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An assessment of alterations to human sperm methylation patterns in coronavirus disease 2019 infected and healthy control males 对 COVID-19 感染者和健康对照组男性精子甲基化模式变化的评估
Pub Date : 2024-02-01 DOI: 10.1016/j.xfss.2023.12.003
Isaac Stirland B.S. , Murilo Racy Soares Ph.D. , Cristiana Libardi Miranda Furtado Ph.D. , Rosana Maria Dos Reis Ph.D. , Kenneth I. Aston Ph.D. , R. Parker Smith , Timothy G. Jenkins Ph.D.

Objective

To determine whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection affects male reproductive health, considering the many potential factors that contribute to declines in male fertility on a semiglobal scale.

Design

In total, 64 human semen samples—32 treatment and 32 control—were laboratory processed and bioinformatically analyzed to assess differences in DNA methylation patterns. Implementing multiple bioinformatic tools, the analyses conducted will elicit between-group differences with respect to epigenetic age, epigenetic instability, semiglobal, and regional methylation, in addition to methylation patterns as a function of time since infection.

Setting

University hospital.

Patients

The study cohort of 64 individuals was drawn from a larger population of 94 volunteer participants recruited at the Human Reproduction Center at the Clinical Hospital of the Ribeirao Preto Medical School—University of São Paulo between June 2021 and January 2022 as well as in accordance with the ethical guidelines established by the Declaration of Helsinki.

Intervention

Exposure to SARS-CoV-2.

Main Outcome Measure(s)

Effects on male reproductive health were reported as differences in DNA methylation measured using an array. Mean β values at key regulatory loci for human spermatocytes were analyzed and compared between groups. Further analysis of β values using epigenetic age, instability, semiglobal, and regional methylation tools provided an analysis with substantial breadth and depth.

Results

In all analyses, there were no differences between groups. Considering these results, it can be inferred that infection with SARS-CoV-2 does not alter the epigenome of human spermatocytes in significant and/or persistent ways. Tangentially, these data also suggest that human male reproductive health is minimally altered by the virus, or that it is altered in a way that is independent of epigenetic programming.

Conclusion

Infection with SARS-CoV-2 has been reportedly associated with alterations in male fertility. This study asserts that such alterations do not have an epigenetic basis but are likely a result of concomitant symptomatology, i.e., fever and inflammation. Across the multiple bioinformatic analyses conducted, the results of this test did not detect any differences in DNA methylation patterns between coronavirus disease 2019 and noncoronavirus disease semen donor groups.

目的 考虑到导致半全球范围内男性生育能力下降的许多潜在因素,确定严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)感染是否会影响男性生殖健康。设计共对 64 份人类精液样本(32 份治疗样本和 32 份对照样本)进行了实验室处理和生物信息学分析,以评估 DNA 甲基化模式的差异。利用多种生物信息学工具进行分析,除了甲基化模式与感染后时间的关系外,还将得出组间在表观遗传年龄、表观遗传不稳定性、半全局和区域甲基化方面的差异。患者64人的研究队列是从圣保罗大学里贝拉普雷图医学院临床医院人类生殖中心于2021年6月至2022年1月期间招募的94名志愿者中抽取出来的。对人类精母细胞关键调控位点的平均β值进行分析,并进行组间比较。使用表观遗传学年龄、不稳定性、半全局性和区域甲基化工具对β值进行进一步分析,提供了具有广度和深度的分析结果。考虑到这些结果,可以推断感染 SARS-CoV-2 并不会显著和/或持续改变人类精母细胞的表观遗传组。从另一个角度看,这些数据也表明,病毒对人类男性生殖健康的改变微乎其微,或者说这种改变与表观遗传学程序无关。本研究认为,这种改变并不具有表观遗传学基础,而很可能是伴随症状(即发烧和炎症)的结果。在进行的多项生物信息学分析中,该检测结果未发现2019年冠状病毒病与非冠状病毒病精液供体组之间的DNA甲基化模式存在任何差异。
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引用次数: 0
Role of tenascin C in lesion formation in early peritoneal endometriosis tenascin-C在早期腹膜子宫内膜异位症病灶形成中的作用。
Pub Date : 2024-02-01 DOI: 10.1016/j.xfss.2023.12.004
Maako Moriyama M.D. , Kazuomi Nakamura Ph.D. , Hiroki Nagata M.D. , Ikumi Wada M.D. , Kei Nagira M.D., Ph.D. , Yukihiro Azuma M.D., Ph.D. , Eri Sato M.D., Ph.D. , Tasuku Harada M.D., Ph.D. , Fuminori Taniguchi M.D., Ph.D.

Objective

To identify cytokines or extracellular matrix components that contribute to adhesion to, and invasion of, the peritoneum, proximal to lesions in the early phase of endometriosis.

Design

Laboratory-based study.

Setting

University Hospital and Laboratory of Animal Science.

Patients and Animals

Five women with ovarian endometrioma, 138 wild-type (WT) C57BL/6N mice, and 48 Tenascin C (Tnc) knockout (TncKO) mice.

Interventions

To establish a murine endometriosis model, 20 pieces of minced uterine tissue fragments from each horn were administered intraperitoneally to syngeneic mice. Three days later, endometriotic lesions and peritoneal tissues were collected. Separately, we transfected human peritoneal mesothelial cells (HMrSV5) or human endometrial stromal cells (hESCs) with Tnc small interfering ribonucleic acid.

