bluuebunny/crossref_metadata_2025_split huggingface dataset:
Elucidation of the Binding Regions of PAI-1 Neutralizing Antibodies Using Chimeric Variants of Human and Rat PAI-1
Authors: Bijnens A. P. , Ngo T. H. , Gils A. , Dewaele J. , Knockaert I. , Stassen J. M. , Declerck P. J. 
Published: March 2018

SummaryIncreased levels of plasminogen activator inhibitor-1 (PAI-1), the main physiological inhibitor of tissue-type plasminogen activator (t-PA) in plasma, are a known risk factor for thromboembolic and cardiovascular diseases. The elucidation of the binding site of inhibitory monoclonal antibodies may contribute to the rational design of PAI-1 modulating therapeutics. In this study, homolog-scanning mutagenesis was used to identify the binding region of a variety of human PAI-1 inhibitory antibodies, lacking cross-reactivity with rat PAI-1. Therefore, eight chimeric human/rat PAI-1 variants, containing rat PAI-1 substitutions at the N-terminal or C-terminal end with splicing sites at positions 26, 81, 187, 277 or 327, were generated and purified. Biochemical characterization revealed that all chimeras were folded properly. Subsequently, surface plasmon resonance was used to determine the affinity of various monoclonal antibodies for these chimera. Comparative analysis of the affinity and ELISA data allowed the identification of the major binding region of the inhibitory antibodies MA-8H9D4, MA-33B8F7, MA-44E4, MA-42A2F6 and MA-56A7C10. Thus, three segments in human PAI-1 containing each at least one site involved in the neutralization of PAI-1 activity could be identified, i.e. (1) the segment from residue 81 to residue 187 (comprising -helices hD, hE and hF, -strands s4C, s3A, s2A and s1A and the loops connecting these elements), (2) the segment between residues 277 and 327 (hI, thIs5A, s5A and s6A) and (3) the region C-terminal from amino acid 327, including the reactive site loop. The current data, together with previous data, indicate that PAI-1 contains at least four different regions that could be considered as putative targets to modulate its activity.
7E3 F(ab’)2, an Effective Antagonist of Rat αIIbβ3 and αvβ3, Blocks In Vivo Thrombus Formation and In Vitro Angiogenesis
Authors: Sassoli Patricia , Emmell Eva , Tam Susan , Trikha Mohit , Zhou Zhao , Jordan Robert , Nakada Marian 
Published: March 2018

SummaryAbciximab (c7E3 Fab, ReoPro®) blocks GPIIb/IIIa and αvβ3 and inhibits thrombotic and proliferative events only in humans and non-human primates. The bivalent F(ab’)2 fragment is an effective anti-thrombotic agent in canine models. In the present study, 7E3 F(ab’)2 was also found to bind to rat GPIIb/IIIa (KD = 27 ± 4 g/mL) and αvβ3 (KD = 9 ± 8 μg/mL), to block in vitro rat platelet aggregation (IC50 = 16 ± 6 μg/mL), and to inhibit αvβ3-mediated microvessel sprout formation in a rat aortic ring assay. Following administration of 7E3 F(ab’)2 (4 mg/kg) to rats, platelet aggregation was completely blocked for up to 6 h and thrombus formation in response to a rat abdominal aorta double crush injury was prevented. Effective chronic dosing was achieved with 6 mg/kg daily I.P. injections. In vitro mixing experiments indicated that 7E3 F(ab’)2 redistributed to unlabeled platelets in 2 h. Ex vivo, 7E3 F(ab’)2 was detected on platelets for up to 4 days after a single 4-mg/kg injection. These data suggest that 7E3 F(ab’)2 may be a useful agent to study the effects of GPIIb/IIIa and αvβ3 blockade in rat models of thrombosis and vascular disease.
Increased ICAM-1 and PECAM-1 Transcription Levels in the Heart of Apo-E Deficient Mice in Comparison to Wild Type (C57BL6).
Authors: Zibara Kazem , Chettab Kamel , McGregor Brigitte , Poston Robin , McGregor John 
Published: March 2018

