bluuebunny/crossref_metadata_2025_split huggingface dataset:
The K+-channel opener NS1619 increases endothelial NO-synthesis involving p42/p44 MAP-kinase
Authors: Carl Trümper Jan Rasmus , Abdallah Yaser , Lüdders Dörte , Schaefer Christian , Most Astrid , Backenköhler Ulrich , Neumann Thomas , Walther Sabine , Piper Hans , Tillmanns Harald , Erdogan Ali , Kuhlmann Christoph Rüdiger 
Published: October 2004

SummaryCa2+-activated K+ channels with large conductance (BKCa) have been shown to play an important role in the regulation of vascular tone. We examined the role of the p42/p44 MAP-kinase (p42/p44MAPK) on nitric oxide (NO) production in human endothelial cells induced by the BKCa-opener NS1619. Using DiBAC-fluorescence imaging a concentration-dependent (2.5-12.5 µM) hyperpolarization induced by NS1619 was observed. A significant increase of intracellular Ca2+-concentration by NS1619 was seen using Fura-2-fluorescence-imaging, which was blocked by 2-APB, or reduction of extracellular Ca2+ (n=30; p<0.05). A cGMP-radioimmunoassay was used to examine NO synthesis. NS1619 significantly increased cGMP levels which was inhibited by LNMMA, iberiotoxin, BAPTA, 2-APB, reduction of extracellular Ca2+, PD 98059, or U0126 (cGMP (pmol/mg protein): NS1619 3.25 ± 0.85; NS1619 + L-NMMA 0.86 ± 0.02; NS1619 + iberiotoxin 0.99 ± 0.09; NS1619 + BAPTA 0.93 ± 0.29; NS1619 + 2-APB 0.99 ± 0.31; NS1619 + Ca2+-reduction 1.17 ± 0.06; NS1619 + PD98059 1.06 ± 0.49; NS1619 + U0126 1.10 ± 0.24; n=10; p<0.05). The phosphorylation of eNOS and p42/p44MAPK was examined by immunocytochemistry. Phosphorylation of p42/p44MAPK was significantly increased after 10 minutes of NS1619 stimulation, whereas eNOS phosphorylation was not changed over a period of 1 to 30 minutes. NS1619-induced hyperpolarization was not affected by treatment with PD 98059 or U0126. Additionally, NS1619 inhibited endothelial proliferation involving a NO-dependent mechanism. Our data demonstrate that NS1619 causes a transmembrane Ca2+-influx leading to an increased NO production involving p42/p44MAPK. This rise of NO formation is responsible for the NS1619 induced reduction of endothelial cell growth.
Combined effects of a factor Xa inhibitor YM466 and a GPIIb/IIIa antagonist YM128 on thrombosis and neointima formation in mice
Authors: Iwatsuki Yoshiyuki , Hayashi Kazumi , Moritani Yumiko , Nii Tomoko , Miyata Keiji , Kawasaki Tomihisa 
Published: October 2004

SummaryThrombosis and neointima formation limit the efficacy of coronary angioplasty. Factor Xa inhibitors and GPIIb/IIIa antagonists have shown to be effective on acute thrombosis and late neointima formation, however, their combined effects remain to be elucidated. Vascular injury was induced by FeCl3 in the carotid artery in mice. For thrombosis studies, the test drug was orally administered 1 hour before vascular injury. For neointima studies, the test drug was orally administered 1 hour before and twice daily for 1 week after vascular injury, and then histological analysis was performed 3 weeks after vascular injury. YM466 inhibited thrombotic occlusion at 30 mg/kg with prolongation of prothrombin time (PT), and tail transection bleeding time (BT) was affected at 100 mg/kg. YM466 also inhibited neointima formation at 10 mg/kg. YM128 inhibited thrombotic occlusion and neointima formation at 10 and 30 mg/kg, respectively, with inhibition of platelet aggregation and prolongation of BT. In contrast, the combination of 10 mg/kg YM466 and 3 mg/kg YM128 inhibited thrombotic occlusion and neointima formation without affecting PT, platelet aggregation and BT. Concomitant inhibition of factor Xa and GPIIb/IIIa may provide a safer and more effective therapeutic regimen for treatment of coronary angioplasty.
Cyclooxygenase COX-2a, a novel COX-2 mRNA variant, in platelets from patients after coronary artery bypass grafting
Authors: Censarek Petra , Freidel Kerstin , Udelhoven Michael , Ku Sun-Jung , Hohlfeld Thomas , Meyer-Kirchrath Jutta , Schrör Karsten , Weber Artur-Aron 
Published: October 2004

