Comparative Analysis of Two Enoxaparin Sodium Drugs by Heptest.

The optimal conditions for analysis of low-molecular-weight heparin (LMWH) drugs by the Heptest and the metrological characteristics of the Heptest method were determined. The effects of two LMWH preparations of enoxaparin sodium, the domestic drug Enoxaparin sodium (Technologiya Lekarstv, Russia) and the original drug Clexane (Sanofi, France), on the clotting time of human plasma in the Heptest were comparatively analyzed. The mean blood clotting times were calculated for each dilution of each series of Enoxaparin sodium and Clexane based on the results of the Heptest. Graphs of the dependence of the plasma clotting time on the degree of dilution of these drugs were plotted. The shapes of the curves of the dependence of the blood plasma clotting time on the activity of enoxaparin sodium were shown to be similar for both Enoxaparin sodium and Clexane. The Heptest method showed that Enoxaparin sodium (Technologiya Lekarstv, Russia) was comparable to the original drug Clexane (Sanofi, France).

[Whether fibrinogen concentrates are necessary in Russia?].

PURPOSE OF THE STUDY: To analyze an efficiency of hypofibrinogenemia treatment. In the Scientific Center for Hematology (Moscow) significant hypofibrinogenemia occurs in 3% of patients with hemoblastosis. 1000 doses of cryoprecipitate are used for a hypofibrinogenemia treatment every year (21-23 doses for each patient). Containing of fibrinogen in a one cryoprecipitate dose is from 108 mug to 711 mug (M = 276 mug). Volume of one dose is from 8 to 90 ml (M = 24 ml). Hypofibrinogenemia occurred in all patients required a cryoprecipitate transfusion (M = 1 g L(-1), min 0.5 g L(-1), max 2 g L(-1)). We fixed an increasing of fibrinogen level in plasma by 0.7 +/- 0.2 g L(-1) after the cryoprecipitate transfusion. We analyzed a world experience of the use of fibrinogen containing blood components. CONCLUSIONS: Fresh frozen plasma transfusion cannot be a choice method of treatment for hypofibrinogenemia. Fibrinogen's concentrate has the same effectiveness as a cryoprecipitate both for congenital and acquired deficit of fibrinogen. The frequency of complications due to fibrinogen's concentrate is low. Currently clinical studies of recombinant fibrinogen are conducted. Ways of implementation of fibrinogen preparations in Russia are discussed.

[Comparative study of osteoplastic materials based on chitosan, alginate or fibrin with tricalcium phosphate].

The study presents comparative analysis of porous composites made of chitosan, alginate, fibrin with beta-tricalcium phosphate. Histological findings on Wistar rat condyles showed that fibrin-beta-TCP-based composite had the most effective positive biological response.

[Assessment of thromboplastins according to the WHO guidelines and the results of external quality control].

The study was undertaken to assess commercial thromboplastins for compliance to the WHO guidelines and to substantiate the validity of the results of a prothrombin test carried out using these thromboplastins. The test thromboplastins were shown to meet the WHO guidelines for assessment of thromboplastins. Over 5-8 years of the authors'participation in two external quality control programs (Federal External Quality Control System, Russia; 72 trials; NEQAS, United Kingdom; 60 trials), the international normalized ratio derived through the use of assessed thromboplastins did not differ from that established due to the interlaboratory consensus value of both external quality control systems. It is concluded that correct thromboplastin assessment provide accurate results of determination of prothrombin time.

[Monitoring the efficiency of heparin anticoagulation in patients on programmed hemodialysis].

The activated partial thromboplastin time (APTT) test is widely used to monitor heparin anticoagulation. The relationship of the APTT to the activity of administered unfractionated heparin was defined in 23 patients on programmed hemodialysis, by using six APTT reagents. A calibration curve was plotted for each reagent. The heparinized and nonheparinized plasma values of APTT depended on the reagent being used. During clinical heparin therapy, an APTT-heparin concentration relationship was also defined by the used reagent. The reagents showed varying heparin sensitivity both in vitro and in vivo studies. APTT was estimated for the therapeutic application of heparin for all the reagents in the examinees. The authors consider it necessary to define an APTT-heparin concentration relationship for each reagent being used.

[Approbation of a new test system determining the activity of a plasmin inhibitor in patients with sepsis].