SummaryAdhesion molecules and chemoattractants are thought to play a critical role in the homing of leukocytes to sites of vascular lesions. Apo-E deficiency in mice creates an atherosclerotic model that mimics vascular lesions in man. Little is known on the effect of Apo-E deficiency on expression of adhesion molecules in the hearts of these animals. In this study, male C57BL6 and Apo-E deficient mice were fed a chow diet over periods of time (0 to 20 weeks). The transcription levels of major adhesion molecules (ICAM-1, PECAM-1), present in the heart, were followed by northern blots. Immunohistochemistry was used to localize these adhesion molecules in the heart. Results show a significant increase in gene transcription levels of ICAM-1 and PECAM-1 in Apo-E animals, but not wild type, at 16 and 20 weeks of chow diet. Such increase in levels of transcription was not observed in younger Apo-E and C57BL6 animals (0, 6 weeks of diet). ICAM-1 and PECAM-1 were strongly expressed in the endocardium and heart microvessels. In contrast, VCAM-1 was poorly stained, with only an occasional expression on the endocardium and arterioles. Enhanced gene expression levels of heart ICAM-1 and PECAM-1 observed in Apo-E deficient mice, but not in control animals, appears to induce the initial stages of an inflammatory reaction. Such observations, not previously reported, may induce heart vascular remodeling.
Management von komplexen neurovaskulären Interventionen mithilfe eines telemedizinischen Netzwerks
Authors: Hüttemann K. , Müller R. , Dabitz R. , Fuhry L. , Janssen H. , Widder B. , Dieterich M. , Pfefferkorn T. , Hamann G. F. , Adamczyk C. 
Published: March 2018

ZusammenfassungDie Forderung nach einem flächendeckenden Zugang zur endovaskulären mechanischen Thrombektomie stellt die telemedizinischen Schlaganfallnetzwerke vor eine neue Herausforderung. NEVAS, das neurovaskuläre Versorgungsnetzwerk für Südwestbayern, ist so strukturiert, dass neben den klassischen Leistungen eines telemedizinischen Schlaganfallnetzwerkes auch moderne Interventionsverfahren besser in die Versorgung verankert werden. NEVAS unterscheidet sich von anderen telemedizinischen Netzwerken durch eine regionale Organisation der Kooperations- und Zentrumskliniken, eine bedarfsorientierte Nutzung der Telemedizin, eine obligate Gefäßdarstellung mittels Schnittbildverfahren im Rahmen der primären Notfalldiagnostik und die Definition sowie Etablierung individueller Verlegungs- und Behandlungsabläufe für jeden Standort. NEVAS kann als Blaupause dienen, wie moderne Schlaganfallversorgung ubiquitär verfügbar gemacht werden kann.
Do concepts of childhood trauma and dissociation provide an epistemological linkage between schizophrenia and PTSD?
Authors: Peña-Salazar C. , Arrufat Nebot F. X. , Alvarez M. J. , Fontseré H. Masramón , Coll-Negre M. , Roura-Poch P. , Valdés-Stauber J. 
Published: March 2018

