bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasma Levels of Total and Free Tissue Factor Pathway Inhibitor (TFPI) as Individual Pharmacological Parameters of Various Heparins
Authors: Gastpar Robert , Alban Susanne
Published: March 2018

The release of circulating tissue factor pathway inhibitor (TFPI) into plasma by heparins is thought to contribute to their overall anti-thrombotic activity. In the presented study in healthy volunteers, we measured the heparin-induced increase of circulating total and free TFPI antigen and the aXa- and aIIa activity after subcutaneous (s.c.) injection of 9000 aXa-U of four different heparins: unfractionated heparin (UFH) (13.0 kDa), a medium molecular weight (MW) heparin with a narrow MW range (HF) (10.5 kDa), certoparin (6.0 kDa) and enoxaparin (4.5 kDa). Based on the administration of equi-active aXa doses, certoparin induced the highest increase in total TFPI determined as AUC (p <0.01). The lowest effect was observed for UFH (p <0.0001). However, the AUC of released free TFPI significantly increased in the order: enoxaparin < UFH < certoparin < HF, showing MW dependency with the exception of UFH. Comparing the effects of equi-gravimetric heparin doses, the MW dependency becomes even more pronounced. The mismatch of UFH may be due to its poor bioavailability, which becomes obvious from its low ex vivo aXa activity. In contrast to the TFPI releasing potency, the ex vivo aXa activity continuously decreased with increasing MW. Although the ex vivo aIIa activity of the heparins increased in the same order like the release of free TFPI, there was no clear correlation. This is attributed to the fact that the aIIa activity of heparin is not only dependent on the MW, but, in contrast to its TFPI releasing effect, also on the percentage of material with high affinity to AT. In conclusion, besides the aXaand aIIa activity, the TFPI releasing effect of heparins is an additional parameter of their individual pharmacological profile.
The NFY Transcription Factor Mediates Induction of the von Willebrand Factor Promoter by Irradiation
Authors: Bertagna Angela , Jahroudi Nadia
Published: March 2018

Ionizing irradiation in patients is proposed to cause thrombus formation. An increase in von Willebrand factor secretion in response to irradiation is a major contributing factor to thrombus formation. We have previously reported that the increased VWF secretion in response to irradiation is mediated at the transcriptional level. The VWF core promoter fragment (sequences –90 to +22) was shown to contain the necessary cis-acting element(s) to mediate the irradiation response of the VWF gene. Here we report that a CCAAT element in the VWF promoter is the cis-acting element necessary for irradiation induction and that the NFY transcription factor interacts with this element. These analyses demonstrate that inhibition of NFY’s interaction with the CCAAT element abolishes the irradiation induction of the VWF promoter. These results provide a novel role for NFY and add this factor to the small list of irradiation-responsive transcription factors. Coimmunoprecipitation experiments demonstrated that NFY is associated with the histone acetylase P/CAF in vivo and that irradiation resulted in an increased association of NFY with coactivator P/CAF. We propose that irradiation induction of the VWF promoter involves a mechanism resulting in increased recruitment of the coactivator P/CAF to the promoter via the NFY transcription factor.
Factor XIII of Blood Coagulation as a Nuclear Crosslinking Enzyme
Authors: Antal Miklós , Módis László , Sárváry Attila , Szücs Sándor , Balogh Imre , Ádány Róza , Bárdos Helga
Published: March 2018

Intracellular localization and distribution of Factor XIII subunit A (FXIIIA) was investigated in association with monocyte-macrophage differentiation in a long term culture of human monocytes by light-and electron microscopical as well as biochemical and immunobio-chemical techniques. To allow the detection of FXIIIA in cells with well-preserved ultrustructure, immunosera against glutaraldehydederivatized recombinant FXIIIA were developed in rabbits, then characterized and used in this study. In the early phase of macrophage differentiation intranuclear accumulation of FXIIIA was detected as a transient phenomenon in cells of the 2nd day culture by optical sectioning with 0,7 m steps in laser scanning confocal microscopy and immunoblotting technique. FXIIIA could be detected by immuno-electron microscopic postembedding staining over electrodense DNA-containing areas. Fluoresceinated monodansylcadaverine incorporation assay was used to demonstrate that FXIIIA is not only present in the nuclei, but also expresses its transglutaminase activity. Our finding of the nuclear accumulation of FXIIIA in differentiating human macrophages is also unique in that a blood clotting factor has, for the first time, been localized in nuclei and has been shown to be an intracellular crosslinking enzyme. The possible role of nuclear FXIIIA in association with cellular processes involving chromatin structure remodeling, such as cell death, cell differentiation or cellular proliferation requires further in-depth investigation.
Mouse Protein Z-Dependent Protease Inhibitor cDNA
Authors: Zhang Jing , Broze George
Published: March 2018