Main Outcome Measures

We employed a polymerase chain reaction array to profile gene expression in the murine peritoneum, in both peritoneum distal to lesions and peritoneum surrounding lesions (PSL). The expression of upregulated genes in the PSL was verified in the peritoneal samples by real-time reverse transcription-polymerase chain reaction. TncKO mice were used to investigate the role of Tnc in the development of endometriosis. We evaluated the proliferative activity or inflammatory state of lesions by Ki67 or CD3 immunostaining. Intraperitoneal distribution of macrophages was assessed by fluorescence-activated cell sorting. Using Tnc small interfering ribonucleic acid, we examined the invasive capacity of hESCs in a coculture system with HMrSV5.

Results

Tnc gene expression was significantly higher in PSL than in peritoneum distal to lesions. The weight and number of TncKO lesions in TncKO hosts were lower than those of WT lesions in WT hosts. In contrast, the weight and number of nonattached TncKO lesions in TncKO hosts were higher than those of nonattached WT lesions in WT hosts. We observed decreased Ki67-positive cells or H-scores for CD3, a lower proportion of M1 macrophages, and a higher proportion of M2 macrophages in TncKO lesions in TncKO recipients. Silencing of Tnc expression in hESCs and HMrSV5 diminished the invasivity of hESCs.

Conclusion

Tnc may be a crucial factor in the development of early peritoneal endometriosis.

目的确定有助于子宫内膜异位症早期病变近端粘附和侵入腹膜的细胞因子或细胞外基质(ECM)成分:设计:实验室研究患者和动物:五名患有卵巢子宫内膜异位症的妇女:5名患有卵巢子宫内膜异位症的妇女、138只野生型(WT)C57BL/6N小鼠和48只Tenascin C(Tnc)基因敲除(TncKO)小鼠:干预措施:为了建立小鼠子宫内膜异位症模型,给合成小鼠腹腔注射每个子宫角的20块碎子宫组织片段。三天后,收集子宫内膜异位症病灶和腹膜组织。另外,我们用Tnc小干扰RNA(siRNA)转染人腹膜间皮细胞(HMrSV5)或人子宫内膜基质细胞(hESCs):我们采用PCR阵列分析了小鼠腹膜中的基因表达,包括病变远端腹膜(PDL)和子宫内膜异位症病变周围腹膜(PSL)。通过实时 RT-PCR 在腹膜样本中验证了 PSL 中上调基因的表达。我们利用 TncKO 小鼠来研究 Tnc 在子宫内膜异位症发病过程中的作用。我们通过 Ki-67 或 CD3 免疫染色法评估了病灶的增殖活性或炎症状态。通过荧光激活细胞分拣(FACS)评估了巨噬细胞在腹腔内的分布。我们使用 Tnc siRNA 检测了 hESCs 与 HMrSV5 共培养系统中的侵袭能力:结果:Tnc基因在PSL中的表达明显高于PDL。TncKO宿主中TncKO病灶的重量和数量低于WT宿主中WT病灶的重量和数量。相反,TncKO宿主体内未附着的TncKO病变的重量和数量高于WT宿主体内未附着的WT病变。我们观察到,在TncKO受体的TncKO病灶中,Ki-67阳性细胞或CD3的H-scores减少,M1巨噬细胞比例降低,M2巨噬细胞比例升高。抑制Tnc在hESCs和HMrSV5中的表达可降低hESCs的侵袭性:结论:Tnc可能是早期腹膜子宫内膜异位症发生的关键因素。
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引用次数: 0
Oocyte-derived growth differentiation factor 9 suppresses the expression of CYP17A1 and androgen production in human theca cells 源于卵母细胞的生长分化因子 9 可抑制 CYP17A1 的表达和人类乳头状瘤细胞中雄激素的产生
Pub Date : 2024-02-01 DOI: 10.1016/j.xfss.2023.10.005
Xi Guo Ph.D. , Yiping Zhong M.Phil. , Yang Liu M.Phil. , Rihan Wu M.Phil. , Ling Huang Ph.D. , Chuan Huang M.Phil. , Minghui Chen Ph.D.

Objective

To investigate the direct effect of growth differentiation factor 9 (GDF9) on androgen production in human theca cells.

Design

Experimental study.

Setting

Tertiary hospital-based research laboratory.

Patient(s)

Women who underwent in vitro fertilization and intracytoplasmic sperm injections at our clinic were included in this study.

Intervention(s)

Primary cultured human theca cells from women undergoing in vitro fertilization and intracytoplasmic sperm injection treatment were treated with GDF9, an activin receptor-like kinase 5 (ALK5) inhibitor, and a SMAD4 agonist.

Main Outcome Measure(s)

The expression of androgen synthesis-related genes StAR, CYP17A1, and LHCGR, levels of androstenedione and testosterone, phosphorylation of SMAD2/3, and the interaction between bone morphogenic protein-activated type II receptor and ALK5 were evaluated using reverse transcription-quantitative polymerase chain reaction, Western blot, enzyme-linked immunosorbent assays, and coimmunoprecipitation assays, respectively.

Result(s)

Growth differentiation factor 9 decreased StAR, CYP17A1, and LHCGR expression levels in human theca cells, which was prevented by treatment with the ALK5 inhibitor, and suppressed production of androgen in human theca cells. Growth differentiation factor 9 increased SMAD2/3 phosphorylation, and the ALK5 inhibitor also suppressed this effect. Bone morphogenic protein-activated type II receptor and ALK5 bound to each other after GDF9 stimulation. The SMAD4 agonist kartogenin also decreased messenger RNA levels of StAR and CYP17A1 and protein levels of StAR in human theca cells.

