bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of 11 members of the BCL‐2 family of apoptosis regulatory molecules during human preimplantation embryo development and fragmentation
Authors: Metcalfe Anthony D. , Hunter Helen R. , Bloor Debra J. , Lieberman Brian A. , Picton Helen M. , Leese Henry J. , Kimber Susan J. , Brison Daniel R. 
Published: March 2004

AbstractApoptosis during preimplantation development has received much interest because of its potential role in eliminating defective cells. Although development in humans is characterised by a high degree of genetic abnormality, little is known of the regulation of apoptosis in embryos. By PolyA PCR we analysed expression of 11 BCL‐2 genes in individual human embryos representative of normal development and in severely fragmented embryos. We demonstrate constitutive expression of BAX in virtually all embryos at all stages of development, and variable expression of BCL2, BCL‐XL, BCL‐W, MCL‐1 BAK, BAD, BOKL, BID, BIK, and BCL‐XS. The frequency of expression of pro‐ and anti‐apoptotic BCL‐2 members was similar throughout development, except at the two‐cell stage where pro‐apoptotic genes predominated. Protein expression was confirmed for BCL‐2, MCL‐1, BCL‐X, BAX, BAD, and activated caspase 3. BCL‐2 protein was associated with mitochondria but expressed inconsistently in the blastocyst inner cell mass. Consistent differences between morphologically intact and fragmented embryos included the expression of BAK in fragmented but not intact four‐cell embryos. Our study addresses the importance of examining single human embryos representative of the viable population for a large number of genes, in order to establish meaningful expression profiles and provide information on overlapping function in a large gene family. Mol. Reprod. Dev. 68: 35–50, 2004. © 2004 Wiley‐Liss, Inc.
“I couldn't think that far”: Infertile women's decision making about multifetal reduction
Authors: Collopy Kate Sullivan 
Published: March 2004

AbstractIn this phenomenological study women's experiences regarding their decisions to undergo or forgo multifetal reduction of their higher‐order multiple pregnancies were explored. Seven women who had conceived higher‐order multiple pregnancies as the result of in vitro fertilization were interviewed. Four participants accepted reduction, whereas three participants declined. Three themes were discerned: (a) the presence of infertility as a barrier to contemplating hyperfertility; (b) multiple‐birth pregnancy as yet another form of loss for infertile women; and (c) the lasting effects of having made the decision. © 2004 Wiley Periodicals, Inc. Res Nurs Health 27:75–86, 2004
Modeling health insurance expansions: Effects of alternate approaches
Authors: Remler Dahlia K. , Zivin Joshua Graff , Glied Sherry A. 
Published: March 2004

AbstractEstimates of the costs and consequences of many types of public policy proposals play an important role in the
development and adoption of particular policy programs. Estimates of the same, or similar, policies that employ
different modeling approaches can yield widely divergent results. Such divergence often undermines effective
policymaking. These problems are particularly prominent for health insurance expansion programs. Concern focuses on
predictions of the numbers of individuals who will be insured and the costs of the proposals. Several different
simulation‐modeling approaches are used to predict these effects, making the predictions difficult to
compare. This paper categorizes and describes the different approaches used; explains the conceptual and
theoretical relationships between the methods; demonstrates empirically an example of the (quite
restrictive) conditions under which all approaches can yield quantitatively identical predictions; and
empirically demonstrates conditions under which the approaches diverge and the quantitative extent of that
divergence. All modeling approaches implicitly make assumptions about functional form that impose restrictions on
unobservable heterogeneity. Those assumptions can dramatically affect the quantitative predictions made. © 2004
by the Association for Public Policy Analysis and Management.
Identification of human whole saliva protein components using proteomics
Authors: Vitorino Rui , Lobo Maria João C. , Ferrer‐Correira António J. , Dubin Joshua R. , Tomer Kenneth B. , Domingues Pedro M. , Amado Francisco M. L. 
Published: March 2004

