bluuebunny/crossref_metadata_2025_split huggingface dataset:
Class II major histocompatibility complex–peptide tetramer staining in relation to functional avidity and T cell receptor diversity in the mouse CD4+ T cell response to a rheumatoid arthritis–associated antigen
Authors: Falta Michael T. , Fontenot Andrew P. , Rosloniec Edward F. , Crawford Frances , Roark Christina L. , Bill Jerome , Marrack Philippa , Kappler John , Kotzin Brian L. 
Published: June 2005

AbstractObjectiveAlthough studies have suggested that human cartilage (HC) gp‐39 may be an antigen recognized by autoreactive CD4+ T cells in rheumatoid arthritis, we previously failed to identify specific CD4+ T cells in patients' synovial fluid or blood using a class II major histocompatibility complex–peptide tetramer composed of the immunodominant HC gp‐39263–275 epitope covalently linked to DR4. We undertook this study to better understand the parameters for specific binding of this tetramer.MethodsDR4‐transgenic mice were immunized with the HC gp‐39 peptide, and a set of peptide‐responsive hybridomas was derived. Hybridomas were stained with the DR4–gp‐39 tetramer and cultured with increasing amounts of peptide in the presence of DR4‐expressing antigen‐presenting cells to determine functional avidity.ResultsGreat variability was apparent in the ability of the tetramer to stain the hybridomas, and there was a strong correlation between the intensity of tetramer staining and functional avidity. Importantly, nearly 30% of the hybridomas did not stain with tetramer, and these cells exhibited relatively low functional avidity. Although the addition of an anti–T cell receptor (anti‐TCR) monoclonal antibody during the staining procedure enhanced binding of the tetramer to a number of the hybridomas, a significant percentage remained unstainable. Analysis of TCR expression showed that >90% of the hybridomas expressed the same TCR β‐chain variable region (Vβ10), and sequencing of the TCR junctional regions showed diversity in the third complementarity‐determining region.ConclusionThese results suggest that immune responses dominated by relatively low‐affinity TCR interactions, such as those that may occur in autoimmune disease, will be difficult to detect using standard tetramer techniques.
Interleukin‐4 can be a key positive regulator of inflammatory arthritis
Authors: Ohmura Koichiro , Nguyen Linh T. , Locksley Richard M. , Mathis Diane , Benoist Christophe 
Published: June 2005

AbstractObjectiveDevelopment of arthritis in the K/BxN mouse model depends on the induction of high titers of antibodies against the enzyme glucose‐6‐phosphate isomerase (GPI), promoted by CD4+ T cells expressing a GPI‐specific transgenic T cell receptor (TCR). This study was undertaken to determine whether this strong autoantibody response depends on T cell differentiation to the Th1 or Th2 phenotype.MethodsThe roles of Th cell–biasing cytokines were investigated by introducing the interleukin‐4 (IL‐4) and IL‐12–specific subunit p35 (IL‐12p35)–knockout mutations into the K/BxN model and evaluating the impact of these deficiencies on disease. The IL‐4–expressing cell types in K/BxN mice were revealed by crossing in a knockin alteration, which resulted in green fluorescent protein expression controlled by endogenous IL‐4 gene–regulatory elements. Transfer experiments permitted the identification of the IL‐4–producing cell type required for arthritis, and quantitative reverse transcriptase–polymerase chain reaction allowed for determination of the cytokine profile of K/BxN T cells.ResultsWhile IL‐12p35 appeared dispensable for the development of arthritis, IL‐4 was crucial for full development of disease. The GPI‐reactive TCR of standard K/BxN mice induced the transcriptional activation of the IL‐4 locus in CD4+ T cells and eosinophils, and CD4+ T cells were the obligatory source of IL‐4 for disease. However, the cytokine profile of K/BxN T cells revealed that K/BxN arthritis is not a “pure” Th2 disease.ConclusionThe K/BxN model, although not a classic Th2 disease, depends critically on IL‐4. The potential of IL‐4 to promote inflammatory arthritis should be considered when proposing therapies for rheumatoid arthritis aimed at biasing T cells toward IL‐4 production.
Diminished transforming growth factor β2 production leads to increased expression of a profibrotic procollagen α2 type I messenger RNA variant in embryonic fibroblasts of UCD‐200 chickens, a model for systemic sclerosis
Authors: Prelog Martina , Scheidegger Paul , Peter Silvia , Gershwin M. Eric , Wick Georg , Sgonc Roswitha 
Published: June 2005