Plasmin inhibitor (PI) determination is an essential diagnostic method. The purpose of the study was to develop an amidolytic assay for estimating PI activity, by applying the test system made by RENAM (Moscow). The new system is based on purified plasmin (human plasma) with the activity attested by the international standards. The developed method shows precision and accuracy with low and normal PI activity. The pilot clinical trial in patients with sepsis had demonstrated that the PI activity determined by this method is associated with some hemostatic parameters (prothrombin index, thrombin generation) and a patient's status (septic shock, hepatic dysfunction).

[Fibrinogen determination by the Clauss method. Reagents and analyzers].

The Clauss method was recommended by the WHO to measure plasma fibrinogen levels. The purpose of this study was to assess the application of test systems made by the RENAM Research-and-Production Association (Moscow) on different types of clinical laboratory analyzers to measure fibrinogen by the Clauss method. The calibration line is shown to be of major value in obtaining correct results on each coagulometer. The findings suggest the accuracy and reproducibility of the determination of fibrinogen concentrations by the Clauss when the REHAM test systems are used to measure fibrinogen concentrations on various analyzers if accurate calibration lines are obtained.

[Analysis of lupus anticoagulant. International external quality control].

The paper describes a reagent kit and standardized methods for lupus anticoagulant (LA) laboratory diagnosis. LA could be determined on the basis of screen/confirm assays and the reagents produced by RENAM, Moscow. The external quality control by ECAT Foundation proved the correctness of the authors' LA determining procedure. A pilot clinical study has shown that the developed approach has a high LA specificity for differential approach in thrombophilic patients.

[Standardization of coagulologic technique for antithrombin III estimation and its clinical application].

A standard assay procedure for estimating antithrombin III activity was derived from the Abildgaard technique as follows: 1) to decrease errors at the preanalytical stage, by using preliminarily defibrinated lyophilized plasma-calibrator and 2) to increase measurement accuracy, by using the concentration of a fibrinogen preparation and simplifying the analytical procedure. The estimation of plasma antithrombin activity in patients with pulmonary thromboembolism or sepsis points to the changes in antithrombin III activity.

[Modified capillary blood prothrombin-time test for the calculation of an international normalized ratio].

Prothrombin time (PT) is usually measured in citrated venous blood. This test is undesirable in pediatric practice and in patients taking anticoagulants due to frequent venepunctures. Capillary blood PT test has no adverse impact. The authors have developed a standardized modified method for PT measurement in capillary blood. The adequacy of the test is due to the fact that analyzed test packed cell volumes are kept in mind. The modified test has shown that the values of normal plasma PT may be used to calculate an international normalized ratio (INR). It makes it possible to determine capillary blood PT in the range of packed cell volumes from 0.15 to 0.7 in the examinees and to monitor anticoagulant therapy. The results of the modified test closely correlated with those when determining PT and INR in venous plasma (r = 0.99) and applying the CoaguChek test (r = 0.97).

[The determination of activity of low-molecular heparins against activated factor Xa by means of the diagnostic kit "Reachrom-Heparin" (scientific-production association "Renam")].

It is possible to successfully determine both the activity against activated factor Xa in animals plasma after heparins administration and specific activity against activated factor Xa of new anticoagulants by means of the domestic diagnostic kit "Reachrom-Heparin" developed by the company "Renam". It was compared to the similar kit "Berichrom heparin" (Dade Behring). No statistically significant differences were detected.

[Mutations of genes associated with thromboses in ischemic stroke in patients with primary antiphospholipid syndrome].

AIM: To study factor V Leiden, prothrombin (G1691A), 5, 10-methylenetetrahydrofolate reductase (MTHFR, C677T) mutations in patients with primary antiphospholipid syndrome (PAPS) and cerebrovascular disease (CVD). MATERIAL AND METHODS: We studied 44 patients (38 female, 6 male, mean age 41.6 +/- 11.6 years) with PAPS and CVD. Detection of mutations was carried out using polymerase chain reaction. RESULTS: Heterozygous factor V Leiden mutation was found in 11% patients, heterozygous prothrombin mutation--in 9%, heterozygous and homozygous MTHFR mutation--in 50% and 9%, respectively. The severity of CVD, frequency of clinical manifestations related to non-cerebral arterial and venous thrombosis did not differ between the patients with and without mutations or there was a tendency to less frequent occurrence of these manifestations in patients with mutations. Patients with heterozygous factor V Leiden mutation, heterozygous prothrombin mutation or homozygous MTHFR mutation less frequently developed recurrent ischemic stroke than patients without these mutations (8% versus 44%, p < 0.02). CONCLUSION: It is suggested that mutations studied do not play a significant role in development of cerebral and systemic thrombosis in patients with PAPS. The leading role belongs to antiphospholipid antibodies (aPL). Sometimes these mutations may protect from thrombogenic aPL action. This could underlie less frequent development of recurrent ischemic stroke in patients with mutation.

