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Antigen unmasking does not improve the visualization of phospholipase C zeta in human spermatozoa

Asian Journal of Andrology 2022年第4期24卷 345-352页

作者： Xin Meng Celine Jones Pedro Melo Caroline Ross Ginny Mounce Tim Child kevin coward Nuffield Department of Women's and Reproductive Health University of OxfordOxford OX39DUUK The Women's Centre Oxford University Hospitals NHS Foundation TrustOxford OX39DUUK xford Fertility Oxford Business Park NorthOxford OX42HWUK.

Phospholipase C zeta(PLCζ)is a sperm-specific protein that triggers oocyte *** analysis of PLCζexpression in human spermatozoa can be used as a diagnostic marker for oocyte activation *** laboratory has previously optimized a standard"in-house"assay to determine PLCζexpression in human ***,one study has suggested that an antigen unmasking method(AUM)would be more efficient in visualizing PLCζin human *** study aimed to compare our established assay and AUM(involving HCl,acidic Tyrode's solution[AT],and heat).The mean relative fluorescence(RF)intensity of PLCζin frozen-thawed spermatozoa from fourteen fertile donors stained with the in-house method was significantly higher than three other AUM groups(in-house[mean±standard error of mean]:18.87±2.39 arbitrary units[a.u.]vs non-AUM:11.44±1.61 a.u.,AT-AUM:12.38±1.89 a.u.,and HCl-AUM:12.51±2.16 a.u.,P<0.05,one-way analysis of variance).The mean RF intensity of PLCζin AT-and HCl-treated spermatozoa from 12 infertile males was not significantly different from that of the non-AUM ***,the in-house method resulted in the highest RF intensity(12.11±1.36 a.u.,P<0.01).Furthermore,specificity testing of antibody-antigen binding indicated that the in-house method showed more specific binding than spermatozoa treated by the *** conclusion,our in-house method showed superior visualization and reliability than the AUM,thus supporting the continued use of our in-house assay for clinical research screening.

关键词： antigen unmasking immunofluorescence staining male infertility phospholipase C zeta

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Advancing male age differentially alters levels and localization patterns of PLCzeta in sperm and testes from different mouse strains

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Asian Journal of Andrology 2021年第2期23卷 178-187页

作者： Junaid Kashir Bhavesh V Mistry Maha Adel Gumssani Muhammad Rajab Reema Abu-Dawas Falah AlMohanna Michail Nomikos Celine Jones Raed Abu-Dawud Nadya Al-Yacoub kevin coward F Anthony Lai Abdullah M Assiri College of Medicine Alfaisal UniversityRiyadh 11533Saudi Arabia Department of Comparative Medicine King Faisal Specialist Hospital and Research CentreRiyadh 11211Saudi Arabia College of Medicine QU HealthQatar UniversityDoha PO Box 2713Qatar Nuffield Department of Women’s and Reproductive Health University of OxfordLevel 3Women’s CentreJohn Radcliffe HospitalOxford OX39DUUnited Kingdom Biomedical Research Centre Qatar UniversityDoha PO Box 2713Qatar

Sperm-specific phospholipase C zeta(PLCζ)initiates intracellular calcium(Ca2+)transients which drive a series of concurrent events collectively termed oocyte *** investigations have linked abrogation and absence/reduction of PLCζwith forms of male infertility in humans where oocyte activation ***,very few studies have examined potential relationships between PLCζand advancing male age,both of which are increasingly considered to be major effectors of male *** efforts in humans may be hindered by inherent PLCζvariability within the human population,alongside a lack of sufficient controllable ***,utilizing immunoblotting,immunofluorescence,and quantitative reverse transcription PCR(qRT-PCR)we examined for the first time PLCζprotein levels and localization patterns in sperm,and PLCζmRNA levels within testes,from mice at 8 weeks,12 weeks,24 weeks,and 36 weeks of age,from two separate strains of mice,C57BL/6(B6;inbred)and CD1(outbred).Collectively,advancing male age generally diminished levels and variability of PLCζprotein and mRNA in sperm and testes,respectively,when both strains were ***,advancing male age altered the predominant pattern of PLCζlocalization in mouse sperm,with younger mice exhibiting predominantly post-acrosomal,and older mice exhibiting both post-acrosomal and acrosomal populations of PLCζ.However,the specific pattern of such decline in levels of protein and mRNA was ***,our results demonstrate a negative relationship between advancing male age and PLCζlevels and localization patterns,indicating that aging male mice from different strains may serve as useful models to investigate PLCζin cases of male infertility and subfertility in humans.

