[Monitoring of low-molecular-weight heparins in pregnant women with inherited thrombophilic disorders].

Women with thrombophilic disorders have higher risk for venous thromboembolism during pregnancy, as well as risk for vascular damages and complications, including preeclampsia and fetal loss. It is supposed that thrombosis prevention with low-molecular-weight heparins (LMWH) directed against recurrent fetal loss carries antiinflamatory and immunomodulation effects besides its well-known anticoagulation action. This study presents results from a retrospective analysis of 120 pregnant women on antithrombotic therapy with prophylactic doses of LMWH, during the period from 2010 to 2013. Women were recruited on the basis of genotyping done in advance in National Genetic Laboratory, Sofia and Laboratory "Genica". Allele frequencies of 12% of the mutation R506Q Factor V Leiden, 7% of G20210A in Prothrombin gene, 53% of 4G/4G in PAI and 28% of the mutation T677 in MTHFR were found (39% single defects, 50% double and 11% combained mutations). Majority of women studied (60%) had embryo loss (< 13th g.w.), 38 out of them were with one episode, 28 with two, and 4--with three pregnancy losses. Level of anti-Xa and monitoring of LMWH was done by chromogenic assay (Berichrom Heparin, Siemens) using coagulometer C 2000i. Anti-Xa inhibition was 0.35 +/- 0.11 IU anti-Xa/ml (0.18-0.60; n = 50) in response to Fraxiparine in dose of 0.3 ml/d, 0.45 +/- 0.09 IU anti-Xa/ml (0.26-0.60; n = 33) with 0.4 ml/d Fraxiparine, and 0.49 +/- 0.13 IU anti-Xa/ml (0.3-0.72; n = 37) with Clexane 40 mg/d. Five out of 120 women (4%) had values below 0.2 IU anti-Xa/ml and required increased doses of LMWH during the third trimester. Slight decrease of anti-Xa (14-20%) was observed during the third trimester. Five women suffered oedema, redness and pain oversensitivity at LMWH application, one woman demonstrated increased eosinophil count, and three other women had mild bleeding during the first weeks of anticoagulation therapy with higher levels of anti-Xa inhibition. Monitoring of LMWH is of help in the rare cases of bleeding during the first trimester of pregnancy, as well as in high risk pregnancies with insufficient anticoagulation response.

Pharmacogenetics of acenocoumarol: CYP2C9, CYP2C19, CYP1A2, CYP3A4, CYP3A5 and ABCB1 gene polymorphisms and dose requirements.

BACKGROUND AND OBJECTIVE: Acenocoumarol (AC) is a coumarin derivative, vitamin K antagonist anticoagulant drug. It has a narrow therapeutic index and shows large pharmacokinetic and pharmacodynamic interindividual variability. Our objective was to investigate the association between AC dose requirements to achieve a target level of anticoagulation and genetic polymorphisms of genes possibly associated with its metabolism (CYP1A2, CYP2C9, CYP2C19, CYP3A4, CYP3A5) and transport (ABCB1). METHODS: Ninety-six Bulgarian patients treated orally with AC for at least 3 months were included. They were separated into three groups according to their AC dose requirement, i.e. low, medium and high. RESULTS AND DISCUSSION: CYP2C9*1/*3 (associated with an intermediate CYP2C9 activity), CYP2C9*2/*2, and CYP2C9*2/*3 genotypes (associated with a low CYP2C9 activity) were more prevalent in the group with low dose requirement of AC compared with the other two groups (P = 0.003). The frequency of CYP2C9*1/*1 genotype, which is associated with an extensive CYP2C9 activity, was higher in the group of patients with high dose requirements (79%), compared with the groups of the medium and low dose requirements (67% and 21% respectively). In addition, the ABCB1 2677GG/3435CC haplotype was associated with use of lower AC dose, whereas the 2677TT/3435TT and 2677GT/3435TT haplotypes were associated with use of higher AC dose (P = 0.03). The distribution of polymorphisms of other genes did not show significant differences between the three groups. CONCLUSION: In vivo, cytochromes P450 isoforms other than CYP2C9 [DOSAGE ERROR CORRECTED] were not significantly associated with dose requirement of AC. In our Bulgarian patients, the presence of CYP2C9*2 or/and CYP2C9*3 alleles, as well as the ABCB1 2677GG/3435CC haplotype were associated with low dose requirement of AC.

Persistence and in vivo effects of Yersinia enterocolitica 0:3 endotoxin in rats.

In vivo effects of Yersinia enterocolitica 0:3 lipopolysaccharide (prepared from bacteria grown at 25 degrees C and 37 degrees C) were investigated after intraperitoneal (i.p.) and intraarticular (i.a.) injection in rats during 30 days of examination. The persistence of endotoxin in the peritoneal and the synovial cavities was demonstrated by the immunofluorescence technique. Peritoneal and synovial exudative cell infiltration, as well as changes in some parameters (glycolytic and acid phosphatase activity, and killing ability of peritoneal cells; lactate dehydrogenase concentration in synovial fluid) were studied. The results indicated that endotoxin could persist longer in the synovial than in the peritoneal cavity.

