The congenital Factor VII deficiency (FVIID) is a rare hemorrhagic disorderwith an autosomal recessive pattern of inheritance and a prevalence of 1:500,000. In 1994 the International Greifswald Registry of congenital FVII deficiency was initiated [1]. We analyzed the phenotype and genotype of subjects,who presented with reduced FVII activities.

Immunosuppression has recently been proposed for low-risk myelodysplastic syndromes (MDS) to reverse bone marrow failure by inhibiting intramedullary secretion of proapoptotic cytokines. We treated 35 MDS patients (24 refractory anaemia (RA), 10 RA with excess blasts and one chronic myelomonocytic leukaemia) with either horse antithymocyte globulin 15 mg/kg/day or rabbit antithymocyte globulin 3.75 mg/kg/day, each for 5 days. Median age was 63 years (range: 41-75). After 1 to 34+ months of follow-up (mean: 15+), four patients experienced complete haematological responses (CR), six good responses (GR) and two minor responses. All CRs and GRs occurred in patients with RA, in whom both horse and rabbit ATG yielded five responses out of 12 (42%). Time to response varied between 1 and 10 (mean: 3) months. The median duration of response was 9+ (1-17+) months; five patients are in continuing response. In all, 23 patients suffered side effects > degrees II WHO (the degree of toxicity encountered according to the internationally accepted WHO toxicity grading); one patient died 2 weeks after rabbit ATG from rhinocerebral mucormycosis. Parameters that correlated with response were duration of disease and RA subgroup. In our experience, immune-modulating therapy with either horse or rabbit ATG is feasible in patients with RA and short duration of disease.

The aim of the study was to look at the concentrations of pro-inflammatory cytokines and soluble cell surface receptors including tissue factor (TF) in the drainage fluid and in the serum after retransfusion. TF is a membranous glycoprotein from the surface of fibroblasts or smooth muscle cells of vessels that functions as a receptor for the coagulation factor VII/VIIa (Kobayashi 1998) ELISAs were used in twelfve patients to measure the concentrations of interleukins (IL-1β, IL-2, IL-6, IL-8), tumor necrosis factor-a as well as of soluble cell surface receptors (sIL-2Ra, sHLA-DR) and of TF. All pro-inflammatory cytokines were released into the drainage fluid at increasing concentrations, IL-6 and IL-8 thereby exceeding postoperative systemic blood levels by 200-fold or 80-fold, respectively. Reinfusion of the collected shed blood led to a further increase in both the IL-6 and IL-8 serum concentrations, which were found to be elevated already postoperatively. sHLA-DR was found in an about 100-fold excess vs. serum. The concentration of TF in the drainage fluid was five times higher (158±71 pg/ml) than in serum. There was no increase of the serum of sHLA-DR or TF levels following the retransfusion. IL-6 and IL-8 seem to be responsible for potential febrile reactions. The 100-fold elevated concentration of sHLA-DR in shed blood could therefore represent a physiological tissue level. The high TF-levels in the shed blood could be related to a local tissue damage. Dilution effects of fluid retransfused may be responsible for minor or no changes of cytokines, soluble cell surface receptors and TF in the circulation

