bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of the human urinary proteome: A method for high‐resolution display of urinary proteins on two‐dimensional electrophoresis gels with a yield of nearly 1400 distinct protein spots
Authors: Pieper Rembert , Gatlin Christine L. , McGrath Andrew M. , Makusky Anthony J. , Mondal Madhu , Seonarain Michael , Field Erin , Schatz Courtney R. , Estock Marla A. , Ahmed Nasir , Anderson Norman G. , Steiner Sandra 
Published: March 2004

AbstractThe abundance profile of the human urinary proteome is known to change as a result of diseases or drug toxicities, particularly of those affecting the kidney and the urogenital tract. A consequence of such insults is the ability to identify proteins in urine, which may be useful as quantitative biomarkers. To succeed in discovering them, reproducible urine sample preparation methods and good protein resolution in two‐dimensional electrophoresis (2‐DE) gels for parallel semiquantitative protein measurements are desirable. Here, we describe a protein fractionation strategy enriching proteins of molecular masses (Mr) lower than 30 kDa in a fraction separate from larger proteins. The fraction containing proteins with Mrs higher than 30 kDa was subsequently subjected to immunoaffinity subtraction chromatography removing most of the highly abundant albumin and immunoglobulin G. Following 2‐DE display, superior protein spot resolution was observed. Subsequent high‐throughput mass spectrometry analysis of ca. 1400 distinct spots using matrix‐assisted laser desorption/ionization‐time of flight peptide mass fingerprinting and liquid chromatography‐electrospray ionization tandem mass spectrometry lead to the successful identification of 30% of the proteins. As expected from high levels of post‐translational modifications in most urinary proteins and the presence of proteolytic products, ca. 420 identified spots collapsed into 150 unique protein annotations. Only a third of the proteins identified in this study are described as classical plasma proteins in circulation, which are known to be relatively abundant in urine despite their retention to a large extent in the glomerular blood filtration process. As a proof of principle that our urinary proteome display effort holds promise for biomarker discovery, proteins isolated from the urine of a renal cell carcinoma patient were profiled prior to and after nephrectomy. Particularly, the decrease in abundance of the kininogen 2‐DE gel spot train in urine after surgery was striking.
A combination of proteomics, principal component analysis and transcriptomics is a powerful tool for the identification of biomarkers for macrophage maturation in the U937 cell line
Authors: Verhoeckx Kitty C. M. , Bijlsma Sabina , de Groene Els M. , Witkamp Renger F. , van der Greef Jan , Rodenburg Richard J. T. 
Published: March 2004

AbstractThe monocyte‐like human histiocytic lymphoma cell line U937 can be induced by phorbol 12‐myristate 13‐acetate (PMA) to undergo differentiation into a macrophage‐like phenotype. We have used two‐dimensional gel electrophoresis (2‐DE), oligonucleotide microarrays and principal component analysis (PCA) to characterize the U937 cell line as a model system for the differentiation of monocytes into macrophages. A total of 226 differentially expressed proteins were found, of which 41 were selected by PCA for identification using matrix‐assisted laser desorption/ionization tandem mass spectrometry. Based on the PCA results, three marker proteins were selected for confirmation of differential expression using Western blot and quantitative real time‐PCR. The selected marker proteins were: gamma interferon inducible lysosomal thiol reductase, cathepsin D and adipocyte‐fatty acid binding protein. All three proved to be good differentiation markers for macrophage maturation of U937 cells as well as peripheral blood‐derived macrophages. The transcriptomics data revealed a large number of additional putative differentiation markers in U937 macrophages, many of which are known to be expressed in peripheral blood‐derived macrophages. These include osteospontin, matrix metalloproteinase 9, and HC‐gp39. Our results show that the characteristics of U937 macrophages resemble those of inflammatory (exudate) macrophages, exemplified by the down‐regulation of 5' nucleotidase and the up‐regulation of leucine aminopeptidase mRNAs. In conclusion, using the powerful combination of transcriptomics, 2‐DE and PCA, our results show that U937 cells differentiated by PMA treatment are an excellent model system for monocyte derived macrophage generation from blood.
Quantification of C‐reactive protein in the serum of patients with rheumatoid arthritis using multiple reaction monitoring mass spectrometry and 13C‐labeled peptide standards
Authors: Kuhn Eric , Wu Jiang , Karl Johann , Liao Hua , Zolg Werner , Guild Brad 
Published: March 2004