AbstractThe determination of salivary biomarkers as a means of monitoring general health and for the early diagnosis of disease is of increasing interest in clinical research. Based on the linkage between salivary proteins and systemic diseases, the aim of this work was the identification of saliva proteins using proteomics. Salivary proteins were separated using two‐dimensional (2‐D) gel electrophoresis over a pH range between 3–10, digested, and then analyzed by matrix assisted laser desorption/ionization‐time of flight (MALDI‐TOF)‐TOF mass spectrometry (MS) and tandem mass spectrometry (MS/MS). Proteins were identified using automated MS and MS/MS data acquisition. The resulting data were searched against a protein database using an internal Mascot search routine. Ninety spots give identifications with high statistical reliability. Of the identified proteins, 11 were separated and identified in saliva for the first time using proteomics tools. Moreover, three proteins that have not been previously identified in saliva, PLUNC, cystatin A, and cystatin B were identified.
Reversible labeling of cysteine‐containing peptides allows their specific chromatographic isolation for non‐gel proteome studies
Authors: Gevaert Kris , Ghesquière Bart , Staes An , Martens Lennart , Van Damme Jozef , Thomas Grégoire R. , Vandekerckhove Joël 
Published: March 2004

AbstractWe report upon a novel procedure to specifically isolate cysteine‐containing peptides from a complex peptide mixture. Cysteines are converted to hydrophobic residues by mixed disulfide formation with Ellman's reagent. Proteins are subsequently digested with trypsin and the generated peptide mixture is a first time fractionated by reverse‐phase high‐performance liquid chromatography. Cysteinyl‐peptides are isolated out of each primary fraction by a reduction step followed by a secondary peptide separation on the same column, performed under identical conditions as for the primary separation. The reducing agent removes the covalently attached group from the cysteine side chain, making cysteine‐peptides more hydrophilic and, thereby, such peptides can be specifically collected during the secondary separation and are finally used to identify their precursor proteins using automated liquid chromatography tandem mass spectrometry. We show that this procedure efficiently isolates cysteine‐peptides, making the sample mixture less complex for further analysis. This method was applied for the analysis of the proteomes of human platelets and enriched human plasma. In both proteomes, a significant number of low abundance proteins were identified next to extremely abundant ones. A dynamic range for protein identification spanning 4–5 orders of magnitude is demonstrated.
Proteome analysis of hepatocellular carcinoma cell strains, MHCC97‐H and MHCC97‐L, with different metastasis potentials
Authors: Ding Shi‐Jian , Li Yan , Shao Xiao‐Xia , Zhou Hu , Zeng Rong , Tang Zhao‐You , Xia Qi‐Chang 
Published: March 2004

AbstractTo better understand the mechanism underlying hepatocellular carcinoma (HCC) metastasis and to search for potential markers for HCC prognosis, differential proteome analysis on two HCC cell strains with high and low metastatic potentials, MHCC97‐H and MHCC97‐L, was conducted using two‐dimensional (2‐D) gel electrophoresis followed by matrix‐assisted laser desorption/time of flight mass spectrometry and liquid chromatography ion trap mass spectrometry. Image analysis of silver‐stained 2‐D gels revealed that 56 protein spots showed significant differential expression in MHCC97‐H and MHCC97‐L cells (Student's t‐test, P < 0.05) and 4 protein spots were only detected in MHCC97‐H cells. Fourteen protein spots were further identified using in‐gel tryptic digestion, peptide mass fingerprinting and tandem mass spectrometry. The expressions of pyruvate kinase M2, ubiquitin carboxy‐terminal hydrolase L1, laminin receptor 67 kDa, S100 calcium‐binding protein A4, thioredoxin and cytokeratin 19 were elevated in MHCC97‐H cells. However, manganese superoxide dismutase, calreticulin precursor, cathepsin D, lactate dehydrogenase B, non‐metastatic cell protein 1, cofilin 1 and calumenin precursor were down‐regulated in MHCC97‐H cells. Intriguingly, most of these identified proteins have been reported to be associated with tumor metastasis. The functional implications of alterations in the levels of these proteins are discussed.
Screening protein refolding using surface plasmon resonance
Authors: Jones Daniel B. , Hutchinson Matthew H. , Middelberg Anton P. J. 
Published: March 2004