关键词： ageing fertilization male infertility oocyte activation phospholipase C zeta sperm

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Recovery of spermatogenesis following testosterone replacement therapy or anabolic-androgenic steroid use

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Asian Journal of Andrology 2016年第3期18卷 373-380,I0007页

作者： J Abram McBride Robert M coward Department of Urology University of North Carolina School of Medicine Chapel Hill NC 27599-7235 USA.

The use of testosterone replacement therapy （TRT） for hypogonadism continues to rise, particularly in younger men who may wish to remain fertile. Concurrently, awareness of a more pervasive use of anabolic-androgenic steroids （AAS） within the general population has been appreciated. Both TRT and AAS can suppress the hypothalamic-pituitary-gonadal （HPG） axis resulting in diminution of spermatogenesis. Therefore, it is important that clinicians recognize previous TRT or AAS use in patients presenting for infertility treatment. Cessation of TRT or AAS use may result in spontaneous recovery of normal spermatogenesis in a reasonable number of patients if allowed sufficient time for recovery. However, some patients may not recover normal spermatogenesis or tolerate waiting for spontaneous recovery. In such cases, clinicians must be aware of the pathophysiologic derangements of the HPG axis related to TRT or AAS use and the pharmacologic agents available to reverse them. The available agents include injectable gonadotropins, selective estrogen receptor modulators, and aromatase inhibitors, but their off-label use is poorly described in the literature, potentially creating a knowledge gap for the clinician. Reviewing their use clinically for the treatment of hypogonadotropic hypogonadism and other HPG axis abnormalities can familiarize the clinician with the manner in which they can be used to recover spermatogenesis after TRT or AAS use.

关键词： anabolic steroids hypogonadism infertility spermatogenesis testosterone testosterone replacement therapy vasectomy reversal

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The modern testicular prosthesis:patient selection and counseling,surgical technique,and outcomes

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Asian Journal of Andrology 2020年第1期22卷 64-69页

作者： Solomon Hayon Jamie Michael R Matthew coward Department of Urology University of North Carolina School of MedicineChapel HillNC 27500-7235USA University of North Carolina School of Medicine Chapel HillNC 27500-7235USA

The testicular prosthesis can be an afterthought for providers when performing an orchiectomy for testicular cancer,torsion,atrophic testis,or ***,data suggest that patients find the offer of a testicular prosthesis and counseling regarding placement to be extremely important from both a pragmatic and a psychosocial *** two-thirds of men undergoing orchiectomy are offered an implant at the time of orchiectomy and of those offered about one-third move forward with prosthesis *** relatively low acceptance rate is in stark contrast with high patient satisfaction and low complication rates for those who undergo the *** most common postoperative patient concerns are minor and involve implant positioning,size,and ***,we provide an up-to-date review of modern preoperative evaluation,patient selection,expectation management,surgical technique,and expected outcomes for testicular prostheses.

关键词： anorchia orchiectomy patient satisfaction prostheses and implants testicular neoplasms testis

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Diagnosis and management of testosterone deficiency

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Asian Journal of Andrology 2015年第2期17卷 177-186,I0006页

作者： James A McBride Culley C Carson Robert M coward Department of Urology University of North Carolina School of Medicine NC USA.

Testosterone supplementation therapy （TST） use has dramatically increased over the past decade, due to the availability of newer agents, aggressive marketing, and an increasing incidence of testosterone deficiency （TD）. Despite the increase in TST, a degree of ambiguity remains as to the exact diagnostic criteria of TD, and administration and monitoring of TST. One explanation for this phenomenon is the complex role testosterone plays in multiple physiologic pathways. Numerous medical co-morbidities and medications can alter testosterone levels resulting in a wide range of nonspecific clinical signs and symptoms of TD. The diagnosis is also challenging due to the lack of a definitive serum total testosterone level that reliably correlates with symptoms. This observation is particularly true in the aging male and is exacerbated by inconsistencies between different laboratory assays. Several prominent medical societies have developed guideline statements to clarify the diagnosis, but they differ from each other and with expert opinion in several ways. Aside from diagnostic dilemmas, there are numerous subtle advantages and disadvantages of the various testosterone agents to appreciate. The available TST agents have changed significantly over the past decade similar to the trends in the diagnosis of TD. Therefore, as the usage of TST increases, clinicians will be challenged to maintain an up-to-date understanding of TD and TST. The purpose of this review is to provide a clear description of the current strategies for diagnosis and management of TD.

