Shotgun Proteomic Analysis of Vaginal Fluid From Women in Late Pregnancy

Klein, Laura L.; Jonscher, Karen R.; Heerwagen, Margaret J.; Gibbs, Ronald S.; McManaman, James L.

doi:10.1177/1933719107311189

Cited by 38 publications

(41 citation statements)

References 51 publications

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“…This may explain why statistical significance was not observed in the validation cohort. FABP5 has been identified in the CVF by at least eight other groups but the existence of different isoforms could not be described using the technologies employed (Venkataraman et al 2005, Shaw et al 2007, Tang et al 2007, Klein et al 2008, Shah et al 2009, Zegels et al 2009). The significance of the different isoforms therefore remains unclear.…”

Section: Discussionmentioning

confidence: 99%

“…The CVF proteome has been comprehensively characterised in pregnant and non-pregnant women , Di Quinzio et al 2007, Shaw et al 2007, Tang et al 2007, Klein et al 2008, Shah et al 2009, Zegels et al 2009, Heng et al 2010a. Using two-dimensional PAGE (2D-PAGE) proteomic analysis of human CVF, we have demonstrated temporal protein changes in association with impending spontaneous term labour (Di Quinzio et al 2008, Heng et al 2010a.…”

Section: Introductionmentioning

confidence: 99%

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Proteomic analysis of human cervicovaginal fluid collected before preterm premature rupture of the fetal membranes

et al. 2013

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A significant obstetric complication facing contemporary materno-fetal medicine is preterm premature rupture of the fetal membranes (preterm PROM), which occurs in 30% of all preterm births. The objective of this study was to identify differentially expressed proteins in the cervicovaginal fluid of asymptomatic women before the clinical manifestation of preterm PROM. The preterm PROM group comprised of women with samples collected 6-23 days before PROM, who subsequently delivered preterm (nZ5). Women who spontaneously delivered at term served as gestation-matched controls (nZ10). Two-dimensional difference in-gel electrophoresis was used to distinguish differential expression between the pooled groups and fold changes were subsequently confirmed by two-dimensional PAGE of individual samples. Spots of interest were identified by mass spectrometry. Proteins that were significantly reduced with impending preterm PROM included the following: thioredoxin (2.7-fold), interleukin 1 receptor antagonist (1.7-fold), fatty acid-binding protein 5 (2.1-fold), cystatin A (dimer; 1.9-fold), monocyte/neutrophil elastase inhibitor (1.6-fold), squamous cell carcinoma antigen-1 (2.1-fold) and g-glutamyl cyclotransferase (3.0-fold). By contrast, annexin A3 (3.7-fold) and vitamin D binding protein (3.9-fold) were significantly increased with impending preterm PROM. Western blot analysis was also performed on an independent cohort of preterm PROM and control samples to validate these candidate biomarkers. These proteins have known biological functions in oxidative balance, anti-inflammatory activity, metabolism or protease inhibition that may facilitate membrane rupture.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Proteomic analysis of human cervicovaginal fluid collected before preterm premature rupture of the fetal membranes

et al. 2013

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“…We developed a protocol for processing the residual Pap test fluid so that peptides can be analyzed by MS/MS and proteins identified from the Human Uniprot database. Finally, we found extensive overlap between the proteins that we define as our “Normal Pap test Core Proteome” and lists of cervical-vaginal fluid proteins identified by others using different sampling methods [10,11,13-15,17-22]. …”

Section: Introductionmentioning

confidence: 87%

A feasibility study to identify proteins in the residual Pap test fluid of women with normal cytology by mass spectrometry-based proteomics