AbstractSurface plasmon resonance (SPR) measurements were used to screen refolding conditions to identify a physicochemical environment which gives an acceptable refolding yield for samples of glutathione‐S‐transferase (GST) denatured in 6 M guanidine hydrochloride and 32 mM dithiothreitol. The SPR measurements were performed on carboxymethylcellulose coated chips that could accommodate two separate flow paths. One side of the chip was derivatized with immobilized glutathione and the other with goat anti‐GST antibody. This created a dual‐derivatized chip capable of showing both the presence of GST and providing a measure of enzyme activity. The dual‐derivatized chip could be regenerated using a two‐step washing procedure and reused to anaylze multiple samples from a screening study of protein refolding conditions. SPR measurements have been shown to be suitable for screening protein refolding conditions due to the high sensitivity, ease of chip regeneration and the ability to incorporate a control in the experimental design. The combination of such advantages with the high‐throughput automated SPR systems currently available may be a valuable approach to determine conditions suitable for protein refolding following insoluble expression in a bacterial host.
Differential proteomic analysis of Aeromonas salmonicida outer membrane proteins in response to low iron and in vivo growth conditions
Authors: Ebanks Roger O. , Dacanay Andrew , Goguen Michel , Pinto Devanand M. , Ross Neil W. 
Published: March 2004

AbstractAeromonas salmonicida subsp. salmonicida is the etiological agent of furunculosis, a serious infectious disease of salmonids. Bacterial phenotypes are known to change in vivo compared to the in vitro state. Proteomic analysis of in vivo phenotypes is usually not possible due to insufficient biomass. Using an in vivo growth chamber model, the pathogenic fish bacterium A. salmonicida was cultured in pure culture in vivo in its host, the Atlantic salmon, to obtain sufficient biomass to allow proteomic analysis. Growth of A. salmonicida under in vitro iron‐restricted conditions resulted in the expression of outer membrane proteins of 73, 76 and 85 kDa, which were not present when grown under in vitro iron‐replete conditions. Mass spectrometry analysis identified the 73 kDa protein as a colicin receptor, the 76 kDa protein as an outer membrane heme receptor, and the 85 kDa protein as a ferric siderophore receptor. When cultured in vivo, A. salmonicida up‐regulated the identical 73, 76 and 85 kDa proteins. The results of this study also suggest, at least with respect to the outer membrane proteins, that the in vitro iron‐restricted growth model largely reproduces the results obtained from growth of A. salmonicida within the peritoneal cavity of salmon.
Gas‐phase thermolysis of benzotriazole derivatives: part 1—synthesis of α‐N(1)‐ andN(2)‐benzotriazolyl ketones and kinetics and mechanism of their gas‐phase pyrolysis
Authors: Dib Hicham H. , Al‐Awadi Nouria A. , Ibrahim Yehia A. , El‐Dusouqui Osman M. E. 
Published: March 2004

Abstract1‐(Benzotriazol‐1‐yl)propan‐2‐one (1) and 3‐(benzotriazol‐1‐yl)butan‐2‐one (2), and their respective phenyl analogues 2‐(benzotriazol‐1‐yl)‐1‐phenylethanone (3) and 2‐(benzotriazol‐1‐yl)‐1‐phenylpropan‐1‐one (4), together with 1‐cyanobenzotriazole (5) and 1‐(benzotriazol‐2‐yl)propan‐2‐one (6), were synthesized and then subjected to gas‐phase thermolysis. The products of complete pyrolysis of1–4were found to be aniline in addition to either 2‐substituted indoles or 2,3‐disubstituted indoles. The kinetics of the gas‐phase thermal elimination reactions of the present compounds were also studied, the first such investigation on benzotriazoles. The Arrhenius pre‐exponential factor (A/s−1) and activation energy (Ea/kJ mol−1) are, respectively: 10.03 and 156.7 for (1), 9.93 and 158.0 for (2), 12.68 and 169.9 for (3), 9.45 and 144.4 for (4) and 15.40 and 163.6 for (5). Compound6appeared to have isomerized by a thermally allowed 1,5‐group migration into1prior to thermolysis. The results of the kinetic and product analysis are rationalized with reference to a mechanism involving extrusion of a stable (N2) molecule and formation of a reactive 1,3‐biradical intermediate, which in turn rearranges into the final products of reaction. Copyright © 2004 John Wiley & Sons, Ltd.
Extending the concept of kinetic stability: toward a paradigm for life
Authors: Pross Addy , Khodorkovsky Vladimir 
Published: March 2004