The fate of Yersinia pseudotuberculosis lipopolysaccharide (LPS) in intraperitoneally and intraarticularly infected rats.

Persistence and in vivo effects of Yersinia pseudotuberculosis serotype III LPS (prepared from bacteria grown at 25 degrees and 37 degrees C) in rats were investigated after intraperitoneal and intraarticular injection during the 30 day period of examination. Localization and persistence of LPS in the peritoneal and synovial cavities were demonstrated by using the immunofluorescence technique. Peritoneal and synovial exudative cell infiltration as well as changes in some parameters (glycolytic and acid phosphatase activities, killing ability of peritoneal cells, lactate-dehydrogenase concentration in synovial fluid) were studied. The results indicated that LPS expressed at 37 degrees C induced stronger peritoneal cell response (increased "killing" ability and elevation of glycolytic and acid phosphatase activities) in comparison to that synthesized at 25 degrees C. It was also found, that LPS persisted longer in the synovial rather than in the peritoneal cavity of rats and induced long-lasting synovial inflammation.

Cell wall-deficient forms (L-forms) of Listeria monocytogenes in experimentally infected rats.

Experimental infections were induced with different bacterial forms of Listeria monocytogenes: parental (S-forms), protoplastic (L-forms) and combined inoculum of both forms by i.p. injection of rats. The parental bacterial forms (S-forms) were isolated up to 7 days after challenge from the peritoneal cavity and the liver, while the L-forms were isolated up to 60 days from the peritoneal cavity. Continuous adhesion of L-forms on the peritoneal macrophage surface was found by scanning-electron microscopy. Erythrocyte and leucocyte count as well as some clinical chemistry parameters were measured during infections. They showed different dynamics in the three experimental groups. Histomorphological changes in the liver (microabscesses and mononuclear cellular granulomas) of infected animals were observed. They were less intensive and appeared later in rats infected with L-forms. The experiments demonstrated that infections caused by parental bacterial forms and by combined inoculum took an acute course, while the infection caused by L-forms could be distinguished as a prolonged and persistent one.

Influence of blood transfusion on bactericidal activity of human leukocytes and sera against Yersinia enterocolitica and Salmonella typhimurium.

Patients undergoing joint surgery and blood transfusion were studied. Serum and leukocyte bactericidal tests in vitro against Salmonella typhimurium and Yersinia enterocolitica were carried out preoperatively as well as on the 1st, 3rd and 7th days after the operation. The serum complement (C3 and C4) concentrations were determined at the same intervals. It was found that after blood transfusion the bactericidic activity of sera and the serum C3 complement concentrations were increased. In contrast the killing ability of leukocytes was suppressed.

[Acid lysosomal hydrolase activity in the serum and synovial fluid of rheumatoid arthritis patients]. / Aktivnost na kiseli lizozomni khidrolazi v serum i sinovialna technost na bolni s revmatoiden artrit.

Five acid lysosome hydrolases were studied in 20 patients with rheumatoid arthritis (RA), active stage in serum (S) and synovial fluid (SF) in parallel, obtained from the knee joint. The enzyme activity was fluorimetrically determined, making use of substrates specific for each enzyme, derivatives of 4-methyl-umbelliferone. The serum lysosome activity of the patients with RA, compared with that of a referent group of healthy subjects, showed a significant increase in three of the hydrolases studied. With the juxtaposition of enzyme activity between SF and serum of the patients with RA, the activity of all acid hydrolases in the synovial fluid studied proved to be several times higher and with high statistical significance (p less than 0.01-0.001). Our data support the valuable informative value of the study on acid lysosome hydrolases in the synovial fluid. A sensitive indicator of the activity of the joint process in RA is the coefficient of lysosome enzymatic activity SF/S, substantially surpassing 1.0.

[Bactericidal effect of the preparation trozin]. / Prouchvane vurkhu bakteritsidnoto deistvie na preparata trozin.

Tested was the bactericidal action of the combined cleansing and disinfecting preparation trozin under laboratory and production conditions. It was found that trozin meets the requirements in terms of rapid disinfection, killing the asporogenous test organisms. The presence of at least 2 per cent milk protein lowers the bactericidal action of the detergent. The cleansing and disinfection of milking units supplied with a milk-can, resp., with a central milk pipeline leads to 97.00, resp., 98.00 per cent reduction, on an average, of microflora. The bacterial count of the udder is likewise reduced by about 96.00 per cent.

[Dynamics of lysosomal enzymes in patients with allergic dermatoses before and after high altitude climate therapy]. / Dynamik lysosomaler Enzyme bei Patienten mit Allergodermatosen vor und nach Hochgebirgsklimatherapie.

Lysosome enzymes were measured in patients with allergic dermatosis before and after high altitude climate therapy. The activity of five acid lysosomal hydrolases in the serum of patients with allergic dermatoses before and after 30 days were investigated. In all cases a reduction of the activity of alpha-glucosidase was observed. A similar reduction of the activity of beta-glucuronidase and beta-galactosidase was observed.