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Immunomagnetic removal of cryo-damaged human spermatozoa

Asian Journal of Andrology 2005年第1期7卷 61-69,页

作者： uwe paasch Sonja Grunewald Katja Wuendrich Tbrsten Jope Hans-Jurgen Glander Division of Andrology Department of Dermatology University of Leipzig Leipzig D-04103 Germany

Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMR,Δψ_m) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3 % ± 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P < 0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.

关键词： spermatozoa cryopreservation immunomagnetic separation plasma membrane annexin V apoptosis caspases mitochondria

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Expression of toxin-related human mono-ADP-ribosyltransferase 3 in human testes

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Asian Journal of Andrology 2006年第3期8卷 281-287,386页

作者： MaikFriedrich AndreasGrahnert uwe paasch Andrea Tannapfel Friedrich Koch-Nolte Sunna Hauschildt Institute Division of Andrology Department of Dermatology Institute of Pathology University of LeipzigD-04103 LeipzigGermany Institute of Immunology University HospitalD-20246 HamburgGermany

Aim:To investigate wether the corresponding protein of mono-ADP-ribosyltransferase 3(ART3)mRNA is ex- pressed in human testes and,if so,whether the expression is cell ***:ART3 mRNA was deter- mined in human testes and sperm by reverse transcription-polymerase chain reaction(RT-PCR).The glycosyl- phosphatidylinositol linkage of ART3 was shown by treating ART3-transfected HEK-293-T cells with phospholipase *** activated cell sorter(FACS)-analyses were used to detect ART3 on mature spermatozoa and immuno- histological studies to detect the protein in ***:ART3 protein was shown to be present in *** was found on spermatocytes *** was absent from spermatogonia,spermatids and *** absence of ART3 from spermatozoa was confirmed by ***3 protein was detected neither within a seminoma nor on Leydig ***:Here we show for the first time that ART3 protein is expressed in testes in particular on spermatocytes,indicating that ART3 exerts a specific function only required at a particular stage of spermatogenesis.

关键词： mono-ADP-ribosyltransferase 3 testes human spermatocyte spermatogenesis

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Stability of fluorochrome based assays to measure subcellular sperm functions

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Asian Journal of Andrology 2008年第3期10卷 455-459页

作者： Sonja Grunewald Manja Rasch Martin Reinhardt Thomas Baumann uwe paasch Hans-Juergen Glander Department of Dermatology/Andrology Unit European Academy of Andrology （EAA ） Training Center University of Leipzig Leipzig 04103 Germany

Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used to perform the following assays：（i） detection of active caspase-3 （n = 17）; （ii） integrity of the mitochondrial membrane potential （MMP）（n = 17）; （iii） externalization of phosphatidylserine （EPS）（n = 16）; and （iv） detection of intact acrosomes via CD46 （n = 37）. After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14. Results： Differences of up to ± 5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA^TM showed mean differences 〈 5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences 〉 5% at day 3. The CD46-FITC labeling displayed absolute differences 〈 5% CD46-positive spermatozoa at days 3, 7, 10 and 14. Conclusion： Although immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day.

关键词： human spermatozoa apoptosis assays caspase-3 mitochondrial membrane potential integrity CD46 semen analysis

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Probability to retrieve testicular spermatozoa in azoospermic patients

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Asian Journal of Andrology 2000年第3期2卷 199-205页

作者： H.-J.Glanden L.-C.Horn W.Dorschner U.paasch J.Kratzsch Department of Dermatology/Andrology Unit Department of Dermatology Department of PathologyDepartment of Dermatology Department of UrologyDepartment of Dermatology/Andrology UnitDepartment of Clinical Chemistry and Pathological Biochemistry University of Leipzig D-04103 Leipzig Germany

Aim: The degree of probability to retrieve spermatozoa from testicular tissue for intracytoplasmic sperm injection intooocytes is of interest for counselling of infertility patients. We investigated the relation of sperm retrieval to clinical dataand histological pattern in testicular biopsies from azoospermic patients. Methods: In 264 testicular biopsies from142 azoospermic patients, the testicular tissue was shredded to separate the spermatozoa, histological semi-thin sec-tions of which were then evaluated using Johnsen score. Results: The retrieval of spermatozoa correlated signifi-cantly (P 18 U/L, tesficular volume < 5mL, mean Johnsen score< 5, and maximum Johnsen score < 7.