AbstractObjectiveA procollagen α2(I) messenger RNA (mRNA) variant, with a 115‐bp band and an expected band of 180 bp, was found to be increased during early, acute scleroderma‐like disease in UCD‐200 chickens. The present study investigated the influence of cytokines on the expression of these 2 proα2(I) mRNA variants.MethodsEmbryonic fibroblasts of UCD‐200 chickens (UCD‐200‐CEF) and normal white leghorns (NWL‐CEF) were grown in 3‐dimensional collagen gels. Procollagen mRNA expression was analyzed by RNase protection assay, and proliferation was determined by 3H‐thymidine incorporation. Transforming growth factor β1 (TGFβ1) and TGFβ2 were measured in culture supernatants by enzyme‐linked immunosorbent assay.ResultsCompared with NWL‐CEF, UCD‐200‐CEF expressed 7.2 times more of the smaller profibrotic proα2(I) mRNA variant. TGFβ1 stimulated the proliferation of UCD‐200‐CEF, but not NWL‐CEF. The 115 bp:180 bp ratio was increased by TGFβ1 in both NWL‐CEF and UCD‐CEF. TGFβ2 and TGFβ3 reduced the expression of the profibrotic proα2(I) mRNA in UCD‐200‐CEF to the same levels observed in healthy control NWL‐CEF. In culture supernatants, NWL‐CEF produced 4.1 times more TGFβ2 than that produced by UCD‐CEF. Inhibition of endogenous TGFβ2 in NWL‐CEF resulted in the same 115 bp:180 bp ratio as seen in untreated UCD‐CEF.ConclusionTGFβ2 reduces the expression of a profibrotic proα2(I) mRNA variant in UCD‐200‐CEF. The constitutive overproduction of this proα2(I) mRNA variant and the diminished synthesis of TGFβ2 in untreated UCD‐200‐CEF suggest that TGFβ2 can act as an antifibrotic cytokine and might be a key player during fibrosis onset. These results shed light on the contradictory observations regarding the role of TGFβ2 in human systemic sclerosis.
Tumor necrosis factor inhibits conversion of dehydroepiandrosterone sulfate (DHEAS) to DHEA in rheumatoid arthritis synovial cells: A prerequisite for local androgen deficiency
Authors: Weidler Claudia , Struharova Sona , Schmidt Martin , Ugele Bernhard , Schölmerich Jürgen , Straub Rainer H. 
Published: June 2005

AbstractObjectiveUse of anti–tumor necrosis factor (anti‐TNF) antibody therapy in rheumatoid arthritis (RA) has expanded our understanding of possible mechanisms by which this treatment reduces inflammation. Beyond its effects on local immune responses, anti‐TNF treatment may also modulate the local hormone supply. Because androgens are thought to inhibit immune responses, their presence in inflamed tissue is an additional important antiinflammatory factor.MethodsWe investigated conversion of the ubiquitous dehydroepiandrosterone sulfate (DHEAS), the biologically inactive precursor of DHEA, to the androgen DHEA in mixed synovial cells from patients with RA and patients with osteoarthritis (OA), making use of thin‐layer chromatography and phosphorimaging. Using immunohistochemical analysis, we detected the key enzyme, steroid sulfatase.ResultsDHEAS‐to‐DHEA conversion in synovial cells from patients with RA was significantly lower than that in synovial cells from patients with OA (mean ± SEM 3.3 ± 0.5% versus 6.0 ± 0.9% of applied 3H‐DHEAS per 106 synovial cells; P = 0.042). In RA, but not in OA, the level of converted 3H‐DHEA was inversely correlated with the density of synovial macrophages (for RA, Rrank = −0.725, P = 0.005; for OA, Rrank = 0.069, P not significant [NS]) and T cells (for RA, Rrank = −0.621, P = 0.024; for OA, Rrank = 0.247, P NS). Double immunohistochemistry analysis revealed that steroid sulfatase was located mainly in synovial macrophages but was also observed in fibroblasts. Neutralization of TNF largely up‐regulated the conversion of DHEAS to DHEA in RA, but not in OA. A similar neutralizing effect was observed with polyclonal human immunoglobulins; this effect is most probably mediated via TNF neutralization at low TNF concentrations.ConclusionThese data indicate that TNF inhibits the conversion of DHEAS to DHEA in RA synovial cells. Because androgens are antiinflammatory mediators, TNF‐induced inhibition of the local androgen supply is a supplementary proinflammatory factor. Consequently, anti‐TNF strategies may also exert their positive effects by increasing tissue androgens.
Empirical and theoretical dosimetry in support of whole body radio frequency (RF) exposure in seated human volunteers at 220 MHz
Authors: Allen Stewart J. , Adair Eleanor R. , Mylacraine Kevin S. , Hurt William , Ziriax John 
Published: June 2005

