率、退化率、裂解率、囊胚率、妊娠率、临床妊娠率、植入率和活产率进行了估算。两组的受精率有明显差异（VITRI 组：71.92%±20.29%；对照组：80.65%±15.22%），而由新鲜卵母细胞或玻璃化卵母细胞获得的胚胎的退化率、分裂率、囊胚率、妊娠率、临床妊娠率、持续妊娠率、植入率和活产率则无明显差异。延时分析表明，任何关键时间参数都没有显著差异。然而，在研究动态参数时，第一个细胞周期（CC1）（t2 - tPB2：从第二个极体挤出（tPB2）到 2 个细胞（t2））显示出显著差异，而 CC1a（t2 - tPNf：从原核消退（tPNf）到 2 个细胞（t2））则处于显著性临界值。相反，与新鲜卵母细胞相比，玻璃化卵母细胞中 CC1a 的进展较快。值得注意的是，这些暂时的偏差对后续发育的影响微乎其微。尽管玻璃化组的临床结果有所下降，但均未达到统计学意义。缺乏显著性可能是由于研究的样本量有限。

{"title":"Embryos from vitrified vs. fresh oocytes in an oocyte donation program: a comparative morphokinetic analysis","authors":"Mary Karagianni M.Sc. , Maria Ioanna Papadopoulou M.Sc. , Chara Oraiopoulou M.Res. , Nikolaos Christoforidis M.D. , Achilleas Papatheodorou Ph.D. , Alexia Chatziparasidou M.Sc.","doi":"10.1016/j.xfss.2024.03.002","DOIUrl":"10.1016/j.xfss.2024.03.002","url":null,"abstract":"<div><h3>Objective</h3><p>To compare the morphokinetic patterns of human embryos originating from vitrified oocytes (VITRI group) with those derived from freshly collected oocytes (CONTROL group) in oocyte donation cycles.</p></div><div><h3>Design</h3><p>This is a retrospective observational study.</p></div><div><h3>Setting</h3><p>Embryolab Fertility Clinic, Embryology Lab, Thessaloniki, Greece.</p></div><div><h3>Patient(s)</h3><p>The study included embryos from 421 vitrified oocytes from 58 oocyte donation cycles and 196 fresh oocytes from 23 oocyte donation cycles.</p></div><div><h3>Intervention(s)</h3><p>None.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Key time parameters, dynamic events, fertilization rates, degeneration rates, cleavage rates, blastocyst rates, pregnancy rates, clinical pregnancy rates, implantation rates, and live birth rates were estimated.</p></div><div><h3>Results</h3><p>The mean survival rate of vitrified oocytes was 92.58% (±7.42%). Fertilization rates were significantly different between the 2 groups (VITRI group: 71.92% ± 20.29% and CONTROL group: 80.65% ± 15.22%) whereas the degeneration, cleavage, blastocyst, pregnancy, clinical pregnancy, ongoing pregnancy, implantation, and live birth rates were not significantly different between embryos derived from fresh or vitrified oocytes. Time-lapse analysis showed no significant difference in any key time parameter. However, when examining dynamic parameters, first cell cycle (CC1) (t2 − tPB2: from the second polar body extrusion (tPB2) up to 2 cells (t2)) showed a significant difference whereas CC1a (t2 − tPNf: from fading of the pronuclei (tPNf) up to 2 cells (t2)) was at the threshold of significance.</p></div><div><h3>Conclusion(s)</h3><p>CC1 in vitrified oocytes exhibited a comparatively slower progression in contrast to fresh oocytes. Conversely, CC1a in vitrified oocytes demonstrated faster progression compared with fresh oocytes. It is worth noting that these temporary deviations had minimal impact on the subsequent development. Despite the clinical outcomes showing a decrease in the vitrified group, none of them reached statistical significance. This lack of significance could be attributed to the limited sample size of the study.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140759714","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}

引用次数: 0

Transcriptomic changes in eutopic endometrium and ectopic lesions during endometriosis progression in a mouse model 小鼠模型中异位子宫内膜和异位病灶在子宫内膜异位症发展过程中的转录组变化。

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2024.02.001

Rong Li Ph.D. , Dinh Nam Tran Ph.D. , Bruce A. Lessey M.D., Ph.D. , Steven L. Young M.D., Ph.D. , Tae Hoon Kim Ph.D. , Jae-Wook Jeong Ph.D.

Objective

To identify the transcriptomic changes of ectopic lesions and eutopic endometrial tissues during the progression of endometriosis, we performed transcriptomic analysis in the eutopic endometrium and ectopic lesions.

Design

Laboratory study.

Setting

Academic medical center.

Animals

Four fertile and 4 subfertile Pgr^cre/+Rosa26^mTmG/+ mice with endometriosis, and 4 sham mice for each group of endometriosis mice as control. These mice underwent either surgery to induce endometriosis or sham surgery. Fertile sham and mice with endometriosis were used 1 month after surgery, whereas subfertile ones were used 3 months after surgery.

Interventions

Early and chronic effects of endometriosis on transcriptomics of ectopic lesions and eutopic endometrium.

Main Outcome Measures

RNA-sequencing analysis and identification of differentially expressed genes and pathways in the ectopic lesions and eutopic uteri from mice with endometriosis and sham mice at day 3.5 of pregnancy.

Results

Our mouse model recapitulates the transcriptomic changes of ectopic lesions in humans. RNA-sequencing analysis was performed in ectopic lesions and eutopic uteri from mice with or without endometriosis during the progression of the disease. Estrogen activity, inflammation, angiogenesis, and fibrosis pathways were consistently elevated in all the ectopic lesions compared with eutopic endometrium. Cholesterol/glucose synthesis and stem cell pluripotency pathways were more enhanced in ectopic lesions from subfertile mice compared with their eutopic endometrium. Dysregulation of infiltration of macrophage, dendritic, T and B cells was validated with the use of immunohistochemistry in ectopic lesions. Multiple ligand–receptor pairs between the ectopic and eutopic endometrium were altered compared with the sham endometrium. Suppressed WNT and EGF pathways were only found in the eutopic endometrium from subfertile not fertile mice compared with sham.

Conclusions

Our mouse endometriosis model recapitulates the transcriptomics of ectopic lesions in humans. Our transcriptomic analysis during endometriosis progression in our mouse model will help us understand the pathophysiology of endometriosis.

目的确定异位病灶和异位子宫内膜组织在子宫内膜异位症进展过程中的转录组变化：我们的假设是，子宫内膜异位症的发展和进展会改变异位内膜和异位病灶的转录组：学术医学中心四只可育和四只亚可育的 Pgrcre/+Rosa26mTmG/+ 子宫内膜异位症小鼠，每组子宫内膜异位症小鼠有四只假小鼠作为对照。这些小鼠要么接受了诱发子宫内膜异位症的手术，要么接受了假手术。肥育假小鼠和子宫内膜异位症小鼠在手术后一个月使用，亚肥育小鼠在手术后三个月使用：干预措施：子宫内膜异位症对异位病灶和异位内膜转录组学的早期和慢性影响：结果：我们的小鼠模型再现了子宫内膜异位症对异位病灶和异位子宫内膜转录组学的早期和慢性影响：我们的小鼠模型再现了人类异位病变的转录组变化。我们对患有或未患有子宫内膜异位症的小鼠异位病灶和异位子宫在疾病进展过程中的转录组变化进行了RNA-seq分析。与异位子宫内膜相比，所有异位病灶中的雌二醇、炎症、血管生成和纤维化通路都持续升高。与异位子宫内膜相比，亚肥育小鼠异位病灶中的胆固醇/葡萄糖合成和干细胞多能性途径更强。异位病灶中巨噬细胞、树突状细胞、T 细胞和 B 细胞的浸润失调通过免疫组化得到了验证。与假性子宫内膜相比，异位和异位子宫内膜的多种配体-受体对发生了改变。与假性内膜相比，仅在亚肥育而非肥育小鼠的异位内膜中发现了受抑制的 WNT 和 EGF 通路：我们的小鼠子宫内膜异位症模型再现了人类异位病变的转录组学。结论：我们的小鼠子宫内膜异位症模型再现了人类异位病变的转录组学，我们在小鼠模型中进行的子宫内膜异位症进展过程中的转录组学分析将有助于了解子宫内膜异位症的病理生理学。