Protein Z-dependent protease inhibitor (ZPI) is plasma proteinase inhibitor in the serpin superfamily that produces rapid inhibition of factor Xa in the presence of phospholipids, Ca++ and protein Z (PZ). Mouse ZPI cDNA was isolated and cloned from mouse liver RNA using RT-PCR. The cDNA contains 100 nucleotides 5’ of a translation initiation codon and an open reading frame of 1344 nucleotides followed by a 163 nucleotide 3’ untranslated sequence with a poly (A) tail. The cDNA predicts a signal peptide containing 21 amino acids and a mature protein of 427 residues with 8 potential sites for N-linked glycosylation. The oligonucleotide and predicted amino acid sequences of mouse ZPI are 72% and 81% homologous with those of human ZPI. Like human ZPI, mouse ZPI contains tyrosine-serine (P1-P1’) at its reactive center in contrast to the rat molecule which contains tyrosinecysteine. By Northern analysis, mouse ZPI mRNA is 1.6 kb in size and, similar to both human and rat, it is detectable in liver, but not in heart, brain, spleen, lung, kidney, skeletal muscle or testes.
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

Increased 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

Abciximab (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

Adhesion 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

Die 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

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

Motorische 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

Das 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

We 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

We 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

Platelets 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

We 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

A 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

We 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

To 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

The 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.
A novel radiographic method to facilitate measurement of the tibial plateau angle in dogs
Authors: Cook J. , Helphrey M. , Crouch D. , Fox D. , Schultz L. , Cook C. , Kunkel J. , Headrick J.
Published: March 2018

The tibial plateau levelling osteotomy (TPLO) is commonly performed for treatment of cranial cruciate ligament deficiency in dogs. In order to be performed as described, this procedure relies on consistent measurement of the tibial plateau angle (TPA) on radiographs. This prospective study compared two radiographic methods for subsequent TPA measurement with respect to measured angle and ease of determining landmarks for measurement as determined by four observers. One method was the accepted standard radiographic protocol outlined in the TPLO training seminars. The other method involved a novel split image radiographic protocol not yet described in the literature. Participants' subjective scores as to ease of identifying landmarks and determining TPA on radiographs for each method were evaluated. Inter-observer TPA measurement variability was also assessed for each method. The novel radiographic method was judged to be significantly better in terms of ease of measuring TPA. Inter-observer measurement variability was considered appropriate for recommending use of this novel method for radiographing patients for TPA measurements.
Contact mechanics of the ovine stifle during simulated early stance in gait
Authors: Lee-Shee N. K. , Hurtig M. B. , Dickey J. P.
Published: March 2018

Ovine stifle joint contact pressures and contact areas were measured in vitro using a six degree-of-freedom (DOF) robotic system. The robot generated static joint loads of 1.875 times body weight (BW) compression, 0.15 BW medial shear and 0.625 BW cranial shear at 6.5o of flexion for four specimens, simulating the early stance phase of gait (walking). This condition represents a period of intense loading and was implemented as a worst-case loading scenario for the joint at this gait. We determined that the medial and lateral compartments bore 5.5 ± 0.9 MPa and 4.4 ± 1.1 MPa of mean pressure, respectively, on 107.7 ± 28.7 mm2 and 60.8 ± 56.3 mm2 of area, respectively. The unique contribution of this study is that stifle contact pressures and areas were determined during loading which simulated physiological levels (early stance phase of gait). This information is important to our understanding of the stresses that must be borne by repair tissues/constructs that are implanted into human and animal tibio-femoral joints.
Diagnostic findings in a lame-free dog with complete rupture of the biceps brachii tendon
Authors: van Ryssen B. , Gielen I. , Taeymans O. , van Bree H. , Wiemer P.
Published: March 2018