关键词： androgens erectile dysfunction hormones hypogonadism testosterone

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Sperm retrieval and intracytoplasmic sperm injection outcomes in men with cystic fibrosis disease versus congenital bilateral absence of the vas deferens

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Asian Journal of Andrology 2021年第2期23卷 140-145页

作者： J Abram McBride Taylor P Kohn Daniel J Mazur Larry I Lipshultz R Matthew coward Scott Department of Urology Baylor College of MedicineHoustonTX 77030USA Department of Urology UNC School of MedicineChapel HillNC 27599USA UNC Fertility RaleighNC 27617USA

Recent data suggest that cystic fibrosis transmembrane conductance regulator(CFTR)gene alterations negatively impact male fertility beyond *** sought to compare gene alterations,sperm retrieval rates,and intracytoplasmic sperm injection(ICSI)outcomes among men with cystic fibrosis(CF)disease and congenital bilateral absence of the vas deferens(CBAVD)*** retrospectively evaluated all men who underwent surgical sperm retrieval at two academic,high-volume andrology centers from 2010 to *** men with documented CFTR alterations and obstructive azoospermia from either CBAVD or CF were *** between groups for CFTR abnormality,sperm retrieval,and ICSI outcomes were statistically ***,39 patients were included with 10 in the CF and 29 in the CBAVD *** sperm retrieval rates were significantly lower in the CF group for sperm concentration(14.8×10^(6)ml^(-1)vs 61.4×10^(6)ml^(−1),P=0.02)and total motile sperm count(2.9 million vs 11.4 million,P=0.01).This difference was only predicted by homozygous delta F508 CFTR mutations(P<0.05).The CF group also demonstrated a significantly higher rate of rescue testicular sperm extraction(70.0%vs 27.6%,P<0.03)and lower fertilization rate with ICSI(32.5%vs 68.9%,P<0.01).In conclusion,those with CF demonstrated lower sperm quality,greater difficulty with sperm retrieval,and worse ICSI outcomes compared with CBAVD-only *** delta F508 CFTR mutations appear to significantly impair spermatogenesis and sperm function.

关键词： congenital bilateral absence of the vas deferens cystic fibrosis intracytoplasmic sperm injection outcomes male infertility obstructive azoospermia

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The effects of advanced paternal age on fertility

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Asian Journal of Andrology 2013年第6期15卷 723-728,I0005页

作者： Jason R Kovac Josephine Addai Ryan P Smith Robert M coward Dolores J Lamb Larry I Lipshultz Scott Department of Urology Department of Molecular and Cellular Biology The Center for Reproductive Medicine Baylor College of Medicine Houston TX 77030 USA

Modern societal pressures and expectations over the past several decades have resulted in the tendency for couples to delay conception. While women experience a notable decrease in oocyte production in their late thirties, the effect of age on spermatogenesis is less well described. While there are no known limits to the age at which men can father children, the effects of advanced paternal age are incompletely understood. This review summarizes the current state of knowledge regarding advanced paternal age and its implications on semen quality, reproductive success and offspring health. This review will serve as a guide to physicians in counseling men about the decision to delay paternity and the risks involved with conception later in life.

关键词： aging male fertility outcomes male infertility paternal age

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中国西北河西走廊酒西盆地:前陆构造特征及油气远景

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国外油气勘探 1993年第6期 677-686页

作者： Q.M.Wangh M.P.coward 王冰

位于中国西北河西走廊西部的酒西(酒泉以西)盆地属于早侏罗世以后发育的前陆盆地。其形成是由于沿2000km长的阿尔金断层的左行剪切作用,北祁连冲断带逐渐向北迁移的结果。盆地中的沉积层不但受前陆负荷和冲断作用控制,而且受与冲断带以... 详细信息