{"title":"Transcriptomic changes in eutopic endometrium and ectopic lesions during endometriosis progression in a mouse model","authors":"Rong Li Ph.D. , Dinh Nam Tran Ph.D. , Bruce A. Lessey M.D., Ph.D. , Steven L. Young M.D., Ph.D. , Tae Hoon Kim Ph.D. , Jae-Wook Jeong Ph.D.","doi":"10.1016/j.xfss.2024.02.001","DOIUrl":"10.1016/j.xfss.2024.02.001","url":null,"abstract":"<div><h3>Objective</h3><p>To identify the transcriptomic changes of ectopic lesions and eutopic endometrial tissues during the progression of endometriosis, we performed transcriptomic analysis in the eutopic endometrium and ectopic lesions.</p></div><div><h3>Design</h3><p>Laboratory study.</p></div><div><h3>Setting</h3><p>Academic medical center.</p></div><div><h3>Animals</h3><p>Four fertile and 4 subfertile <em>Pgr</em><sup>cre/+</sup><em>Rosa26</em><sup>mTmG/+</sup> mice with endometriosis, and 4 sham mice for each group of endometriosis mice as control. These mice underwent either surgery to induce endometriosis or sham surgery. Fertile sham and mice with endometriosis were used 1 month after surgery, whereas subfertile ones were used 3 months after surgery.</p></div><div><h3>Interventions</h3><p>Early and chronic effects of endometriosis on transcriptomics of ectopic lesions and eutopic endometrium.</p></div><div><h3>Main Outcome Measures</h3><p>RNA-sequencing analysis and identification of differentially expressed genes and pathways in the ectopic lesions and eutopic uteri from mice with endometriosis and sham mice at day 3.5 of pregnancy.</p></div><div><h3>Results</h3><p>Our mouse model recapitulates the transcriptomic changes of ectopic lesions in humans. RNA-sequencing analysis was performed in ectopic lesions and eutopic uteri from mice with or without endometriosis during the progression of the disease. Estrogen activity, inflammation, angiogenesis, and fibrosis pathways were consistently elevated in all the ectopic lesions compared with eutopic endometrium. Cholesterol/glucose synthesis and stem cell pluripotency pathways were more enhanced in ectopic lesions from subfertile mice compared with their eutopic endometrium. Dysregulation of infiltration of macrophage, dendritic, T and B cells was validated with the use of immunohistochemistry in ectopic lesions. Multiple ligand–receptor pairs between the ectopic and eutopic endometrium were altered compared with the sham endometrium. Suppressed WNT and EGF pathways were only found in the eutopic endometrium from subfertile not fertile mice compared with sham.</p></div><div><h3>Conclusions</h3><p>Our mouse endometriosis model recapitulates the transcriptomics of ectopic lesions in humans. Our transcriptomic analysis during endometriosis progression in our mouse model will help us understand the pathophysiology of endometriosis.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139718103","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}

引用次数: 0

From the Editor-in-Chief 主编的话

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2024.04.001

William H. Catherino M.D., Ph.D.

引用次数: 0

Assessment of ovarian tissue and follicular integrity after cryopreservation via slow freezing or vitrification followed by in vitro culture 通过缓慢冷冻或玻璃化冷冻保存卵巢组织和卵泡后进行体外培养的完整性评估

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2023.10.004

Juliana I. Candelaria M.S., Anna C. Denicol D.V.M., Ph.D.

Objective

To evaluate ovarian tissue and follicle integrity before and after slow freezing or vitrification and postthawing in vitro culture.

Design

A laboratory study using bovine ovarian cortical tissue.

Setting

Academic laboratory.

Animals

Ovaries from healthy cattle.

Interventions

Bovine ovarian cortical tissue was subjected to either slow freezing or vitrification and subsequent in vitro culture. Tissue and follicle integrity were assessed before and after cryopreservation and culture.

Main Outcome Measures

Hematoxylin and eosin staining was used to assess follicle stages, morphology, and stromal cell density. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining was used to examine apoptosis, and Masson’s trichrome staining was used to evaluate collagen content in the stromal environment. Immunofluorescent labeling was used to localize and quantify connexin 37 (CX37) and Ki67 expression.

Results

Regardless of previous cryopreservation, ovarian tissue culture resulted in a decreased percentage of primordial follicles and an increased percentage of primary follicles compared with fresh tissue, indicating that follicle activation was not negatively affected by cryopreservation. However, both culture and cryopreservation followed by culture decreased the percentage of normal preantral follicles compared with fresh tissue that had not been cultured. Culture and/or cryopreservation did not impact stromal cell number, but there was increased cell apoptosis in tissue that was cultured after vitrification compared with tissue that was not cultured. Tissue culture, regardless of cryopreservation, resulted in decreased collagen deposition. There were fewer follicles expressing CX37 in vitrified and thawed tissue compared with all other treatments. Cryopreservation and/or culture of ovarian tissue did not change the percentage of follicles that contained Ki67-positive granulosa cells or the percentage of Ki67-positive granulosa cells within those follicles.

Conclusion

Based on these data, we conclude that tissue cryopreservation followed by culture does not affect follicle activation and growth, but it decreases the proportion of viable follicles within the tissue. Slow freezing was superior to vitrification as indicated by a higher proportion of follicles with normal morphology, lower stromal cell apoptosis, and maintenance of CX37 expression postthawing and after culture.

目的评估缓慢冷冻或玻璃化以及解冻后体外培养前后卵巢组织和卵泡的完整性。主要结果测量血红素和伊红染色用于评估卵泡阶段、形态和基质细胞密度。末端脱氧核苷酸转移酶dUTP缺口标记染色用于检查细胞凋亡，马森氏三色染色用于评估基质环境中的胶原蛋白含量。结果无论之前是否进行过冷冻保存，与新鲜组织相比，卵巢组织培养后原始卵泡的比例下降，初级卵泡的比例上升，这表明卵泡的活化并没有受到冷冻保存的负面影响。然而，与未经培养的新鲜组织相比，培养和冷冻后再培养都会降低正常前胚乳卵泡的比例。培养和/或冷冻对基质细胞数量没有影响，但与未培养的组织相比，玻璃化后培养的组织细胞凋亡率增加。无论冷冻保存与否，组织培养都会导致胶原沉积减少。与所有其他处理方法相比，玻璃化和解冻组织中表达 CX37 的卵泡较少。卵巢组织的冷冻保存和/或培养不会改变含有 Ki67 阳性颗粒细胞的卵泡的百分比，也不会改变这些卵泡中 Ki67 阳性颗粒细胞的百分比。缓慢冷冻优于玻璃化冷冻，这表现在具有正常形态的卵泡比例更高、基质细胞凋亡率更低、解冻后和培养后 CX37 表达保持不变。