A case of a complete rupture of the biceps brachii tendon in a lame-free three-year-old Labrador Retriever is described. The dog had a history of chronic lameness following a trauma, but had become lame-free shortly before presentation. Clinical examination, biceps tendon test and shoulder instability tests under general anaesthesia were inconspicuous for shoulder instability. Routine radiographs, arthrography and ultrasound were suspicious for bicipital tendopathy. The definite diagnosis of a complete rupture of the biceps brachii tendon was made upon arthroscopic inspection of the glenohumeral joint.
Schwere systemische juvenile idiopathische Arthritis und familiäres Mittelmeerfieber mit homozygoter Mutation im MEFV-Gen
Authors: Geikowski T. , Peters D. , Peitz J. , Horneff G. , Wintrich S.
Published: March 2018

Die systemische juvenile idiopathische Arthritis (sJIA) und das familiäre Mittelmeerfieber (FMF) zeigen mit Fieber, Arthritis, Exanthem, generalisierter Lymphadenopathie und Sero sitis viele Gemeinsamkeiten. Therapieoptio-nen bei der sJIA sind neben Glukokortikoi-den, nichtsteroidalen Antirheumatika und Methotrexat, Biologika wie Interleukin-1-und Interleukin-6-Antagonisten. Firstline Therapie des FMF ist Colchicin. Bei Therapie resistenz oder -unverträglichkeit können aber auch bei dieser Erkrankung Interleukin-1-und Interleukin-6-Antagonisten erfolgreich eingesetzt werden.

Berichtet wird über einen elf Jahre alten Jungen, geboren in Armenien. Im Alter von zwei Jahren kam es zu anhaltendem Fieber mit ausgeprägter Arthritis. Die Diagnose sJIA wurde gestellt und das Vorliegen eines FMF genetisch nachgewiesen. Unter Therapie mit Kortikosteroiden, Sulfasalazin und Colchicin entwickelte der Patient eine deutliche Dystro phie und ausgeprägte Destruktionen der Hüft-, Knie- und Sprunggelenke mit resultie-render Laufunfähigkeit. Die Halswirbelsäule war durch Ankylosen kyphotisch fixiert. La borchemisch waren die Entzündungszeichen deutlich erhöht. Röntgen- und CT-Aufnah-men zeigten das Ausmaß der Gelenkzer -störung. Nach einem Jahr unter intravenöser Tocilizumab-Therapie liegen CRP, SAA und BSG im Normbereich. Bestehende Kontraktu ren sind unter intensiver Physiotherapie rück-läufig. Der Patient ist mit eigenem Rollstuhl mobil.

Bei diesem Patient liegt sowohl eine systemische JIA als auch ein familiäres Mittelmeerfieber vor. Tocilizumab ist bereits seit mehreren Jahren für die Behandlung der systemischen JIA zugelassen. Unser Patient zeigt unter Tocilizumab eine Remission beider Erkrankungen.
Perikoxale Schmerzsyndrome – nicht immer einfach zu diagnostizieren
Authors: Guttke T. , Arnold I.
Published: March 2018