Zusammenfassung
Hintergrund: Seit Bleuler und Janet wurde der Betrachtung von Schizophrenie als dissoziatives Phänomen wenig Beachtung geschenkt. Eine strikte Trennung zwischen Schizophrenie als ausschließlich biologischen Prozess und dissoziativen Störungen als alleinige Folge von traumatischen Erlebnissen in der Kindheit ist inzwischen überwunden. Methode: Die Autoren wählen eine narrative Literaturübersicht, die mögliche Zusammenhänge zwischen Kindheitstraumata, dissoziativen Störungen und psychotischen Symptomen in der Allgemeinbevölkerung als auch bei an Schizophrenie leidenden Menschen empirisch untersuchen. Wissenschaftstheoretische Partikularitäten postulierter Zusammenhangsannahmen werden diskutiert. Ergebnisse: Die untersuchte Literatur legt die Annahme nahe, dass schizophrene Menschen, die gleichzeitig dissoziative Störungen aufweisen, unter stärkeren positiven psychotischen Symptomen, häufiger Komorbidität und höherer Krankheitsbelastung leiden. Andererseits weisen Patienten mit „strukturellen dissoziativen Störungen“ phänomenologische Ähnlichkeiten mit schizophrenen Patienten. Gemeinsame biologische als auch psychologische Erklärungsansätze werden diskutiert. Schlussfolgerungen: Anhand der hier diskutierten empirischen Ergebnissen darf beim heutigen Wissensstand davon ausgegangen werden, dass sowohl posttraumatische Belastungsstörungen (PTSD) als auch schizophrene Psychosen auf biologischen als auch auf eine das Selbst vor unerträglichen Bedrohungen schützende Abwehrfunktionen basieren, jedoch klinisch und phänomenologisch unterschiedlich zum Ausdruck kommen.
Münchner Anti-Freezing-Training (MAFT)
Authors: Ceballos-Baumann A. O. , Schroeteler F. E. 
Published: March 2018

ZusammenfassungMotorische Blockaden während des Gehens (engl. freezing of gait) behindern die motorische Selbstständigkeit und sind Ursache für ein gesteigertes Sturzrisiko bei Parkinson-Syndromen. Sie kommen während des Gehens in fünf Situationen vor und kennen drei Schweregrade. Motorische Blockaden sind therapeutisch schwer anzugehen, da sie in vielen Fällen unabhängig von L-Dopa-Gaben auftreten. Für diese als On-Freezing bezeichneten motorischen Blockaden ist das Münchner Anti-Freezing-Training (MAFT) konzipiert. Signifikante Resultate in der Reduktion von Freezing konnten nach zweiwöchigem Training gezeigt werden. MAFT trainiert standardisiert und intensiv die individuell Freezing auslösenden Situationen. Das Training umfasst fünf Komponenten: Motorische FOG-Evaluation, Einsatz von individuell zugeschnittenem Cueing, Schulung von Bewegungsstrategien, hochrepetitives Training von Schrittfolgen und systematische Steigerung der Trainingsanforderung.
Das Internationale Komitee vom Roten Kreuz
Authors: Kesselring J. 
Published: March 2018

ZusammenfassungDas Internationale Komitee vom Roten Kreuz (IKRK) war Initiant der gesamten Rotkreuzbewegung und wurde 1863 in Genf gegründet. Heute ist es weltweit im Einsatz mit über 15 000 Mitarbeitern weltweit vor allem in bewaffneten Konflikten und als Hüter des Humanitären Völkerechtes. Seine Tätigkeit ist eine praktische Anwendung des Gedankengutes der Resilienz, d. h. der aufrichtenden Kräfte, die in verschiedenen Dimensionen so wichtig sind.
Evidence of a Founder Effect for the Protein C Gene 3363 Inserted C Mutation in Thrombophilic Pedigrees of French Origin
Authors: Bovill Edwin , Demers Christine , Delage Robert , Scott Bruce , Valliere Julia , Callas Peter , Jomphe Michèle , Rosendaal Frits , Aiach Martine , Long George , Couture Patrick 
Published: March 2018

