[Personification of the standard treatment of patients with hemophilia A (results of a multicenter Russian open-labeled prospective study evaluating the use of haemoctin)].

AIM: To evaluate the clinical efficiency, tolerability, safety, and immunogenicity of replacement therapy with haemoctin SDH (Biotest Pharma GmbH, Germany) in pretreated patients with hemophilia A (HA). SUBJECTS AND METHODS: The pretreated patients (n = 140) with varying GA from 12 regions of the Russian Federation received haemoctin replacement therapy (for prophylaxis and if needed) during a year. The levels of coagulation factor VIII, its inhibitor, HIV-1/HIV-2, the markers of hepatic and renal diseases, clinical blood test were determined and a virological study was made thrice during this period at an interval of 6 months. The therapy was considered to be effective if the pain syndrome regressed within 24 hours after the last administration of the drug and/or bleeding stopped. The results of a one-year follow-up of 106 patients receiving haemoctin monotherapy were analyzed. RESULTS: The history data suggest that there are problems in the diagnosis of hereditary coagulopathies (the mean age of diagnosis verification of 3.5 years; no family history data in 67% of cases), there is a need for the guaranteed provision of patients with adequate quantities of the drug due to the fact that a third of patients had life-threatening bleedings/hemorrhages and that the patients (46%) need additional examination and treatment for comorbidity according to the standards of therapy. Compliance of haemoctin doses and regimens with their timely individual correction ensures a 3-fold reduction on average in the incidence of the hemorrhagic syndrome, an increase in quality of life, and a decrease in the frequency of analgesic use (from 21% to 0). Treatment is satisfactorily tolerated. There is evidence for the safety and low immunogenicity of the drug. CONCLUSION: To enhance the efficiency of therapeutic-and-prophylactic care to patients with GA and to optimize their quality of life, a personification approach to treatment is recommended, by individually choosing drugs, their doses and regimens.

[Thromboembolism of the pulmonary artery in Willebrand's disease].

AIM: To ascertain risk factors of thromboembolism of the pulmonary artery (TPA) in Willebrand's disease (WD). MATERIAL AND METHODS: We made a retrospective analysis of hospitalizations of WD patients for 10 years. We analysed causes of the patients' admission, interventions, registered maximal levels of factor VIII (FVIII) and Willebrand's factor (FW) in which the interventions were made, cases of TPA. RESULTS: Thirty four patients with WD were hospitalized 45 times. Three patients were treated conservatively because of gastrointestinal bleeding, the rest patients received surgical therapy. All the patients were given FVIII concentrates, cryoprecipitate, fresh-frozen plasma. In the course of the treatment, FW and FVIII levels were determined in 38% cases, FW--in 23%, FVIII--in 27%, coagulation was studied in 12% without test for FVIII and FW levels. Maximal concentration of FW was 72.1 +/- 11.8%, FVIII--125 +/- 15.8%. TPA developed in 2 (4.4%) of 45 patients. In both cases we observed a marked rise of plasmic concentration of FVIII due to therapy (250 and 240%). CONCLUSION: In patients with WD thromboembolic complications risk factors are age, obesity, surgical interventions, immobilization, etc. Simultaneous administration of several drugs containing FW and FVIII was also among the risk factors. Overdosage of FVIII is one of the causes of thrombotic complications in WD. FW and FVIII correlations in FVIII preparations must be considered. Prophylactic heparin therapy is recommended in patients with a high risk of thrombotic complications upon achievement of normal hemostasis.

[Prevention of new cases of the disease in families of hemophilia patients in Moscow using a genetic register]. / Profilaktika povtornykh sluchaev zabolevaniia v sem'iakh bol'nykh gemofiliei v Moskve s ispol'zavaniem geneticheskogo registra.

The genetic familial register in question contains information on 441 hemophilia patients. Two-thirds of the patients suffered from moderate or severe hemophilia, 81.3% had hemophilia A. Fifty-two families with the latter disease were examined with molecular methods which confirmed heterozygous carriage in 40% of the female examinees. The prenatal diagnosis in two families led to delivery of healthy boys. The genetic register provides effective protection for hemophiliac families.

[Study of the affinity characteristics of monoclonal antibodies by solid-phase immunoenzyme analysis]. / Issledovanie affinnykh kharakteristik monoklonal'nykh antitel v metode tverdofaznogo immunofermentnogo analiza.