Pertrochantäre Schmerzen stellen ein häufiges klinisches Problem dar. Auch wenn perikoxale Schmerzsyndrome bereits früh beschrieben worden sind, hat die Weiterent-wicklung der arthroskopischen Techniken unter Einbezug der extrakapsulären Kompartimente die Sensibilität für diese häufig fehl -interpretierten Erkrankungsformen erhöht. Voos und Mitarbeiter veröffentlichten 2007 sehr früh ihre ersten Ergebnisse im Hinblick auf endoskopische Interventionen in diesem Zusammenhang (1). Andere Erkrankungsformen wie das “low anterior inferior iliac spine impingement” wurden sogar erst durch die Hüftarthroskopie der Beschreibung zugänglich. Zu den pertrochantären Pathologien, die das laterale Kompartiment betreffen, zählen Bursitiden und Insertionstendinosen der Glutealmuskulatur bis hin zur Ruptur und das Tractus-iliotibialis-Syndrom einschließlich der externen Schnapphüfte. Projektionsschmerzen von der proximalen Lendenwirbelsäule sind in dieser Übersicht nicht berücksichtigt, verursachen aber in bis zu 30 % pertrochantäre Beschwerden. Auf nervaler Ebene können Irritationen des inferioren Glutealnervs mit radikulärem Ursprung L5–S2 (Innervation des M. gluteus maximus) oder des N. gluteus superior (Wurzeln L4–S1; Innervation des M. gluteus medius und minimus, M. tensor fasciae latae und vordere Partien des Hüft gelenks) Trochanter-bezogene Schmerzen imitieren. Im ventralen extraartikulären Kompartiment kann eine Friktion der Sehne des M. iliopsoas ein internes Impingement und Schnappen verant-worten. Natürlich sind differenzialdiagnostisch auch Arthrose-assoziierte muskulotendinöse Kontrakturen oder auch Gangasymmetrien bzw. Störungen im lumbosakralen Übergang mit konsekutiven muskulären Dysbalancen auszuschließen. Aber auch nach Hüftgelenkersatz stellen unzureichend azetabulär-knöchern gedeckte Pfannenkomponenten eine Disposition für eine Irritation der Hüftbeuger dar. Anatomisch dem posterioren Bereich zugeordnet sind die Strukturen um den Nervus ischiadicus (z. B. Entrapmentphänomene durch fibrinöse Stränge), die Hüftaußenrotatoren und explizit der M. piriformis. Aber auch Insertionstendinopathien der Hamstrings sind differenzialdiagnostisch perikoxalen Schmerzsyndromen zuzuordnen.
Thrombophilia in the young
Authors: Kurnik K. , Krümpel A. , Stoll M. , Nowak-Göttl U.
Published: March 2018

Venous thromboembolism (VTE) is a rare disease that is being increasingly diagnosed and recognized in paediatrics in the past decade, usually as a secondary complication of primary severe underlying diseases. Apart from acquired thrombophilic risk factors, such as lupus anticoagulants, inherited thrombophilias (IT) have been established as risk factors for venous thromboembolic events in adults. In children with idiopathic VTE and in paediatric populations in which thromboses were associated with underlying medical diseases, IT have been described as additional prothrombotic risk factors. Follow-up data for VTE recurrence in children are available and suggest a recurrence rate of approximately 3% in neonates and 8% in other children. Here we present a review of the impact of IT on early onset of VTE and recurrence in children. Statistically significant associations between the IT traits investigated, e.g. factor V G1691A, factor II G20210A, protein C-, protein S-, antithrombin deficiency, elevated lipoprotein (a), combined IT and VTE onset were reported. In addition, statistically significant associations with recurrent VTE were calculated for protein S-, antithrombin-deficiency, and the factor II variant and combined IT. The absolute risk increase for VTE recurrence associated with IT ranged from 9.8 % for children carrying the factorII variant to 26% and 29% in children with combined IT and protein S-deficiency, respectively. Data obtained gave evidence that the detection of IT is clinically meaningful in children with VTE and underlines the importance of a paediatric thrombophilia screening program. Based on these data treatment algorithms have to be discussed.
Role of coagulation and fibrinolysis in lung and renal fibrosis
Authors: Ruppert C. , Markart P. , Wygrecka M. , Preissner K. T. , Günther A.
Published: March 2018

Elevated procoagulant and suppressed fibrinolytic activities are regularly encountered in different forms of clinical and experimental fibrosis of the lungs and the kidneys. Although primarily serving to provide a provisional matrix of repair largely consisting of fibrin and fibronectin, the involved procoagulant serine proteases and protease inhibitors may also exert distinct cellular downstream signaling events modifying the fibrotic reponse.

In this review, evidence for an impaired regulation of coagulation and fibrinolysis factors in clinical and experimental lung and renal fibrosis is provided and the role of PAR (protease activated receptor) induced profibrotic and HGF (hepatocyte growth factor) elicited antifibrotic cellular events is worked out. In view of experiments obtained in animal models of lung and renal fibrosis, the potential therapeutic usefulness of anticoagulant or profibrinolytic strategies is discussed.
Pulmonary embolism
Authors: Humenberger M. , Lang I. M.
Published: March 2018

Die akute Pulmonalembolie ist Teil des Spektrums venöser Thromboembolien. Die tiefe Beinvenenthrombose, der akute Thrombus in Transit durch das rechte Herz, die akute Pulmonalembolie selbst und die chronisch thromboembolische pulmonale Hypertension (CTEPH) gehören dazu. Pulmonalembolie kann rezidivieren und schwere Spätfolgen (z. B. postthrombotisches Syndrom, CTEPH) nach sich ziehen.

