BACKGROUND & AIMS: In the spermatozoa of some species creatine kinase (CKE.C. 2.7.3.2) is involved in shuttling energy, in the form of creatine phosphate, between the mid-piece mitochondria and flagellum. In this study, the effects of the CK inhibitor dinitrofluorobenzene (DNFB) on human sperm CK activity, motility and ATP concentrations were assessed with different energy substrates. There was a dose-dependent inhibition of CK activity by DNFB but inhibition was incomplete and there was no effect on the percentage of flagellating cells, irrespective of substrate. However, when lactate alone supported the cells DNFB decreased velocities and increased amplitude of head displacement (fewer progressively motile forms were observed), whereas ATP concentrations in spermatozoa were unaltered. Demembranated sperm models could be reactivated by ADP plus creatine phosphate, but not to the extent caused by ATP, and were able to be inhibited by myokinase inhibitors. Increased velocities, linearity (LIN) and beat cross frequency (BCF) were demonstrated for spermatozoa incubated with lactate, in contrast to glucose as sole energy source, and higher velocities and BCF were generated in the presence of both substrates. This suggests that the production of ATP by glycolysis and respiration are independent and complementary. CK is not obligatory for sperm motility but supplements energy provision under certain conditions.

背景与目标： 在某些物种的精子中，肌酸激酶（CKE.C. 2.7.3.2）以中枢线粒体和鞭毛之间的肌酸磷酸形式参与穿梭能。在这项研究中，使用不同的能量底物评估了CK抑制剂二硝基氟苯（DNFB）对人精子CK活性，运动性和ATP浓度的影响。 DNFB对CK的活性有剂量依赖性的抑制作用，但是抑制作用是不完全的，并且对鞭毛细胞的百分比没有影响，而与底物无关。然而，当单独的乳酸支持细胞时，DNFB降低了速度，增加了头部移位的幅度（观察到逐渐减少的运动形式），而精子中的ATP浓度未改变。去膜的精子模型可以被ADP加磷酸肌酸再活化，但不能达到由ATP引起的程度，并且能够被肌动酶抑制剂抑制。与葡萄糖作为唯一能源相比，用乳酸孵育的精子显示出更高的速度，线性（LIN）和拍频（BCF），与之相比，葡萄糖和葡萄糖是唯一的能量来源，在两种底物均存在的情况下，产生了更高的速度和BCF。这表明通过糖酵解和呼吸作用产生的ATP是独立的和互补的。 CK对精子活力不是强制性的，而是在某些条件下补充能量供应。

BACKGROUND & AIMS: Selectins are a family of adhesive molecules, involved in the interactions between leukocytes and endothelium and in platelet adhesion. P-selectin, one of the members of this family, is stored in alpha-granules and dense granules of platelets as well as in Weibel-Palade bodies of endothelial cells, and it is rapidly redistributed to the cell surface after activation. It recognizes carbohydrate structures as ligands, in particular sialyl-Lewis(x), which is part of the CD15 antigen. In this work we studied P-selectin expression on gametes. While zona-free human and hamster oocytes did not react with a monoclonal antibody directed against P-selectin, oocytes from both species displayed a reactivity with this antibody following their contact with human spermatozoa, as demonstrated both by covasphere binding and indirect immunofluorescence. Artificial activation of zona-intact human oocytes by means of the calcium lonophore A23187 induced the expression on the oolemma of a moiety reacting with anti-P-selectin antibody as well. P-selectin also appeared to be expressed on the sperm surface following the acrosome reaction, as demonstrated by a flow cytometric study of reactivity of spermatozoa with the anti-P-selectin antibody, using the expression of CD46 as a marker of the acrosome reaction. The localization of the P-selectin moiety on the equatorial region of the plasma membrane of acrosome reacted spermatozoa was confirmed by transmission electron microscopy using immunogold labelling. We suggest that P-selectin might be involved in gamete interactions.