Conclusion(s)

Growth differentiation factor 9 can activate the bone morphogenic protein-activated type II receptor-ALK5-SMAD2/3 signaling pathway, suppress CYP17A1 expression, and decrease androgen production in human theca cells.

目的 探讨生长分化因子9(GDF9)对人类绒毛膜细胞雄激素生成的直接影响.设计实验研究.设置以三级医院为基础的研究实验室.患者包括在本诊所接受体外受精和卵胞浆内单精子注射的女性.干预来自接受体外受精和卵胞浆内单精子注射治疗的女性的原代培养人类绒毛膜细胞.用GDF9和活化因子受体样激酶5(activin receptor-like kinase 5,GDF9)处理绒毛膜细胞.干预措施:用GDF9、活化素受体样激酶5(ALK5)抑制剂和SMAD4激动剂处理接受体外受精和胞浆内精子注射治疗的妇女的原代培养人theca细胞。主要结果指标)采用逆转录-定量聚合酶链反应、Western印迹、酶联免疫吸附试验和共沉淀试验分别评估雄激素合成相关基因StAR、CYP17A1和LHCGR的表达、雄烯二酮和睾酮的水平、SMAD2/3的磷酸化以及骨形态发生蛋白激活的II型受体与ALK5之间的相互作用。结果 生长分化因子9降低了人乳头状瘤细胞中StAR、CYP17A1和LHCGR的表达水平,而ALK5抑制剂可阻止这种降低,并抑制了人乳头状瘤细胞中雄激素的产生。生长分化因子 9 增加了 SMAD2/3 磷酸化,ALK5 抑制剂也抑制了这种效应。骨形态发生蛋白激活的II型受体和ALK5在GDF9刺激后相互结合。结论:生长分化因子9能激活骨形态发生蛋白激活的II型受体-ALK5-SMAD2/3信号通路,抑制CYP17A1的表达,并减少人乳头状瘤细胞中雄激素的产生。
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引用次数: 0
Embryo ranking agreement between embryologists and artificial intelligence algorithms 胚胎学家和人工智能算法之间的胚胎排名协议。
Pub Date : 2024-02-01 DOI: 10.1016/j.xfss.2023.10.002
Nikica Zaninovic M.S., Ph.D. , Jose T. Sierra Ph.D. , Jonas E. Malmsten D.P.S. , Zev Rosenwaks M.D.

Objective

To evaluate the degree of agreement of embryo ranking between embryologists and eight artificial intelligence (AI) algorithms.

Design

Retrospective study.

Patient(s)

A total of 100 cycles with at least eight embryos were selected from the Weill Cornell Medicine database. For each embryo, the full-length time-lapse (TL) videos, as well as a single embryo image at 120 hours, were given to five embryologists and eight AI algorithms for ranking.

Intervention(s)

None.

Main Outcome Measure(s)

Kendall rank correlation coefficient (Kendall’s τ).

Result(s)

Embryologists had a high degree of agreement in the overall ranking of 100 cycles with an average Kendall’s tau (K-τ) of 0.70, slightly lower than the interembryologist agreement when using a single image or video (average K-τ = 0.78). Overall agreement between embryologists and the AI algorithms was significantly lower (average K-τ = 0.53) and similar to the observed low inter-AI algorithm agreement (average K-τ = 0.47). Notably, two of the eight algorithms had a very low agreement with other ranking methodologies (average K-τ = 0.05) and between each other (K-τ = 0.01). The average agreement in selecting the best-quality embryo (1/8 in 100 cycles with an expected agreement by random chance of 12.5%; confidence interval [CI]95: 6%–19%) was 59.5% among embryologists and 40.3% for six AI algorithms. The incidence of the agreement for the two algorithms with the low overall agreement was 11.7%. Agreement on selecting the same top two embryos/cycle (expected agreement by random chance corresponds to 25.0%; CI95: 17%–32%) was 73.5% among embryologists and 56.0% among AI methods excluding two discordant algorithms, which had an average agreement of 24.4%, the expected range of agreement by random chance. Intraembryologist ranking agreement (single image vs. video) was 71.7% and 77.8% for single and top two embryos, respectively. Analysis of average raw scores indicated that cycles with low diversity of embryo quality generally resulted in a lower overall agreement between the methods (embryologists and AI models).

Conclusion(s)

To our knowledge, this is the first study that evaluates the level of agreement in ranking embryo quality between different AI algorithms and embryologists. The different concordance methods were consistent and indicated that the highest agreement was intraembryologist agreement, followed by interembryologist agreement. In contrast, the agreement between some of the AI algorithms and embryologists was similar to the inter-AI algorithm agreement, which also showed a wide range of pairwise concordance. Specifically, two AI models showed intra- and interagreement at the level expected from random selection.