AbstractThe physico‐chemical relationship between living and non‐living systems remains a question of intense debate. This paper introduces the concept of dynamic kinetic stability as a means of incorporating living systems within a conventional physico‐chemical framework. Its essence, all replicating systems, both animate and inanimate, represent elements of a replicator space. In contrast to the world of non‐replicating systems (all inanimate), where selection is fundamentally thermodynamic, selection within replicator space is effectively kinetic. Driven by mutagenic autocatalysis selection in replicator space leads from kinetically less stable systems to kinetically more stable systems. Our kinetic approach suggests that all living systems may be thought of as manifesting a kinetic state of matter (as opposed to the traditional thermodynamic states), and allows the translation of key Darwinian concepts, such as fitness and natural selection, into traditional physico‐chemical terms that are applicable at the molecular level. Copyright © 2004 John Wiley & Sons, Ltd.
Small aggregates of benzoporphyrin molecules observed in water–organic solvent mixtures
Authors: Simplicio Fernanda I. , Maionchi Florângela , Filho Ourides Santin , Hioka Noboru 
Published: March 2004

AbstractBenzoporphyrins, chlorin‐type compounds, are efficient new drugs in photodynamic therapy. In this work, some aspects of the benzoporphyrin‐1,3‐diene dimethyl ester (DiesterB) aggregation were investigated in water–organic solvent mixtures, using visible spectrophotometry. The effects of solvent mixtures, water with acetonitrile, dioxane, methanol and ethanol, were compared taking the calculated aggregation equilibrium constant (K) and the aggregation number. The K dependence on temperature, DiesterB concentration and water content suggest that, in water with acetonitrile, the predominant species is a dimer, whereas in dioxane, methanol and ethanol systems, there are multiple equilibria, with a pre‐equilibrium between the monomer and dimer (small amount), followed by trimer formation. Not only the solvent polarity but also the nature of the organic solvent are important for the aggregation process, driving the aggregate size and the magnitude of the equilibrium constant. Copyright © 2004 John Wiley & Sons, Ltd.
Synthesis of new heterocyclic compounds: 4‐(3,3,3‐trifluoro‐2‐trifluoromethyl‐1‐propenyl)‐1,3‐thiazoles: structural and conformational study of 2‐phenyl‐4‐(3,3,3‐trifluoro‐2‐trifluoromethyl‐1‐propenyl)‐1,3‐thiazole
Authors: Buceta N. N. , Coyanis E. M. , Védova C. O. Della , Haas A. , Schettino M. , Winter M. 
Published: March 2004

AbstractThe synthesis of 4‐(3,3,3‐trifluoro‐2‐trifluoromethyl‐1‐propenyl)‐1,3‐thiazole derivatives from 5,5,5‐trifluoro‐4‐(trifluoromethy)‐3‐penten‐2‐one is described. Attention is mainly focused on 2‐N‐phenylamino‐4‐(3,3,3‐trifluoro‐2‐trifluoromethyl‐1‐propenyl)‐1,3‐thiazole. Structural and conformational studies using several spectroscopic methods (FT‐IR, NMR, Raman, x‐ray, MS) were performed. Geometry optimization and prediction of vibrational and magnetic properties were carried out using ab initio Hartree–Fock (HF) and density functional theory (DFT) calculations to assist the spectroscopic analysis. Copyright © 2004 John Wiley & Sons, Ltd.
Bone microenvironment‐related growth factors modulate differentially the anticancer actions of zoledronic acid and doxorubicin on PC‐3 prostate cancer cells
Authors: Tenta Roxane , Tiblalexi Despina , Sotiriou Evangelia , Lembessis Peter , Manoussakis Menelaos , Koutsilieris Michael 
Published: March 2004