背景与目标： 选择素是粘附分子家族，参与白细胞和内皮之间的相互作用以及血小板粘附。 P-选择蛋白是该家族的成员之一，存储在血小板的α颗粒和致密颗粒中以及内皮细胞的Weibel-Palade体中，并在激活后迅速重新分布到细胞表面。它识别碳水化合物结构作为配体，特别是唾液酸化-Lewis（x），它是CD15抗原的一部分。在这项工作中，我们研究了配子上P选择素的表达。尽管无透明带的人和仓鼠卵母细胞不与针对P-选择素的单克隆抗体反应，但两种卵母细胞与人精子接触后均表现出与该抗体的反应性，这已通过卵泡球结合和间接免疫荧光证明。借助钙离子载体A23187人工激活完整完整的人卵母细胞，也会诱导与抗P-选择素抗体反应的部分在血肿中表达。 P-选择素似乎在顶体反应后在精子表面表达，如流式细胞术研究表明，使用CD46的表达作为顶体反应的标记，精子与抗P-选择素抗体发生反应。通过使用免疫金标记的透射电子显微镜确认了P-选择蛋白部分在顶体反应的精子的质膜的赤道区域上的定位。我们建议P-选择素可能参与配子的相互作用。

BACKGROUND & AIMS: :A mathematical model is presented to study the motion of the spermatozoa in the cervical canal by considering the transverse waves along its tail and the transverse and longitudinal motions of the cervical wall. In an attempt to control fertility by reducing the speed of sperm, the transverse waves have been considered in the direction opposite to the motion of the spermatozoa. It has been shown that by having appropriate transverse wave motion and longitudinal velocity, the sperm may not be able to move towards the oviduct even if it could continue to have its own propelling velocity. A particular case of the motion of a thin plane sheet in a channel under peristaltic motion of its walls has also been obtained and studied.

背景与目标： ：提出了一个数学模型，通过考虑沿精子尾巴的横波以及子宫颈壁的横向和纵向运动来研究子宫颈管中精子的运动。为了通过降低精子的速度来控制生育力，已经在与精子运动相反的方向上考虑了横波。已经显示出，通过具有适当的横向波运动和纵向速度，即使精子可以继续具有自己的推进速度，它也可能无法向输卵管移动。还已经获得并研究了在其壁的蠕动下通道中薄板的运动的特殊情况。

BACKGROUND & AIMS: :Sermatozoa from two brothers who are not twins were found to be straight and immotile. Examinations of the sperm showed that oxygen consumption and lactic acid production were normal; viability tests showed that the percentage of dead sperm was not increased. The ultrastructural appearance of the sperm tail was normal except for a complete lack of dynein arms and some irregularities in the arrangement of the accessory fibers and the longitudinal columns of the fibrous sheath. The mitochondrial apparatus and the sperm head conform to the conventional model. According to the sliding-filament hypothesis first proposed by Afzelius (1959. J. Biophys. Biochem. Cytol. 5:269.), the arms are responsible for the bending movements of the tail. The simplest explanation for the simultaneous lack of arms and sperm motility appears to be that the two brothers have a genetic disorder involving production, assembly, or attachment of the dynein arms.

背景与目标： ：来自不是孪生兄弟的两个兄弟的Sermatozoa被发现是直挺的，没有动静。精子检查显示耗氧量和乳酸生成是正常的。生存力测试表明，死亡的精子百分比没有增加。精子尾巴的超微结构是正常的，除了完全缺乏动力蛋白的臂和附属纤维和纤维鞘纵列的排列有些不规则。线粒体设备和精子头符合常规模型。根据Afzelius最初提出的滑动丝假说（1959. J. Biophys。Biochem。Cytol。5：269。），手臂负责尾巴的弯曲运动。同时缺乏手臂和精子活动力的最简单的解释似乎是两兄弟患有涉及动力蛋白的生产，组装或附着的遗传性疾病。

