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Biomedical subjects

J L Boucher

Publications and source records attributed to J L Boucher.

17 recordsLinked to original sources
PubMed1995

Nitric oxide formation during the cytochrome P-450-dependent reductive metabolism of 18-nitro-oxyandrostenedione.

18-Nitro-oxyandrostenedione (18-ONO2A), a potential mechanism-based inhibitor of the last steps of aldosterone biosynthesis, is well recognized by different cytochrome P-450s, which are able to metabolize it reductively into nitric oxide (NO) and 18-hydroxyandrostenedione. Rat liver microsomal P-450s are able to carry out this reaction with increased efficiency under anaerobic conditions. P-450 3A isozymes induced upon treatment of rats with dexamethasone or troleandomycin were best able to bind and metabolize 18-ONO2A. This reaction was shown to occur in the presence of dioxygen as well, suggesting that it may be of physiological relevance. The formation of NO was detected as a transient P-450-Fe(II)NO complex by UV-visible and EPR spectroscopy. In addition, steroidogenic tissues containing cytochrome P-450s such as bovine adrenal mitochondria or human placental microsomes also were capable of binding and metabolizing 18-ONO2A as judged by the formation of an Fe(II)NO complex. This recognition of a steroid nitrate, a potential antialdosterone and its subsequent metabolism under reductive conditions to generate NO both in hepatic and steroidogenic tissues, can be of pharmacological interest, because NO has been demonstrated to modulate steroidogenesis in addition to other processes such as vascular relaxation, neurotransmission or cytostasis. A nitrate derivative of a steroid could perhaps act as a vectorized NO precursor in which the steroid moiety is targeted specifically to steroid receptors or steroidogenic tissues, thus leading to localized NO liberation.

Adrenal Glands↗
PubMed1994

N omega-hydroxyl-L-arginine, an intermediate in the L-arginine to nitric oxide pathway, is a strong inhibitor of liver and macrophage arginase.

N omega-Hydroxy-L-arginine (L-NOHA) is a potent inhibitor of the hydrolysis of L-arginine (L-Arg) to L-ornithine (L-Orn) catalyzed by purified bovine liver arginase (BLA). It appears as one of the most powerful arginase inhibitors reported so far (Ki = 150 microM). The other products of NO synthase are either without effect (NO2-, NO3-) or much weaker inhibitors (L-citrulline (L-Cit) and NO) of BLA. Products derived from a possible hydrolysis of L-Arg (L-Orn and urea) or of L-NOHA (L-Cit, hydroxyurea and hydroxylamine) are also inactive toward BLA at concentrations up to 2 mM. The configuration of L-NOHA is important as D-NOHA is much less active, and its free -COOH and alpha-NH2 functions are required for recognition of BLA. L-NOHA is also a potent inhibitor of the arginase activity of rat liver homogenates and of murine macrophages (IC50 of 150 and 450 microM, respectively). These remarkable properties of L-NOHA could play a role in the modulation of the biosynthesis of the biological mediator NO by increasing local L-Arg concentrations.

Animals↗
PubMed1994

Dehydration of alkyl- and arylaldoximes as a new cytochrome P450-catalyzed reaction: mechanism and stereochemical characteristics.

The Z isomers of benzaldoxime and 4-(hexyloxy)benzaldoxime were dehydrated into the corresponding nitriles in the presence of rat liver microsomes and NADPH or dithionite. Their E isomers remained unchanged under identical conditions. Alkylaldoximes, like phenylacetaldoxime and heptanaldoxime, are also dehydrated under these conditions, the alkylaldoximes being more rapidly transformed than the arylaldoximes. A genetically well-defined P450 expressed in yeast, P450 3A4, the major P450 isozyme in human liver, was also found to be catalytically active for dehydration of (Z)-benzaldoxime. All these reactions were found to be catalyzed by P450 Fe(II) as they required the use of intact microsomes in the presence of NADPH or dithionite and were strongly inhibited by O2 and CO as well as by classical P450 inhibitors. A P450 complex characterized by a Soret peak at 442 nm was detected during these reactions; its disappearance was found to be concomitant with the consumption of the aldoxime and the formation of the corresponding nitrile. (E)-benzaldoximes and all the studied ketoximes failed to give such complexes with P450 Fe(II). On the basis of these results, a possible mechanism for this new P450 reaction is proposed. It involves a P450 Fe(II)<--N(OH)=CHR complex as a key intermediate and a charge transfer from P450 Fe(II) to the aldoxime C=N bond which results in a cleavage of the aldoxime N-O bond.