AbstractOBJECTIVESWe analyzed the actions of zoledronic acid (10–250 μM) and doxorubicin (10–250 nM) on PC‐3 prostate cancer cells using both continuous (48–96 hr) and pulsatile exposures (15 min/day for up to three consecutive days).RESULTSThe proliferation of PC‐3 cells was inhibited by either continuous or pulsatile exposures of zoledronic acid in a dose‐dependent manner. In contrast, pulsatile exposures of doxorubicin failed to inhibit the growth of PC‐3 cells. In addition, the inhibition of PC‐3 cells by zoledronic acid was partially neutralized by exogenous administration of geranylgeranyl pyrophosphate (GGPP), however, not by farnesyl pyrophosphate (FPP). Furthermore, exogenous administration of transforming growth factor beta 1 (TGF‐β1), interleukin 6 (IL‐6), basic fibroblast growth factor (bFGF), and more potently, insulin‐like growth factor 1 (IGF‐1) inhibited the doxorubicin‐induced apoptosis of PC‐3 cells. Under identical experimental conditions, these growth factors failed to alter the cytotoxicity of PC‐3 cells induced by zoledronic acid.CONCLUSIONSThese data suggest that (i) repetitive and pulsatile (15 min/day) exposure to zoledronic acid inhibited the growth of PC‐3 cells, (ii) this anticancer action of zoledronic acid was partially mediated by the attenuation of GGPP production, and (iii) bone microenvironment‐related growth factors do not alter the anticancer actions of zoledronic acid on PC‐3 cells. © 2004 Wileey‐Liss, Inc.
1α,25‐dihydroxyvitamin D3 induced growth inhibition of PC‐3 prostate cancer cells requires an active transforming growth factor beta signaling pathway
Authors: Murthy Shalini , Weigel Nancy L. 
Published: March 2004

AbstractBACKGROUNDProstate cancer growth inhibition by 1α,25‐dihydroxyvitamin D3 (1,25(OH)2D3) is best characterized in the androgen dependent LNCaP cell line, where treatment with this hormone causes cell cycle arrest and apoptosis. 1,25(OH)2D3 also inhibits the growth of PC‐3 prostate cancer cells, but not through the induction of G1 arrest or apoptosis. In this study, we have sought to elucidate the mechanism/s involved in PC‐3 cell growth inhibition by 1,25(OH)2D3.EXPERIMENTAL METHODSWe determined the effect of transforming growth factor beta (TGFβ) blocking antibodies on 1,25(OH)2D3 mediated growth inhibition of PC‐3 cells. In addition, we also studied the effects of 1,25(OH)2D3 on TGFβ signaling and receptor expression. Finally, we assessed the role of TGFβ signaling in the induction of the growth inhibitory protein, insulin like growth factor binding protein 3 (IGFBP‐3), by 1,25(OH)2D3.RESULTSWe find that 1,25(OH)2D3 action in PC‐3 cells is mediated through at least two distinct pathways, the TGFβ pathway and the IGFBP‐3 pathway. We show that 1,25(OH)2D3 treatment elevates TGFβ production and signaling, as well as receptor levels, in PC‐3 cells. Further, using a blocking antibody against TGFβ substantially reduces 1,25(OH)2D3 mediated growth inhibition without affecting IGFBP‐3 induction, suggesting that IGFBP‐3, alone, is insufficient to inhibit the growth of PC‐3 cells. © 2004 Wiley‐Liss, Inc.
Expression of serotonin receptors and role of serotonin in human prostate cancer tissue and cell lines
Authors: Dizeyi N. , Bjartell A. , Nilsson E. , Hansson J. , Gadaleanu V. , Cross N. , Abrahamsson P.‐A. 
Published: March 2004