{"title":"Assessment of ovarian tissue and follicular integrity after cryopreservation via slow freezing or vitrification followed by in vitro culture","authors":"Juliana I. Candelaria M.S., Anna C. Denicol D.V.M., Ph.D.","doi":"10.1016/j.xfss.2023.10.004","DOIUrl":"10.1016/j.xfss.2023.10.004","url":null,"abstract":"<div><h3>Objective</h3><p>To evaluate ovarian tissue and follicle integrity before and after slow freezing or vitrification and postthawing in vitro culture.</p></div><div><h3>Design</h3><p>A laboratory study using bovine ovarian cortical tissue.</p></div><div><h3>Setting</h3><p>Academic laboratory.</p></div><div><h3>Animals</h3><p>Ovaries from healthy cattle.</p></div><div><h3>Interventions</h3><p>Bovine ovarian cortical tissue was subjected to either slow freezing or vitrification and subsequent in vitro culture. Tissue and follicle integrity were assessed before and after cryopreservation and culture.</p></div><div><h3>Main Outcome Measures</h3><p>Hematoxylin and eosin staining was used to assess follicle stages, morphology, and stromal cell density. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining was used to examine apoptosis, and Masson’s trichrome staining was used to evaluate collagen content in the stromal environment. Immunofluorescent labeling was used to localize and quantify connexin 37 (CX37) and Ki67 expression.</p></div><div><h3>Results</h3><p>Regardless of previous cryopreservation, ovarian tissue culture resulted in a decreased percentage of primordial follicles and an increased percentage of primary follicles compared with fresh tissue, indicating that follicle activation was not negatively affected by cryopreservation. However, both culture and cryopreservation followed by culture decreased the percentage of normal preantral follicles compared with fresh tissue that had not been cultured. Culture and/or cryopreservation did not impact stromal cell number, but there was increased cell apoptosis in tissue that was cultured after vitrification compared with tissue that was not cultured. Tissue culture, regardless of cryopreservation, resulted in decreased collagen deposition. There were fewer follicles expressing CX37 in vitrified and thawed tissue compared with all other treatments. Cryopreservation and/or culture of ovarian tissue did not change the percentage of follicles that contained Ki67-positive granulosa cells or the percentage of Ki67-positive granulosa cells within those follicles.</p></div><div><h3>Conclusion</h3><p>Based on these data, we conclude that tissue cryopreservation followed by culture does not affect follicle activation and growth, but it decreases the proportion of viable follicles within the tissue. Slow freezing was superior to vitrification as indicated by a higher proportion of follicles with normal morphology, lower stromal cell apoptosis, and maintenance of CX37 expression postthawing and after culture.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666335X23000587/pdfft?md5=b50d78d37663973eb5ae0caaf11c031b&pid=1-s2.0-S2666335X23000587-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136152554","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}

引用次数: 0

Urine microbes and predictive metagenomic profiles associate with abnormalities in sperm parameters: implications for male subfertility 尿液微生物和预测性元基因组图谱与精子参数异常有关：对男性不育症的影响。

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2024.01.002

Vadim Osadchiy M.D. , Andre Belarmino M.D. , Reza Kianian M.D. , John T. Sigalos M.D. , Thiago P. Furtado M.D. , Jacob S. Ancira B.S. , Trisha Kanie A.S. , Sarah F. Mangum M.S. , Craig D. Tipton Ph.D. , Tung-Chin M. Hsieh M.D. , Jesse N. Mills M.D. , Sriram V. Eleswarapu M.D., Ph.D.

Objective

To explore the taxonomic and predicted functional relationship between the urine microbiome and alterations of semen analysis (SA) parameters.

Design

Cross-sectional study.

Setting

Academic medical center.

Patient(s)

Men presenting for fertility evaluation or men presenting for vasectomy consultation with proven biological paternity were recruited and stratified on the basis of alterations, or lack thereof, in SA parameters.

Main Outcome Measure

Changes in the functional and taxonomic urine microbiome profiles of participants with or without alterations in SA parameters.

Results

Seventy-three participants were included in our study. Men with abnormal sperm motility (N = 27) showed a nearly 50-fold higher abundance of Dialister micraerophilus compared with those with normal sperm motility (N = 46). This relationship persisted on canonical correlational analysis (r = 0.439). Men with abnormal sperm concentration (N = 20) showed a lower abundance of Enterococcus faecalis and Staphylococcus aureus, compared with those with normal sperm concentration (N = 53). The urine of participants with impaired sperm motility demonstrated dramatic differences in predictive functional profiles in pathways involved in oxidation–reduction balance and cell longevity.

Conclusions

Our findings underscore differences in the urinary microbiome and abnormalities in semen parameters, especially sperm motility. By incorporating predictive functional profiling, we also highlight possible mechanisms that may drive the observed differences in sperm parameters.

目的：探讨尿液微生物组与精液分析参数变化之间的分类和预测功能关系：探索尿液微生物组与精液分析（SA）参数改变之间的分类和预测功能关系：横断面研究：研究对象招募前来进行生育力评估的男性或前来进行输精管切除术咨询并已证明生物学父子关系的男性，并根据精液分析参数的改变或缺乏改变进行分层：主要结果测量：尿液微生物组参数发生或未发生变化的参与者尿液微生物组功能和分类概况的变化：我们的研究共纳入了 73 名参与者。与精子活力正常的男性（N=46）相比，精子活力异常的男性（N=27）的嗜酸性粒细胞（Dialister micraerophilus）含量高出近50倍（P=0.032）。这种关系在典型相关分析中依然存在（r=0.439，p=0.014）。精子浓度异常的男性（20 人）与精子浓度正常的男性（53 人）相比，粪肠球菌（p=0.012）和金黄色葡萄球菌（p=0.037）的含量较低。精子活力受损者的尿液在涉及氧化还原平衡和细胞寿命的通路的预测功能特征方面存在巨大差异（所有 p 结论：我们的研究结果强调了尿液微生物组的差异和精液参数的异常，尤其是精子活力。通过预测性功能图谱分析，我们还强调了精子参数差异的可能驱动机制。

{"title":"Urine microbes and predictive metagenomic profiles associate with abnormalities in sperm parameters: implications for male subfertility","authors":"Vadim Osadchiy M.D. , Andre Belarmino M.D. , Reza Kianian M.D. , John T. Sigalos M.D. , Thiago P. Furtado M.D. , Jacob S. Ancira B.S. , Trisha Kanie A.S. , Sarah F. Mangum M.S. , Craig D. Tipton Ph.D. , Tung-Chin M. Hsieh M.D. , Jesse N. Mills M.D. , Sriram V. Eleswarapu M.D., Ph.D.","doi":"10.1016/j.xfss.2024.01.002","DOIUrl":"10.1016/j.xfss.2024.01.002","url":null,"abstract":"<div><h3>Objective</h3><p>To explore the taxonomic and predicted functional relationship between the urine microbiome and alterations of semen analysis (SA) parameters.</p></div><div><h3>Design</h3><p>Cross-sectional study.</p></div><div><h3>Setting</h3><p>Academic medical center.</p></div><div><h3>Patient(s)</h3><p>Men presenting for fertility evaluation or men presenting for vasectomy consultation with proven biological paternity were recruited and stratified on the basis of alterations, or lack thereof, in SA parameters.</p></div><div><h3>Main Outcome Measure</h3><p>Changes in the functional and taxonomic urine microbiome profiles of participants with or without alterations in SA parameters.</p></div><div><h3>Results</h3><p>Seventy-three participants were included in our study. Men with abnormal sperm motility (N = 27) showed a nearly 50-fold higher abundance of <em>Dialister micraerophilus</em> compared with those with normal sperm motility (N = 46). This relationship persisted on canonical correlational analysis (<em>r</em> = 0.439). Men with abnormal sperm concentration (N = 20) showed a lower abundance of <em>Enterococcus faecalis</em> and <em>Staphylococcus aureus</em>, compared with those with normal sperm concentration (N = 53). The urine of participants with impaired sperm motility demonstrated dramatic differences in predictive functional profiles in pathways involved in oxidation–reduction balance and cell longevity.</p></div><div><h3>Conclusions</h3><p>Our findings underscore differences in the urinary microbiome and abnormalities in semen parameters, especially sperm motility. By incorporating predictive functional profiling, we also highlight possible mechanisms that may drive the observed differences in sperm parameters.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666335X24000132/pdfft?md5=80d6e20265146e82c2932ede911b0a42&pid=1-s2.0-S2666335X24000132-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139713487","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}