SummaryWe have previously reported that the 3363 inserted (Ins) C mutation in exon 6 of the protein C gene was present in four unrelated French patients and in four French Canadian families with type I protein C deficiency as well as in a large Vermont protein C deficient kindred of French Canadian origin. The present study was designed to investigate the likelihood of the existence of a founder effect for this mutation in protein C deficient individuals of French origin living in France, Québec and Vermont. In order to demonstrate a possible founder effect for the 3363 InsC mutation, we have previously constructed a high-resolution genetic map to locate several highly polymorphic markers close to the protein C locus. Thereafter, the markers D2S347, D2S2339, D2S383, D2S2271 and D2S2215 were genotyped in 117 heterozygotes from France (n = 7), Québec (n = 36) or Vermont (n = 74). The allelic frequency distribution of these five markers was also determined in fifty control French Canadian subjects and thirty-two unaffected members of the Vermont kindred with normal protein C levels and compared with their frequency in our cohort of heterozygotes. Our data suggest that patients from Québec and Vermont carry a common haplotype at the protein C locus. Moreover, in order to study the evolutionary history of the 3363 InsC mutation, we traced back the ascending genealogy of one proband in each of the families with this mutation. These results showed that the 3363 InsC mutation was most probably introduced in North America by a couple of French settlers who established themselves in 1669 on Isle d‘Orleans located near Québec City. All heterozygotes for the 3363 InsC mutation living in North America are related to these founders within 10 generations. Thus, these families afford a unique opportunity to evaluate the role of the protein C system in thrombophilia due to the high degree of linkage disequilibrium at the protein C gene, which in essence holds that variable more constant than in a more heterogeneous population.
Homozygous Antithrombin Deficiency Type II (99 Leu to Phe Mutation) and Childhood Thromboembolism
Authors: Kuhle Stefan , Lane David , Jochmanns Kristin , Male Christoph , Quehenberger Peter , Lechner Klaus , Pabinger Ingrid 
Published: March 2018

SummaryWe report 5 children from 3 families with homozygous antithrombin deficiency type II affecting the heparin binding site (99 Leu to Phe mutation). Four children had severe spontaneous thromboembolic events (deep leg or caval vein thrombosis, ischaemic stroke) at one week, 3 months, 13 and 14 years of age. The fifth patient, a 17 year-old boy was asymptomatic. Early manifestation of homozygous deficiency calls for prompt and accurate diagnosis. In doubtful cases genetic analysis is required. Long-term oral anticoagulation should be considered in affected individuals.
Both the High Affinity Thrombin Receptor (GPIb-IX-V) and GPIIb/IIIa Are Implicated in Expression of Thrombin-induced Platelet Procoagulant Activity
Authors: Pedicord Donna , Seiffert Dietmar , Jamieson G.A. , Greco Nicholas , Dicker Ira 
Published: March 2018

SummaryPlatelets activated by -thrombin express surface procoagulant activity (PCA) that accelerates the conversion of prothrombin to α-thrombin. Following activation with 10 nM α-thrombin, the PCA of normal platelets was approximately five-fold higher than that of Bernard-Soulier platelets (lacking GPIb). Normal platelet PCA was inhibited ~50 % by activation in the presence of the anti-GPIb MoAbs LJIb10 or TM60. Moreover, normal platelet PCA was completely abrogated in the presence of a combination of both LJIb10 and c7E3, a MoAb directed against αIIbβ3 (GPIIb/IIIa). In contrast, PCA expressed by Bernard Soulier or Glanzmann platelets was not inhibited by either LJIb10 or c7E3 MoAb. The platelet activating peptide SFLLRN at 10 μM, a concentration which fully activates platelet aggregation and Ca2+ mobilization, generated PCA activity one fifth of that generated by α-thrombin at 10 nM but anti-PAR1 antibodies did not affect thrombin-induced PCA expression. These results demonstrate that GPIb mediates, at least in part, the thrombin-induced activation of platelets that leads to PCA, and that αIIbβ3 is also involved in PCA generation, but these results do not support a major role for PAR1 in this activation.
Oxidation of β2-glycoprotein I (β2GPI) by the Hydroxyl Radical Alters Phospholipid Binding and Modulates Recognition by anti- β2GPI Autoantibodies
Authors: Regnault V. , Hachulla E. , Darnige L. , Berthou F. , Youinou P. , Arvieux J. 
Published: March 2018