目的:评价胚胎学家与8种人工智能算法在胚胎排序上的一致性。设计:回顾性研究患者:从威尔康奈尔医学数据库中选择总共100个周期,至少有8个胚胎。对于每个胚胎,将全长延时(TL)视频以及120小时的单个胚胎图像交给五名胚胎学家和八种人工智能算法进行排名。干预措施:无主要结果测量(S):肯德尔秩相关系数(Kendall’Sτ)结果:胚胎学家在100个周期的总体排名中高度一致,平均肯德尔τ(K-τ)为0.70,略低于使用单个图像或视频时胚胎学家之间的一致性(平均K-τ=0.78)。胚胎学家和人工智能算法之间的总体一致性显著较低(平均K-τ=0.53),与观察到的人工智能算法间的低一致性(平均K-τ=0.47)相似。值得注意的是,八种算法中有两种算法与其他排序方法的一致性很低(平均K-τ=0.05),彼此之间的一致性也很低(K-τ=0.01)。胚胎学家在选择最优质胚胎方面的平均一致性(100个周期中的1/8,随机概率为12.5%;CI95:6-19%)为59.5%,六种AI算法为40.3%。总体一致性较低的两种算法的一致性发生率为11.7%。胚胎学家对选择相同的前两个胚胎/周期的一致性(随机机会的预期一致性对应于25.0%;CI95:17-32%)为73.5%,人工智能方法中为56.0%,排除了两种不一致的算法,平均一致性为24.4%,通过随机机会达成一致的预期范围。胚胎学家对单个和前两个胚胎的排名一致性(单个图像与视频)分别为71.7%和77.8%。对平均原始分数的分析表明,胚胎质量多样性较低的周期通常会导致方法(胚胎学家和人工智能模型)之间的总体一致性较低。结论:据我们所知,这是第一项评估不同人工智能算法和胚胎学家在胚胎质量排序方面一致性水平的研究。不同的一致性方法是一致的,表明最高的一致性是胚胎内一致性,其次是胚胎间一致性。相比之下,一些人工智能算法和胚胎学家之间的一致性类似于人工智能算法间的一致性,也显示出广泛的成对一致性。具体而言,两个人工智能模型在随机选择的预期水平上显示出内部和内部一致性。
{"title":"Embryo ranking agreement between embryologists and artificial intelligence algorithms","authors":"Nikica Zaninovic M.S., Ph.D. ,&nbsp;Jose T. Sierra Ph.D. ,&nbsp;Jonas E. Malmsten D.P.S. ,&nbsp;Zev Rosenwaks M.D.","doi":"10.1016/j.xfss.2023.10.002","DOIUrl":"10.1016/j.xfss.2023.10.002","url":null,"abstract":"<div><h3>Objective</h3><p>To evaluate the degree of agreement of embryo ranking between embryologists and eight artificial intelligence (AI) algorithms.</p></div><div><h3>Design</h3><p>Retrospective study.</p></div><div><h3>Patient(s)</h3><p>A total of 100 cycles with at least eight embryos were selected from the Weill Cornell Medicine database. For each embryo, the full-length time-lapse (TL) videos, as well as a single embryo image at 120 hours, were given to five embryologists and eight AI algorithms for ranking.</p></div><div><h3>Intervention(s)</h3><p>None.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Kendall rank correlation coefficient (Kendall’s τ).</p></div><div><h3>Result(s)</h3><p>Embryologists had a high degree of agreement in the overall ranking of 100 cycles with an average Kendall’s tau (K-τ) of 0.70, slightly lower than the interembryologist agreement when using a single image or video (average K-τ = 0.78). Overall agreement between embryologists and the AI algorithms was significantly lower (average K-τ = 0.53) and similar to the observed low inter-AI algorithm agreement (average K-τ = 0.47). Notably, two of the eight algorithms had a very low agreement with other ranking methodologies (average K-τ = 0.05) and between each other (K-τ = 0.01). The average agreement in selecting the best-quality embryo (1/8 in 100 cycles with an expected agreement by random chance of 12.5%; confidence interval [CI]95: 6%–19%) was 59.5% among embryologists and 40.3% for six AI algorithms. The incidence of the agreement for the two algorithms with the low overall agreement was 11.7%. Agreement on selecting the same top two embryos/cycle (expected agreement by random chance corresponds to 25.0%; CI95: 17%–32%) was 73.5% among embryologists and 56.0% among AI methods excluding two discordant algorithms, which had an average agreement of 24.4%, the expected range of agreement by random chance. Intraembryologist ranking agreement (single image vs. video) was 71.7% and 77.8% for single and top two embryos, respectively. Analysis of average raw scores indicated that cycles with low diversity of embryo quality generally resulted in a lower overall agreement between the methods (embryologists and AI models).</p></div><div><h3>Conclusion(s)</h3><p>To our knowledge, this is the first study that evaluates the level of agreement in ranking embryo quality between different AI algorithms and embryologists. The different concordance methods were consistent and indicated that the highest agreement was intraembryologist agreement, followed by interembryologist agreement. In contrast, the agreement between some of the AI algorithms and embryologists was similar to the inter-AI algorithm agreement, which also showed a wide range of pairwise concordance. Specifically, two AI models showed intra- and interagreement at the level expected from random selection.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 50-57"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41222069","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
Pub Date : 2024-02-01 DOI: 10.1016/j.xfss.2024.01.001
William H. Catherino M.D., Ph.D.
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引用次数: 0
Simvastatin induces degradation of the extracellular matrix in human leiomyomata: novel in vitro, in vivo, and patient level evidence of matrix metalloproteinase involvement 辛伐他汀诱导人平滑肌瘤细胞外基质的降解:新的体外、体内和患者水平的证据表明基质金属蛋白酶参与。
Pub Date : 2024-02-01 DOI: 10.1016/j.xfss.2023.11.005
Joy Britten M.D. , Jaime A. Roura-Monllor M.D., M.S. , Minnie Malik Ph.D. , Sean Moran Ph.D. , Anthony DeAngelis M.D., Ph.D. , Paul Driggers Ph.D. , Sadia Afrin Ph.D. , Mostafa Borahay M.D., Ph.D. , William H. Catherino M.D., Ph.D.