引用次数: 0

A novel sorting method for the enrichment of early human spermatocytes from clinical biopsies 从临床活检样本中富集人类早期精母细胞的新型分选方法。

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2024.02.002

Meghan Robinson B.Sc. , Kevin Zhou B.Sc. , Sonia H.Y. Kung M.Sc. , Fatih Karaoğlanoğlu M.Sc. , Andrew Golin M.D. , Armita Safa M.Sc. , Charley Cai B.Sc. , Luke Witherspoon M.D. , Faraz Hach Ph.D. , Ryan Flannigan M.D.

Objective

To determine if early spermatocytes can be enriched from a human testis biopsy using fluorescence-activated cell sorting (FACS).

Design

Potential surface markers for early spermatocytes were identified using bioinformatics analysis of single-cell RNA–sequenced human testis tissue. Testicular sperm extraction samples from three participants with normal spermatogenesis were digested into single-cell suspensions and cryopreserved. Two to four million cells were obtained from each and sorted by FACS as separate biologic replicates using antibodies for the identified surface markers. A portion from each biopsy remained unsorted to serve as controls. The sorted cells were then characterized for enrichment of early spermatocytes.

Setting

A laboratory study.

Patients

Three men with a diagnosis of obstructive azoospermia (age range, 30–40 years).

Intervention

None.

Main Outcome Measures

Sorted cells were characterized for RNA expression of markers encompassing the stages of spermatogenesis. Sorting markers were validated by their reactivity on human testis formalin-fixed paraffin-embedded tissue.

Results

Serine protease 50 (TSP50) and SWI5-dependent homologous recombination repair protein 1 were identified as potential surface proteins specific for early spermatocytes. After FACS sorting, the TSP50-sorted populations accounted for 1.6%–8.9% of total populations and exhibited the greatest average-fold increases in RNA expression for the premeiotic marker stimulated by retinoic acid (STRA8), by 23-fold. Immunohistochemistry showed the staining pattern for TSP50 to be strong in premeiotic undifferentiated embryonic cell transcription factor 1⁻/doublesex and Mab-3 related transcription factor 1⁻/STRA8⁺ spermatogonia as well as SYCP3⁺/protamine 2⁻ spermatocytes.

Conclusion

This work shows that TSP50 can be used to enrich early STRA8-expressing spermatocytes from human testicular biopsies, providing a means for targeted single-cell RNA sequencing analysis and in vitro functional interrogation of germ cells during the onset of meiosis. This could enable investigation into details of the regulatory pathways underlying this critical stage of spermatogenesis, previously difficult to enrich from whole tissue samples.

目的确定能否利用荧光激活细胞分拣技术（FACS）从人类睾丸活检组织中筛选出早期精母细胞：设计：通过对单细胞 RNA 测序（scRNAseq）人类睾丸组织进行生物信息学分析，确定早期精母细胞的潜在表面标志物。将 3 名精子发生正常的参与者的睾丸精子提取（TESE）样本消化成单细胞悬浮液并冷冻保存。从每个样本中提取 200-400 万个细胞，并使用针对已确定表面标记物的抗体进行 FACS 分选，作为单独的生物重复。每份活检样本中仍有一部分细胞未分选，作为对照。然后对分选的细胞进行表征，以富集早期精母细胞：实验室研究：干预措施：无：干预措施：无：主要结果测量：对分选的细胞进行特征描述，以确定精子发生各阶段标记物的 RNA 表达。通过对人类睾丸福尔马林固定石蜡包埋（FFPE）组织的反应验证分选标记物：结果：丝氨酸蛋白酶50（TSP50）和SWI5依赖性同源重组修复蛋白1（SFR1）被鉴定为早期精母细胞的潜在特异性表面蛋白。经 FACS 分选后，TSP50 分选群体占总群体的 1.6-8.9%，其减数分裂前标记物维甲酸刺激（STRA8）的 RNA 表达平均增加了 23 倍。免疫组化显示，TSP50在减数分裂前期未分化胚胎细胞转录因子1（UTF1）-/双性和Mab-3相关转录因子1（DMRT1）-/STRA8+精原细胞以及SYCP3+/Protamine 2（PRM2）-精母细胞中的染色模式很强：这项研究表明，TSP50可用于从人类睾丸活检组织中富集早期表达STRA8的精母细胞，为减数分裂开始阶段的定向scRNAseq分析和生殖细胞体外功能检测提供了一种方法。这将有助于研究精子发生这一关键阶段的调控通路细节，以前很难从整个组织样本中进行富集。

{"title":"A novel sorting method for the enrichment of early human spermatocytes from clinical biopsies","authors":"Meghan Robinson B.Sc. , Kevin Zhou B.Sc. , Sonia H.Y. Kung M.Sc. , Fatih Karaoğlanoğlu M.Sc. , Andrew Golin M.D. , Armita Safa M.Sc. , Charley Cai B.Sc. , Luke Witherspoon M.D. , Faraz Hach Ph.D. , Ryan Flannigan M.D.","doi":"10.1016/j.xfss.2024.02.002","DOIUrl":"10.1016/j.xfss.2024.02.002","url":null,"abstract":"<div><h3>Objective</h3><p>To determine if early spermatocytes can be enriched from a human testis biopsy using fluorescence-activated cell sorting (FACS).</p></div><div><h3>Design</h3><p>Potential surface markers for early spermatocytes were identified using bioinformatics analysis of single-cell RNA–sequenced human testis tissue. Testicular sperm extraction samples from three participants with normal spermatogenesis were digested into single-cell suspensions and cryopreserved. Two to four million cells were obtained from each and sorted by FACS as separate biologic replicates using antibodies for the identified surface markers. A portion from each biopsy remained unsorted to serve as controls. The sorted cells were then characterized for enrichment of early spermatocytes.</p></div><div><h3>Setting</h3><p>A laboratory study.</p></div><div><h3>Patients</h3><p>Three men with a diagnosis of obstructive azoospermia (age range, 30–40 years).</p></div><div><h3>Intervention</h3><p>None.</p></div><div><h3>Main Outcome Measures</h3><p>Sorted cells were characterized for RNA expression of markers encompassing the stages of spermatogenesis. Sorting markers were validated by their reactivity on human testis formalin-fixed paraffin-embedded tissue.</p></div><div><h3>Results</h3><p>Serine protease 50 (TSP50) and SWI5-dependent homologous recombination repair protein 1 were identified as potential surface proteins specific for early spermatocytes. After FACS sorting, the TSP50-sorted populations accounted for 1.6%–8.9% of total populations and exhibited the greatest average-fold increases in RNA expression for the premeiotic marker stimulated by retinoic acid (STRA8), by 23-fold. Immunohistochemistry showed the staining pattern for TSP50 to be strong in premeiotic undifferentiated embryonic cell transcription factor 1<sup>−</sup>/doublesex and Mab-3 related transcription factor 1<sup>−</sup>/STRA8<sup>+</sup> spermatogonia as well as SYCP3<sup>+</sup>/protamine 2<sup>−</sup> spermatocytes.</p></div><div><h3>Conclusion</h3><p>This work shows that TSP50 can be used to enrich early STRA8-expressing spermatocytes from human testicular biopsies, providing a means for targeted single-cell RNA sequencing analysis and in vitro functional interrogation of germ cells during the onset of meiosis. This could enable investigation into details of the regulatory pathways underlying this critical stage of spermatogenesis, previously difficult to enrich from whole tissue samples.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139900996","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}