SummaryWe investigated whether β2-glycoprotein I (β2GPI), the key antigen in the antiphospholipid syndrome, is susceptible to oxidative modifications by the hydroxyl radical (°OH) that may influence its lipid-binding and antigenic properties. The effects on human and bovine β2GPI of °OH free radicals generated by γ-radiolysis of water with 137Cs were studied. Radiolytic °OH caused a dose-dependent loss of tryptophan, production of dityrosine and carbonyl groups, dimerization and/or extensive aggregation of β2GPI. It ensued a reduction in affinity binding to cardiolipin liposomes and loss of β2GPI-dependent autoanti-body binding to immobilized cardiolipin. Patient anti- β2GPI antibodies (n = 20) segregated into two groups based on the effect in the β2GPIELISA of β2GPI pretreatment with °OH: enhancement (group A, n = 10) or suppression (group B, n = 10) of IgG binding. The avidities of group A antibodies for fluid-phase β2GPI were low but increased in a dose-dependent manner upon β2GPI irradiation, in relation to protein crosslinking. Distinguishing features of group B antibodies included higher avidities for fluid-phase β2GPI that was no longer recognized after °OH treatment, and negative anticardiolipin tests suggesting epitope location near the phospholipid binding site. The °OH scavengers thiourea and mannitol efficiently protected against all above changes. Therefore, oxidative modifications of β2GPI via °OH attack of susceptible amino acids alter phospholipid binding, and modulate recognition by autoantibodies depending on their epitope specificities. These findings may be of clinical relevance for the generation and/or reactivity of anti- β2GPI antibodies.
A Novel Tetrameric Venom Protein, Agglucetin from Agkistrodon acutus , Acts as a Glycoprotein Ib Agonist
Authors: Wang Wen-Jeng , Huang Tur-Fu 
Published: March 2018

SummaryA novel platelet agglutination inducer, agglucetin, was purified from the Formosan Agkistrodon acutus snake venom. It migrated as a single band with an apparent molecular mass of 58.8 kDa and two distinct bands of 16.2/14.5 kDa under non-reducing and reducing conditions by SDS-PAGE, respectively. Further confirmed by FPLC, electrospray ionization mass spectrometry and 2D-PAGE, native agglucetin exists as a tetramer composed of disulfide-linked α1, α2, β1 and β2 subunits. Partial N-terminal sequence of agglucetin subunit showed a high degree of homology to those of C-type lectin-like glycoprotein (GP) Ib binding proteins. Functional studies showed that agglucetin, in the absence of von Willebrand factor (vWF), dose-dependently induced platelet agglutination and caused a negligible elevation of intracellular Ca+2 mobilization and thromboxane B2 formation in human platelet suspensions. Anti-GP Ib monoclonal antibodies (mAbs), AP1 or LJ-Ib1, specifically inhibited agglucetin-induced platelet agglutination in a dose-dependent manner. However, EDTA, arietin (a long chain RGD-containing disintegrin), 7E3 (an anti-GP IIb/IIIa mAb), heparin, hirudin, PGE1, or indomethacin exhibited no inhibitory effect on agglucetin-induced platelet agglutination. Furthermore, flow cytometric analysis revealed that FITC-agglucetin dose-dependently bound to human formalin-fixed platelets in a saturable manner, and its binding was specifically blocked by anti-GP Ib mAb. It is concluded that agglucetin, acts specifically on an epitope of platelet membrane GP Ib overlapping with that of AP1, causing platelet agglutination in a Ca+2- and GP IIb/IIIa-independent manner.
Uremic Medium Disturbs the Hemostatic Balance of Cultured Human Endothelial Cells
Authors: Serradell Mireia , Cases Aleix , Zurbano María , Aznar-Salatti José , López-Pedret José , Ordinas Antonio , Escolar Ginés , Díaz-Ricart Maribel 
Published: March 2018