The protein C (PC)/protein S (PS) pathway represents one fundamental endogenous anticoagulant pathway that is of crucial importance to prevent thrombosis. Important components are thrombomodulin, PC, endothelial cell protein C receptor (EPCR), and PS [ 1 Esmon C.T. The protein C anticoagulant pathway. Arterioscler. Thromb. 1992; 12: 135-145 Crossref PubMed Google Scholar , 2 Esmon C.T. The endothelial cell protein C receptor. Thromb. Haemost. 2000; 83: 639-643 PubMed Google Scholar ]. Thrombomodulin, a transmembrane protein, is of importance in initiating and activating the PC/PS pathway [ 3 Sadler J.E. Thrombomodulin structure and function. Thromb. Haemost. 1997; 78: 392-395 PubMed Google Scholar , 4 Dittman W.A. Majerus P.W. Structure and function of thrombomodulin: a natural anticoagulant. Blood. 1990; 75: 329-336 Crossref PubMed Google Scholar ]. In man, the intronless thrombomodulin gene exhibits a length of 3.6 kb and maps on chromosome 20 [ 5 Jackman R.W. Beeler D.L. Fritze L. Soff G. Rosenberg R.D. Human thrombomodulin gene is intron depleted: nucleic acid sequences of the cDNA and gene predict protein structure and suggest sites of regulatory control. Proc. Natl. Acad. Sci. U. S. A. 1987; 84: 6425-6429 Crossref PubMed Scopus (95) Google Scholar , 6 Shirai T. Shiojiri S. Ito H. Yamamoto S. Kusumoto H. Deyashiki Y. et al. Gene structure of human thrombomodulin, a cofactor for thrombin-catalyzed activation of protein C. J. Biochem. 1988; 103: 281-285 Crossref PubMed Scopus (45) Google Scholar ]. It is most conceivable that mutations within the thrombomodulin gene may affect the PC/PS anticoagulant cascade resulting in clinical manifestation of thrombosis. Several thrombomodulin gene point mutations [nt −33: G→A, nt 127: G→A, (Ala25→Thr), nt 236: G→C (Gly61→Ala), nt 543: G→A, (Glu163→Glu), nt 1456: G→T (Asp468→Tyr), nt 1502: C→T (Pro483→Leu)] were identified in patients with venous thrombosis [ 7 Norlund L. Zöller B. Öhlin A.-K. A novel thrombomodulin gene mutation in a patient suffering from sagittal sinus thrombosis. Thromb. Haemost. 1997; 78: 1164-1166 PubMed Google Scholar , 8 Le Flem L. Picard V. Emmerich J. Gandrille S. Fiessinger J.N. Aich M. et al. Mutations in promoter region of thrombomodulin and venous thromboembolic disease. Arterioscler. Thromb. Vasc. Biol. 1999; 19: 1098-1104 Crossref PubMed Scopus (67) Google Scholar , 9 Öhlin A.-K. Norlund L. Marlar A.R. Thrombomodulin gene variations and thromboembolic disease. Thromb. Haemost. 1997; 78: 396-400 PubMed Google Scholar , 10 Öhlin A.-K. Marlar R.A. The first mutation identified in the thrombomodulin gene in a 45-year-old man presenting with thromboembolic disease. Blood. 1995; 85: 330-336 PubMed Google Scholar ]. However, the association between these mutations and thromboembolic disease remains unclear, because family studies are missing or do not demonstrate perfect co-segregation [ 7 Norlund L. Zöller B. Öhlin A.-K. A novel thrombomodulin gene mutation in a patient suffering from sagittal sinus thrombosis. Thromb. Haemost. 1997; 78: 1164-1166 PubMed Google Scholar , 11 Öhlin A.-K. Marlar A.R. Thrombomodulin gene defects in families with thromboembolic disease—a report on four families. Thromb. Haemost. 1999; 81: 338-344 PubMed Google Scholar ]. Furthermore, the transition C→T at position 1418 (Ala455→Val) was shown to be not associated with thrombophilia [ [12] Van der Velden P.A. Krommenheok-van Es.T. Allaart C.F. Bertina R.M. Reitsma P.H. A frequent thrombomodulin amino acid dimorphism is not associated with thrombophilia. Thromb. Haemost. 1991; 65: 511-513 PubMed Google Scholar ].

BACKGROUND AND OBJECTIVES:Chronic refractory autoimmune thrombocytopenic purpura (AITP) is an autoimmune disorder due to autoantibodies against platelet glycoproteins (GP). Human platelet alloantigenic (HPA) systems are distributed to different platelet GPs. We carried out genotyping of diallelic HPA-1, -2, -3, and -5 systems to clarify potential associations between HPA alleles and the development of chronic refractory AITP.PATIENTS AND METHODS:DNA was isolated from 33 unrelated German patients with chronic refractory AITP and from 80 randomly selected German blood donors to determine the phenotype and allele frequencies for the HPA-1, -2, -3, and -5 systems. Fragments carrying the polymorphic sequences corresponding to those alleles were amplified by the polymerase chain reaction and further characterized by restriction analysis.RESULTS:Whereas HPA-1, -3, and -5 allele frequencies were identical in 33 patients with chronic refractory AITP and in controls, HPA-2 allele frequencies showed a statistically significant difference (p = 0.017). In our group of patients, the HPA-2a allele frequency was 100%, but HPA-2b was not seen. In contrast, the allele frequency of HPA-2a in the control group was 92% (n = 147), and in HPA-2b it was 8% (n = 13).CONCLUSION:This study suggests an association between the HPA-2a allele and chronic refractory AITP. The HPA-2a allele may be involved in the formation of an AITP-specific autoepitope.

This open-label prospective multicenter study was conducted to investigate the ex vivo biological effect of a high purity factor VIII:C/vWF complex concentrate, IMMUNATE STIM plus, manufactured by IMMUNO, Vienna, Austria [2]. The protocol used for the study was in conformity with the respective recommendations of the ISTH Scientific Subcommittee on von Willebrand Factor (vWF) studies [1].