Diese Übersicht fasst aktuelle Konzepte zur Pathophysiologie, Epidemiologie, Diagnose und Therapie dieser häufigen Erkrankung zusammen.
Thrombophilia in the young
Authors: Kurnik K. , Krümpel A. , Stoll M. , Nowak-Göttl U.
Published: March 2018

Venous thromboembolism (VTE) is a rare disease that is being increasingly diagnosed and recognized in paediatrics in the past decade, usually as a secondary complication of primary severe underlying diseases. Apart from acquired thrombophilic risk factors, such as lupus anticoagulants, inherited thrombophilias (IT) have been established as risk factors for venous thromboembolic events in adults. In children with idiopathic VTE and in paediatric populations in which thromboses were associated with underlying medical diseases, IT have been described as additional prothrombotic risk factors. Follow-up data for VTE recurrence in children are available and suggest a recurrence rate of approximately 3% in neonates and 8% in other children. Here we present a review of the impact of IT on early onset of VTE and recurrence in children. Statistically significant associations between the IT traits investigated, e.g. factor V G1691A, factor II G20210A, protein C-, protein S-, antithrombin deficiency, elevated lipoprotein (a), combined IT and VTE onset were reported. In addition, statistically significant associations with recurrent VTE were calculated for protein S-, antithrombin-deficiency, and the factor II variant and combined IT. The absolute risk increase for VTE recurrence associated with IT ranged from 9.8 % for children carrying the factor II variant to 26% and 29% in children with combined IT and protein S-deficiency, respectively. Data obtained gave evidence that the detection of IT is clinically meaningful in children with VTE and underlines the importance of a paediatric thrombophilia screening program. Based on these data treatment algorithms have to be discussed.
Niedrigdosierte direkte Katheterlyse venöser Thrombosen des Schultergürtels
Authors: van Werde M. , Timmermann J. , Lachhein M.
Published: March 2018

Bei venösen Thrombosen des Schultergürtels bietet sich eine direkte Applikation thrombolytischer Substanzen in das verschlossene Venensegment an, da man von einer geringen Spontanrekanalisationsrate und der Wahrscheinlichkeit chronischer Beschwerden ausgehen muß.

Bei 14 Patienten mit nachgewiesenen ausgeprägten Schultervenenthrombosen (Anamnesedauer 2-35 d, Mittelwert 11 d) wurde eine direkte venöse Katheterlyse durchgeführt. Über einen Lysekatheter wurde das Fibrinolytikum (20 mg rt-PA in 6 h/d) mit suffizienter Heparingabe (PTT 1,5fach) in das thrombosierte Venensegment appliziert. Die Therapiedauer betrug 2 bis maximal 5 Tage. Zusätzlich führten wir nach der Lysetherapie bei 5 Patienten eine Ballondilatation und ergänzend zweimal eine Stentimplantation bestehender venöser Stenosen im ehemals thrombosierten Gefäßsegment durch.

Eine vollständige Rekanalisation wurde bei 8 Patienten erreicht, bei 3 Patienten eine Teilrekanalisation; 3 Fälle waren nicht zu eröffnen. Lebensbedrohliche Komplikationen traten nicht auf. In der Langzeitbeobachtung bis 30 Monate nach der Therapie wiesen 12 Patienten keine Beschwerden mehr auf, 2 Patienten klagten über Funktionseinschränkungen und Schwellneigung.
Response to DDAVP in children with von Willebrand disease type 2
Authors: Budde U. , Beutel K. , Hassenpflug W.-A. , Hauch H. , Obser T. , Oyen F. , Schneppenheim S. , Schrum J. , Schneppenheim R.
Published: March 2018