[Anti-phosphatidylethanolamine antibodies in patients with Sneddon's syndrome].

Anti-phosphatidylethanolamine antibodies (aPE) belong to the group of anti-phospholipid antibodies (aPL) and are directed against neutral phospholipid, connected with co-factor protein, while cardiolipin antibodies (aKL) are directed against negative phospholipid. The paper presents a study of prevalence and clinical significance of IgG aPE in 28 patients (22 women and 6 men, mean age 47.6 +/- 11.6 years) with Sneddon's syndrome (SS), which consists in cerebrovascular disturbances and extensive livedo reticularis. IgG aPE were detected by immune-enzyme assay. The upper normal limit, calculated as mean + 3SD after studying 19 healthy donors, was 0.303 optic density units. aPE were found in 15 (54%), aKL and/or lupus anticoagulant (LA)--in 6 (21%) patients with SS. aPE were found in 10 (46%) out of 22 aKL- and LA-negative patients. Among the aPE-positive patients there was a higher incidence of cortic dementia (53% vs. 8%, p = 0.02), the widening of cortical sulci, detected by means of computed tomography and magnetic resonance imaging (73% vs. 31%, p = 0.05), and mild renal syndrome (73% vs. 16%, p = 0.03). Besides, they displayed a higher rate of headaches (87% vs. 62%), chorea (33% vs. 8%), epilepsy (27% vs. 8%), non-carrying of pregnancy (91% vs. 50%), peripheral venous thrombosis (27% vs. 15%), coronary heart disease (47% vs. 31%), cardiac valvular thickening, detected by means of EchoCG (93% vs. 69%), arterial hypertension (87% vs. 54%), thrombocytopenia (20% vs. 0), anemia (40% vs. 15%); however, the difference was not significant. The results show that aPE detection, performed in addition to detection of classic immunological antiphospholipid syndrome markers (aKL and LA), increases the portion of aPE-positive patients with SS by 33%. aPE are often (in 46% of cases) found in aKL- and LA-negative patients with SS. aPE is likely to be the most significant factor of thrombosis in small arteries of the brain cortex and kidneys, which could explain their association with dementia and renal syndrome.

[Clotting factor VIII in Sneddon syndrome]. / VIII faktor svertyvaniia krovi pri Sneddon sindrome.

Hyperactivity of coagulation factor VIII (fVIII) marks hypercoagulation. FVIII enhances activity of factor IX and their combination activates factor X, which is of primary importance in prothrombin transformation into thrombin, on the phospholipid membrane. The activity of fVIII was studied in 28 patients (26 women, 2 men, mean age 49.6 +/- 7.8 years) with Sneddon's syndrome (SS). SS manifests clinically similarly to primary antiphospholipid syndrome (PAS). The leading of them are ischemic disorders of cerebral circulation (IDCC) and advanced livedo present in all the examinees. Hyperactivity of fVIII was registered in 21 (75%) of 28 patients. Most of thrombosis-related symptoms occurred more frequently in patients with high than normal activity of fVIII: ischemic strokes (91% vs 57%, p > 0.05), repeated strokes (71% vs 0%, p = 0.0014), transient IDCC (76% vs 57%, p > 0.05), vascular dementia (43% vs 0%, p > 0.05), ischemic heart disease (43% vs 0%, p > 0.05), thickening of heart valves according to echocardiography (91% vs 57%, p > 0.05), peripheral venous thromboses (24% vs 0%, p > 0.05). In high fVIII activity cardiolipin antibodies occurred more rarely (24% vs 43%, p > 0.05) but lupus anticoagulant was seen more often (47% vs 14%, p > 0.05). High fVIII activity was in 8 of 12 aPL-negative patients. It is demonstrated that elevated fVIII activity is an essential mechanism of thrombosis development in SS. The cause of this enhanced activity is suggested to be special aPL in interaction with which fVIII becomes insensitive to inactivation with protein C. The activity of protein C was normal in all the cases.