BACKGROUND & AIMS: :Variation in localization and distribution of saccharides on the sperm surface of a marsupial, the brushtail possum, Trichosurus vulpecula, was compared between spermatozoa from the caput and cauda epididymides. Spermatozoa were subjected to the following treatments: (i) unfixed and fixed spermatozoa were stained with fluorescein-labelled lectins; (ii) unfixed spermatozoa were incubated with lectins for determination of agglutination; and (iii) spermatozoa were incubated with detergent to remove the plasmalemma, the glycoproteins were separated on SDS-PAGE and western blots were stained with biotinylated lectins. Many of the fluorescein isothiocyanate (FITC)-labelled lectins bound selectively to the sperm surface, and marked differences were found in lectin staining affinity between caput and cauda epididymal spermatozoa. Incubation of spermatozoa from the cauda epididymidis with neuraminidase reversed many of the differences in staining of the cauda epididymal spermatozoa, indicating masking of some terminal saccharides by sialic acid. Agglutination of spermatozoa from the caput epididymidis occurred after incubation with Concanavalin A (ConA) and soybean agglutinin (SBA), but agglutination was less extensive for spermatozoa from the cauda epididymidis. Western blot analysis indicated several ConA-positive bands in caput sperm extracts, but fewer positive bands in the cauda sperm extracts, whereas SBA stained four bands from caput but none from the cauda epididymal spermatozoa. These results demonstrate extensive glycosylation of the surface proteins of spermatozoa from the caput epididymidis and significant differences in spermatozoa from the cauda epididymidis. In general, the findings indicate similar glycosylation of the surface of marsupial spermatozoa to those from eutherian mammals despite marked differences in their morphology and early divergence of marsupials from eutherian mammals. It would appear that this situation differs markedly from that in sub-mammalian vertebrates.

背景与目标： ：比较了有帽动物的精子和附睾马尾精子中，有袋动物的小头负鼠Trichosurus vulpecula的精子表面上的糖的定位和分布的变化。对精子进行以下处理：（i）用荧光素标记的凝集素对未固定和固定的精子进行染色； （ii）将未固定的精子与凝集素一起温育以测定凝集； （iii）将精子与去污剂一起温育以去除质膜，在SDS-PAGE上分离糖蛋白，并用生物素化的凝集素对蛋白质印迹进行染色。许多荧光素异硫氰酸酯（FITC）标记的凝集素选择性地结合到精子表面，并且在凝块和附睾附睾精子之间的凝集素染色亲和力方面发现了显着差异。用神经氨酸酶从附睾马尾培养精子逆转了附睾马尾染色的许多差异，表明唾液酸掩盖了某些末端糖。与伴刀豆球蛋白A（ConA）和大豆凝集素（SBA）孵育后，来自附睾附睾的精子凝集发生，但来自附睾马尾的精子的凝集作用较弱。 Western印迹分析表明，在子cap精子提取物中有多个ConA阳性条带，但在马尾精子提取物中的阳性条带较少，而SBA染色了来自子ut的四个条带，但没有染上马尾附睾精子的条带。这些结果证明了来自附子附睾的精子表面蛋白的广泛糖基化和来自附睾的精子的显着差异。总的来说，这些发现表明有袋类动物精子表面的糖基化程度与来自真人哺乳动物的相似，尽管有形动物和有色动物的早期差异明显。看来这种情况与亚哺乳动物脊椎动物的情况明显不同。

BACKGROUND & AIMS: :Spermatozoa produced in the testis undergo maturation in the epididymis which secretes an osmolyte-rich fluid that bathes the sperm cells. These cells need to maintain their volume after ejaculation when they first encounter hypo-osmolal environments of accessory gland fluids and later within the female tract. If they do not, they experience swelling that is manifested in flagellar angulation that prevents their passage through cervical mucus or the uterotubal junction and they never reach the oocytes. This is a cause of male infertility in domestic species and certain infertile transgenic mice in which flagellar angulation has been shown to indicate cell swelling as a consequence of reduced epididymal provision of osmolytes. The reduced volume regulating ability of spermatozoa from subfertile boars and bulls has prompted study of volume regulation of spermatozoa as a possible cause of human male infertility. Understanding this process may make its manipulation possible and could suggest better sperm handling and storage techniques and thus provide therapy for infertile patients. On the other hand, volume regulation is a potential target for contraception if mimicking the conditions expressed by the "sterile studs" were possible. The evidence for the presence of ion channels probably responsible for regulatory volume decreases in spermatozoa is reviewed here that implicate voltage-gated potassium channels (especially Kv1.5 (KCNA5), minK (KCNE1) and TASK2 (KCNK5)) and the chloride channels CLCN3 and CLNS1A. The involvement of ion co-transporters in volume regulation of spermatozoa is also discussed.