AbstractBACKGROUNDIncrease in the number of serotonin (5‐HT) releasing neuroendocrine (NE) cells has been shown to be correlated with tumor progression, loss of androgen dependence, and poor prognosis. Serotonin is a well‐known mitogen which mediates a wide variety of physiological effects via multiple receptors, of which receptor subtype 1 (5‐HTR1) has been identified in prostate cancer (PC) cell lines. Recently, 5‐HT has been found to show growth‐promoting activity and to be functionally related to oncogenes.MATERIALS AND METHODSLocalization, protein content, and mRNA expression of 5‐HTR subtype 1A, 1B, and 1D was studied in prostatic tissue (35 patients), metastases, PC cell lines, a benign prostatic stromal cell line (human prostate cell preparation (hPCP)), and xenografts of PC‐3 cells by immunohistochemistry (IHC), Western blotting, and RT‐PCR, respectively. The growth‐inhibition effect of a 5‐HT1A antagonist (NAN‐190) on PC cell lines was studied using a bromodeoxyuridine (BrdU) assay.RESULTSA strong immunoreaction of 5‐HTR1A and 1B was demonstrated in high‐grade tumor cells (35/35) and a small number of BPH cells, whereas 5‐HTR1D was confined to vascular endothelial cells. 5‐HTR1A was also demonstrated in PC cells metastasized to lymph node and bone, PC‐3, DU145, LNCaP, and in xenografts of PC‐3 cells and hPCP. Western blot analysis gave strong bands from PC tissue extracts compared to BPH tissue. Using RT‐PCR, 5‐HTR1A mRNA was demonstrated in all PC cell lines. An antagonist of 5‐HTR1A (NAN‐190) inhibited the growth of PC‐3, DU145, and LNCaP cells but not of hPCP cells.CONCLUSIONSThis is the first study demonstrating an overexpression of 5‐HTR subtypes 1A and 1B in PC cells, especially in high‐grade tumors. Moreover, 5‐HT stimulates proliferation of PC cells and 5‐HTR1A antagonists inhibit proliferation. Thus, we propose that 5‐HT has an important role in tumor progression, especially in the androgen‐independent state of the disease. The design of specific antagonists for this type of receptor might be useful for the growth control of androgen‐independent tumors. © 2004 Wiley‐Liss, Inc.
Imaging of bioluminescent LNCaP‐luc‐M6 Tumors: A new animal model for the study of metastatic human prostate cancer
Authors: Scatena Caroline D. , Hepner Mischa A. , Oei Yoko A. , Dusich Joan M. , Yu Shang‐Fan , Purchio Tony , Contag Pamela R. , Jenkins Darlene E. 
Published: March 2004

AbstractBACKGROUNDAnimal experiments examining hormone‐sensitive metastatic prostate cancer using the human LNCaP cell line have been limited to endpoint analyses. To permit longitudinal studies, we generated a luciferase‐expressing cell line and used bioluminescent imaging (BLI) to non‐invasively monitor the in vivo growth of primary LNCaP tumors and metastasis.METHODSLNCaP.FGC cells were transfected to constitutively express firefly luciferase. LNCaP‐luc‐M6 cells were tested for bioluminescent signal intensity and hormone responsiveness in vitro. The cells were implanted in subcutaneous and orthotopic sites in SCID‐bg mice and imaged over time.RESULTSThe LNCaP‐luc‐M6 cells formed subcutaneous and orthotopic tumors in SCID‐bg mice, and nearly all tumor‐bearing animals developed pulmonary metastases. Early detection and temporal growth of primary tumors and metastatic lesions was successfully monitored by BLI.CONCLUSIONSThe LNCaP‐luc‐M6 cell line is a bioluminescent, hormone‐sensitive prostate cancer cell line applicable for BLI studies to non‐invasively monitor subcutaneous and orthotopic prostate tumor growth and metastasis in vivo. © 2004 Wiley‐Liss, Inc.