AbstractA general method for the quantification of proteins in human serum was developed using mass spectrometry (MS) and stable isotope‐labeled synthetic peptides as internal standards. Using this approach, C‐reactive protein (CRP), a diagnostic marker of rheumatoid arthritis (RA), was detected in serum samples taken from patients with either erosive or nonerosive RA and compared to healthy individuals. Small volumes of serum samples were enriched for low‐abundance proteins through the selective removal of human serum albumin (HSA), immunoglobulin G (IgG), and haptoglobin. After depletion of abundant proteins, the complexity of the protein mixture was further simplified using size exclusion chromatography (SEC) to fractionate denatured proteins into discrete molecular weight ranges. Fractions of interest containing CRP, Mr = 25 000, were pooled, digested with trypsin, and then fixed quantities of the synthetic peptides were added to the mixture. The mixture of tryptic peptides was subsequently analyzed by nanoflow chromatography‐tandem MS (nanoLC‐MS/MS) using multiple‐reaction monitoring (MRM) on a triple quadrupole mass spectrometer (TQ‐MS). The ratio of transition ions derived from the endogenous and isotope‐labeled peptides provided a quantitative measure of CRP in the original samples as assessed by independent measurement of CRP in the same patient samples using an immunoassay. The use of isotope‐labeled synthetic peptides and MRM is a powerful analytical method for the prescreening of candidate protein biomarkers in human serum prior to antibody and immunoassay development.
Organic solvent extraction of proteins and peptides from serum as an effective sample preparation for detection and identification of biomarkers by mass spectrometry
Authors: Chertov Oleg , Biragyn Arya , Kwak Larry W. , Simpson John T. , Boronina Tatiana , Hoang Van M. , Prieto DaRue A. , Conrads Thomas P. , Veenstra Timothy D. , Fisher Robert J. 
Published: March 2004

AbstractA method to extract peptides and low molecular weight proteins from serum under denaturing conditions using acetonitrile containing 0.1% trifluoroacetic acid has been developed. The extraction procedure precipitates large, abundant proteins to simplify subsequent mass spectral analysis. This sample preparation method provides an efficient way to extract serum peptides, enabling them to be compared and identified using different mass spectrometry approaches. Surface‐enhanced laser desorption/ionization‐time of flight mass spectrometry analysis of mouse blood serum samples prepared by this method allowed detection of two markers which were significantly reduced in mice with B cell lymphoma tumor. One of these markers has been identified as apolipoprotein A‐II.
Contribution of the antibodies response induced by a low virulent Candida albicans strain in protection against systemic candidiasis
Authors: Fernández‐Arenas Elena , Molero Gloria , Nombela César , Diez‐Orejas Rosalía , Gil Concha 
Published: March 2004

AbstractA low virulent Candida albicans mutant, CNC13, deleted in the Mitogen Activated Protein (MAP) kynase HOG1 was used to immunize BALB/c mice. Hog1p is essential for the oxidative stress and hyperosmolarity responses. Several doses and immunization procedures were employed. The protection capacity of the different sera generated was analyzed in a murine model of systemic candidiasis. Using a proteomic approach (two‐dimensional gel electrophoresis followed by Western blotting), we were able to distinguish two categories of serum: protective and nonprotective, which showed different titres of total Immunoglobulins (Igs) and IgG2a (analyzed by enzyme‐linked immunosorbent assay). The levels of Igs and IgG2a in protective sera were significantly higher compared to nonprotective sera. The pattern of a “nonprotective” profile was composed of enolase (Eno1p), transketolase, heat shock protein and methionine synthase. Only antibodies against enolase are the IgG2a isotype. The pattern of a “protective” sera, on the other hand, was composed of antibodies against the following antigens: several isoforms of Eno1p, pyruvate decarboxylase, pyruvate kynase, a protein of the 40S ribosomal subunit, triosephosphate isomerase, DL‐glycerol phosphatase and fructose‐bisphosphate aldolase. All these antibodies are the IgG2a isotype. The proteins described in the protective sera might be useful for future vaccine development.
Loss of BMP2, Smad8, and Smad4 expression in prostate cancer progression
Authors: Horvath Lisa G. , Henshall Susan M. , Kench James G. , Turner Jennifer J. , Golovsky David , Brenner Phillip C. , O'Neill Gordon F. , Kooner Raji , Stricker Phillip D. , Grygiel John J. , Sutherland Robert L. 
Published: March 2004