Animals↗
PubMed1993

Particular ability of liver P450s3A to catalyze the oxidation of N omega-hydroxyarginine to citrulline and nitrogen oxides and occurrence in no synthases of a sequence very similar to the heme-binding sequence in P450s.

Liver microsomes from rats pretreated with various inducers of P450 isoforms exhibit very different abilities to catalyze the oxidation of N omega-hydroxy-L-arginine (NOHA) by NADPH and O2 with formation of citrulline and nitrogen oxides. Treatment of rats with dexamethasone, a classical inducer of P450 3A, leads to a spectacular 7-fold increase of the activity found for untreated rats, while induction by phenobarbital causes a much lower increase of this activity and induction by 3-methylcholanthrene or clofibrate decreases it. Specific inhibitors of P450s3A as troleandomycin and dihydroergotamine strongly inhibit NOHA oxidation whereas metyrapone, an inhibitor of other P450 subfamilies, was without effect. These data show the particular ability of P450s of the 3A subfamily to catalyze the second step of the oxidation of L-arginine by NO synthases (NOS). This analogy between NOSs and P450s3A is further substantiated by a protein sequence comparison which shows that a 9-amino acid segment present in all NOSs exhibits a strong similarity with the sequence mainly responsible for heme binding in P450s3A which is well conserved in all P450s. This segment contains all the structural factors which are thought to be crucial for heme binding in P450s.

Amino Acid Oxidoreductases↗
PubMed1993

EPR characterization of molecular targets for NO in mammalian cells and organelles.

Nitric oxide is synthesized in mammalian cells from L-arginine or from pharmaceutical drugs. It forms paramagnetic complexes with some metalloproteins, inhibiting key enzymes in DNA synthesis, mitochondrial respiration, iron metabolism, etc. This article reviews how electron paramagnetic resonance spectroscopy helps to detect unambiguously such specific molecular targets for NO in mammalian whole cells and organelles. EPR has also been used for the detection of spin adducts of free NO by spin-trapping methods.

Animals↗
PubMed1993

Lack of effect of phenobarbital on ex vivo leukocyte oxidative metabolism in healthy volunteers.

Oxidative metabolism in activated human polymorphonuclears catabolizes leukotriene B4 by a cytochrome P450 omega-hydroxylase and procainamide by a myeloperoxidase. The percentage of leukotriene B4 metabolized by activated human polymorphonuclears and the apparent enzymatic parameters of procainamide metabolism were studied ex vivo in six healthy volunteers before and after phenobarbital intake (100 mg/day) for 10 days and in six healthy control volunteers. No differences were found between groups for the difference in percentage of leukotriene B4 metabolized between day 11 and day 1. The apparent enzymatic parameters, Km and Vm, of procainamide oxidation did not differ significantly between the groups both on day 1 and day 11. These results do not show any evidence of inducibility of leukotriene B4 and procainamide metabolism in human polymorphonuclears. However, a positive correlation between 6 beta OH-cortisol excretion and percentage of leukotriene B4 metabolized was observed on day 11. This study suggests that human polymorphonuclears cytochrome P450 leukotriene B4 omega-hydroxylase and procainamide metabolism is not a useful method to study cytochrome P450 induction in clinical pharmacology.

17-Hydroxycorticosteroids↗
PubMed1992

Cytochrome P450 catalyzes the oxidation of N omega-hydroxy-L-arginine by NADPH and O2 to nitric oxide and citrulline.