背景与目标： ：睾丸中产生的精子在附睾中成熟，分泌出富含渗透压的液体，使精子细胞沐浴。这些细胞在射精后首先遇到辅助腺体液体的低渗环境时，随后在雌性道中时，需要保持其体积。如果没有，它们会经历鞭毛成角的肿胀，从而阻止它们通过宫颈粘液或子宫输卵管交界处，而且它们永远不会到达卵母细胞。这是导致家庭物种和某些不育转基因小鼠中男性不育的原因，其中鞭毛成角表明已表明由于溶菌附睾提供减少而导致细胞肿胀。来自不育公猪和公牛精子的体积调节能力降低，促使人们对精子体积调节作为人类男性不育症的可能原因进行了研究。了解此过程可能使其操作成为可能，并可能建议使用更好的精子处理和储存技术，从而为不育患者提供治疗。另一方面，如果有可能模仿“不育螺柱”所表示的状况，则进行量调节是避孕的潜在目标。本文回顾了可能导致精子调节体积减少的离子通道的证据，这些证据暗示电压门控性钾通道（尤其是Kv1.5（KCNA5），minK（KCNE1）和TASK2（KCNK5））和氯离子通道CLCN3和CLNS1A。还讨论了离子共转运蛋白在精子体积调节中的作用。

BACKGROUND & AIMS: :Expression of olfactory receptors (ORs) in non-olfactory tissues has been widely reported over the last 20 years. Olfactory marker protein (OMP) is highly expressed in mature olfactory sensory neurons (mOSNs) of the olfactory epithelium. It is involved in the olfactory signal transduction pathway, which is mediated by well-conserved components, including ORs, olfactory G protein (Golf), and adenylyl cyclase 3 (AC3). OMP is widely expressed in non-olfactory tissues with an apparent preference for motile cells. We hypothesized that OMP is expressed in compartment-specific locations and co-localize with an OR, Golf, and AC3 in rat epididymal and human-ejaculated spermatozoa. We used immunocytochemistry to examine the expression patterns of OMP and OR6B2 (human OR, served as positive olfactory control) in experimentally induced modes of activation and determine whether there are any observable differences in proteins expression during the post-ejaculatory stages of spermatozoal functional maturation. We found that OMP was expressed in compartment-specific locations in human and rat spermatozoa. OMP was co-expressed with Golf and AC3 in rat spermatozoa and with OR6B2 in all three modes of activation (control, activated, and hyperactivated), and the mode of activation changed the co-expression pattern in acrosomal-reacted human spermatozoa. These observations suggest that OMP expression is a reliable indicator of OR-mediated chemoreception, may be used to identify ectopically expressed ORs, and could participate in second messenger signaling cascades that mediate fertility.