位于中国西北河西走廊西部的酒西(酒泉以西)盆地属于早侏罗世以后发育的前陆盆地。其形成是由于沿2000km长的阿尔金断层的左行剪切作用,北祁连冲断带逐渐向北迁移的结果。盆地中的沉积层不但受前陆负荷和冲断作用控制,而且受与冲断带以高角度相交的犁式正断层控制。两组断裂交汇处堆积了巨厚的富含有机质的沉积物和储集层。第三纪和第四纪期间,冲断层沿北祁连断层下盘扩展,削截了早期形成的油藏和源岩,因而给石油勘探带来了巨大的困难。根据沉积相、钻探和地震资料、构造分析和横剖面恢复的综合研究,本文提出了盆地演化模式。自上新世以来,北祁连山前缘带向北运动的平均速率估计每年约为8mm。因此,大约1/2的侏罗纪—白垩纪盆地可能掩埋于老君庙和北祁连边缘断层之下,实际上并未被探井揭露。盆地中的油源岩为晚侏罗世到早白垩世时期的黑色湖相页岩,在青西拗陷,最大厚度达1.2km。液态烃的生成始于晚白垩世或中渐新世;本文描述了自志留系到中新统的7套地层储集层;背斜、断层、"潜山"构造是最重要的圈闭。本文详述了个别拗陷的石油潜力,并且提出了潜在的区域石油远景区。

关键词：酒西盆地前陆构造构造演化油气勘探远景评价生油岩

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涂层卷材的应用

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轻合金加工技术 1989年第5期17卷 26-30页

作者： A.W.Peabhey E.P.Kent-coward 张境安〔英国〕

本文说明了涂层钢卷材和涂层铝卷材在过去二十年内是如何进入涂层加工市场的,并从工厂、设备和人员的角度,概述了它们对涂层板用户的重要作用。随着将来完全自动化处理不可避免的增长,人们会坚决相信,涂层的带卷和板材合乎情理的发展应... 详细信息

本文说明了涂层钢卷材和涂层铝卷材在过去二十年内是如何进入涂层加工市场的,并从工厂、设备和人员的角度,概述了它们对涂层板用户的重要作用。随着将来完全自动化处理不可避免的增长,人们会坚决相信,涂层的带卷和板材合乎情理的发展应该是消除强体力劳动操作和改善工作环境。本文回顾了迄今为止所获得的成功(特别是在建筑业),讨论了卷材涂层机、涂料制造机、冷压加工成形和工程技术人员诸因素综合在一起构成全新建筑业概念这一事实。由于在汽车、包装、自动化和通用工程部门取得的可喜成功,涂层卷材的使用已进入一般的工业领域。

关键词：卷材涂层板材材料

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薄皮构造带的应变和构成史——穿过苏格兰西北部阿辛特区的莫因逆冲断层带上的几个实例

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宝石和宝石学杂志(中英文) 1986年第4期 53-61页

作者： W.P.coward 王豪

穿过阿辛特区的莫因逆冲断层带是个向前陆扩展的薄皮逆冲断层带的典型实例,对于研究与逆冲断层有关的褶皱、变形作用产生的结构和应变,以及研究复杂的增量应变的历史是理想的实例。离散的运动方向、斜向褶皱、双重构造(duplex)带和垂直... 详细信息

穿过阿辛特区的莫因逆冲断层带是个向前陆扩展的薄皮逆冲断层带的典型实例,对于研究与逆冲断层有关的褶皱、变形作用产生的结构和应变,以及研究复杂的增量应变的历史是理想的实例。离散的运动方向、斜向褶皱、双重构造(duplex)带和垂直于主要运动方向的拉伸应变等,全都可以用逆冲断层的扩展来解释,扩展结果引起了斜断坡至断坡的发育。大多数逆冲断层都沿运动方向从基底往上切断盖层,但是还有逆冲席体的局部的拉伸性塑流与变薄。在阿辛特北部,此逆冲断层带包括有一条宽阔的、直立的左旋剪切带,这个剪切带范围内前逆冲断层、后逆冲断层以及伴生的褶皱、劈理等都与总的运动方向斜交。本文提出,在该剪切带以北,那些逆冲断层都运动得更远,很可能是盖在较厚层的下面,而在此剪切带以南。则运动是更加断断续续的,很可能是盖在较薄盖层的下面。这种变化很可能归因于逆冲断层的几何特征。在莫因逆冲断层带以东的莫因系地层里的改变,导致了整个莫因逆冲断层线上重力位能的变化。

关键词：逆冲断层几何特征冲断层剪切带剪切破裂带断裂带褶皱地质构造逆冲带运移方向双重构造重力断层地质断层阿辛侧断坡盖层地层叠瓦构造逆断层特区苏格兰

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