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Human Seminal Fluid (human + seminal_fluid)
Selected AbstractsIntegrated selective enrichment target , a microtechnology platform for matrix-assisted laser desorption/ionization-mass spectrometry applied on protein biomarkers in prostate diseasesELECTROPHORESIS, Issue 21-22 2004Simon Ekström Abstract The performance of a miniaturized sample processing platform for matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS), manufactured by silicon microfabrication, called integrated selective enrichment target (ISET) technology was evaluated in a biological context. The ISET serves as both sample treatment device and MALDI-MS target, and contains an array of 96 perforated nanovials, which each can be filled with 40 nL of reversed-phase beads. This methodology minimizes the number of sample transfers and the total surface area available for undesired adsorption of the analytes in order to provide high-sensitivity analysis. ISET technology was successfully applied for characterization of proteins coisolated by affinity chromatography of prostate-specific antigen (PSA) from human seminal fluid. The application of ISET sample preparation enabled multiple analyses to be performed on a limited sample volume, which resulted in the discovery that prolactin inducible protein (PIP) was coisolated from the samples. [source] Heparin-binding proteins of human seminal plasma: purification and characterizationMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 12 2008Vijay Kumar Abstract Human seminal plasma (HuSP) contains several proteins that bind heparin and related glycosaminoglycans. Heparin binding proteins (HBPs) from seminal plasma have been shown to participate in modulation of capacitation or acrosome reaction and thus have been correlated with fertility in some species. However, these have not been studied in detail in human. The objective of this study was to purify major HBPs from HuSP in order to characterize these proteins. HBPs were isolated by affinity,chromatography on Heparin,Sepharose column, purified by reverse-phase high-performance liquid chromatography (RP-HPLC) and Size-exclusion chromatography and checked for purity on sodium-dodecyl PAGE (SDS,PAGE). Identification of HBPs was done by matrix-assisted laser desorption-ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS). Here we report the purification and identification of seven HBPs in seminal fluid. The major HBPs are lactoferrin and its fragments, semenogelin I fragments, semenogelin II, prostate specific antigen, homolog of bovine seminal plasma-proteins (BSP), zinc finger protein (Znf 169) and fibronectin fragments. In this study we are reporting for the first time the purification and identification of BSP-homolog and Znf 169 from HuSP and classified them as HBPs. Here we report the purification of seven clinically important proteins from human seminal fluid through heparin affinity chromatography and RP-HPLC, in limited steps with higher yield. Mol. Reprod. Dev. 75: 1767,1774, 2008. © 2008 Wiley-Liss, Inc. [source] Fast and novel purification method to obtain the prostate specific antigen (PSA) from human seminal plasmaTHE PROSTATE, Issue 10 2006Boris Acevedo Abstract Background Prostate specific antigen (PSA) is a relevant antigen in diagnosis; follow-up, and therapeutic approaches for fighting the prostate cancer. Several methods have been published previously to obtain a high purity preparation of PSA. In general, these methods are expensive, time-consuming, laborious, and in some cases produce low yields. Methods Based on a panel of 7 anti-PSA Mab's we carried on binding and elution experiments of PSA antigen in 96-well plates. The selected Mab were immobilized in a Sepharose CL-4B activated matrix with the purpose of purify PSA from human seminal fluid. In order to optimize the purification procedure, we test several washing and elution conditions (chaotropic agents, high ionic strength solution, and extreme pH). Results We selected a high ionic strength solution (2 M MgCl2) as elution condition, and a previous washing step with a mix of two ionic solutions (2.5 M NaCl pH 8/1 M MgCl2 pH 5.5) in order to purify PSA. Using such conditions we obtained a PSA preparation with 90% of purity and 50% of recovery. Conclusion In this article, we report a simple, quickly, and non-expensive procedure to obtain free-PSA from human seminal plasma at high purity levels. Prostate 66: 1029,1036, 2006. © 2006 Wiley-Liss, Inc. [source] The in vitro effects of melatonin on human sperm function and its scavenging activities on NO and ROSANDROLOGIA, Issue 2 2010S. S. Du Plessis Summary Various systems of antioxidants exist endogenously in the body to help protect it against free radical damage by scavenging excessive ROS and RNS. Melatonin, a hormone secreted by the pineal gland, and responsible for controlling the circadian rhythm, is one such endogenous antioxidant. Melatonin has been reported to be present in human seminal fluid, but its antioxidant activities in semen are rather contradictory. This study aimed at establishing the effects of melatonin treatment on human spermatozoa. Spermatozoa were incubated with 2 mm melatonin (120 min, 37 °C, 5% CO2) after which motility parameters were measured by computer aided motility analysis, while cell viability (PI), intracellular NO (DAF-2/DA) and ROS (DCFH-DA) were assessed using flow cytometry. In vitro melatonin treated samples (n = 12) showed a significantly higher percentage of motile, progressive motile and rapid cells, while simultaneously reducing the number of nonviable spermatozoa when compared with the control. Endogenous NO was significantly decreased, but no effect was observed on ROS levels. From these results, it can be concluded that melatonin was able to directly or indirectly scavenge NO, as indicated by the reduction in 4,5-diaminofluorescein-2/diacetate fluorescence. Future studies will indicate whether melatonin treatment during sperm preparation techniques could protect spermatozoa from excessive NO production. [source] | |||||||||||||