Objectives

To assess the effect of simvastatin on uterine leiomyoma growth and extracellular matrix (ECM) deposition.

Design

Laboratory analysis of human leiomyoma cell culture, xenograft in a mouse model, and patient tissue from a clinical trial.

Setting

Academic research center.

Patient(s)

Tissue culture from human leiomyoma tissue and surgical leiomyoma tissue sections from a placebo-controlled randomized clinical trial.

Intervention(s)

Simvastatin treatment.

Main Outcome Measure(s)

Serum concentrations, xenograft volumes, and protein expression.

Results

Mice xenografted with 3-dimensional human leiomyoma cultures were divided as follows: 7 untreated controls; 12 treated with activated simvastatin at 10 mg/kg body weight; and 15 at 20 mg/kg body weight. Simvastatin was detected in the serum of mice injected at the highest dose. Xenograft volumes were significantly smaller (mean 53% smaller at the highest concentration). There was dissolution of compact ECM, decreased ECM formation, and lower collagen protein expression in xenografts. Membrane type 1 matrix metalloproteinase was increased in vitro and in vivo. Matrix metalloproteinase 2 and low-density lipoprotein receptor-related protein 1 were increased in vitro.

Conclusions

Simvastatin exhibited antitumoral activity with ECM degradation and decreased leiomyoma tumor volume in vivo. Activation of the matrix metalloproteinase 2, membrane type 1 matrix metalloproteinase, and low-density lipoprotein receptor-related protein 1 pathway may explain these findings.

目的:探讨辛伐他汀对子宫平滑肌瘤生长及细胞外基质(ECM)沉积的影响。设计:实验室分析人类平滑肌瘤细胞培养,小鼠模型中的异种移植物和临床试验中的患者组织。环境:学术研究中心。患者:取自人体平滑肌瘤组织的组织培养和取自安慰剂对照随机临床试验的手术平滑肌瘤组织切片。干预措施:辛伐他汀治疗。主要结局指标:血清浓度、异种移植物体积和蛋白表达。结果:移植三维人平滑肌瘤培养物的小鼠分为:对照组7只,10 mg/kg体重辛伐他汀活化组12只,20 mg/kg体重辛伐他汀活化组15只。在注射最高剂量小鼠血清中检测到辛伐他汀。异种移植物体积明显变小(最高浓度时平均小53%,p < 0.05)。异种移植物中致密ECM溶解,ECM形成减少,胶原蛋白表达降低。膜1型基质金属蛋白酶(MT1-MMP)在体内外均升高。基质金属蛋白酶2 (MMP2)和低密度脂蛋白受体相关蛋白1 (LRP1)升高。结论:辛伐他汀在体内表现出抗肿瘤活性,细胞外基质降解并减少平滑肌瘤肿瘤体积。MMP2/MT1-MMP/LRP1通路的激活可以解释这些发现。
{"title":"Simvastatin induces degradation of the extracellular matrix in human leiomyomata: novel in vitro, in vivo, and patient level evidence of matrix metalloproteinase involvement","authors":"Joy Britten M.D. ,&nbsp;Jaime A. Roura-Monllor M.D., M.S. ,&nbsp;Minnie Malik Ph.D. ,&nbsp;Sean Moran Ph.D. ,&nbsp;Anthony DeAngelis M.D., Ph.D. ,&nbsp;Paul Driggers Ph.D. ,&nbsp;Sadia Afrin Ph.D. ,&nbsp;Mostafa Borahay M.D., Ph.D. ,&nbsp;William H. Catherino M.D., Ph.D.","doi":"10.1016/j.xfss.2023.11.005","DOIUrl":"10.1016/j.xfss.2023.11.005","url":null,"abstract":"<div><h3>Objectives</h3><p>To assess the effect of simvastatin<span> on uterine leiomyoma<span> growth and extracellular matrix (ECM) deposition.</span></span></p></div><div><h3>Design</h3><p><span>Laboratory analysis of human leiomyoma<span> cell culture, xenograft in a mouse model, and patient tissue from a </span></span>clinical trial.</p></div><div><h3>Setting</h3><p>Academic research center.</p></div><div><h3>Patient(s)</h3><p>Tissue culture from human leiomyoma tissue and surgical leiomyoma tissue sections from a placebo-controlled randomized clinical trial.</p></div><div><h3>Intervention(s)</h3><p>Simvastatin treatment.</p></div><div><h3>Main Outcome Measure(s)</h3><p>Serum concentrations, xenograft volumes, and protein expression.</p></div><div><h3>Results</h3><p><span>Mice xenografted with 3-dimensional human leiomyoma cultures were divided as follows: 7 untreated controls; 12 treated with activated simvastatin at 10 mg/kg body weight; and 15 at 20 mg/kg body weight. Simvastatin was detected in the serum of mice injected at the highest dose. Xenograft volumes were significantly smaller (mean 53% smaller at the highest concentration). There was dissolution of compact ECM, decreased ECM formation, and lower collagen protein expression in xenografts. </span>Membrane type 1 matrix metalloproteinase<span> was increased in vitro and in vivo. Matrix metalloproteinase 2 and low-density lipoprotein receptor-related protein 1 were increased in vitro.</span></p></div><div><h3>Conclusions</h3><p>Simvastatin exhibited antitumoral activity with ECM degradation and decreased leiomyoma tumor volume in vivo. Activation of the matrix metalloproteinase 2, membrane type 1 matrix metalloproteinase, and low-density lipoprotein receptor-related protein 1 pathway may explain these findings.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"5 1","pages":"Pages 80-91"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138479718","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
Progesterone-primed cycles result in slower embryos without compromising implantation potential and with the advantages of oral administration and potential cost reduction 以孕酮为前体的周期会导致胚胎发育较慢,但不会影响植入潜力,而且具有口服给药和潜在成本降低的优势
Pub Date : 2024-02-01 DOI: 10.1016/j.xfss.2023.12.001
Daniela Paes de Almeida Ferreira Braga D.V.M., Ph.D. , Amanda Setti M.Sc. , Edward Carrilho M.D. , Patrícia Guilherme M.Sc. , Assumpto Iaconelli Jr. M.D. , Edson Borges Jr. M.D., Ph.D.