背景与目标： 在过去的20年中，已经广泛报道了嗅觉受体（ORs）在非嗅觉组织中的表达。嗅觉标记蛋白（OMP）在嗅觉上皮的成熟嗅觉感觉神经元（mOSNs）中高表达。它参与嗅觉信号转导途径，该途径由保守性良好的成分介导，包括OR，嗅觉G蛋白（Golf）和腺苷酸环化酶3（AC3）。 OMP在非嗅觉组织中广泛表达，对运动细胞具有明显的偏爱。我们假设OMP在大鼠附睾和人类射精的精子中在特定的区室表达并与OR，Golf和AC3共同定位。我们使用免疫细胞化学检查了在实验诱导的激活模式下OMP和OR6B2（人类OR，用作阳性嗅觉对照）的表达模式，并确定了在精子功能成熟的射精后阶段蛋白表达是否存在可观察的差异。我们发现OMP在人类和大鼠的精子中特定于区室的位置表达。 OMP在大鼠精子中与Golf和AC3共表达，并在所有三种激活模式（对照，激活和超激活）中与OR6B2共表达，并且激活模式改变了在顶体反应的人精子中的共表达模式。这些观察结果表明，OMP表达是OR介导的化学感受的可靠指标，可用于鉴定异位表达的OR，并可参与介导生育力的第二信使信号级联反应。

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BACKGROUND & AIMS: :Studies on the flagellar movement of carp spermatozoa induced by dilution in distilled water allowed us to describe a sequence of early, rapid morphological and kinetic changes which begin at the very tip of the flagellum. They cause the progressive folding of the axoneme which ends stuck to the head within 90-120 seconds after the initiation of motility. However, the axonemal machinery remains functional as the folding can be reversed after transfer back into a high osmolality medium and partially folded flagella were able to propagate efficient waves along the non-folded proximal portion of the axoneme. The data also revealed that the membrane area of the terminal piece exhibits strong sensitivity to hypotonicity. These results suggest that in the normal freshwater medium, the brief swimming period allowing fertilization of oocytes is limited by the osmotic stress induced coiling of the carp sperm tail and not by ATP stores.

背景与目标： ：关于在蒸馏水中稀释引起的鲤鱼精子鞭毛运动的研究，我们可以描述一系列从鞭毛尖端开始的早期，快速的形态学和动力学变化。它们导致轴突逐渐折叠，在运动开始后的90-120秒内，轴突最终粘在头上。然而，由于在折叠回到高渗透压介质后，折叠可以逆转，因此轴突的机械装置仍然起作用，并且部分折叠的鞭毛能够沿着轴突的未折叠的近端部分传播有效的波。数据还显示，端子片的膜面积对低渗性表现出很强的敏感性。这些结果表明，在正常的淡水培养基中，允许卵母细胞受精的短暂游泳时期受到渗透压诱导的鲤鱼精子尾巴卷曲的限制，而不是受ATP贮藏的限制。

BACKGROUND & AIMS: :Serum amyloid P component (SAP) belongs to the pentraxin family of proteins, members of which are characterized by radial pentameric structure and calcium-dependent ligand binding. SAP is present in all types of amyloidosis and has been shown to bind to several ligands, but the physiological function of this protein has not been fully elucidated. The present study identified and characterized SAP in human semen and immunolocalized it to the male reproductive tract. SAP was also detected in seminal plasma by immunoblotting and purification by affinity chromatography followed by mass spectrometry. According to electroimmunoassay, the concentration of SAP in semen is approximately 2 mg/L, and flow cytometry revealed SAP attached to the surface of spermatozoa. Moreover, immunohistochemistry showed positive staining of spermatozoa, subsets of epithelial cells, and the stroma of accessory male genital glands and testis. Presence of mRNA supports local production of SAP, as shown with reverse transcription polymerase chain reaction. We identified SAP in a new setting - the human male reproductive system. SAP was detected on ejaculated spermatozoa, in seminal plasma and in tissue sections from the male reproductive tract. Further functional studies are needed to explain the role of SAP in human reproduction.