Simple collagen-related peptides (CRPs) containing a repeat Gly-Pro-Hyp sequence are highly potent platelet agonists. Like collagen, they must exhibit tertiary (triple-helical) and quaternary (polymeric) structure to activate platelets. Platelet signaling events induced by the peptides are the same as most of those induced by collagen. The peptides do not recognize the alpha 2 beta 1 integrin. To identify the signaling receptor involved, we have evaluated the response to the CRP, Gly-Lys-Hyp(Gly-Pro-Hyp)10-Gly-Lys-Hyp-Gly of platelets with defined functional deficiencies. These studies exclude a primary recognition role for CD36, von Willebrand factor (vWF), or glycoprotein (GP) IIb/IIIa. Thus, both CD36 and vWF-deficient platelets exhibited normal aggregation, normal fibrinogen binding, and normal expression of CD62 and CD63, measured by flow cytometry, in response to the peptide, and there was normal expression of CD62 and CD63 on thrombasthenic platelets. In contrast, GPVI-deficient platelets were totally unresponsive to the peptide, indicating that this receptor recognizes the Gly-Pro-Hyp sequence in collagen. GPVI-deficient platelets showed some fibrinogen binding in response to collagen but failed to aggregate and to express CD62 and CD63. Collagen, but not CRP-XL, contains binding sites for alpha 2 beta 1. Therefore, it is possible that collagen still induces some signaling via alpha 2 beta 1, leading to activation of GPIIb/IIIa. Our findings are consistent with a two-site, two-step model of collagen interaction with platelets involving recognition of specific sequences in collagen by an adhesive receptor such as alpha 2 beta 1 to arrest platelets under flow and subsequent recognition of another specific collagen sequence by an activatory receptor, namely GPVI.

Im Jahr 1994 bzw. 1995 wurden die Gene für die Gerinnungsfaktoren VIII:C und IX:C isoliert und charakterisiert (Gietschier et al. 1984; Wood et al. 1984; Toole et al. 1985; Anson et al. 1984; Yoshitake et al. 1985). Damit waren die Voraussetzungen für die genomische Diagnostik zur Konduktorinnenbestimmung (Carrierdiagnostik) und pränatalen Diagnostik gegeben.

Ziel dieser Studie sind die Erfassung und Verlaufskontrolle von entzündlichen Mundschleimhautalterationen bei Leukämiepatienten unter Zytostatikatherapie. Bei 13 Patienten mit einer akuten myeloischen bzw. lymphatischen Leukämie erfolgte vor, während und nach 4-bis 6wöchiger Zytostatikatherapie eine exfoliativzytologische Befunderhebung im Bereich der Gingiva, der Wangen- und Gaumenschleimhaut. Der prätherapeutische Befundvergleich mit der Normalpopulation ergab bei den Leukämiepatienten eine statistisch signifikante Zunahme der Zellen aus tieferen Schichten und ein Bsinken des Keratiniationsindexes in allen untersuchten Regionen als Folge einer allgemeinen entzündlichen Schleimhautalteration. Während der Zytostatikatherapie bestand dieser Befund mit Ausnahme der Gaumenregion unverändert. Unmittelbar nach Therapieende zeigte die Gingiva zytologisch keine und die Wangenregion eine leichte, statistisch nicht signifikante Normalisierungstendenz. Allein die entzündlichen Befunde der Gauenschleimhaut wiesen zu diesem Zeitpunkt im Zytogramm eine deutliche, statistisch signifikante Rückbildungstendenz auf. Daraus folgt, daß bei Patienten mit akuten Leukämien nach Abschluß der Zytostatikatherapie selbst bei klinisch unauffälligem Schleimhauthefund eine weitere intensive orale Nachsorge geboten ist, da sich an stark beanspruchten bzw. vulnerablen Schleimhautregionen weiter subklinisch Entzündungsreaktionen abspielen.

We report the unusual complication of focal necrotizing skin lesions accompanied by moderate thrombocytopenia in a female patient undergoing thromboprophylaxis with low-molecular-weight heparin after total knee arthroplasty. Heparin-induced thrombocytopenia was suspected and confirmed using the heparin-induced platelet activation assay. The skin lesions improved gradually after the discontinuation of heparin application. In addition to the description of this exceptionally rare adverse effect of low-molecular-weight heparin, a brief discussion of previously reported cases is provided.