AbstractBACKGROUNDThe role of the bone morphogenetic protein (BMP) pathway in prostate cancer (PC) is unclear. This study aimed to characterize aspects of the BMP pathway in PC by assessing BMP2, Smad8, and Smad4 expression in normal, hyperplastic, and malignant prostate tissue, and to correlate findings with progression to PC.METHODSRadical prostatectomy (RP) specimens from 74 patients with clinically localized PC (median follow‐up 51 months, range 15–152), 44 benign prostatic hypertrophy (BPH) lesions, and 4 normal prostates (NPs) were assessed for BMP2, Smad8, and Smad4 expression using immunohistochemistry.RESULTSBoth BMP2 (P < 0.001) and nuclear Smad4 (P < 0.0001) expression were significantly decreased in PC compared to benign prostate tissue. Nuclear Smad8 was present in normal/benign prostate tissue but absent in PC and adjacent hyperplasia. Furthermore, loss of BMP2 (P < 0.001) and decreased nuclear Smad4 (P = 0.05) expression correlated with increasing Gleason score.CONCLUSIONSThese data suggest that decreased BMP2, nuclear smad8 and nuclear Smad4 expression are associated with the progression to PC, and in particular loss of BMP2 and Smad4 are related to progression to a more aggressive phenotype. © 2004 Wiley‐Liss, Inc.
The specific role of isoflavones in reducing prostate cancer risk
Authors: Kumar Nagi B. , Cantor Alan , Allen Kathy , Riccardi Diane , Besterman‐Dahan Karen , Seigne John , Helal Mohamad , Salup Raoul , Pow‐Sang Julio 
Published: March 2004

AbstractAIMSTo evaluate the effectiveness of supplementing a group of early stage prostate cancer patients, with 60 mg of soy isoflavones in producing a change in hormonal and proliferative risk parameters that are implicated in prostate cancer promotion.METHODSSeventy six eligible prostate cancer patients with a Gleason score of 6 or below, between ages 50 and 80 were admitted and supplemented with soy isoflavones or placebo for a 12 week period and changes in PSA and steroid hormones were analyzed at baseline and post intervention.RESULTSFifty‐nine patients completed the 12‐week intervention. Serum free testosterone was reduced or showed no change in 61% of subjects in the isoflavone group compared to 33% in the placebo group. Serum total PSA decreased or was unchanged in 69% of the subjects in the isoflavone treated group compared to 55% in the placebo group. However, we did not see an increase in SHBG levels. Nineteen percent of subjects receiving soy isoflavones reduced total PSA by two points or more during the intervention period.CONCLUSIONSThese data suggest that supplementing early stage prostate cancer patients with soy isoflavones, even in a study of short duration, altered surrogate markers of proliferation such as serum PSA and free testosterone in a larger number of subjects in the isoflavone supplemented group than the group receiving placebo. The study establishes the need to explore further the effects of prolonged and consistent soy consumption, which could potentially delay onset of histologic disease in this patient population. © 2004 Wileey‐Liss, Inc.
Modulation of anti‐adhesion molecule MUC‐1 is associated with arctiin‐induced growth inhibition in PC‐3 cells
Authors: Huang Dong‐Ming , Guh Jih‐Hwa , Chueh Shih‐Chieh , Teng Che‐Ming 
Published: March 2004

AbstractBACKGROUNDLignans have been reported to possess anti‐tumor activity in various cancer cells. However, their anticancer effects in human prostate cancer have not been well established. Here, we examine the effect of arctiin, a lignan compound, on growth regulation in prostate cancer PC‐3 cells. We postulated that arctiin modulates the attachment/detachment of PC‐3 cells and we investigated the role of arctiin on MUC‐1 expression.METHODSThe effect of arctiin on PC‐3 cell growth was examined using an MTT assay method and cell number was calculated by means of a standard regression line. The expressions of MUC‐1 and integrins α2, α5, and β1 were detected using FACScan flow cytometric analysis. Levels of MUC‐1 mRNA were determined using reverse transcriptase PCR (RT‐PCR).RESULTSTreatment of PC‐3 cells with arctiin decreased the cell number in a concentration‐ and time‐dependent manner in serum‐containing condition. Arctiin preferentially induced cell detachment, but did not have anti‐proliferation or cytotoxic effects in PC‐3 cells. The arctiin‐induced effect was inhibited by cycloheximide, indicating that protein synthesis was required. FACScan flow cytometric analysis demonstrated that arctiin increased the expression of the anti‐adhesion mucin MUC‐1, but did not affect integrin expression in PC‐3 cells. The arctiin‐induced increase in MUC‐1 protein expression was due to up‐regulation of mRNA, as revealed by RT‐PCR analysis.CONCLUSIONSArctiin significantly induces cell detachment and decreases the cell numbers via the up‐regulation of MUC‐1 mRNA and protein in PC‐3 cells. © 2004 Wileey‐Liss, Inc.
Androgen receptor CAG repeat length correlates with semen PSA levels in adolescence
Authors: Giwercman Yvonne Lundberg , Richthoff Jonas , Lilja Hans , Anderberg Camilla , Abrahamsson Per‐Anders , Giwercman Aleksander 
Published: March 2004