SummaryThere are two principal cyclooxygenase isoforms referred to as COX-1 and COX-2. Recently, COX-3 has been identified. We have demonstrated the expression of COX-2 in platelets from patients after coronary artery bypass grafting (CABG). Careful biochemical analysis revealed that, when compared to recombinant COX-2, platelet COX-2 had a slightly higher electrophoretic mobility. Two COX-2 sequences (∼1.8 kb, ∼1.7 kb) were cloned from platelet mRNA. The ∼1.7 kb sequence, designated COX-2a, differed from the human COX-2 sequence only in a deletion from position +458 to +567. Similar to the human COX-3, there is a frame shift in the COX-2a sequence resulting in a TAA stop codon at position +490. Thus, the expression of a COX-2a protein corresponding to the 67 kDa COX-2 protein is not clear. However, the marked shifting from COX-2 to COX-2a in platelets from some patients after CABG is a striking finding.
Percutaneous endoluminal arterial cryoenergy improves vascular remodelling after angioplasty
Authors: Geoffroy Pascale , Dorval Jean-François , Sirois Martin , Tanguay Jean-François 
Published: October 2004

SummaryThis study aimed to investigate the effect of percutaneous endoluminal arterial cryoenergy immediately after balloon angioplasty on vascular remodeling. Restenosis, the main complication after coronary artery angioplasty, is a complex phenomenon in which vascular remodeling plays a prominent role. Observations of reduced scarring in freeze-induced wounds suggest potential value for cryoenergy in the prevention of restenosis. Juvenile swine underwent a first oversized balloon injury in both carotid arteries (CA) (3 injury sites/artery) and a second injury at day 14. At that time, one CA was randomly assigned to endoluminal cryoenergy of the sequential segments (proximal, medial, distal) at −15ºC, −30ºC, and −50ºC for 120 sec, and the other CA was used as control. Animals were sacrificed 28 days after the second procedure. Compared with intact reference segments, angioplasty reduced both the luminal (LA) and the external elastic lamina (EEL) areas from 6.66±0.59 to 3.13±0.54 mm2 (p<0.05) and from 8.81±0.81 to 6.48±0.52 mm2 (p<0.05), respectively. Cryoenergy, at the temperature with maximal benefits (-50°C), caused a temperature-related protection, as the LA was maintained (6.79±0.89 versus 7.18± 0.78 mm2 for reference) and the EEL area increased from 9.12±0.78 to 12.98±1.07 mm2, p<0.05. Moreover, cryoenergy increased the density of collagen III (p<0.05) which correlated with the maintenance of the LA (r=0.8045, p<0.009). Cryoenergy prevents late luminal loss after double-injury angioplasty by improving vascular remodeling, and is an interesting new therapeutic approach for the prevention of restenosis after angioplasty. The increased synthesis of collagen III appears to be involved in this phenomenon and could be a potential method of stabilizing the vulnerable plaque.
Prostaglandin E2 activates channel-mediated calcium entry in human erythrocytes: an indication for a blood clot formation supporting process
Authors: Tabellion Wiebke , Lipp Peter , Bernhardt Ingolf , Kaestner Lars 
Published: October 2004

SummaryProstaglandin E2 (PGE2) is released from platelets when they are activated. Using fluorescence imaging and the patch-clamp technique, we provide evidence that PGE2 at physiological concentrations (10−10 M) activates calcium rises mediated by calcium influx through a non-selective cation-channel in human red blood cells. The extent of calcium increase varied between cells with a total of 45% of the cells responding. It is well known that calcium increases elicited the calcium-activated potassium channel (Gardos channel) in the red cell membrane. Previously, it was shown that the Gardos channel activation results in potassium efflux and shrinkage of the cells. Therefore, we conclude that the PGE2 responses of red blood cells described here reveal a direct and active participation of erythrocytes in blood clot formation.
On requesting conference papers electronically
Authors: Hartley James 
Published: October 2004

Aims. The aims of this study were (i) to determine the success rate for obtaining conference papers by using e-mail, and (ii) to compare the results with those found previously for reprints of published papers. Method. 100 requests for copies of papers delivered at a conference in the USA were made by e-mail from the UK. A reminder was sent to the appropriate authors if no paper had been received within three weeks. Results. 54 papers were received within six weeks. Most came by return or within one day, and all came electronically. Few of these papers could be described as conference papers, however, as the majority of them were copies of papers prepared for publication elsewhere. In addition, many lacked the information necessary for them to be adequately cited in other publications.
Quality of Life in Ovarian Cancer Patients: Comparison of Paclitaxel Plus Cisplatin, With Cyclophosphamide Plus Cisplatin in a Randomized Study
Authors: Bezjak Andrea , Tu Dongsheng , Bacon Monica , Osoba David , Zee Benny , Stuart Gavin , Roy Josee-Anne , Piccart Martine , Eisenhauer Elizabeth 
Published: October 2004

