Citosol (thiamylal sodium) is one of generally used anesthetic–sedative agents for clinical patients, and it has not been reported to show induction of cytotoxic effects in cancer cells, especially ...in mice leukemia RAW 264.7 cells in vitro. In the present study, we investigated the cytotoxic effects of citosol on mice leukemic RAW 264.7 cells, including the effects on protein and gene expression levels which are determined by Western blotting and DNA microarray methods, respectively. Results indicated that citosol induced cell morphological changes, cytotoxic effect, and induction of apoptosis in RAW 264.7 cells. Western blotting analysis demonstrated that citosol promoted the levels of Fas, cytochrome c, caspase 9 and 3 active form and Bax levels, but it suppressed Bcl-xl protein level that may lead to apoptotic death in RAW 264.7 cells. Furthermore, DNA microarray assay indicated that citosol significantly promoted the expression of 5 genes (Gm4884, Gm10883, Lce1c, Lrg1, and LOC100045878) and significantly inhibited the expression of 24 genes (Gm10679, Zfp617, LOC621831, Gm5929, Snord116, Gm3994, LOC380994, Gm5592, LOC380994, LOC280487, Gm4638, Tex24, A530064D06Rik, BC094916, EG668725, Gm189, Hist2h3c2, Gm8020, Snord115, Gm3079, Olfr198, Tdh, Snord115, and Olfr1249). Based on these observations, citosol induced cell apoptosis and influenced gene expression in mice leukemia RAW 264.7 cells in vitro.
The enzymatic study and transport of N in the xylem sap was carried out with a view to observing the influence of different nitrate levels and growth stages of the plant in chemically treated mutants ...of Lupinus albus. Several stresses induce a reduction in plant growth, resulting in the accumulation of free amino acids, amides or ureides, not only in the shoot, but also in the roots and nodules. Although enzyme activity is decisive in avoiding products that inhibit nitrogenase by ammonium, little is known about the mechanism by which the xylem carries these products. However, this process may be the key to the function of avoiding the accumulation of amino acids in the cells of infected nodules. The behaviour of the enzymes nitrate reductase (NR), phosphoenolpyruvate carboxylase (PEPC), glutamine synthetase (GS) and nitrogen compounds derived from fixation, such as N-a-amino, N-ureides and N-amide in mutant genotypes were observed. The NR enzyme activity was highly influenced by the application of nitrate showing much higher values than those in the non-application of nitrate, independently of genotype, being that the NR, the best evaluation period was in the tenth week. The L-62 genotype characterized with nitrate- resistance, clearly showed that the enzyme PEPC is inhibited by presence of nitrate. The L-135 genotype (nod- fix-) showed GS activity extremely low, thus demonstrating that GS is an enzyme highly correlated with fixation. With regard to the best growth stage for GS, Lupinus albus should be evaluated in the seventh week.
O estudo enzimático e o transporte de N na seiva do xilema foi realizado visando observar a influência de diferentes níveis de nitrato e estágios de crescimento da planta em mutantes tratadas quimicamente. Vários estresses induzem a redução no crescimento da planta da qual resulta na acumulação de aminoácidos livres, amidas ou ureídos, tanto na parte aérea como nas raízes e nódulos. A atividade de enzimas é decisiva em evitar produtos inibidores da nitrogenase pelo amônio, porém poucas são as informações deste mecanismo com o qual o xilema carrega estes produtos, em compensação, este processo, pode ser a chave da função em evitar acumulação de aminoácidos nas células dos nódulos infectados. O comportamento das enzimas nitrato redutase (NR), fosfoenolpiruvato carboxilase (PEPC), glutamina sintetase (GS) e dos compostos nitrogenados derivados da fixação como: N-a-amino, N-ureídos e N-amida em genótipos mutados foi observado. A atividade da enzima nitrato redutase foi altamente inflenciada pela aplicação de nitrato apresentando valores bastante superiores em relação a não aplicação do nitrato, independentemente dos genótipos, sendo que o melhor período de avaliação foi na décima semana. O genótipo L-62, com caracteríticas de resistência ao nitrato, mostrou de forma bem evidenciada que a enzima PEPC é inibida pela presença de nitrato. O genótipo L-135 (nod- fix-) apresentou atividade da GS extremamente baixo, mostrando desta forma que a GS é uma enzima altamente correlacionada com a fixação. Em relação ao melhor estágio de crescimento para a GS, o Lupinus albus, deve ser avaliado na sétima semana.
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