AbstractBACKGROUNDAndrogens exert their action through the androgen receptor (AR). The length of the AR CAG repeat is inversely correlated to receptor function and short CAG length might be a risk factor for development of prostate cancer. Our aim was to investigate whether CAG repeat number might have an impact on prostate function in adolescence.METHODSAR genotyping was performed by direct sequencing of leukocyte DNA from 274 military conscripts. All men underwent endocrine evaluation and semen analysis.RESULTSPSA in seminal plasma, total sperm count and motility all are inversely correlated with CAG numbers (rho = −0.128, P = 0.038; rho = −0.156, P = 0.010; rho = −0.158, P = 0.011), whereas serum levels of free testosterone (rho = 0.132; P = 0.029) and luteinizing hormone (rho = 0.126; P = 0.037) are positively correlated to CAG length. No correlation between seminal PSA and serum testosterone, neither free nor total, was found.CONCLUSIONSIn adolescence, AR genotype, but not serum testosterone, is associated with the level of seminal PSA. © 2004 Wileey‐Liss, Inc.
Murine prostate cancer inhibits both in vivo and in vitro generation of dendritic cells from bone marrow precursors
Authors: Tourkova Irina L. , Yamabe Kazuo , Foster Barbara , Chatta Gurkamal , Perez Lori , Shurin Galina V. , Shurin Michael R. 
Published: March 2004

AbstractBACKGROUNDThere is increasing evidence to suggest that dendritic cells (DC) are functionally impaired in tumor bearing hosts. However there is little or no data on the effects of murine prostate cancer (CaP) on DC generation from bone marrow precursors.METHODSFlow cytometry, mixed leukocyte reactions (MLR), and immunohistochemical analyses were used to characterize DC in CaP.RESULTSDC generated in the presence of CaP cell lines RM1 and the cell line C2 from the transgenic adenocarcinoma of the mouse prostate (TRAMP) mouse in a Transwell system expressed significantly lower levels of DC differentiation markers. This effect was confirmed when TK‐neo‐transfected RM1 cells were directly added to DC cultures and eliminated 5 days later using gancyclovir (GCV). Furthermore, co‐incubation of DC with CaP cells resulted in a decrease in the stimulatory capacity of DC to induce T cell proliferation in the MLR assay. These results were further confirmed in vivo in two different murine models of CaP: i) DC generated from mice intrafemorally injected with TK‐neo‐transfected RM1 cells; and ii) in DC generated from TRAMP mice.CONCLUSIONSThe generation and function of DC are significantly suppressed in the CaP microenvironment in both in vivo and in vitro murine models. © 2004 Wileey‐Liss, Inc.
Canine prostate stimulates osteoblast function using the endothelin receptors
Authors: LeRoy Bruce E. , Sellers Rani S. , Rosol Thomas J. 
Published: March 2004

AbstractBACKGROUNDBone metastases are common in humans and dogs with late‐stage prostate cancer. A unique feature of prostate cancer metastases is new bone formation at metastatic sites (“osteoblastic metastases”). Many carcinomas that metastasize to bone cause bone destruction, not new bone formation. The mechanisms by which prostate cancer induces bone formation at sites of bone metastasis are not well understood. We hypothesized that stimulation of osteoblasts by prostate tissue at metastatic sites was due to the paracrine actions of growth factors produced by prostate epithelial cells.METHODSWe have previously shown that normal canine prostate tissue induced new bone formation when implanted adjacent to the calvarium of nude mice. To complement this in vivo model, we developed an in vitro system of prostate‐stimulated osteoblast function to investigate mechanisms of prostate‐induced new bone formation.RESULTSWe found that treatment of cultured rat calvaria for 24 hr with proteins from normal dog prostate stimulated alkaline phosphatase activity in a dose‐dependent manner 4–6 fold compared to controls. Stimulation began approximately 8 hr after treatment, and was diminished after 72 hr. Calvaria treated with homogenates of normal dog salivary gland, kidney, bladder, and muscle did not increase ALP activity. Pretreatment of the calvaria for 1 hr with endothelin antagonists, but not anti‐parathyroid hormone‐related protein (PTHrP) antibody or indomethacin, abrogated the stimulation of ALP.CONCLUSIONSOur results indicated that osteoblast activation by canine prostate occurs via an endothelin‐dependent mechanism, and that PTHrP or prostaglandin synthase‐mediated pathways are likely not involved. This is a reliable, reproducible assay for determining the roles of molecules important in the activation of osteoblasts by the prostate. © 2004 Wileey‐Liss, Inc.
Overexpressing leptin genetic polymorphism (−2548 G/A) is associated with susceptibility to prostate cancer and risk of advanced disease
Authors: Ribeiro Ricardo , Vasconcelos André , Costa Sandra , Pinto Daniela , Morais Antonio , Oliveira Jorge , Lobo Francisco , Lopes Carlos , Medeiros Rui 
Published: March 2004