Purpose Formal quality-of-life (QOL) assessments may contribute important information on patient symptoms. Despite many trials of systemic chemotherapy in ovarian cancer, reports of its effect on QOL are few. Patients and Methods QOL was assessed in an Intergroup randomized trial comparing paclitaxel plus cisplatin to cyclophosphamide plus cisplatin in women with advanced ovarian cancer. One hundred fifty-two eligible patients accrued in Canada completed the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C30 and a trial-specific checklist at baseline (after surgical debulking) and at regular intervals during and after chemotherapy. Mean change scores over time in the two arms were calculated. Results Compliance with QOL questionnaire completion was excellent (81% to 93%). In general, deterioration was seen in the QOL domains immediately after chemotherapy (day 8 of cycle 1), followed by clinically meaningful improvements compared with baseline (change scores ≥ 10) in both arms during the treatment period in a number of domains and items, including global QOL, emotional function, social function, fatigue, pain, sleep, constipation, appetite, abdominal swelling, and abdominal cramps. Improvements in global QOL persisted for the duration of follow-up. More neurosensory effects and myalgia were documented in the paclitaxel arm; however, this did not adversely affect global or other domains of QOL and improved once chemotherapy was completed. Conclusion Improvement from baseline in QOL measures was seen in both treatment arms. The greater neurologic and muscle toxicity of paclitaxel did not adversely influence QOL. QOL data can contribute useful information on the experience of symptoms and their time course, which may assist patients and physicians in their discussion about the anticipated effects of therapy.
Hybridization of Surface-modified Semiconductor Nanoparticles and a Resin
Authors: Matsumoto Kazuaki , Tsuji Ryotaro , Yonemushi Yoshiharu , Yoshida Tatsushi 
Published: October 2004

Abstract
A mercapto-terminated linear polymer, which was prepared by a reversible addition-fragmentation chain transfer (RAFT) technique, was used to modify semiconductor nanoparticle surfaces. This modification resulted in an easier dispersion of the nanoparticles in the resins by a simple mixing process, as confirmed by transmission electron microscopy.
Monoallelic Expression of Pax5: A Paradigm for the Haploinsufficiency of Mammalian Pax Genes?
Authors: Nutt S.L. , Busslinger M. 
Published: October 2004

AbstractIt is generally assumed that most mammalian genes are transcribed from both alleles. Hence, the diploid state of the genome offers the advantage that a loss-of-function mutation in one allele can be compensated for by the remaining wild-type allele of the same gene. Indeed, the vast majority of human disease syndromes and engineered mutations in the mouse genome are recessive, indicating that recessiveness is the ‘default’ state. However, a minority of genes are semi-dominant, as heterozygous loss-of-function mutation in these genes leads to phenotypic abnormalities. This condition, known as haploinsufficiency, has been described for five of the nine mammalian
Adeno-Associated Viral Vectors for Gene Transfer and Gene Therapy
Authors: Büeler H. 
Published: October 2004

AbstractAdeno-associated virus (AAV) is a defective, non-pathogenic human parvovirus that depends for growth on coinfection with a helper adenovirus or herpes virus. Recombinant adeno-associated viruses (rAAVs) have attracted considerable interest as vectors for gene therapy. In contrast to other gene delivery systems, rAAVs lack all viral genes and show long-term gene expression
Histidyl-tRNA Synthetase
Authors: Freist W. , Verhey J.F. , Rühlmann A. , Gauss D.H. , Arnez J.G. 
Published: October 2004