Rat liver microsomes catalyze the oxidative denitration of N omega-hydroxy-L-arginine (NOHA) by NADPH and O2 with formation of citrulline and nitrogen oxides like NO and NO2-. Besides NO2- and citrulline, whose simultaneous formation is linear for at least 20 min, the formation of NO could be detected under the form of its P450 and P420-Fe(II) complexes by UV-visible and EPR spectroscopy. Classical inhibitors of NO-synthases, like N omega-methyl-and N omega-nitro-arginine, fail to inhibit the microsomal oxidation of NOHA to citrulline and NO2-. On the contrary classical inhibitors of hepatic cytochromes P450 like CO, miconazole, dihydroergotamine and troleandomycin, strongly inhibit this monooxygenase reaction. These results show that the oxygenation of NOHA by NADPH and O2 with formation of citrulline and NO can be efficiently catalyzed by cytochromes P450 (with rates up to 1.5 turnovers per min for the cytochromes of the 3A subfamily).

Animals↗
PubMed1992

Formation of nitric oxide by cytochrome P450-catalyzed oxidation of aromatic amidoximes.

Rat liver microsomes catalyze the oxidation of para-hexyloxy-benzamidoxime 1 to the corresponding arylamide 2 and NO2-, by NADPH and O2. Involvement of cytochromes P450 as catalysts of this reaction was shown by the strong inhibitory effects of CO and miconazole and the spectacular increase of the activity upon treatment of rats with dexamethasone, a specific inducer of cytochromes P450 of the 3A subfamily. Formation of NO during oxidation of 1 was shown by detection of the formation of cytochrome P450- and cytochrome P420-Fe(II)-NO complexes by visible and EPR spectroscopy. The formation of these complexes should be responsible, at least in part, for the fast decrease of the rate of microsomal oxidation of 1 with time. These results suggest that exogenous compounds containing amidine or amidoxime functions could act as precursors of NO in vivo after in situ oxidation by cytochromes P450.

Animals↗
PubMed1992

Formation of nitrogen oxides and citrulline upon oxidation of N omega-hydroxy-L-arginine by hemeproteins.

HRP catalyzes the oxidation of N omega-hydroxy-L-arginine (NOHA) by H2O2 with formation of citrulline and NO2- with initial rates of about 0.7 and 0.2 nmol per nmol HRP per min. In the same manner, cytochromes P450 from rat liver microsomes catalyze the oxidation of NOHA to citrulline and NO2- by cumylhydroperoxide. Inhibitors of these hemeproteins (N3- and CN- for HRP and miconazole for P450) strongly inhibit both citrulline and NO2- formation. Rates of NOHA oxidation by these hemeproteins markedly decrease with time presumably because of their denaturation by nitrogen oxides and of the formation of hemeprotein-iron-NO complexes. These results suggest that NO (and other nitrogen oxides) could be formed from oxidation of NOHA by other enzymes than NO-synthases.

Animals↗
PubMed1992

Knowledge and motor performance.

Traditionally, motor skill acquisition has implied that the performance of a given individual on a particular skill is dependent on the amount of prior practice of that skill. However, concepts such as schema theory, or kinetic formulae, or the strategic allocation of resources imply that, even when practising specific skills, performers gain knowledge about their own motor performance which can be used or applied to related or novel situations. An attempt was made to relate the performance of a complex psychomotor task to differing levels of motor skill expertise or knowledge (athlete and nonathlete). 20 subjects performed (1600 responses) on a novel pursuit or tracking task. Analysis indicated that the athletes performed significantly better. Their main advantage appeared to be more in their ability to control and produce fast, accurate movements than in their decision-making. Accepting Henry and Rogers' 1960 proposition that there is no such thing as a general motor ability or coordination factor does not imply that the only alternative is for all motor skills to be specific. It is argued that the differences in the present study arose from the athletes' greater knowledge (schema, kinetic formulae) related to their understanding of their own motor capabilities.

Adult↗
PubMed1991

Metabolism of lipoxins A4 and B4 and of their all-trans isomers by human leukocytes and rat liver microsomes.