[Procalcitonin--marker of infectious inflammation: clinical value and application]. / Prokal'tsitonin--marker infektsionnogo vospaleniia: klinicheskoe znachenie i oblast' primeneniia.

The development of septic shock is accompanied by an essential increase of the level of Procalcitonin (PCT). The growth of the PCT level in blood is often associated with bacteremia. A-->2 ng/ml PCT increase in patients with agranulocytosis makes it possible to distinguish between the pneumonia as a local focus of infection and the pneumonia in sepsis. A high PCT level in bacterial and fungus infections makes it possible to distinguish them from viral and non-infection diseases.

[Stable thrombin preparations for the Klauss method of determination of thrombin time and fibrinogen level]. / Stabil'nye preparaty trombina dlia opredeleniia trombinovogo vremeni i kontsentratsii fibrinogena po Klaussu.

A method was elaborated enabling an isolation of a stable and highly active thrombin to be used in determining the thrombin time and fibrinogen concentration in plasma. The stability of the thus obtained preparations was studied, which showed no changes in the activity of their liquid variations for as long as 28 days at 2-8 degrees C.

[Study of storage stability of the commercial series of thromboplastin preparations]. / Issledovanie stabil'nosti promyshlennykh serii preparatov tromboplastina pri khranenii.

The stability of commercial series of the thromboplastin preparation (obtained from rabbit brain) with calcium (TKS, Researach-and-Production Enterprise "Renam" MICH 1.1-1.2), of TKS "Tromborel" and of "Immunoplastin" (Technoclon) was comparatively studied. TKS "Renam" preserved its activity and sensitivity to factor VII in the liophilizated state for as long as 6 months. The preparations of "Renam", TKS, preserved the stable activity in the reconstructed liquid state, under the conditions of continuous storage, at 37 degrees C for 2 days, at 18-20 degrees C--for 2-3 days, and at 2-8 degrees C--for 5-7 days, which was accompanied by a stable high sensitivity to factor VII. The stability of the "Renam", TKS, was found to be similar to that of imported preparations.

[Tumor necrosis factor, interleukin-6, endotoxin and procalcitonin in septic shock in patients with hematologic neoplasms]. / Faktor nekroza opukholi, interleikin-6, éndotoksin i prokal'tsitonin pri septicheskom shoke u bol'nykh s opukholevymi zabolevaniiami sistemy krovi.

AIM: To study changes in proinflammatory markers and mediators in septic shock in patients with hematologic malignancy (HM). MATERIAL AND METHODS: The examination of 33 patients with HM and septic shock included measurement of plasma concentrations of tumor necrosis factor (TNF), interleukine-6 (IL-6), endotoxin, procalcitonin (PCT) 12-24 hours before and each 12 hours after shock; registration of central hemodynamics parameters, the condition severity by APACHE II. RESULTS: Out of 33 patients 18 died of refractory shock, 15 survived the shock. Within the first shock hour TNF fell from 571.2 +/- 195 to 115.8 +/- 71.1 pg/ml (p < 0.02), later being stable. In those who died and survived TNF was the same. IL-6 fall was seen 36 hours after shock and was observed in the survivors; in those who died IL-6 was unchanged. Endotoxin in the blood was detected in 21 of 33 patients. In the survivors endotoxinemia declined after 2 days of treatment. 72 hours after beginning of the shock the survivors had no endotoxin. In shock APACH II severity of the patient's condition was graver in patients with endotoxinemia than without it (31.6 +/- 1.6 and 28.1 +/- 1.6 scores, p < 0.05). Blood endotoxin levels and APACHE II scores correlated (r = 0.24, p < 0.05) positively and negatively with deficiency of buffer bases (r = -0.29, p < 0.05) and blood pH) r = -0.3, p < 0.05), left ventricular contractility index (r = -0.46, p < 0.01) and right ventricle (r = -0.52, p < 0.01), mean AP (r = -0.22, p < 0.03). PCT concentration was lower before shock than on its hour 1 (4.2 +/- 2.9 and 6.9 +/- 1.1 ng/ml, p < 0.05). No significant changes in PCT were found later. CONCLUSION: PCT is a specific marker of a severe infection. Rapid elimination from the blood of TNF and IL-6 makes them inadequate in sepsis diagnosis. Endotoxinemia aggravates the patients condition. Positive LAL-test results were obtained in gram-negative and fungal infections.