AbstractHistidyl-tRNA synthetase (HisRS) is responsible for the synthesis of histidyl-transfer RNA, which is essential for the incorporation of histidine into proteins. This amino acid has uniquely moderate basic properties and is an important group in many catalytic functions of enzymes.A compilation of currently known primary structures of HisRS shows that the subunits of these homodimeric enzymes consist of 420–550 amino acid residues. This represents a relatively short chain length among aminoacyl-tRNA synthetases (aaRS), whose peptide chain sizes range from about 300 to 1100 amino acid residues.The crystal structures of HisRS from two organisms and their complexes with histidine, histidyl-adenylate and histidinol with ATP have been solved. HisRS fromThe aminoacylation reaction follows the standard two-step mechanism. HisRS also belongs to the group of aaRS that can rapidly synthesize diadenosine tetraphosphate, a compound that is suspected to be involved in several regulatory mechanisms of cell metabolism. Many analogs of histidine have been tested for their properties as substrates or inhibitors of HisRS, leading to the elucidation of structure-activity relationships concerning configuration, importance of the carboxy and amino group, and the nature of the side chain.HisRS has been found to act as a particularly important antigen in autoimmune diseases such as rheumatic arthritis or myositis. Successful attempts have been made to identify epitopes responsible for the complexation with such auto-antibodies.
Dequalinium TM Vesicles Form Stable Complexes with Plasmid DNA which Are Protected from DNase Attack
Authors: Lasch J. , Meye A. , Taubert H. , Koelsch R. , Mansa-ard J. , Weissig V. 
Published: October 2004

AbstractUpon sonication, the antimicrobial and antineoplastic compound dequaliniumWe hypothesize that the DQA/DNA complexes are well-suited for ‘DQAsomal gene transfer’
On the Role of Thymopoietins in Cell Proliferation. Immunochemical Evidence for New Members of the Human Thymopoietin Family
Authors: Weber P.J.A. , Eckard C.P. , Gonser S. , Otto H. , Folkers G. , Beck-Sickinger A.G. 
Published: October 2004

AbstractThymopoietins (TMPOs) are a group of ubiquitously expressed nuclear proteins. They are suggested to play an important role in nuclear envelope organization and cell cycle control, as has been shown for lamina-associated polypeptides 2 α and β, which are the rat homologs of human TMPOα and TMPOβ, respectively. The recent isolation and characterization of seven mouse TMPO mRNA transcripts named TMPO-α, β, β', γ, ε, δ, and ζ, suggest that more than the three previously reported transcripts, α, β, and γ forms, may exist in humans. Here we report on the demonstration of putative human TMPOδ and ε by immunoblotting of human cell lines using a newly prepared polyclonal antiserum against the common N-terminal region of TMPO. Furthermore, we prepared the first truly TMPO-β-specific, affinity-purified polyclonal antiserum, using a part of the human analog of the β-specific domain of mouse TMPO 220–259 for immunization. We showed that human TMPOβ is highly expressed in all cancerous cells tested, while hardly any cross-reactivities with other proteins could be detected. In contrast to the high expression of human TMPOβ in the cancer-derived neuroblastoma cell lines SK-N-MC and SMS-KAN, we found very low expression of human TMPOβ in low-proliferative nerve tissue. These data led us to the assumption that expression of TMPOβ may correlate with the occurrence of cancer, and therefore may serve as a new tumor marker, or even as a new target for cancer therapy.
A Non-Radioactive Method for Inexpensive Quantitative RT-PCR
Authors: Maggiolini M. , Donzé O. , Picard D. 
Published: October 2004

Abstract
We present a novel method for quantitative RT-PCR that involves direct incorporation of digoxigenin-11-dUTP (DIG-dUTP) during amplification of cDNAs, separation of RT-PCR products by agarose gel electrophoresis, Southern transfer to a nylon membrane, and chemiluminescent detection with an anti-DIG antibody. The whole procedure can be done in about a day and has the following advantages: It is highly sensitive, specificity is confirmed by monitoring the size of the RT-PCR product, it is non-radioactive, quantitative, and does not require expensive specialized equipment.
Activation of the Transcription Factor ISGF3 by Interferon-gamma
Authors: Matsumoto M. , Tanaka N. , Harada H. , Kimura T. , Yokochi T. , Kitagawa M. , Schindler C. , Taniguchi T. 
Published: October 2004

AbstractThe interferon-stimulated gene factor 3 (ISGF3) transcription factor has been extensively studied in the context of the type I interferon (IFN-α/β)-mediated antiviral response; it consists of the major DNA-binding component p48, and the signal transducers and activators of transcription (Stat)1 and Stat2. We show here that type II IFN (IFN-γ) can also invoke the activation of ISGF3 in mouse primary embryonic fibroblasts. In fact, the two Stat proteins were tyrosine phosphorylated in IFN-γ stimulated cells. Our present findings reveal an additional mechanism by which these two distinct types of cytokines, IFN-α/β and -γ, can commonly elicit antiviral activities.
Human Fc gamma Receptor IIb Expressed in <I>Escherichia coli</I> Reveals IgG Binding Capability
Authors: Sondermann P. , Jacob U. 
Published: October 2004