AbstractThis study reports the dosimetry performed to support an experiment that measured physiological responses of seated volunteer human subjects exposed to 220 MHz fields. Exposures were performed in an anechoic chamber which was designed to provide uniform fields for frequencies of 100 MHz or greater. A vertical half‐wave dipole with a 90° reflector was used to optimize the field at the subject's location. The vertically polarized E field was incident on the dorsal side of the phantoms and human volunteers. The dosimetry plan required measurement of stationary probe drift, field strengths as a function of distance, electric and magnetic field maps at 200, 225, and 250 cm from the dipole antenna, and specific absorption rate (SAR) measurements using a human phantom, as well as theoretical predictions of SAR with the finite difference time domain (FDTD) method. A NBS (National Bureau of Standards, now NIST, National Institute of Standards and Technology, Boulder, CO) 10 cm loop antenna was positioned 150 cm to the right, 100 cm above and 60 cm behind the subject (toward the transmitting antenna) and was read prior to each subject's exposure and at 5 min intervals during all RF exposures. Transmitter stability was determined by measuring plate voltage, plate current, screen voltage and grid voltage for the driver and final amplifiers before and at 5 min intervals throughout the RF exposures. These dosimetry measurements assured accurate and consistent exposures. FDTD calculations were used to determine SAR distribution in a seated human subject. This study reports the necessary dosimetry to precisely control exposure levels for studies of the physiological consequences of human volunteer exposures to 220 MHz. Bioelectromagnetics 26:440–447, 2005. Published 2005 Wiley‐Liss, Inc.
Bemessung von Dübeln am Bauteilrand unter Querlast: Ingenieurmäßiger Ansatz zur Berücksichtigung der Lastrichtung
Authors: Mallée Rainer , Pusill‐Wachtsmuth Peter 
Published: May 2005

AbstractEs wird ein allgemeingültiger Bemessungsansatz für Dübel unter Querlast am Bauteilrand bei der Versagensart Betonkantenbruch abgeleitet. Die Richtung der Querlast wird bei der Lastverteilung auf die einzelnen Dübel einer Gruppe berücksichtigt. Damit kann das Modell für Anwendungen ohne und mit Lochspiel sowie für Querlasten und für Torsionsmomente angewendet werden.
Anspruchsvolles Sichtbetonbauwerk: Neubau der Universität Brixen
Authors: Taferner Josef , Sodeikat Christian , Eltze Rainer 
Published: May 2005

AbstractWegen der vorhandenen Komplexität sind bei der Planung und Herstellung von Sichtbeton erfahrene Sonderfachleute gefragt, die den dazugehörigen Willen und das erforderliche Engagement zur Vorbereitung, Koordination und baubegleitenden Kontrolle und Steuerung bei der Herstellung besitzen. Sehenswerte Ergebnisse lassen sich nur dann erreichen, wenn Wissensvermittlung, Informationsfluß, Fachkenntnisse, Erfahrungen und Engagement aller Beteiligten zusammenwirken und wenn der Bauherr bereit ist, diese planerischen und ausführungstechnischen Leistungen entsprechend zu vergüten. Am Beispiel des Baues der Universität Brixen soll gezeigt werden, wie trotz hoher architektonischer Anforderungen an das Tragwerk mit filigranen Bauteilen und bauphysikalischen Randbedingungen ein alle Seiten zufriedenstellendes Sichtbetonergebnis erzielt werden kann. Dabei waren besondere Überlegungen in statisch‐konstruktiver, betontechnologischer und ausführungstechnischer Hinsicht erforderlich.
Applications of Binary Segmentation to the Estimation of Quantal Response Curves and Spatial Intensity
Authors: Yang Tae Y. , Swartz Tim B. 
Published: June 2005

AbstractThis paper explores the use of binary segmentation procedures in two applications. The first application is concerned with the estimation of nonparametric quantal response curves. With Bernoulli data and an assumed monotone increasing curve, this gives rise a change‐point model where the change points are determined using a sequence of nested hypothesis tests of whether a change point exists. The second application concerns cluster identification and inference for spatial data where the shape of the clusters and the number of clusters is unknown. The procedure involves a sequence of nested hypothesis tests of a single cluster versus a pair of distinct clusters. Examples of both applications are provided. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Combination of Dsb coexpression and an addition of sorbitol markedly enhanced soluble expression of single‐chain Fv in Escherichia coli
Authors: Sandee Duanpen , Tungpradabkul Sumalee , Kurokawa Yoichi , Fukui Kiichi , Takagi Masahiro 
Published: June 2005