An approach is proposed for measuring the binding constant (Kb) for monoclonal antibodies (MA) interacting with an immobilized antigen in indirect ELISA. This approach allows the measurement of optical density (A405) in the peroxidase reaction initiated by the conjugate at different concentrations (C0) of antibodies. Using the Scatchard plots, the dependence of A405/C0 = f (A405) for the whole range of MA concentrations was examined, and the tangential of the slope (tg alpha = Kb) of the linear portion of the antigen molecule was calculated. Analysis of MA affinity parameters by using this approach may find wide use in immunodiagnostic studies aimed at measuring antigen and antibody concentrations in biological fluids as well as for estimating the efficiency of vector drugs in which the diagnostic or therapeutic component is conjugated with the vector (MA or F(ab) fragment) responsible for the drug transport to target cells. The method proposed was used for testing mouse (BALB/C) monoclonal antibodies (IgG1) to pig insulin produced by various hybridomas as well as for estimating the effect on MA of pH, temperature and hydrophobization. The minimal detectable concentration (method sensitivity) was found to depend on Kb.

[Detection of carriers of hemophilia A by testing for HindIII polymorphism in the factor VIII gene by PCR]. / Vyiavlenie nositelei gemofilii A pri pomoshchi testirovaniia polimorfnogo saita HindIII v gene faktora VIII metodom PCR.

Representatives of 62 families from Moscow and Leningrad with haemophilia A observed in the pedigree were tested for HindIII polymorphism in the factor VIII gene. The proposed scheme of investigation was based on intron 19 of the FVIII gene amplification by the PCR technique followed by restriction analysis with the inner control of hydrolysis. 207 unrelated X-chromosomes were analysed, the frequency of the incidence of the polymorphic HindIII site in the given population found to be 0.29. The frequency of incidence of the HindIII heterozygotes calculated according to Hardy-Weinberg equation was 0.41. This value evidences for relatively high informativity of this polymorphism for carrier detection and prenatal diagnosis of haemophilia A. 23 families (37%) out of 62 examined in the study were informative for this criteria. The new scheme proved to be effective in testing HindIII polymorphism for haemophilia A carrier detection and prenatal diagnosis. The whole procedure takes one day, the radiolabelled probes are not used. The scheme described was inculcated in the All-Union Research Center for Haematology, Ministry of Health, USSR, Moscow, Research Institute for Obstetrics and Gynecology, Leningrad, Institute of Medical Genetics, Greifswald, DDR.

[Appearance of factor VIII inhibitors in various diseases]. / Sluchai poiavleniia ingibitorov k faktoru VIII pri razlichnykh zabolevaniiakh.

Cases of hemorrhagic syndrome development due to the appearance of factor VIII inhibitors have been described in varying diseases. Clinical signs for the diagnosis and methods of combined therapy of such cases have been presented that are of great importance for practical use.

[Serological markers of viral infections and various indicators of immunity in patients with hemophilia]. / Serologicheskie markery virusnykh infektsii i nekotorye pokazateli immuniteta u bol'nykh gemofiliei.

The authors have studied the incidence rate for markers of viruses of AIDS (HIV), hepatitis B (HBV) and cytomegalia (CMV), as well as certain parameters of antiinfectious defense in hemophilia patients. A high incidence rate of HBV and CMV markers was established in the investigated patients, and among them no seropositive subjects, by antibodies to HIV, were detected. In the presence of a low incidence rate of chronic HBsAg-carriership in this category of patients, a significant number of immune subjects was observed, that correlated with a relative stability of such immunity parameters as concentration of serum immunoglobulin G and the level of circulating immune complexes.

[Detection of hemophilia A carriers by testing polymorphic Bcl I and HINDIII sites using the PCR method with internal splitting control]. / Vyiavlenie nositelei gemofilii A testirovaniem polimorfnykh Bcl I i Hind III saitov metodom PCR s vnutrennim kontrolem rasshchepleniia.

A new variant of the PCR test system is discussed which allows one to detect Bcl I and Hind III polymorphic sites of FVIII gene. It can be used for rapid and effective diagnosis of hemophilia A, especially, in combination with the blot-hybridization technique that detects other polymorphic variants of FVIII gene. The method proposed is highly accurate, reliable and simple. It allows one to analyze submicrogram quantities of DNA without using radiolabeled probes. The whole procedure takes several hours. The variant discussed can, possibly, be the part of the general scheme of hemophilia diagnosis completely based on the effective PCR test.

[Adenine phosphoribosyltransferase and hypoxanthine phosphoribosyltransferase of blood platelets in hereditary coagulopathies]. / Adeninfosforiboziltransferaza i gipoksantin-guaninfosforiboziltransferaza trombotsitov krovi pri nasledstvennykh koagulopatiiakh.

Activities of adenine phosphoribosyltransferase (EC 2.4.2.7 APRT) and hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8 HGPRT) were studied in thrombocytes of healthy donors, patients with hemophilia A and B and of women--heterozygote carriers of the pathologic gene. The data obtained suggest that HGPRT test may be used as a genetic marker of hemophilia as well as to detect the heterozygote carriers; estimation of APRT activity is suitable test for differentiation of hemophilia forms.