AbstractFcγ receptors (FcγR) are expressed on immunologically active cells where they trigger B and T cell responses and are responsible for the clearance of immunocomplexes. They occur as type I transmembrane proteins and also in soluble forms (sFcR) comprising only the ecto domains of the receptors. State-of-the-art research has generated demand for highly pure and homogeneous sFcγR preparations: first, studies of the immunoregulative potential of the soluble FcγRs have been hampered by co-purified growth factors. Second, they are needed for crystallographic analyses to solve questions such as the exact location of the binding site for IgG on the receptor, and the graded affinities of the receptors for different IgG subclasses. This has been unsuccessful due to limitations in availability and homogeneity of sFcγR expressed in eukaryotic cells. In order to address these problems we expressed the extracellular part of the human Fc-γRIIb inE. coli. The protein was refolded, purified in a three-step procedure and characterized by SDS-PAGE, mass spectrometry as well as N-terminal sequencing. The unglycosylated FcγRIIb is active because it binds immobilized antibody as well as the IgG Fc-fragment in solution. Finally, the receptor was crystallized in orthorhombic, tetragonal and hexagonal crystal forms that diffracted X-rays to resolutions of 1.7 Å, 2.7 Å and 3.8 Å respectively.
Diphosphoinositol Polyphosphates: The Final Frontier for Inositide Research?
Authors: Safrany S. T. , Caffrey J. J. , Yang X. , Shears S. B. 
Published: October 2004

AbstractThe diphosphoinositol polyphosphates comprise a group of highly phosphorylated compounds which have a rapid rate of metabolic turnover through tightly-regulated kinase/phosphohydrolase substrate cycles. The phosphohydrolases occur as multiple isoforms, the expression of which is apparently carefully controlled. Cellular levels of the diphosphoinositol polyphosphates are regulated by cAMP and cGMP in a protein kinase-independent manner. These inositides can also sense a specific mode of intracellular Ca2+pool depletion. In this review, we will argue that these are characteristics of highly significant cellular molecules.
Differential Distribution of Four Hyperpolarization-Activated Cation Channels in Mouse Brain
Authors: Moosmang S. , Biel M. , Hofmann F. , Ludwig A. 
Published: October 2004

Abstract
Hyperpolarization-activated cation currents, termed Ih, are observed in a variety of neurons. Four members of a gene family encoding hyperpolarization-activated cyclic-nucleotide-gated cation channels (HCN1-4) have been cloned. The regional expression and cellular localization of the four HCN channel types in mouse brain was investigated using in situ hybridization. The expression of HCN1 was restricted to the olfactory bulb, cerebral cortex, hippocampus, superior colliculus and cerebellum. In contrast, HCN2 transcripts were found at high levels nearly ubiquitiously in the brain, and the strongest signals were seen in the olfactory bulb, hippocampus, thalamus and brain stem. HCN3 was uniformly expressed at very low levels throughout the brain. Finally, HCN4 transcripts were prominently expressed selectively in the thalamus and olfactory bulb. Some neurons expressed two or more HCN channel transcripts including hippocampal pyramidal neurons (HCN1, HCN2 and low levels of HCN 4) and thalamic relay neurons (HCN2 and HCN4). Our results demonstrate that each HCN channel transcript has a unique distribution in the brain. Furthermore, they suggest that the heterogeneity of neuronal Ih may be, at least in part, due to the differential expression of HCN channel genes.
Functional Characterisation of Dictyostelium Myosin II with Conserved Tryptophanyl Residue 501 Mutated to Tyrosine
Authors: Batra R. , Manstein D. J. 
Published: October 2004

AbstractWe created aDictyostelium discoideummyosin II mutant in which the highly conserved residue Trp-501 was replaced by a tyrosine residue. The mutant myosin alone, when expressed in aDictyosteliumstrain lacking the functional myosin II heavy chain gene, supported cytokinesis and multicellular development, processes which require a functional myosin inDictyostelium. Additionally, we expressed the W501Y mutant in the soluble myosin head fragment M761-2R (W501Y-2R) to characterise the kinetic properties of the mutant myosin motor domain. The affinity of the mutant myosin for actin was approximately 6-fold decreased, but other kinetic properties of the protein were changed less than 2-fold by the W501Y mutation. Based on spectroscopic studies and structural considerations, Trp-501, corresponding to Trp-510 in chicken fast skeletal muscle myosin, has been proposed to be the primary ATP-sensitive tryptophanyl residue. Our results confirm these conclusions. While the wild-type construct displayed a 10% fluorescence increase, addition of ATP to W501Y-2R was not followed by an increase in tryptophan fluorescence emission.