背景与目标： ：血清淀粉样蛋白P组分（SAP）属于pentraxin家族蛋白，其成员以放射状五聚体结构和钙依赖性配体结合为特征。 SAP存在于所有类型的淀粉样变性病中，并已显示与几种配体结合，但尚未完全阐明该蛋白的生理功能。本研究鉴定并表征了人类精液中的SAP，并将其免疫定位于男性生殖道。还可以通过免疫印迹法在精浆中检测到SAP，然后通过亲和色谱法和质谱法进行纯化。根据电免疫测定，精液中SAP的浓度约为2 mg / L，流式细胞仪显示SAP附着在精子表面。此外，免疫组化显示精子，上皮细胞亚群以及雄性生殖器官和睾丸间质的阳性染色。 mRNA的存在支持SAP的局部生产，如逆转录聚合酶链反应所示。我们在新的环境-人类雄性生殖系统中确定了SAP。在射精的精子，精浆和雄性生殖道的组织切片中检测到SAP。需要进一步的功能研究来解释SAP在人类生殖中的作用。

BACKGROUND & AIMS: :Prostasomes are membranous vesicles present in ejaculated human semen. They are very rich in cholesterol and can interact with spermatozoa. Their physiological roles are still under study. Prostasomes were mixed with liposomes prepared from various lipids, such as N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium (DOTAP), DOTAP/1,2-dipalmytoyl-sn-glycero-3-phosphorylcholine (DPPC, 4:1 molar ratio) and DOTAP/cholesterol (4:1, molar ratio) at different pH values (5-8). The mixing of the lipid phases (fusion) was determined by the relief of octadecyl rhodamine B chloride (R(18)) self-quenching and the radii of the vesicles, by light scattering measurements. The mixing of lipids and the radii of prostasomes were both influenced by the addition of liposome, although in a different manner. The ability of prostasomes (modified by previous treatment with liposomes) to transfer lipid to spermatozoa was also measured. Pretreatment with DOTAP decreased the phenomenon and addition of DPPC abolished it. On the other hand, pretreatment of prostasomes with DOTAP/cholesterol liposomes did not affect the transfer of lipid between prostasome and spermatozoa. Therefore, the ability of vesicles to fuse (or, at least, to exchange the lipid component) was affected by the enrichment in either natural or artificial lipid. This may open new possibilities for the modulation of spermatozoa capacitation and acrosome reaction.

背景与目标： ：prosomesomes是射精的人精液中存在的膜状囊泡。它们富含胆固醇，可以与精子相互作用。它们的生理作用仍在研究中。将前体与由各种脂质（例如N- [1-（2,3-二油酰氧基）丙基] -N，N，N-三甲基铵（DOTAP），DOTAP / 1,2-二巴甲酰基-sn-甘油-在不同的pH值（5-8）下，3-磷酸胆碱（DPPC，摩尔比为4：1）和DOTAP /胆固醇（摩尔比为4：1）。脂质相的混合（融合）是通过减轻十八烷基若丹明B氯化物（R（18））自猝灭和囊泡半径（通过光散射测量）来确定的。脂质的混合和前体半径均受脂质体添加的影响，尽管方式不同。还测量了前体（通过先前用脂质体处理修饰的）将脂质转移至精子的能力。用DOTAP预处理可以减少这种现象，并且添加DPPC可以消除这种现象。另一方面，用DOTAP /胆固醇脂质体对前列腺体进行预处理不会影响脂质在前列腺体与精子之间的转移。因此，天然或人工脂质的富集影响了囊泡融合（或至少交换脂质组分）的能力。这可能为调节精子获能和顶体反应开辟新的可能性。

BACKGROUND & AIMS: OBJECTIVE:To assess fertilization, implantation, and pregnancy rates in patients undergoing ICSI using fresh and cryopreserved sperm from ejaculated semen samples. DESIGN:Retrospective study. SETTING:Academic and private medical centers. PATIENT(S):One hundred fifty-eight patients. INTERVENTION(S):Intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S):Fertilization, implantation, and pregnancy rates were evaluated; 61 cycles of ICSI were performed with cryopreserved and 79 cycles of ICSI were performed with fresh spermatozoa. Also, we divided the outcomes according to the semen characteristics, normozoospermia, oligozoospermia, asthenozoospermia, and oligoasthenozoospermia. RESULT(S):Overall, normal-fertilization rates were higher using fresh sperm (73.8%) compared with cryopreserved sperm (68.7%). Cycles performed in patients with normozoospermia or oligozoospermia had similar fertilization, implantation, and pregnancy rates using fresh or cryopreserved sperm. When asthenozoospermic and oligoasthenozoospermic semen samples were used, the normal-fertilization rate was higher with fresh sperm compared with cryopreserved sperm. However, implantation and pregnancy rates were similar in fresh and cryopreserved sperm samples from patients with asthenozoospermia or oligoasthenozoospermia. CONCLUSION(S):Semen with abnormalities in the motility may be more susceptible to sperm cryopreservation damage, resulting in lower fertilization rates. However, once the oocyte is fertilized, implantation and pregnancy rates are similar to those in patients with oligozoospermia and normozoospermia.