引用次数: 0

Proteomic analysis of human sperm reveals changes in protamine 1 phosphorylation in men with infertility 人类精子的蛋白质组分析揭示了不育症男性体内原胺 1 磷酸化的变化

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2023.12.002

Samantha B. Schon M.D. , Lindsay Moritz Ph.D. , Mashiat Rabbani B.S. , Julia Meguid , Brock R. Juliano Ph.D. , Brandon T. Ruotolo Ph.D. , Kenneth Aston Ph.D. , Saher Sue Hammoud Ph.D.

Objective

To perform a comprehensive assessment of protamine (P) isoforms and modifications in human sperm with the aim of identifying how P modifications and isoforms are altered in men with reduced sperm motility and low sperm count.

Design

Cross-sectional.

Setting

Academic medical center.

Patients

A total of 18 men with prior reported pregnancy and normozoospermia (normal sperm), 14 men from couples with infertility and asthenozoospermia (reduced sperm motility), and 24 men from couples with infertility and oligoasthenoteratozoospermia (low sperm count and motility and abnormal sperm morphology).

Intervention(s)

Not applicable.

Main Outcome Measure(s)

Proteomic assessment using both top-down and bottom-up liquid chromatography mass spectrometry (MS) analysis.

Results

A total of 13 posttranslational modifications were identified on P1 and P2 using bottom-up MS, including both phosphorylation and methylation. Top-down MS revealed an unmodified and phosphorylated isoform of P1 and the 3 major isoforms of P2, HP2, HP3, and HP4. Protamine 1 phosphorylation was overall higher in men with male factor infertility compared with those with normal semen analysis (40.5% vs. 32.6). There was no difference in P posttranslational modifications or isoforms of P2 in men with normal vs. abnormal fertility.

Conclusion

Human protamines bear a number of posttranslational modifications, with alterations in P1 phosphorylation noted in the setting of male factor infertility.

目的对人类精子中的质胺（P）同工酶和修饰进行全面评估，以确定精子活力降低和精子数量少的男性精子中的质胺修饰和同工酶是如何改变的。患者共有 18 名曾报告怀孕且患有正常精子症（正常精子）的男性、14 名不育夫妇中患有无精子症（精子活力降低）的男性以及 24 名不育夫妇中患有少精子症（精子数量少、活力低且精子形态异常）的男性。干预措施：不适用。主要结果测量指标：采用自上而下和自下而上的液相色谱质谱分析进行蛋白质组学评估。结果：采用自下而上的质谱分析在 P1 和 P2 上共发现 13 种翻译后修饰，包括磷酸化和甲基化。自上而下质谱分析发现了 P1 的未修饰和磷酸化同工型以及 P2 的 3 种主要同工型：HP2、HP3 和 HP4。与精液分析正常的男性相比，患有男性因素不育症的男性的原胺1磷酸化率总体较高（40.5% 对 32.6%）。结论人类原胺具有多种翻译后修饰，在男性因素不育的情况下，P1 磷酸化会发生改变。

{"title":"Proteomic analysis of human sperm reveals changes in protamine 1 phosphorylation in men with infertility","authors":"Samantha B. Schon M.D. , Lindsay Moritz Ph.D. , Mashiat Rabbani B.S. , Julia Meguid , Brock R. Juliano Ph.D. , Brandon T. Ruotolo Ph.D. , Kenneth Aston Ph.D. , Saher Sue Hammoud Ph.D.","doi":"10.1016/j.xfss.2023.12.002","DOIUrl":"10.1016/j.xfss.2023.12.002","url":null,"abstract":"<div><h3>Objective</h3><p><span>To perform a comprehensive assessment of protamine (P) isoforms and modifications in human sperm with the aim of identifying how P modifications and isoforms are altered in men with reduced </span>sperm motility and low sperm count.</p></div><div><h3>Design</h3><p>Cross-sectional.</p></div><div><h3>Setting</h3><p>Academic medical center.</p></div><div><h3>Patients</h3><p>A total of 18 men with prior reported pregnancy and normozoospermia (normal sperm), 14 men from couples with infertility and asthenozoospermia (reduced sperm motility), and 24 men from couples with infertility and oligoasthenoteratozoospermia (low sperm count and motility and abnormal sperm morphology).</p></div><div><h3>Intervention(s)</h3><p>Not applicable.</p></div><div><h3>Main Outcome Measure(s)</h3><p><span>Proteomic assessment using both top-down and bottom-up </span>liquid chromatography mass spectrometry (MS) analysis.</p></div><div><h3>Results</h3><p><span>A total of 13 posttranslational modifications were identified on P1 and P2 using bottom-up MS, including both phosphorylation and </span>methylation<span>. Top-down MS revealed an unmodified and phosphorylated isoform of P1 and the 3 major isoforms of P2, HP2, HP3, and HP4. Protamine 1 phosphorylation was overall higher in men with male factor infertility compared with those with normal semen analysis (40.5% vs. 32.6). There was no difference in P posttranslational modifications or isoforms of P2 in men with normal vs. abnormal fertility.</span></p></div><div><h3>Conclusion</h3><p>Human protamines bear a number of posttranslational modifications, with alterations in P1 phosphorylation noted in the setting of male factor infertility.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138607666","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}

引用次数: 0

Comparison of salpingectomy and tubal detorsion procedures after experimental ischemia-reperfusion injury in a rat fallopian tube model: biochemical and histopathological evaluation 大鼠输卵管模型实验性缺血再灌注损伤后输卵管切除术和输卵管剥离术的比较；生化和组织病理学评估。

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2024.03.003

Gizem Nur Koyan Karadeniz M.D. , Ozan Karadeniz M.D. , Eralp Bulutlar M.D. , Bugra Yilmaz M.D. , Asuman Gedikbasi M.D. , Hilal Serap Arslan M.D. , Berna Aslan Cetin M.D. , İbrahim Polat M.D.

Objective

To compare salpingectomy and detorsion procedures and investigate the biochemical and histopathological changes in the fallopian tubes in the experimentally isolated fallopian tube torsion model in rats.

Design

Experimental study.

Setting

Experimental surgery laboratory in a training and research hospital.

Animal(s)

Twenty-seven Sprague-Dawley rats in the reproductive period.

Intervention(s)

Group 1, control group (n = 6); group 2, bilateral total salpingectomy group after 4 hours of tubal ischemia (n = 7); group 3: 4 hours of bilateral tubal ischemia plus 1 week of reperfusion (n = 7); and group 4, 4-hour period of bilateral tubal ischemia plus 30 days of reperfusion (n = 7). A 22-gauge catheter was administered before and after surgery using methylene blue through the uterine horn of the rat to evaluate tubal patency.

Main Outcome Measure(s)

Preoperative and postoperative serum antimüllerian hormone (AMH) levels, histopathological examination of the rat tuba uterine and histopathological damage scores, antioxidant compounds (superoxide dismutase [SOD], catalase, and glutathione peroxidase [GSH-Px]), and oxidative stress end product levels (malondialdehyde [MDA] and 8-hydroxy-2′-deoxyguanosine [8-OHdG]).