SummaryWe have investigated the ability of serum from uremic patients to modify the thrombogenic properties of the endothelium. The effects of uremic medium on the morphology of endothelial cells (ECs), and their resistance to flow was analyzed. The influence of uremic media on the reactivity of the extracellular matrix (ECM) generated by ECs towards normal platelets was evaluated in a parallel-plate perfusion chamber. Exposure of ECs to uremic medium resulted in abnormal cell morphology and signs of an accelerated growth. Detachment of ECs exposed to circulating blood was increased when cells had been grown with media supplemented with uremic serum (21% vs. 14% non exposed). Platelet deposition was significantly elevated on ECMs generated in the presence of uremic media (uremicECMs) (p<0.01 vs. control studies). Effects of uremic serum were not observed at short incubation periods (5 h) but were evident after 24 or 72 h of incubation. Northern blot analysis revealed increased expression of tissue factor (TF) mRNA in ECs exposed to uremic conditions. Immunocytochemical methods detected an augmented expression of TF antigen on uremic ECMs. Incubation of ECMs with an antibody to human tissue factor prevented the increase in platelet deposition observed in uremic ECMs, suggesting that the presence of TF in ECM could be responsible for the enhanced platelet deposition. Results from our study indicate that uremic medium impairs the antithrombotic functions of cultured endothelial cells.
Biogenesis of G-protein Mediated Calcium Signaling in Human Megakaryocytes
Authors: Gorter Gertie , Vuurst Hans van der , Heemskerk Johan , Akkerman Jan-Willem , Dekker Els den 
Published: March 2018

SummaryTo understand how platelet signal transduction pathways develop during megakaryocytopoiesis, we isolated human stem cells from umbilical cord blood and cultured the cells in the presence of thrombopoietin (TPO). Based on the early expression of CD61 and late expression of CD42b, immature (CD61+/CD42blow) and mature (CD61+/ CD42bhigh) megakaryocytes were immunomagnetically purified and, together with stem cells (CD34+), characterized for G -protein expression and agonist-induced [Ca2+]i increases. Megakaryocytopoiesis was accompanied by down-regulation of the 43 kDa and 46 kDa variants of G16α, constant expression of Gsα, and up-regulation of Gqα and Giα1/2. The increase in Gq and Gi 1/2 expression was accompanied by an increase in Ca2+ signaling triggered by thrombin and other agonists known to signal to Ca2+ via these G-proteins in platelets. The prostacyclin analog iloprost and TPO also induced [Ca2+]i increases, and the iloprost-induced Ca2+ response disappeared during maturation. These data reveal sharp changes in Ca2+ regulation during megakaryocytopoiesis.
3D kinematics of the interphalangeal joints in the forelimb of walking and trotting horses
Authors: Sha D. H. , Stick J. A. , Robinson P. , Clayton H. M. 
Published: March 2018

SummaryThe objective was to measure 3D rotations of the distal (DIP) and proximal (PIP) interphalangeal joints at walk and trot. 3D trajectories of markers fixed to the proximal phalanx, middle phalanx and the hoof wall of the right forelimb of four sound horses were recorded at 120 Hz. Joint kinematics were calculated in terms of anatomically-based joint coordinate systems between the bone segments. Ranges of motion were similar at walk and trot. Values for the DIP joint were: flexion/extension: 46 ± 3° at walk, 47 ± 4° at trot; internal/ external rotation: 5 ± 1° at walk, 6 ± 3° at trot; and adduction/abduction: 5 ± 2° at walk, 5 ± 3° at trot. Within each gait, kinematic profiles at the DIP joint were similar between horses with the exception of adduction/abduction during breakover, which may vary depending on the direction of hoof rotation over the toe. Knowledge of the types and amounts of motion at the DIP joint will be useful in understanding the aetiology and treatment of injuries to the soft tissues, which are being recognized more frequently through the use of sensitive imaging techniques. Ranges of motion for the PIP joint were: flexion/extension: 13 ± 4° at walk, 14 ± 4° at trot; adduction/abduction: 3 ± 1° at walk, 3 ± 1° at trot; and internal/external rotation: 3 ± 1° at walk, 4 ± 1° at trot. The PIP joint made a significant contribution to flexion/extension of the digit. During surgical arthrodesis, the angle of fusion may be important since loss of PIP joint extension in late stance is likely to be accommodated by increased extension of the DIP joint.