Objective

To study the impact of the use of progesterone on embryo morphokinetics and on the outcomes of intracytoplasmic sperm injection cycles.

Design

Cohort study.

Setting

Private university–affiliated in vitro fertilization center.

Patient(s)

This study included 236 freeze-all intracytoplasmic sperm injection cycles and the resultant 2,768 injected oocytes cultured in a time-lapse imaging incubation system. Patients were matched by age and divided into groups depending on the protocol used to prevent the luteinizing hormone surge: progestin-primed (144 cycles and 1,360 embryos) and gonadotropin hormone-releasing hormone (GnRH) antagonist (144 cycles and 1,408 embryos) groups.

Intervention(s)

The kinetic recorded markers were time to pronuclear appearance and fading, time to 2–8 cells, time to morulation, time to start of blastulation, and time to blastulation. The durations of cell cycles and time to complete synchronous divisions were calculated. The Known Implantation Data Score ranking was recorded. Morphokinetics and clinical outcomes were compared between the groups.

Main Outcome Measure(s)

Embryo morphokinetics and clinical outcomes.

Results

Slower time to pronuclear appearance, time to 2 cells, time to 7 cells, time to start of blastulation, and time to blastulation were observed in embryos derived from progestin-primed cycles than in those from the GnRH antagonist group. No significant differences were noted in any other morphokinetic milestone. Significantly higher cancellation and implantation rates were observed in the progestin-primed group. However, no significant differences were noted in the pregnancy and miscarriage rates. The expenses for treatment using premature GnRH antagonist and progestins were US$318.18 and US$11.05, respectively.

Conclusions

Exogenous progesterone replaces the GnRH antagonist for the prevention of premature luteinizing hormone surge, in freeze-all cycles, with the advantage of oral administration and potential cost reduction.

研究使用黄体酮对胚胎形态动力学和卵胞浆内单精子显微注射周期结果的影响.设计队列研究.设置私立大学附属体外受精中心.患者本研究包括 236 个冷冻-全卵胞浆内单精子显微注射周期和在延时成像培养系统中培养的 2,768 个注射卵母细胞。患者年龄相匹配,并根据防止黄体生成素激增的方案分为两组:孕激素刺激组(144 个周期和 1,360 个胚胎)和促性腺激素释放激素(GnRH)拮抗剂组(144 个周期和 1,408 个胚胎)。计算细胞周期的持续时间和完成同步分裂的时间。记录已知植入数据得分排名。主要结果指标:胚胎形态动力学和临床结果。结果与 GnRH 拮抗剂组相比,孕激素促排卵周期的胚胎出现原核的时间、发育到 2 个细胞的时间、发育到 7 个细胞的时间、开始胚泡着床的时间和胚泡着床的时间均较短。其他形态动力学指标均无明显差异。孕激素前体组的胚胎流产率和植入率明显更高。然而,在妊娠率和流产率方面没有发现明显差异。结论外源性黄体酮取代了 GnRH 拮抗剂,用于预防全冻周期中黄体生成素过早激增,其优点是口服给药,并有可能降低成本。
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引用次数: 0
The combination of natural compounds Crila and epigallocatechin gallate showed enhanced antiproliferative effects on human uterine fibroid cells compared with single treatments 与单一治疗相比,天然化合物Crila和表没食子儿茶素没食子酸盐(EGCG)的组合对人类子宫肌瘤细胞的抗增殖作用增强。
Pub Date : 2023-11-01 DOI: 10.1016/j.xfss.2023.09.004
Tao Bai Ph.D. , Mohamed Ali Ph.D. , Bernard Somers B.S. , Qiwei Yang Ph.D. , Sue McKinney J.D. , Ayman Al-Hendy M.D., Ph.D.

Objective

To investigate the combined effects of Crila and green tea extract, epigallocatechin gallate (EGCG), compared with single treatments, on human uterine fibroid cells.

Design

Human uterine leiomyoma (HuLM) cells were treated with different concentrations of Crila, alone or in combination with EGCG, and several experiments were employed.

Setting

A laboratory study.

Patientss

N/A.

Interventions

Crila, EGCG.

Main Outcome Measures

Cell proliferation assay, drug synergy using combination index, protein and gene expression analysis of proliferation marker proliferating cell nuclear antigen, and apoptosis marker BAX using western blotting and quantitative polymerase chain reaction, respectively.

Results

Results showed that tested Crila concentrations, when combined with 25 and 50 μM EGCG, exerted synergistic growth inhibitory effects on HuLM viability. This inhibitory effect on HuLM cell viability was because of decreased cell proliferation, as shown by a decrease in the proliferation marker proliferating cell nuclear antigen at messenger RNA and protein levels, rather than inducing apoptosis.

Conclusion

Our study concludes that the utility of natural compounds may provide a safe and cost-effective alternative to currently used short-term hormonal therapies against uterine fibroids.