We have prospectively evaluated the biologic response to desmopressin (DDAVP) in 28 children with type 2 von Willebrand disease (VWD) in correlation with the phenotype and the molecular defect of VWF. The diagnosis of VWD type 2 was mainly based on VWF functional parameters and/or an aberrant VWF multimer pattern. Seventeen different mutations were identified (6 of them novel). No response with respect to the functional parameters VWF:RCo and/or VWF:CB was seen in patients with severe abnormality of the VWF multimer pattern. One patient with VWD type 2A phenotype IIC Miami did not respond with respect to VWF:CB, but showed a good response of VWF:Ag and FVIII:C as expected. Interestingly he showed a persistently high level of VWF:Ag and FVIII:C up to 4 hours after DDAVP infusion. Patients with minor alterations of multimer structure and particular mutations responded well to DDAVP, whereas patients with normal multimer structure but a defect in platelet dependent functional parameters did not respond with VWF:RCo. Conclusion: Children with VWD type 2 show a variable response to desmopressin depending on the mutation that correlates with the functional defect and the presence or absence as well as the half-life of large VWF multimers. Our data emphasize the usefulness of DDAVP testing even in patients with VWD type 2, possibly with the exception of VWD type 2B.
ITI with high-dose FIX and combined immunosuppressive therapy in a patient with severe haemophilia B and inhibitor
Authors: Hauch H. , Rischewski J. , Kordes U. , Schneppenheim J. , Schneppenheim R. , Beutel K.
Published: March 2018

Inhibitor development is a rare but serious event in hemophilia B patients. Management is hampered by the frequent occurrence of allergic reactions to factor IX, low success rates of current inhibitor elimination protocols and the risk of development of nephrotic syndrome. Single cases of immune tolerance induction (ITI) including immunosuppressive agents like mycophenolat mofetil (MMF) or rituximab have been reported. We present a case of successful inhibitor elimination with a combined immune-modulating therapy and high-dose factor IX (FIX). This boy had developed a FIX inhibitor at the age of 5 years and had a history of allergic reactions to FIX and to FEIBA→. Under on-demand treatment with recombinant activated FVII the inhibitor became undetectable but the boy suffered from multiple joint and muscle bleeds. At the age of 11.5 years ITI was attempted with a combination of rituximab, MMF, dexamethasone, intravenous immunoglobulins and high-dose FIX. The inhibitor did not reappear and FIX half-life normalized. No allergic reaction, no signs of nephrotic syndrome and no serious infections were observed.
Severe prekallikrein deficiency due to a compound heterozygosis in the KLKB1-gene
Authors: Kochhan L. , Heuchel P. , Jenderny J. , Maak B.
Published: March 2018

A 14 year old boy was referred to us for a detailed coagulation study because a previously performed aPTT has been found prolonged. The boy had no history of bleeding symptoms and also the family history was negative for bleeding or thrombotic events. The aPTT in the patient was 96 s (reference range: 24–36 s), prothrombin time and thrombin time were both normal.

As the cause for the prolonged aPTT we identified a severe prekallikrein deficiency (prekallikrein activity < 1%). The prekallikrein deficiency results from two mutations in the KLKB 1-gene: first, an insertion of 1 bp in codon 149 in exon 5 and, second, a base exchange Cys 548 (TGC) > Tyr (TAC) in exon 14. The boy inherited the first mutation from his father and the second from his mother. The mutation in the paternal allele was not described before the completion of our study. There are two brothers of the propositus, one with normal prekallikrein activity and no mutations in the KLKB1-gene, the other showed the same constellation as the propositus.
New anticoagulants for secondary haemostasis
Authors: Nowak G.
Published: March 2018

In contrast to heparins and oral anticoagulants, anti IIa inhibitors (thrombin inhibitors) are able to directly inhibit the protease activity of thrombin and can thereby precisely control the blood coagulation process.

Direct thrombin inhibitors are either biosimilars (r-hirudin) or synthetically produced substances (bivalirudin, argatroban, dabigatran). In 1997 r-hirudin was introduced into clinical practice, however due to its narrow therapeutic range and the necessity of drug monitoring it has not gained widespread clinical use by now. Since 2004 and 2005 the synthetic thrombin inhibitors bivalirudin and argatroban, respectively, are available. With dabigatran the first oral synthetic thrombin inhibitor followed in 2008. These four drugs can inhibit even clot bound thrombin and show low plasma protein binding. They differ in respect to route and duration of application as well as elimination from the body, thereby offering a precise inhibition of blood coagulation adjusted to the individual case and without danger of HIT II. These advantages shall be used and advanced by the development of further direct thrombin inhibitors.