AbstractMany eukaryotic proteins have been produced successfully in Escherichia coli. However, not every gene can be expressed efficiently in this organism. Most proteins, especially those with multiple disulfide bonds, have been shown to form insoluble protein or inclusion body in E. coli. An inactive form of protein would require an in vitro refolding step to regain biological functions. In this study, we described the system for soluble expression of a single‐chain variable fragment (scFv) against hepatocellular carcinoma (Hep27scFv) by coexpressing Dsb protein and enhancing with medium additives. The results revealed that overexpression of DsbABCD protein showed marked effect on the soluble production of Hep27scFv, presumably facilitating correct folding. The optimal condition for soluble scFv expression could be obtained by adding 0.5M sorbitol to the culture medium. The competitive enzyme‐linked immunosorbent assay (ELISA) indicated that soluble scFv expressed by our method retains binding activity toward the same epitope on a hepatocellular carcinoma cell line (HCC‐S102) recognized by intact antibody (Ab) (Hep27 Mab). Here, we report an effective method for soluble expression of scFv in E. coli by the Dsb coexpression system with the addition of sorbitol medium additive. This method might be applicable for high‐yield soluble expression of proteins with multiple disulfide bonds. © 2005 Wiley Periodicals, Inc.
Monitoring complex media fermentations with near‐infrared spectroscopy: Comparison of different variable selection methods
Authors: Ferreira Ana P. , Alves Teresa P. , Menezes José C. 
Published: June 2005

AbstractNear‐infrared spectroscopy (NIRS) is known to be a suitable technique for rapid fermentation monitoring. Industrial fermentation media are complex, both chemically (ill‐defined composition) and physically (multiphase sample matrix), which poses an additional challenge to the development of robust NIRS calibration models. We investigated the use of NIRS for at‐line monitoring of the concentration of clavulanic acid during an industrial fermentation. An industrial strain of Streptomyces clavuligerus was cultivated at 200‐L scale for the production of clavulanic acid. Partial least squares (PLS) regression was used to develop calibration models between spectral and analytical data. In this work, two different variable selection methods, genetic algorithms (GA) and PLS‐bootstrap, were studied and compared with models built using all the spectral variables. Calibration models for clavulanic acid concentration performed well both on internal and external validation. The two variable selection methods improved the predictive ability of the models up to 20%, relative to the calibration model built using the whole spectra. © 2005 Wiley Periodicals, Inc.
Raman optical activity of simple alanyl peptides: Backscattering in‐phase dual circular polarization measurements in aqueous solution
Authors: Yu Gu‐sheng , Che Diping , Freedman Teresa B. , Nafie Laurence A. 
Published: May 2005

AbstractWe report the measurement of backscattering in‐phase dual circular polarization (DCPI) Raman optical activity (ROA) for the simple alanyl peptides L‐alanyl‐L‐alanine ([L‐Ala]2), L‐alanylglycine (L‐Ala‐Gly), glycyl‐L‐alanine (Gly‐L‐Ala), and L‐alanyl‐L‐alanyl‐L‐alanine ([L‐Ala]3) in aqueous solutions. Empirical correlations between ROA features and the amino acid composition of the peptides are described. It is shown that the sum of the DCPIROA spectra for L‐Ala‐Gly and Gly‐L‐Ala is nearly the same as the corresponding ROA spectrum for (L‐Ala)2. Additionally, the DCPIROA spectrum for (L‐Ala)2 is very close to that of (L‐Ala)3, as has been observed previously using backscattering unpolarized incident circular polarization (ICPu) ROA. The ROA contributions of the different L‐Ala groups, which occur separately in the two glycyl peptides of alanine and jointly in (L‐Ala)2, are described in terms of the extent of vibrational coupling between peptide subunits. From these results, it appears that ROA has a more local sensitivity to vibrational coupling than is found in VCD or electronic circular dichroism. © 1995 John Wiley & Sons, Inc.
Fluorescence spectroscopy of 13′‐cis‐spheroidene at 170 K: A reason for the natural selection of the all‐trans configuration for the light‐harvesting function
Authors: Koyama Yasushi , Jiang Yue‐shun , Watanabe Yasutaka 
Published: May 2005