AbstractBACKGROUNDLeptin has been consistently associated with angiogenesis and tumoral growth. A G/A single nucleotide polymorphism (SNP) at the −2548 site in leptin gene (LEP) is associated with overexpression of leptin (A‐allele).METHODSWe evaluated DNA samples from 268 (536 alleles) unrelated individuals, 118 healthy controls (HCs) and 150 prostate cancer (PC) patients, for leptin gene (LEP) locus −2548 genotypes.RESULTSWe found an overrepresentation of the A‐allele in PC patients and that there is a significantly higher risk for PC among A carriers (OR = 1.60; confidence interval (CI), 1.13–2.28, P = 0.008). Linear trend analysis showed that quantitative increase of A‐allele presence was associated with significantly higher risk for PC (P = 0.003) in heterozygous (OR = 2.11; CI, 1.20–3.71) and homozygous (OR = 2.93; CI, 1.27–6.75) genotypes. Furthermore, the AA and AG genotypes represent significantly higher risk (OR = 4.67; CI, 1.69–12.88 and OR = 2.58; CI, 1.19–5.58, respectively) for advanced disease.CONCLUSIONSAccording to our results we hypothesize that the polymorphism in LEP gene may be relevant to PC risk and progression, supporting the hypothesis for leptin involvement in cancer ethiopathogenesis. © 2004 Wiley‐Liss, Inc.
USF2 is connected to GAAAATATGATA element and associates with androgen receptor‐dependent transcriptional regulation in prostate
Authors: Kivinen Anne , Patrikainen Lila , Kurkela Riitta , Porvari Katja , Vihko Pirkko 
Published: March 2004

AbstractBACKGROUNDWe have previously identified a GAAAATATGATA binding site (pros) of a transcription factor involved in prostatic and androgen‐dependent gene regulation. We now purified the potential factors interacting with the pros and characterized their co‐operation with the androgen receptor (AR).METHODSSequence‐specific DNA affinity chromatography, mass‐spectrometry, electromobility shift assays, supershifts, glutathione‐S‐transferase pull‐downs, and transient transfections.RESULTSSeveral proteins bound to the pros site, but only upstream stimulatory factor 2 (USF2) was confirmed to be part of the transcription factor complex. Weak interaction was detected between AR and the transcription factor complex. Physical proximity between the androgen response element (ARE) and the pros was shown to be important for their co‐operation. In the presence of pros and androgen, AR achieves its maximal efficiency even at low concentrations.CONCLUSIONSThe protein complex binding to the GAAAATATGATA site does not have a significant independent function, but may interact with AR if GAAAATATGATA is physically close to the ARE and enhances the transactivation function of AR. © 2004 Wiley‐Liss, Inc.
Serum osteoprotegerin (OPG) levels are associated with disease progression and response to androgen ablation in patients with prostate cancer
Authors: Eaton Colby L. , Wells Jason M. , Holen Ingunn , Croucher Peter I. , Hamdy Freddie C. 
Published: March 2004

AbstractBACKGROUNDOsteoprotegerin (OPG) is a tumour and/or bone derived factor that may protect tumour cells from apoptosis. In this study, we have measured serum OPG levels in untreated prostate cancer patients with advanced prostate cancer compared to patients with organ confined disease and in treated patients receiving androgen ablation.METHODSSerum OPG levels were measured by ELISA in samples collected from 104 patients with either newly diagnosed (n = 59) or advanced prostate cancer treated by androgen ablation (n = 45) and compared with levels in serum from patients with benign prostatic hyperplasia (BPH) (n = 10) and young healthy men (n = 10).RESULTSUntreated patients with locally advanced disease had significantly higher OPG levels than those with organ confined disease. Patients with advanced disease responding to androgen ablation (serum PSA < 1 ng/ml) had serum OPG levels that were significantly lower than those with clinically progressing disease (PSA > 10 ng/ml). OPG levels in the latter were not significantly different from levels in patients with early signs of biochemical progression (PSA >1 but <10 ng/ml).CONCLUSIONSOPG is a potential new marker, which is elevated in the serum of patients with advanced prostate cancer and may be an indicator of early disease progression. © 2004 Wiley‐Liss, Inc.