et al. 2014

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BackgroundThe proteomic analysis of body fluids is a growing technology for the identification of protein biomarkers of disease. Given that Papanicolaou tests (Pap tests) are routinely performed on over 30 million women annually in the U.S. to screen for cervical cancer, we examined the residual Pap test fluid as a source of protein for analysis by mass spectrometry (MS). In the liquid-based Pap test, cervical cells are collected from the ectocervix and placed into an alcohol-based fixative prior to staining and pathologic examination. We hypothesized that proteins shed by cells of the female genital tract can be detected in the Pap test fixative by MS-based proteomic techniques. We examined the feasibility of using residual fluid from discarded Pap tests with cytologically “normal” results to optimize sample preparation for MS analysis. The protein composition of the cell-free Pap test fluid was determined by silver staining of sodium dodecyl sulfate -polyacrylamide gels, and the abundance of serum proteins was examined by Western immunoblot using an antibody against human serum albumin. Both pooled and individual samples were trypsin digested and analyzed by two-dimensional MS/MS. Proteins were identified by searching against the Human Uniprot database, and characterized for localization, function and relative abundance.ResultsThe average volume of the residual Pap test fluid was 1.5 ml and the average protein concentration was 0.14 mg/ml. By Western immunoblot we showed that the amount of albumin in each sample was significantly reduced compared to normal serum. By MS/MS, we identified 714 unique proteins in pooled Pap test samples and an average of 431 proteins in individual samples. About 40% of the proteins identified were extracellular or localized to the plasma membrane. Almost 20% of the proteins identified were involved in immunity and defense, characteristic of the healthy cervical-vaginal proteome. By merging the protein sets from the individual and pooled Pap test samples, we created a “Normal Pap test Core Proteome” consisting of 153 proteins.ConclusionsResidual Pap test fluid contains a sufficient amount of protein for analysis by MS and represents a valuable biospecimen source for the identification of protein biomarkers for gynecological diseases.

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“…The CVF proteome was analyzed in humans in various physiological conditions (1,2), mainly during pregnancy (3)(4)(5)(6)(7) or in several pathologies such as human papilloma virus (8) or candida albicans (9) infections. But few studies have investigated the CVF proteome during the cycle (10,11).…”

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confidence: 99%

Proteomes of the Female Genital Tract During the Oestrous Cycle

Soleilhavoup

Riou

Tsikis

et al. 2016

Molecular & Cellular Proteomics

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The success of fertilisation in mammals is linked to the correct migration of spermatozoa in the different compartments of the female genital tract and the adequate timing of their interaction with the female gamete. In many mammalian species including human, the deposition of semen in the vagina is followed by the sperm migration through the cervix, the uterus and then the oviduct before reaching the site of fertilisation. This sperm transit within the female tract includes mechanical and biochemical interactions with the luminal fluids leading to selection of spermatozoa able to fertilize the oocyte.The first physiological barrier the spermatozoa will have to go through is the cervix whose vaginal side is covered by a highly viscous mucus, the cervical vaginal fluid (CVF) 1 . The CVF proteome was analyzed in humans in various physiological conditions (1, 2), mainly during pregnancy (3-7) or in several pathologies such as human papilloma virus (8) or candida albicans (9) infections. But few studies have investigated the CVF proteome during the cycle (10, 11). The amount of CVF increases at the time of ovulation concomitantly with a higher state of hydration and a reduced viscosity, to facilitate sperm migration (12). The mechanical properties of the mucus are essential to select spermatozoa with the highest fertilizing ability, i.e. with normal morphology and efficient mobility. The main structural components of the cervical mucus are mucins, highly glycosylated, high-molecular-weight proteins assembled into a filamentous and viscous mesh (13). The transcription of the mucin genes is increased at oestrus in the bovine cervical epithelium (14). The amount of mucins and their level of glycosylation are expected to contribute to the viscosity of the mucus (11). Therefore, the quantification of proteins such From the ‡INRA, UMR 85 Physiologie de la Reproduction et des Comportements,

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Shotgun Proteomic Analysis of Vaginal Fluid From Women in Late Pregnancy

Cited by 38 publications

References 51 publications

Proteomic analysis of human cervicovaginal fluid collected before preterm premature rupture of the fetal membranes

Proteomic analysis of human cervicovaginal fluid collected before preterm premature rupture of the fetal membranes

A feasibility study to identify proteins in the residual Pap test fluid of women with normal cytology by mass spectrometry-based proteomics

Proteomes of the Female Genital Tract During the Oestrous Cycle

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