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n-Dodecane and fatty acids were good inducers of cytochrome P450 (CYP) and the x-hydroxylase of lauric acid, which is a marker for x-hydroxylation of n-alkanes, in Trichoderma harzianum. A cDNA, containing an ORF of 1520 bp, encoding a CYP52 of 520 amino acids, was isolated by RACE. Another n-alkane-inducible CYP was identified by LLC-MS/MS analysis of a microsomal protein band induced by n-dodecane in a library of T. harzianum. This suggests that T. harzianum has a CYP-dependent conversion of alkanes to fatty acids allowing their incorporation into lipids.

Il presente lavoro, svolto nell’ambito del progetto PRIN 2007FRBK9N_003 intitolato: “Studio della transizione dimorfica di Pichia fermentans mediante approccio proteomico e citogenetico”, si prefiggeva due scopi principali: l’identificazione delle proteine coinvolte nella transizione dimorfica di P. fermentano dallo stato lievitiforme a quello pseudoifale e la caratterizzazione delle componenti morfo-strutturali delle due fasi, mediante approccio immunoistochimico.

Mutagenicity study "in vivo" (Intravenous host mediated assay) confirms suspension test results with cis- and trans -1,2-dichloroethylene: no genetic effects could be detected. Single and repeated doses of trans-1,2-DCE induced liver aminopyrine demethylase and cytochrome P-450 whereas the cis- isomer decreased them. Similar results were obtained after induction with phenobarbital + beta-Naphthoflavone.

1,2-dichloroethylene cis and trans were tested for their ability to induce point mutation, mitotic gene conversion and mitotic recombination in a diploid strain (D7) of the yeast Saccharomyces cerevisiae in a suspension test with and without a mammalian microsomal activation system. In this test 1,2-DCE cis and trans were toxic but not genetically active even with S9 activation.

A short diethyl ether anesthesia before the three injections to induce xenobiotic metabolizing monooxygenase in mice for the preparation of liver S-9 fraction used in mutagenesis test in vitro does not negatively alter the biochemical properties of the activating system and gives unchanged genetic results. Instead, the use of more prolonged ether anesthesia also at the day of the sacrifice, results in less active and less stable enzymes in the S-9 fraction, which can lead to altered genetic results.

The level of cyt.p-450 in the D7 strain of the yeast S.cerevisiae depended on the substrate supporting the growth, on its concentration, on the starting inoculated number of cells. (1) In the yeast grown on D-mannose where fermentation and respiration occurred concomitantly, cytochrome P-450 was also formed. It was detected a maximal concentration during the logarithmic phase when in the cultures there are about 50 . 10(6) cell/ml. We use cells harvested at this moment of the growth for mutagenesis tests. The tested substances were dimethylnitrosamine and styrene. DMNA to probe the sensibility of our cells and styrene that has always given contrasting results but from which the formation is known of genetically active metabolite: styrene oxide(6-7). Styrene gave positive results with our metabolizing yeast cells.

Rat liver S-9 microsomal fractions were fortified with NADP+, G-6-P, and MgCl2 and stored at -80 degrees C and -20 degrees C. At various times aminopyrine demethylase and lipid peroxidation were determined and compared with that of non-fortified fractions. No difference in stability was noted between fortified and not fortified fractions. The known stability pattern of non fortified fractions was confirmed. Lipid peroxidation was found blocked in the fractions stored at -80 degrees C whereas a small release of malonyl dialdehyde proceeded in the frozen state at -20 degrees C, contributing to the loss of activity at this temperature. The use of pre-fortified fractions stored at -80 degrees C is justified.

Aminopyrine-N-demethylase, p-nitroanisole-O-demethylase and lipid peroxidation were determined in liver postmitochondrial fractions (S-9) of mice in the basal state and after induction with beta-naphtoflavone, sodium phenobarbital, or both the inducers. The two activities were similar in their response to all inductions: meanly, 1,2 times with beta-NF, 2,0 times with Na-PB, 5,8 times with Na-PB+ beta-NF. Lipid peroxidation was sensibly more active in basal animals with respect to that with induction. The results are discussed in a partial review of the sector literature, which deserves still much work to be completed.