背景与目标： 目的：使用新鲜精子和冷冻保存的精子样本评估ICSI患者的受精率，着床率和妊娠率。
设计：回顾性研究。
地点：学术和私人医疗中心。
患者：158名患者。
干预：胞浆内精子注射。
主要观察指标：评估受精，着床和妊娠率。冷冻保存ICSI周期为61个周期，新鲜精子进行ICSI周期为79个周期。此外，我们根据精液特征，正常精子症，少精子症，弱精子症和少精子症精子来划分结局。
结果：与冷冻保存的精子（68.7％）相比，新鲜精子（73.8％）的总体正常受精率更高。使用新鲜或冷冻保存的精子，正常精子症或少精子症患者的周期具有相似的受精，着床和妊娠率。当使用精子少精子和精子少精子精子时，新鲜精子的正常受精率要高于冷冻精子。然而，来自弱精子症或少精子症精子症患者的新鲜和冷冻保存的精子样本中的着床率和妊娠率相似。
结论：运动能力较弱的精液更易受精子冷冻保存的损害，导致受精率降低。但是，一旦卵母细胞受精，其植入率和妊娠率与少精症和正常精子症的患者相似。

BACKGROUND & AIMS: :We analyzed the testicular spermatozoa of 23 infertile men after cryopreservation to evaluate their cryosurvival and to scrutinize the effectiveness of freezing protocol used in our institution. An excellent postthaw motility was observed indicating an effective cryosurvival protocol.

背景与目标： ：我们分析了冷冻保存后的23名不育男性的睾丸精子，以评估他们的冷冻存活率，并研究我们机构采用的冷冻方案的有效性。观察到极好的解冻后运动性，表明有效的冷冻生存方案。