Result(s)

Although a significant difference was observed in the tissue SOD, GSH-Px, MDA, and 8-OHdG values, no significant difference was observed between the groups in serum samples. The tissue SOD and tissue GSH-Px levels in group 2 significantly decreased, and a significant increase was observed in the tissue MDA and 8-OHdG values in group 2. Among the histopathological parameters, epithelial changes, vascular congestion, and the total fallopian tube mean damage score of 4 showed a significant decrease in group 4. When the methylene blue transitions before and after ischemia-reperfusion injury were compared, the values of the methylene blue transition after ischemia-reperfusion injury in groups 2–4 significantly decreased. When the serum AMH levels were analyzed, the postoperative AMH value in group 2 significantly increased.

Conclusion(s)

This study reveals that biochemical and histopathological improvement is observed in the fallopian tube tissues gradually when the detorsion procedure is performed for the necrotized tubal tissue instead of salpingectomy. Although there is restoration of epithelial integrity after reperfusion, tubal passage remains absent.

Clinical Trial Registration Number

This study was approved by the Local Ethics Committee for Animal Experiments of the Health Sciences University, Istanbul Hamidiye Medicine Faculty (approval number 27.05.2022-9269). The study followed the ethics standards recommended by the Declaration of Helsinki.

目的比较输卵管切除术和输卵管剥离术，研究实验性分离输卵管扭转模型大鼠输卵管的生化和组织病理学变化。干预措施第1组，对照组（n = 6）；第2组，输卵管缺血4小时后双侧输卵管全切除组（n = 7）；第3组：双侧输卵管缺血4小时加再灌注1周（n = 7）；第4组：双侧输卵管缺血4小时加再灌注30天（n = 7）。手术前后使用亚甲蓝通过大鼠子宫角插入 22 号导管，以评估输卵管的通畅性。主要结果指标：术前和术后血清抗苗勒氏管激素（AMH）水平、大鼠输卵管子宫组织病理学检查和组织病理学损伤评分、抗氧化化合物（超氧化物歧化酶[SOD]、过氧化氢酶和谷胱甘肽过氧化物酶[GSH-Px]）和氧化应激终产物水平（丙二醛[MDA]和 8-羟基-2′-脱氧鸟苷[8-OHdG]）。结果虽然在组织 SOD、GSH-Px、MDA 和 8-OHdG 值上观察到了显著差异，但在血清样本中未观察到组间的显著差异。第 2 组的组织 SOD 和组织 GSH-Px 水平明显降低，而组织 MDA 和 8-OHdG 值则明显升高。比较缺血再灌注损伤前后的亚甲蓝转阴值，2-4 组缺血再灌注损伤后的亚甲蓝转阴值明显下降。结论：本研究表明，对坏死的输卵管组织进行疏通术而非输卵管切除术，可逐渐改善输卵管组织的生化和组织病理学状况。虽然再灌注后上皮的完整性得到了恢复，但输卵管通道仍然缺失。临床试验注册号本研究获得了伊斯坦布尔哈米迪耶医学院健康科学大学当地动物实验伦理委员会的批准（批准号 27.05.2022-9269）。该研究遵循了《赫尔辛基宣言》推荐的伦理标准。

{"title":"Comparison of salpingectomy and tubal detorsion procedures after experimental ischemia-reperfusion injury in a rat fallopian tube model: biochemical and histopathological evaluation","authors":"Gizem Nur Koyan Karadeniz M.D. , Ozan Karadeniz M.D. , Eralp Bulutlar M.D. , Bugra Yilmaz M.D. , Asuman Gedikbasi M.D. , Hilal Serap Arslan M.D. , Berna Aslan Cetin M.D. , İbrahim Polat M.D.","doi":"10.1016/j.xfss.2024.03.003","DOIUrl":"10.1016/j.xfss.2024.03.003","url":null,"abstract":"<div><h3>Objective</h3><p>To compare salpingectomy and detorsion procedures and investigate the biochemical and histopathological changes in the fallopian tubes in the experimentally isolated fallopian tube torsion model in rats.</p></div><div><h3>Design</h3><p>Experimental study.</p></div><div><h3>Setting</h3><p>Experimental surgery laboratory in a training and research hospital.</p></div><div><h3>Animal(s)</h3><p>Twenty-seven Sprague-Dawley rats in the reproductive period.</p></div><div><h3>Intervention(s)</h3><p>Group 1, control group (n = 6); group 2, bilateral total salpingectomy group after 4 hours of tubal ischemia (n = 7); group 3: 4 hours of bilateral tubal ischemia plus 1 week of reperfusion (n = 7); and group 4, 4-hour period of bilateral tubal ischemia plus 30 days of reperfusion (n = 7). A 22-gauge catheter was administered before and after surgery using methylene blue through the uterine horn of the rat to evaluate tubal patency.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Preoperative and postoperative serum antimüllerian hormone (AMH) levels, histopathological examination of the rat tuba uterine and histopathological damage scores, antioxidant compounds (superoxide dismutase [SOD], catalase, and glutathione peroxidase [GSH-Px]), and oxidative stress end product levels (malondialdehyde [MDA] and 8-hydroxy-2′-deoxyguanosine [8-OHdG]).</p></div><div><h3>Result(s)</h3><p>Although a significant difference was observed in the tissue SOD, GSH-Px, MDA, and 8-OHdG values, no significant difference was observed between the groups in serum samples. The tissue SOD and tissue GSH-Px levels in group 2 significantly decreased, and a significant increase was observed in the tissue MDA and 8-OHdG values in group 2. Among the histopathological parameters, epithelial changes, vascular congestion, and the total fallopian tube mean damage score of 4 showed a significant decrease in group 4. When the methylene blue transitions before and after ischemia-reperfusion injury were compared, the values of the methylene blue transition after ischemia-reperfusion injury in groups 2–4 significantly decreased. When the serum AMH levels were analyzed, the postoperative AMH value in group 2 significantly increased.</p></div><div><h3>Conclusion(s)</h3><p>This study reveals that biochemical and histopathological improvement is observed in the fallopian tube tissues gradually when the detorsion procedure is performed for the necrotized tubal tissue instead of salpingectomy. Although there is restoration of epithelial integrity after reperfusion, tubal passage remains absent.</p></div><div><h3>Clinical Trial Registration Number</h3><p>This study was approved by the Local Ethics Committee for Animal Experiments of the Health Sciences University, Istanbul Hamidiye Medicine Faculty (approval number 27.05.2022-9269). The study followed the ethics standards recommended by the Declaration of Helsinki.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140769109","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}

引用次数: 0

Effects of maternal taxane chemotherapy exposure on daughters’ ovarian reserve and fertility potential 母亲接受紫杉类药物化疗对女儿卵巢储备和生育能力的影响

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2023.10.003

Julienne Chaqour B.S. , Meghan C.H. Ozcan M.D. , Payton De La Cruz M.S. , Morgan F. Woodman-Sousa B.S. , Julia N. McAdams B.S. , Kathryn J. Grive Ph.D.

Objective

To investigate the long-term effects of in utero taxane exposure on exposed daughters’ ovarian reserve and reproductive potential.

Design

Pregnant dams were treated with a single, human-relevant animal-equivalent dose of saline, docetaxel, or paclitaxel at embryonic day 16.5. In utero-exposed daughters were aged to multiple postnatal time points for ovarian and endocrine analysis or were bred to assess fertility and fecundity. Granddaughters of treated dams were assessed also for ovarian follicle composition and atresia.