Incubation of lipoxin A4 (LXA4) either with human leukocytes or with rat liver microsomes in the presence of NADPH very selectively led to a more polar metabolite retaining the conjugated tetraenic structure of LXA4. Lipoxin B4 (LXB4) underwent a very similar metabolism into a more polar metabolite, whereas the all-trans isomers of LXA4 and LXB4 were selectively transformed by the same biological systems into metabolites derived from the reduction of one of the double bonds of the conjugated tetraene moiety of the starting compounds. Microsomal metabolism of LXA4 and LXB4 was NADPH-dependent and strongly inhibited by CO and miconazole indicating the involvement of cytochrome P-450 monooxygenase enzymes. Striking similarities between the metabolism of lipoxins and that of leukotriene B4 (LTB4) suggest that LXA4 and LXB4 are mainly hydroxylated, on omega or omega-I position, by human leukocytes and rat and human liver microsomes, whereas their all-trans isomers are mainly reduced into conjugated trienic compounds.

Animals↗
PubMed1991

Sex differences and effects of aging on visuomotor coordination.

The purpose of this study was to examine the effects of aging on men and women in processing information required to reproduce direction and distance on a visuomotor task. Subjects (45 men and 45 women) selected from three age groups (25-34 yr., 45-54 yr., 65-74 yr.) were required to estimate blindly the exact criterion location of a target-point they had identified under one of the following conditions: visual, kinesthetic, and visuokinesthetic. Errors in direction (degrees) and in distance (cm) were recorded. The analysis of the total variability in responding (E) indicated that the women of the 65- to 74-yr. age group were significantly less accurate in estimating distance than were the men of the two older age groups (45 to 54 yr., 65 to 74 yr.) and less accurate than were the women of the 25- to 34-yr. age group. No significant differences in estimating distance were found among the three age groups of men. No significant differences for direction were found between men and women.

Adult↗
PubMed1991

[Spatial representation of a two-dimensional pattern].

The purpose of this study was (a) to determine if vision and kinesthesis contribute differentially to the coding of a specific two-dimensional pattern and (b) to identify the effect of repetition on the spatial representation of this pattern. The reproductions of a specific pattern presented visually were compared with those of a pattern presented kinesthetically. The results showed that vision and kinesthesis had contributed equally to the coding of the directional components of the pattern. However, there was dominance of visual information over kinesthetic information when coding the distance between the intersecting points of the pattern, especially at the beginning of the process. Generally speaking, the visual or kinesthetic repetition, or both, have increased favourably the precision with which a specific pattern was reproduced in distance and direction.

Adolescent↗
PubMed1989

Oxidative metabolism of linoleic acid by human leukocytes.

Upon incubation with human leukocytes, [1-14C] linoleic acid is almost exclusively transformed into 13-hydroxy-9Z, 11E-octadecadienoic acid (13-HODE) if the linoleic acid concentration is lower than 50 microM. Identification of 13-HODE was done by GLC-MS at the level of its methyl ester, trimethylsilyl ether and by comparison with authentic 13-HODE in two different HPLC systems. Analysis of the products by chiral phase HPLC shows that 13(S)-hydroxy-9Z, 11E-octadecadienoic acid is by far the major metabolite formed by human leukocytes. Comparison of reactions performed with intact or lyzed cells suggests that the formation of 13(S)-HODE by human leukocytes occurs in two steps, a dioxygenation catalyzed by a 15-lipoxygenase and a reduction of intermediate 13-HPODE by a glutathione-dependent peroxidase.

Chromatography, High Pressure Liquid↗
PubMed1989

Hydroxylation of leukotriene B4 in leukocytes from various species: identification of a metabolite to authentic 5S,12R,19-trihydroxy-6Z,8E,10E,14Z-eicosatetra enoic acid and relative importance of 19- and 20-hydroxylations.

The major hydroxylated metabolite of leukotriene B4 in rat PMNL was found identical (UV spectrum and retention times in 3 different HPLC systems) to a synthetic compound of known stereochemistry, 19-hydroxy-LTB4. PMNL from various species exhibited 3 different types of behaviour for LTB4 hydroxylation. Human and monkey PMNL showed a high hydroxylating activity and a high regioselectivity with almost exclusive formation of products from 20-hydroxylation. Rat and mini-pig PMNL exhibited a very different regioselectivity with major formation of 19-OH-LTB4 (3:1 ratio). Finally, pig and beef PMNL were found almost devoid of any hydroxylating activity toward LTB4.

Animals↗