BACKGROUND & AIMS: -2

背景与目标： -2

BACKGROUND & AIMS: STUDY QUESTION:Do cell penetrating peptides (CPPs) translocate into spermatozoa and, if so, could they be utilized to deliver a much larger protein cargo? SUMMARY ANSWER:Chemically diverse polycationic CPPs rapidly and efficiently translocate into spermatozoa. They exhibit differential accumulation within intracellular compartments without detrimental influences upon cellular viability or motility but they are relatively ineffective in transporting larger proteins. WHAT IS ALREADY KNOWN:Endocytosis, the prevalent route of protein internalization into eukaryotic cells, is severely compromised in mature spermatozoa. Thus, the translocation of many bioactive agents into sperm is relatively inefficient. However, the delivery of bioactive moieties into mature spermatozoa could be significantly improved by the identification and utility of an efficient and inert vectorial delivery technology. STUDY DESIGN:CPP translocation efficacies, their subsequent differential intracellular distribution and the influence of peptides upon viability were determined in bovine spermatozoa. Temporal analyses of sperm motility in the presence of exogenously CPPs utilized normozoospermic human donor samples. MATERIALS AND METHODS:CPPs were prepared by manual, automated and microwave-enhanced solid phase synthesis. Confocal fluorescence microscopy determined the intracellular distribution of rhodamine-conjugated CPPs in spermatozoa. Quantitative uptake and kinetic analyses compared the translocation efficacies of chemically diverse CPPs and conjugates of biotinylated CPPs and avidin. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) conversion assays were employed to analyse the influence of CPPs upon sperm cell viability and sperm class assays determined the impact of CPPs on motility in capacitated and non-capacitated human samples. MAIN RESULTS:Chemically heterogeneous CPPs readily translocated into sperm to accumulate within discrete intracellular compartments. Mitoparan (INLKKLAKL(Aib)KKIL), for example, specifically accumulated within the mitochondria located in the sperm midpiece. The unique plasma membrane composition of sperm is a critical factor that directly influences the uptake efficacy of structurally diverse CPPs. No correlations in efficacies were observed when comparing CPP uptake into sperm with either uptake into fibroblasts or direct translocation across a phosphatidylcholine membrane. These comparative investigations identified C105Y (CSIPPEVKFNKPFVYLI) as a most efficient pharmacokinetic modifier for general applications in sperm biology. Significantly, CPP uptake induced no detrimental influence upon either bovine sperm viability or the motility of human sperm. As a consequence of the lack of endocytotic machinery, the CPP-mediated delivery of much larger protein complexes into sperm is relatively inefficient when compared with the similar process in fibroblasts. LIMITATIONS, REASONS FOR CAUTION:It is possible that some CPPs could directly influence aspects of sperm biology and physiology that were not analysed in this study. WIDER IMPLICATIONS OF THE FINDINGS:CPP technologies have significant potential to deliver selected bioactive moieties and so could modulate the biology and physiology of human sperm biology both prior- and post-fertilization.

背景与目标： 研究问题：细胞穿透肽（CPP）是否易位进入精子，如果可以，它们是否可以用于运送更大的蛋白质？
总结：化学上多样化的聚阳离子CPP可以快速有效地转移到精子中。它们在细胞内区室中显示出不同的积累，而对细胞生存力或运动性没有不利影响，但是它们在运输较大蛋白方面相对无效。
已经知道：内吞作用，即蛋白质内在化进入真核细胞的普遍途径，在成熟的精子中被严重破坏。因此，许多生物活性剂向精子的转运是相对低效的。然而，通过鉴定和使用有效且惰性的载体递送技术可以显着改善将生物活性部分递送至成熟的精子中。
研究设计：测定牛精子中CPP的转运效率，其随后的差异细胞内分布以及肽对生存力的影响。在外源性CPPs存在下，精子活力的时间分析利用了正常精子人类供体样品。
材料与方法：CPPs是通过手动，自动和微波增强固相合成制备的。共聚焦荧光显微镜确定了若丹明结合的CPP在精子中的细胞内分布。定量吸收和动力学分析比较了化学上多样化的CPP以及生物素化CPP和抗生物素蛋白结合物的转运效率。 3-（4,5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑鎓，内盐（MTS）转化分析用于分析CPPs对精子细胞活力和精子类别测定确定了CPPs对有能力和无能力的人类样品中运动性的影响。
主要结果：化学上异质的CPP易于转移到精子中，从而在离散的细胞内区室中积累。例如，线粒体（INLKKLAKL（Aib）KKIL）专门积聚在精子中期的线粒体内。精子独特的质膜组成是直接影响结构多样的CPP吸收功效的关键因素。比较CPP摄入精子与成纤维细胞摄入或直接转运通过磷脂酰胆碱膜时，没有观察到疗效相关性。这些比较研究确定C105Y（CSIPPEVKFNKPFVYLI）是精子生物学中一般应用的最有效的药代动力学修饰剂。值得注意的是，摄取CPP不会对牛精子的生存力或人类精子的运动性产生不利影响。由于缺乏内吞作用机制，与成纤维细胞中的类似过程相比，CPP介导的将更大的蛋白质复合物送入精子的效率相对较低。
局限性，警告原因：某些CPP可能会直接影响本研究中未分析的精子生物学和生理学方面。
结果的进一步含义：CPP技术具有提供选定的生物活性部分的巨大潜力，因此可以在受精前和受精后调节人类精子生物学的生物学和生理学。