目的:研究红樱桃和绿茶提取物表没食子儿茶素没食子酸酯(EGCG)对人子宫肌瘤细胞的联合作用,并与单次治疗进行比较。设计:用不同浓度的Crila单独或与EGCG联合处理人子宫平滑肌瘤(HuLM)细胞,并进行了几个实验。设置:实验室研究患者(S):不适用。干预措施:Crila,EGCG主要观察指标(S),细胞增殖测定,使用组合指数的药物协同作用,分别使用蛋白质印迹和qPCR对增殖标志物PCNA和凋亡标志物BAX的蛋白质和基因表达进行分析。结果:结果表明,当与25和50μM EGCG联合使用时,测试的Crila浓度对HuLM的活力具有协同生长抑制作用。这种对HuLM细胞活力的抑制作用是由于细胞增殖减少,如增殖标记物PCNA在mRNA和蛋白质水平上的减少所示,而不是诱导细胞凋亡。结论:我们的研究得出结论,天然化合物的效用可能为目前使用的针对不明飞行物的短期激素疗法提供一种安全且具有成本效益的替代方案。
{"title":"The combination of natural compounds Crila and epigallocatechin gallate showed enhanced antiproliferative effects on human uterine fibroid cells compared with single treatments","authors":"Tao Bai Ph.D. ,&nbsp;Mohamed Ali Ph.D. ,&nbsp;Bernard Somers B.S. ,&nbsp;Qiwei Yang Ph.D. ,&nbsp;Sue McKinney J.D. ,&nbsp;Ayman Al-Hendy M.D., Ph.D.","doi":"10.1016/j.xfss.2023.09.004","DOIUrl":"10.1016/j.xfss.2023.09.004","url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the combined effects of Crila and green tea extract, epigallocatechin gallate (EGCG), compared with single treatments, on human uterine fibroid cells.</p></div><div><h3>Design</h3><p>Human uterine leiomyoma (HuLM) cells were treated with different concentrations of Crila, alone or in combination with EGCG, and several experiments were employed.</p></div><div><h3>Setting</h3><p>A laboratory study.</p></div><div><h3>Patientss</h3><p>N/A.</p></div><div><h3>Interventions</h3><p>Crila, EGCG.</p></div><div><h3>Main Outcome Measures</h3><p>Cell proliferation assay, drug synergy using combination index, protein and gene expression analysis of proliferation marker proliferating cell nuclear antigen, and apoptosis marker BAX using western blotting and quantitative polymerase chain reaction, respectively.</p></div><div><h3>Results</h3><p>Results showed that tested Crila concentrations, when combined with 25 and 50 μM EGCG, exerted synergistic growth inhibitory effects on HuLM viability. This inhibitory effect on HuLM cell viability was because of decreased cell proliferation, as shown by a decrease in the proliferation marker proliferating cell nuclear antigen at messenger RNA and protein levels, rather than inducing apoptosis.</p></div><div><h3>Conclusion</h3><p>Our study concludes that the utility of natural compounds may provide a safe and cost-effective alternative to currently used short-term hormonal therapies against uterine fibroids.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":"4 4","pages":"Pages 341-349"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666335X23000526/pdfft?md5=16ad7e27860baaa5dca2d9ad106e25e0&pid=1-s2.0-S2666335X23000526-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41175902","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
The ultrastructural nature of human oocytes’ cytoplasmic abnormalities and the role of cytoskeleton dysfunction 人卵母细胞胞浆异常的超微结构性质和细胞骨架功能障碍的作用。
Pub Date : 2023-11-01 DOI: 10.1016/j.xfss.2023.09.002
Martina Tatíčková M.Sc. , Zuzana Trebichalská M.Sc. , Drahomíra Kyjovská , Pavel Otevřel M.D. , Soňa Kloudová Ph.D. , Zuzana Holubcová Ph.D.

Objective

To investigate the structural bases of human oocytes’ cytoplasmic abnormalities and the causative mechanism of their emergence. Knowledge of an abnormal oocyte’s intracellular organization is vital to establishing reliable criteria for clinical evaluation of oocyte morphology.

Design

Laboratory-based study on experimental material provided by a private assisted reproduction clinic.

Setting

University laboratory and imaging center.

Patients

A total of 105 women undergoing hormonal stimulation for in vitro fertilization (IVF) donated their spare oocytes for this study.

Interventions

Transmission electron microscopy (TEM) was used to analyze the fine morphology of 22 dysmorphic IVF oocytes exhibiting different types of cytoplasmic irregularities, namely, refractile bodies; centrally located cytoplasmic granularity (CLCG); smooth endoplasmic reticulum (SER) disc; and vacuoles. A total of 133 immature oocytes were exposed to cytoskeleton-targeting compounds or matured in control conditions, and their morphology was examined using fluorescent and electron microscopy.

Main Outcome Measures

The ultrastructural morphology of dysmorphic oocytes was analyzed. Drug-treated oocytes had their maturation efficiency, chromosome-microtubule configurations, and fine intracellular morphology examined.

Results

TEM revealed ultrastructural characteristics of common oocyte aberrations and indicated that excessive organelle clustering was the underlying cause of 2 of the studied morphotypes. Inhibition experiments showed that disruption of actin, not microtubules, allows for inordinate aggregation of subcellular structures, resembling the ultrastructural pattern seen in morphologically abnormal oocytes retrieved in IVF cycles. These results imply that actin serves as a regulator of organelle distribution during human oocyte maturation.