Setting

Laboratory study.

Animals

C57BL/6 mice.

Intervention(s)

In utero exposure to saline, docetaxel, or paclitaxel.

Main Outcome Measure(s)

Ovarian follicle composition, rates of follicle atresia, and rates of multioocyte follicles were analyzed in all exposure groups. Serum hormone levels and oocyte retrieval outcomes following ovarian hyperstimulation were also assessed. Finally, animals from all exposure groups were bred with the number of litters, pups per litter, live births, interlitter time interval, and age at the last litter analyzed.

Result(s)

We found that docetaxel and paclitaxel exposure in utero results in ovarian toxicity later in life, significantly affecting folliculogenesis as well as increasing the rate of follicular abnormalities, including follicle atresia and multioocyte follicles. Furthermore, viability staining indicates that the ovaries of daughters exposed to taxanes in utero demonstrate a significantly higher number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive follicles. Hormone measurements also revealed that serum follicle-stimulating hormone concentration was significantly altered in taxane-exposed daughters, with the ratio of luteinizing hormone to follicle-stimulating hormone significantly elevated, specifically after paclitaxel exposure, coincident with the inability of these animals to properly respond to ovarian stimulation. Breeding studies over the course of a year also suggest that these taxane-exposed mice are fertile, although the duration of their fertility is shortened and they produce significantly fewer litters. Finally, ovarian effects are apparent in granddaughters of mice treated with docetaxel, suggesting persistent and multigenerational effects of taxane exposure.

Conclusion(s)

Our studies demonstrate that in utero exposure to taxane-based therapy during late gestation has a significant effect on the long-term reproductive health of exposed daughters (as well as their daughters) and will be instrumental in helping clinicians better understand which chemotherapies for maternal malignancy are least detrimental to a developing fetus.

目的研究子宫内暴露于紫杉类药物对暴露女儿的卵巢储备和生殖潜能的长期影响。设计在胚胎第16.5天对怀孕母鼠进行单次人类相关动物等效剂量的生理盐水、多西他赛或紫杉醇治疗。子宫内暴露的女儿在出生后的多个时间点进行卵巢和内分泌分析，或进行繁殖以评估生育能力。干预措施子宫内暴露于生理盐水、多西他赛或紫杉醇。主要结果测量指标分析所有暴露组的卵泡组成、卵泡闭锁率和多卵泡率。此外，还评估了血清激素水平和卵巢过度刺激后的卵母细胞检索结果。最后，对所有暴露组的动物进行了繁殖，并分析了产仔数、每窝产仔数、活产仔数、产仔间隔时间和最后一窝产仔年龄。此外，存活率染色显示，在子宫内接触过紫杉类药物的女儿的卵巢中，末端脱氧核苷酸转移酶 dUTP 缺口标记阳性卵泡的数量明显增多。激素测定结果还显示，暴露于紫杉类药物的女儿的血清卵泡刺激素浓度发生了显著变化，尤其是在暴露于紫杉醇后，黄体生成素与卵泡刺激素的比率明显升高，这与这些动物无法对卵巢刺激做出适当反应的情况相吻合。为期一年的繁殖研究也表明，这些暴露于紫杉醇的小鼠具有生育能力，但生育期缩短，产仔数明显减少。最后，卵巢效应在接受多西他赛治疗的小鼠的孙女身上也很明显，这表明暴露于紫杉类药物会产生持续的多代效应。结论：我们的研究表明，在妊娠晚期子宫内暴露于紫杉类药物治疗会对暴露女儿（以及它们的女儿）的长期生殖健康产生重大影响，这将有助于临床医生更好地了解哪种母体恶性肿瘤化疗对发育中的胎儿危害最小。

{"title":"Effects of maternal taxane chemotherapy exposure on daughters’ ovarian reserve and fertility potential","authors":"Julienne Chaqour B.S. , Meghan C.H. Ozcan M.D. , Payton De La Cruz M.S. , Morgan F. Woodman-Sousa B.S. , Julia N. McAdams B.S. , Kathryn J. Grive Ph.D.","doi":"10.1016/j.xfss.2023.10.003","DOIUrl":"10.1016/j.xfss.2023.10.003","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the long-term effects of in utero taxane exposure on exposed daughters’ ovarian reserve and reproductive potential.</p></div><div><h3>Design</h3><p>Pregnant dams were treated with a single, human-relevant animal-equivalent dose of saline, docetaxel, or paclitaxel at embryonic day 16.5. In utero-exposed daughters were aged to multiple postnatal time points for ovarian and endocrine analysis or were bred to assess fertility and fecundity. Granddaughters of treated dams were assessed also for ovarian follicle composition and atresia.</p></div><div><h3>Setting</h3><p>Laboratory study.</p></div><div><h3>Animals</h3><p>C57BL/6 mice.</p></div><div><h3>Intervention(s)</h3><p>In utero exposure to saline, docetaxel, or paclitaxel.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Ovarian follicle composition, rates of follicle atresia, and rates of multioocyte follicles were analyzed in all exposure groups. Serum hormone levels and oocyte retrieval outcomes following ovarian hyperstimulation were also assessed. Finally, animals from all exposure groups were bred with the number of litters, pups per litter, live births, interlitter time interval, and age at the last litter analyzed.</p></div><div><h3>Result(s)</h3><p>We found that docetaxel and paclitaxel exposure in utero results in ovarian toxicity later in life, significantly affecting folliculogenesis as well as increasing the rate of follicular abnormalities, including follicle atresia and multioocyte follicles. Furthermore, viability staining indicates that the ovaries of daughters exposed to taxanes in utero demonstrate a significantly higher number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive follicles. Hormone measurements also revealed that serum follicle-stimulating hormone concentration was significantly altered in taxane-exposed daughters, with the ratio of luteinizing hormone to follicle-stimulating hormone significantly elevated, specifically after paclitaxel exposure, coincident with the inability of these animals to properly respond to ovarian stimulation. Breeding studies over the course of a year also suggest that these taxane-exposed mice are fertile, although the duration of their fertility is shortened and they produce significantly fewer litters. Finally, ovarian effects are apparent in granddaughters of mice treated with docetaxel, suggesting persistent and multigenerational effects of taxane exposure.</p></div><div><h3>Conclusion(s)</h3><p>Our studies demonstrate that in utero exposure to taxane-based therapy during late gestation has a significant effect on the long-term reproductive health of exposed daughters (as well as their daughters) and will be instrumental in helping clinicians better understand which chemotherapies for maternal malignancy are least detrimental to a developing fetus.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666335X23000599/pdfft?md5=9eb1c3d8c0989508e651c49d1b3d4dce&pid=1-s2.0-S2666335X23000599-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136152202","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}

引用次数: 0

Effects of melatonin and metformin on the ovaries of rats with polycystic ovary syndrome 褪黑素和二甲双胍对多囊卵巢综合征大鼠卵巢的影响

F&S science

Pub Date : 2024-05-01 DOI: 10.1016/j.xfss.2024.03.001

Leonardo Augusto Lombardi Ph.D. , Leandro Sabará Mattos M.Sc. , Ana Paula Espindula Ph.D. , Ricardo Santos Simões Ph.D. , Gisela Rodrigues da Silva Sasso Ph.D. , Manuel de Jesus Simões Ph.D. , José Maria Soares-Jr Ph.D. , Rinaldo Florencio-Silva Ph.D.

Objective

To study the combined and isolated effects of melatonin and metformin in the ovarian tissue of rats with PCOS.

Design

Experimental study using a rat model of PCOS induced by continuous light exposure.