关键词： sperm extraction testicular histology azoospermia male infertility follicle stimulating hormone

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S-enriched porous polymer derived N-doped porous carbons for electrochemical energy storage and conversion

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Frontiers of Chemical Science and Engineering 2018年第3期12卷 346-357页

作者： Chao Zhang Chenbao Lu Shuai Bi Yang Hou Fan Zhang Ming Cai Yafei He Silvia paasch Xinliang Feng Eike Brunner Xiaodong Zhuang State Key Laboratory of Metal Matrix Composites & Shanghai Key Lab of Electrical Insulation and Thermal Ageing School of Chemistry and Chemical Engineering Shanghai Jiao Tong University Shanghai 200240 China Key Laboratory of Biomass Chemical Engineering of Ministry of Education College 0f Chemical and Biological Engineering Zhejiang University Hangzhou 310027 China Center for Advancing Electronics Dresden (cfaed) Technische Universitat Dresden 01062 Dresden. Germany Chair for Molecular Functional Materials Department of Chemistry and Food Chemistry School of Science Technische Universit~t Dresden Mommensenstr. 4 01069 Dresden Germany

Porous polymers have been recently recog- nized as one of the most important precursors for fabrication of heteroatom-doped porous carbons due to the intrinsic porous structure, easy available heteroatom- containing monomers and versatile polymerization meth- ods. However, the heteroatom elements in as-produced porous carbons are quite relied on monomers. So far, the manipulating of heteroatom in porous polymer derived porous carbons are still very rare and challenge. In this work, a sulfur-enriched porous polymer, which was prepared from a diacetylene-linked porous polymer, was used as precursor to prepare S-doped and/or N-doped porous carbons under nitrogen and/or ammonia atmo- spheres. Remarkably, S content can sharply decrease from 36.3% to 0.05% after ammonia treatment. The N content and specific surface area of as-fabricated porous carbons can reach up to 1.32% and 1508 m^2·g^-1, respectively. As the electrode materials for electrical double-layer capacitors, as-fabricated porous carbons exhibit high specific capacitance of up to 431.6 F·g^-1 at 5 mW·s^-1 and excellent cycling stability of 99.74% capacitance retention after 3000 cycles at 100 mV·s^-1. Furthermore, as the electro- chemical catalysts for oxygen reduction reaction, as- fabricated porous carbons presented ultralow half-wave- potential of 0.78 V versus RHE. This work not only offers a new strategy for manipulating S and N doping features for the porous carbons derived from S-containing porous polymers, but also paves the way for the structure- performance interrelationship study of heteroatoms co- doped porous carbon for energy applications.

关键词： porous polymers porous carbons sulfur and nitrogen doping supercapacitor

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人工种子——无性植物胚的包装

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世界科学 1987年第11期 27-30页

作者： K.Redenbaugh B.D.paasch 倪德祥周中一

苜蓿、芹菜和花椰菜的体细胞胚(SE)被包装成直径约4mm的单胚颗粒,即成为人工种子。苜蓿和芹菜人工种子在沙盘和移植块中发育成植株率为7%和10%。本文就人工种子的生产和研究状况进行较全面的报道,它有新颖性。人工种子有广泛而潜在的实... 详细信息

苜蓿、芹菜和花椰菜的体细胞胚(SE)被包装成直径约4mm的单胚颗粒,即成为人工种子。苜蓿和芹菜人工种子在沙盘和移植块中发育成植株率为7%和10%。本文就人工种子的生产和研究状况进行较全面的报道,它有新颖性。人工种子有广泛而潜在的实用价值。

关键词：人工种子芹菜苜蓿绿叶蔬菜褐藻酸钠植株包装植物

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薄层琼脂方法检测结核分枝杆菌分离物对利福平、氧氟沙星和卡那霉素的耐药性

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国际结核病与肺部疾病杂志 2010年第1期5卷 1-4页

作者： A.Martin F.paasch A.Von Groll K.Fissette P.Almeida F.Varaine F.Portaels J-C.Palomino 夏辉刘宇红 Institute of Tropical Medicine Mycobacteriology UnitAntwerpBelgium Laboratory of Mycobacteriology Universidade Federal do Rio GrandeRio Grande do SulBrazil Medecins Sans Frontieres ParisFrance

背景:在低收入国家迫切需要经济的方法检测结核分枝杆菌对抗结核药物的敏感性。目的:评估薄层琼脂方法(thin-layer agar,TLA)快速检测结核分枝杆菌临床分离物对利福平(RMP)、氧氟沙星(Ofx)、卡那霉素(Km)的耐药性;与金标准比较,评估该... 详细信息

背景:在低收入国家迫切需要经济的方法检测结核分枝杆菌对抗结核药物的敏感性。目的:评估薄层琼脂方法(thin-layer agar,TLA)快速检测结核分枝杆菌临床分离物对利福平(RMP)、氧氟沙星(Ofx)、卡那霉素(Km)的耐药性;与金标准比较,评估该方法的敏感度、特异度以及阳性结果的报告时间。方法:共纳入147份结核结核分枝杆菌临床分离物。进行TLA方法时,采用含有RMP,Ofx及Km的7H11琼脂培养基的四象限平皿。TLA方法RMP耐药性结果与Bactec MGIT960结果进行比较,TLA方法检测Ofx和Km耐药性结果与比例法的耐药性结果进行比较。结果:RMP和Ofx的敏感度和特异度均为100%,KM的敏感度和特异度分别为100%和98.7%。使用TLA四象限平皿方法可以快速(中位数为10 d)检测3种抗结核药物(RMP、Ofx、Km)的耐药性。结论:对于检测所研究的3种药物的耐药性,LTA是1种准确的方法。这种快速的方法操作简便,为资源有限、不能应用复杂技术的国家提供了一种替代的方法。

关键词：结核耐药薄层琼脂

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