AbstractAs a follow‐up to fluorescence and fluorescence‐excitation spectroscopy at 170 K of all‐trans‐spheroidene free in solutionsY. Watanabe, T. Kameyama, Y. Miki, M. Kuki, and Y. Koyama, “The 2′Ag− state and two additional low‐lying electronic states of spheroidene newly identified by fluorescence and fluorescence‐excitation spectroscopy at 170K,” Chem. Phys. Lett. 206, 62–68 (1993).
and bound to the light‐harvesting complex of Rhodobacter sphaeroides 2.4.1,Y. Koyama, Y. Miki, T. Kameyama, R. J. Cogdell, and Y. Watanabe, “Low‐lying electronic levels of spheroidene bound to the light‐harvesting (LH2) complex of Rhodobacter sphaeroides 2.4.1. as determined by fluorescence and fluorescence‐excitation spectroscopy at 170K,” Chem. Phys. Lett. 208, 479–485 (1993).
13′‐cis‐spheroidene in n‐hexane solution has been examined for comparison. All‐trans‐spheroidene exhibits efficient internal conversion from the 3A to the 2A state and fluorescence from both the 2A and B states, while 13‐‐cis‐spheroidene exhibits internal conversion from the “3Ag‐” state to the “B” state and fluorescence only from the “B” state. Thus, all‐trans‐spheroidene can provide two channels of carotenoid‐to‐bacteriochlorophyll singlet‐energy transfer, while 13′‐cis‐spheroidene can provide only one. This must be a reason for the natural selection of the all‐trans configuration for the light‐harvesting function. © 1995 John Wiley & Sons, Inc.
Engineering Substrate Specificity of O6‐Alkylguanine‐DNA Alkyltransferase for Specific Protein Labeling in Living Cells
Authors: Juillerat Alexandre , Heinis Christian , Sielaff India , Barnikow Jan , Jaccard Hugues , Kunz Beatrice , Terskikh Alexey , Johnsson Kai 
Published: June 2005

AbstractFusion proteins of human O6‐alkylguanine‐DNA alkyltransferase (AGT) can be specifically labeled with a wide variety of synthetic probes in mammalian cells; this makes them an attractive tool for studying protein function. However, to avoid undesired labeling of endogenous wild‐type AGT (wtAGT), the specific labeling of AGT fusion proteins has been restricted to AGT‐deficient mammalian cell lines. We present here the synthesis of an inhibitor of wtAGT and the generation of AGT mutants that are resistant to this inhibitor. This enabled the inactivation of wtAGT and specific labeling of fusion proteins of the AGT mutant in vitro and in living cells. The ability to specifically label AGT fusion proteins in the presence of endogenous AGT, after brief incubation of the cells with a small‐molecule inhibitor, should significantly broaden the scope of application of AGT fusion proteins for studying protein function in living cells.
Reinigung jodhaltiger Spülwasser aus der Röntgenkontrastmittelproduktion mittels Nanofiltration: Prozessgestaltung und Modellierung
Authors: Drews A. , Klahm T. , Renk B. , Saygili M. , Baumgarten G. , Kraume M. 
Published: June 2005

AbstractNo abstracts.
The melanoma antigen gene as a surveillance marker for the detection of circulating tumor cells in patients with breast carcinoma
Authors: Kwon Soim , Kang Seok Hyung , Ro Jungsil , Jeon Chang‐Ho , Park Jong‐Wook , Lee Eun Sook 
Published: June 2005

AbstractBACKGROUNDCirculating occult tumors cells could be used for the surveillance of metastases after primary breast carcinoma therapy, but their detection is limited by the lack of specific molecular markers. Melanoma antigen genes (MAGEs), which are expressed in malignant tissues but not in normal tissues (except for placenta and testis), might provide such a marker. To date, however, the use of MAGEs in the detection of occult tumor cells using reverse transcription‐polymerase chain reaction (RT‐PCR) has been limited because of the heterogeneity and low expression of individual MAGEs in tumor tissues.METHODSWe developed multiple MAGE‐recognizing primers (MMRPs) that were capable of binding to the cyclic DNA of 6 MAGE‐A gene subtypes (MAGE‐A1–MAGE‐A6). We assessed the ability of the MMRPs to detect the expression of MAGE‐A gene subtypes in peripheral blood obtained from patients with benign or malignant breast disease.RESULTSMAGE‐A gene expression was not detected in 32 patients with benign disease but was detected in 1 of 31 patients (3%) patients with negative lymph node breast carcinoma, in 10 of 52 patients (19%) with 1–3 positive lymph nodes, in 11 of 53 patients (21%) with ≥ 4 positive lymph nodes, and in 20 of 52 patients (39%) with metastatic disease. The results were statistically significant (P < 0.0001; chi‐square test for linear‐by‐linear association). The results also showed that the detection of MAGE‐A gene expression in the blood predicted tumor progression or recurrence.CONCLUSIONSThe results suggested that MAGE‐A gene expression may be used for the surveillance of circulating breast carcinoma cells after primary therapy by RT‐nested PCR using MMRPs. Cancer 2005. © 2005 American Cancer Society.