Conclusion

The ultrastructural analogy between dysmorphic oocytes and oocytes, in which actin network integrity was perturbed, suggests that dysfunction of the actin cytoskeleton might be implicated in generating common cytoplasmic aberrations. Knowledge of human oocytes’ inner workings and the origin of morphological abnormalities is a step forward to a more objective oocyte quality assessment in IVF practice.

目的:探讨人卵母细胞胞质异常的结构基础及其产生的机制。了解(ab)正常卵母细胞的细胞内组织对于建立可靠的卵母细胞形态学临床评估标准至关重要。设计:对私人辅助生殖诊所提供的实验材料进行实验室研究。设置:大学实验室和影像中心。患者:共有105名接受激素刺激进行体外受精(IVF)的女性为本研究捐献了备用卵母细胞。干预:使用透射电子显微镜(TEM)分析了22个表现出不同类型细胞质不规则的变形IVF卵母细胞的精细形态,即(1)折射体,(2)中央定位的细胞质颗粒(CLCG),(3)滑面内质网(SER)盘和(4)液泡。共有133个未成熟卵母细胞暴露于细胞骨架靶向化合物或在对照条件下成熟,并通过荧光和电子显微镜检查其形态。主要观察指标:分析畸形卵母细胞的超微结构形态。药物处理的卵母细胞具有成熟效率、染色体微管结构和精细的细胞内形态。结果:透射电镜显示了常见卵母细胞畸变的超微结构特征,并表明细胞器过度聚集是两种研究形态类型的根本原因。抑制实验表明,肌动蛋白而非微管的破坏导致亚细胞结构过度聚集,类似于IVF周期中获得的形态异常卵母细胞的超微结构模式。这些结果表明,肌动蛋白在人类卵母细胞成熟过程中是细胞器分布的调节因子。结论:畸形卵母细胞和肌动蛋白网络完整性受到干扰的卵母细胞之间的超微结构相似性表明,肌动蛋白细胞骨架的功能障碍可能与常见的细胞质畸变有关。了解人类卵母细胞的内部运作和形态学异常的起源,是在体外受精实践中更客观地评估卵母细胞质量的一步。
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引用次数: 0
In vitro fertilization outcomes in a mouse model of gender-affirming hormone therapy in transmasculine youth 变性青年性别确认激素治疗小鼠模型的体外受精结果。
Pub Date : 2023-11-01 DOI: 10.1016/j.xfss.2023.08.001
Cynthia Dela Cruz Ph.D. , Abigail Wandoff B.S. , Margaret Brunette M.S. , Vasantha Padmanabhan Ph.D. , Ariella Shikanov Ph.D. , Molly B. Moravek M.D., M.P.H.

Objective

To investigate in vitro fertilization (IVF) outcomes in an adolescent transmasculine mouse model mimicking gender-affirming hormone therapy in prepubertal youth, both on testosterone (T) and after T washout.

Design

Experimental laboratory study using a validated mouse model.

Setting

University-based basic science research laboratory.

Animal(s)

A total of 80 prepubertal 26-day-old C57BL/6N female mice were used in this study.

Intervention(s)

Animals (n = 10/group) were implanted subcutaneously with gonadotropin-releasing hormone agonist at 3.6 mg or received sham surgery. After 21 days, they were implanted with silastic tubing containing either T 10 mg or placebo for 6 weeks. After 6 weeks, a group of animals were superovulated for immediate IVF, and another group had the implant removed and went through superovulation for IVF after 2 weeks (washout IVF). The total number of oocytes yielded, oocyte maturity rate, fertilization rate, and numbers of 2-cell embryos, 4–8-cell embryos, morula, blastocysts, and hatching blastocysts were recorded.

Result(s)

Testosterone treatment negatively impacted IVF outcomes in animals stimulated when receiving T, but not after T washout. Pretreatment with gonadotropin-releasing hormone agonist did not affect IVF outcomes.

Conclusion(s)

Although current T had a negative impact on IVF outcomes compared with controls, animals were still able to produce viable oocytes for fertilization and develop into blastocysts. Future efforts to study the impact of long-term T exposure on oocyte quality, especially aneuploidy rates, pregnancy outcomes, and live birth rates, are necessary.

目的:研究青春期变性小鼠模型的体外受精(IVF)结果,该模型模拟青春期前青年的性别肯定激素治疗,包括睾酮(T)和T清除后。设计:使用经验证的小鼠模型进行实验性实验室研究。设置:大学基础科学研究实验室。动物:本研究共使用80只26日龄的青春期前C57BL/6N雌性小鼠。干预:动物(n=10/组)皮下植入3.6 mg促性腺激素释放激素激动剂或接受假手术。21天后,他们被植入含有T10 mg T或安慰剂的硅胶管6周。6周后,一组动物进行超排,立即进行试管婴儿,另一组动物移除植入物,并在2周后进行超排试管婴儿(冲洗试管婴儿)。记录产生的卵母细胞总数、卵母细胞成熟率、受精率以及2细胞胚胎、4-8细胞胚胎、桑椹胚、胚泡和孵化胚泡的数量。结果:睾酮治疗对接受T刺激的动物的IVF结果产生了负面影响,但在T冲洗后没有。促性腺激素释放激素激动剂预处理不影响IVF结果。结论:尽管与对照组相比,目前的T对体外受精结果有负面影响,但动物仍然能够产生可存活的卵母细胞进行受精并发育成胚泡。未来有必要研究长期T暴露对卵母细胞质量的影响,特别是非整倍体率、妊娠结局和活产率。
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引用次数: 0
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