Intervention(s)

Forty adult female rats were divided into 5 groups: physiological estrus phase (Sham); permanente estrus with PCOS induced by continuous lighting exposure for 60 consecutive days (control); with PCOS treated with melatonin; with PCOS treated with metformin; with PCOS treated with melatonin + metformin. After 60 days of treatments, all rats were killed, and ovaries were collected and processed for paraffin-embedding. Formalin-fixed paraffin-embedded sections were stained with hematoxylin and eosin or subjected to immunohistochemistry for proliferation (Ki-67) and apoptosis (cleaved caspase 3) detection markers.

Setting

Federal University of São Paulo, Brazil.

Animals

Forty adult female Wistar rats (Rattus norvegicus albinus).

Main Outcome Measure(s)

Number of corpus luteum and ovarian cysts, number of ovarian follicles (primary and antral follicles), number of interstitial cells, percentage of ovarian follicles (primary and antral follicles), and of interstitial cells immunostained to cleaved caspase-3 and Ki-67.

Results

Absence of corpus luteum, a higher number of cysts, and increased nuclear volume and area of interstitial cells, along with a decrease in primary and antral follicle numbers, were noticed in the control group compared with the Sham group. Melatonin and metformin treatments attenuated these effects, although the combined treatment did not mitigate the increased number of cysts and ovaries induced by PCOS. An increase in theca interna cell apoptosis was observed in the control group, whereas melatonina and metformin treatments reduced it significantly. A higher percentage of caspase-3-immunostained granulosa cells was noted in the Sham and all treated groups compared with the control group; no aditive effects on ovarian cell apoptosis were observed in the combined treatment. The percentage of Ki-67- immunostained granulosa cells was significantly higher in the control group compared with the Sham group. However, the combined treatment, not melatonin and metformin alone, mitigated this effect. A higher percentage of Ki-67-immunostained interstitial cells was observed in all treated groups compared with the Sham and control groups, whereas no additive effects in that immunoreactivity were observed in the combined treatment.

Conclusions

Melatonin and metformin may improve ovarian function in rats with PCOS. The combined melatonin and metformin treatment is more effective in attenuating excessive granulosa cell proliferation, but it is not more effective in improving ovarian function than these drugs applied alone in

目的研究褪黑素和二甲双胍对多囊卵巢综合征大鼠卵巢组织的联合和单独作用：将40只成年雌性大鼠分为5组：生理性发情期（Sham）；连续60天持续光照诱导的PCOS永久性发情期（Control）；褪黑素治疗PCOS；二甲双胍治疗PCOS；褪黑素+二甲双胍治疗PCOS。治疗 60 天后，对所有大鼠实施安乐死，收集卵巢并进行石蜡包埋处理。经福尔马林固定的石蜡包埋切片用苏木精/伊红染色，或用免疫组化法检测增殖（Ki-67）和凋亡（裂解-天冬酶 3）标记物：结果：与Sham组相比，对照组发现黄体缺失，囊肿数量增加，间质细胞核体积和面积增大，原发卵泡和前卵泡数量减少。褪黑素和二甲双胍治疗可减轻这些影响，但联合治疗并不能减轻多囊卵巢综合征引起的囊肿/卵巢数量的增加。对照组中观察到卵巢间质细胞凋亡增加，而褪黑素和二甲双胍治疗则显著减少了这一现象。与对照组相比，SHAM组和所有治疗组中Caspase-3免疫染色的颗粒细胞比例更高；在联合治疗中未观察到对卵巢细胞凋亡的叠加效应。与 Sham 组相比，对照组 Ki-67 免疫染色颗粒细胞的百分比明显升高。然而，联合治疗（而非单独使用褪黑素和二甲双胍）减轻了这种影响。与Sham组和对照组相比，所有治疗组均观察到较高比例的Ki-67免疫染色间质细胞，而在联合治疗中未观察到该免疫反应的叠加效应：结论：褪黑素和二甲双胍可改善多囊卵巢综合征大鼠的卵巢功能。结论：褪黑素和二甲双胍可改善多囊卵巢综合征大鼠的卵巢功能，褪黑素/二甲双胍联合治疗在抑制颗粒细胞过度增殖方面更有效，但在改善卵巢功能方面并不比单独使用这两种药物更有效。

{"title":"Effects of melatonin and metformin on the ovaries of rats with polycystic ovary syndrome","authors":"Leonardo Augusto Lombardi Ph.D. , Leandro Sabará Mattos M.Sc. , Ana Paula Espindula Ph.D. , Ricardo Santos Simões Ph.D. , Gisela Rodrigues da Silva Sasso Ph.D. , Manuel de Jesus Simões Ph.D. , José Maria Soares-Jr Ph.D. , Rinaldo Florencio-Silva Ph.D.","doi":"10.1016/j.xfss.2024.03.001","DOIUrl":"10.1016/j.xfss.2024.03.001","url":null,"abstract":"<div><h3>Objective</h3><p>To study the combined and isolated effects of melatonin and metformin in the ovarian tissue of rats with PCOS.</p></div><div><h3>Design</h3><p>Experimental study using a rat model of PCOS induced by continuous light exposure.</p></div><div><h3>Intervention(s)</h3><p>Forty adult female rats were divided into 5 groups: physiological estrus phase (Sham); permanente estrus with PCOS induced by continuous lighting exposure for 60 consecutive days (control); with PCOS treated with melatonin; with PCOS treated with metformin; with PCOS treated with melatonin + metformin. After 60 days of treatments, all rats were killed, and ovaries were collected and processed for paraffin-embedding. Formalin-fixed paraffin-embedded sections were stained with hematoxylin and eosin or subjected to immunohistochemistry for proliferation (Ki-67) and apoptosis (cleaved caspase 3) detection markers.</p></div><div><h3>Setting</h3><p>Federal University of São Paulo, Brazil.</p></div><div><h3>Animals</h3><p>Forty adult female Wistar rats (<em>Rattus norvegicus albinus</em>).</p></div><div><h3>Main Outcome Measure(s)</h3><p>Number of corpus luteum and ovarian cysts, number of ovarian follicles (primary and antral follicles), number of interstitial cells, percentage of ovarian follicles (primary and antral follicles), and of interstitial cells immunostained to cleaved caspase-3 and Ki-67.</p></div><div><h3>Results</h3><p>Absence of corpus luteum, a higher number of cysts, and increased nuclear volume and area of interstitial cells, along with a decrease in primary and antral follicle numbers, were noticed in the control group compared with the Sham group. Melatonin and metformin treatments attenuated these effects, although the combined treatment did not mitigate the increased number of cysts and ovaries induced by PCOS. An increase in theca interna cell apoptosis was observed in the control group, whereas melatonina and metformin treatments reduced it significantly. A higher percentage of caspase-3-immunostained granulosa cells was noted in the Sham and all treated groups compared with the control group; no aditive effects on ovarian cell apoptosis were observed in the combined treatment. The percentage of Ki-67- immunostained granulosa cells was significantly higher in the control group compared with the Sham group. However, the combined treatment, not melatonin and metformin alone, mitigated this effect. A higher percentage of Ki-67-immunostained interstitial cells was observed in all treated groups compared with the Sham and control groups, whereas no additive effects in that immunoreactivity were observed in the combined treatment.</p></div><div><h3>Conclusions</h3><p>Melatonin and metformin may improve ovarian function in rats with PCOS. The combined melatonin and metformin treatment is more effective in attenuating excessive granulosa cell proliferation, but it is not more effective in improving ovarian function than these drugs applied alone in ","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140133378","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}

引用次数: 0

首页上一页

1
2
3
4
5
6
7
...
10

下一页尾页