RESUMO
The recombinant producer of Rhodospirillum rubrum L-asparaginase (RrA) was received and purification procedure of RrA was developed. It was shown that RrA has following biochemical and catalytic characteristics: K(m) for L-asn 0.22 MM, pH optimum 9.2; temperature optimum 54 degrees C; pI = 5.1 +/- 0.3; L-gln activity seems to be low-to-negligible. K562, DU145 and MDA-MB-231 cellular lines displayed significant sensitivity towards the enzyme (IC50 = 1.80; 9.19 and 34.62 ME/ml, respectively. In comparison with L-asparaginases from E. coli II type (EcA) and Erwinia carotovora (EwA) cytotoxicity of RrA seems to be higher than EwA, but lower than EcA. 10-fold i.p. RrA administration (4000 ME/kg per day) in L5178y bearing mice showed T/C = 172%. The received results show that RrA belongs to I type cellular L-asparaginases with low L-gln activity and the high antiproliferative effect.
Assuntos
Antineoplásicos/farmacologia , Asparaginase/farmacologia , Proteínas de Bactérias/farmacologia , Proliferação de Células/efeitos dos fármacos , Glutaminase/farmacologia , Rhodospirillum rubrum/enzimologia , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Asparaginase/biossíntese , Asparaginase/química , Asparaginase/genética , Asparaginase/isolamento & purificação , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Ensaios de Seleção de Medicamentos Antitumorais , Glutaminase/biossíntese , Glutaminase/química , Glutaminase/genética , Glutaminase/isolamento & purificação , Humanos , Células K562 , Camundongos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/farmacologia , Rhodospirillum rubrum/genéticaRESUMO
Morphological changes in decellularized allogenic trachea populated with recipient bone marrow stromal (mesenchymal) stem cells and transplanted heterotopically, were examined in 30 C57Bl/6 and Balb/c mice of 22-25 g body mass. The research results have shown the insufficient efficacy of a transplant preparation mode by freezing and thawing method as in this case inflammatory reaction developed in the transplant area and its rejection took place. It was established that the mode of obtaining decellularized tracheal transplant by means of sodium perchlorate (NaClO4) treatment, proposed by the authors, unlike a freezing-thawing mode, allowed to efficiently remove immunocompetent cells that expressed MHC I and II markers. NaClO4 effect did not result in either chondrocyte damage or significant disturbance of tracheal cartilaginous and connective tissue structure in heterotopic transplants. Since transplant population with bone marrow stromal stem cells promoted connective tissue restoration, reduced the formation of granulations in anastomosis area and favored faster transplant epithelization, most promising method of trachea preparation for transplantation apparently seems to be the combination of immune cell removal from this organ by NaClO4 treatment with subsequent bone marrow stromal stem cell population of transplant obtained.
Assuntos
Imunocompetência , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/efeitos dos fármacos , Traqueia/efeitos dos fármacos , Traqueia/transplante , Animais , Transplante de Medula Óssea/métodos , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Imunocompetência/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Percloratos/farmacologia , Compostos de Sódio/farmacologia , Traqueia/citologia , Transplante Heterotópico/métodosRESUMO
AIM: Analysis of LPS and LPS-binding protein (LBP) serum levels and prognostic value evaluation in cancer patients with sepsis. MATERIALS AND METHODS: Blood samples of 17 healthy donors and 46 cancer patients with sepsis were analyzed by using "Hycult biotechnology" kits (The Netherlands). RESULTS: Serum LPS level in cancer patients with sepsis was increased significantly. A substantial decrease of LBP level was detected in blood samples of deceased patients. CONCLUSION: Dynamic increase of LPS serum level and decrease of LPS-binding protein serum level in cancer patients should be considered as an unfavorable factor for sepsis.
