Constitutive expression and localization of cytochrome P-450 1A1 in rat and human brain: presence of a splice variant form in human brain

Cytochrome P-450 function as mono-oxygenases and metabolize xenobiotics. CYP1A1, a cytochrome P-450 enzyme, bioactivates polycyclic aromatic hydrocarbons to reactive metabolite(s) that bind to DNA and initiate carcinogenesis. Northern and immunoblot analyses revealed constitutive expression of Cyp1a1 and CYP1A1 in rat and human brain, respectively. CYP1A1 mRNA and protein were localized predominantly in neurons of cerebral cortex, Purkinje and granule cell layers of cerebellum and pyramidal neurons of CA1, CA2, and CA3 subfields of the hippocampus. RT-PCR analyses using RNA obtained from autopsy human brain samples demonstrated the presence of a splice variant having a deletion of 87 bp of exon 6. This splice variant was present in human brain, but not in the liver from the same individual, and was absent in rat brain and liver. Structural modeling indicated broadening of the substrate access channel in the brain variant. The study demonstrates the presence of a unique cytochrome P-450 enzyme in human brain that is generated by alternate splicing. The presence of distinct cytochrome P-450 enzymes in human brain that are different from well-characterized hepatic forms indicates that metabolism of xenobiotics including drugs could occur in brain by pathways different from those known to occur in liver

An alternatively spliced cytochrome P4501A1 in human brain fails to bioactivate polycyclic aromatic hydrocarbons to DNA-reactive metabolites

CYP1A1, a cytochrome P450 enzyme, metabolizes polycyclic aromatic hydrocarbons to genotoxic metabolite(s) that bind to DNA and initiate carcinogenesis. RT-PCR amplification of the complete open reading frame of CYP1A1 generated an amplicon of 1593 bp having deletion of 87 bp of exon-6 that translated into functional P450 enzyme. Unlike wild type CYP1A1, exon 6 del CYP1A1 did not metabolize polycyclic aromatic hydrocarbons such as, benzo(a)pyrene to genotoxic, ultimate carcinogens that form DNA adducts. Exon 6 del CYP1A1 metabolized ethoxyresorufin (the classical substrate for CYP1A1) less efficiently compared with wild type CYP1A1 while pentoxy and benzyloxyresorufin (classical substrates for CYP2B) were dealkylated more efficiently. In silico docking showed alteration of the substrate access channel in exon 6 del CYP1A1 such that benzo(a)pyrene does not bind in any orientation that would permit the formation of carcinogenic metabolites. Genotyping revealed that the splice variant was not generated due to differences in genomic DNA sequence and the variant was present only in brain but not in liver, kidney, lung, or heart from the same individual. We provide evidence that unique P450 enzymes, generated by alternate splicing in a histiospecific manner can modify genotoxic potential of carcinogens such as benzo(a)pyrene by altering their biotransformation pathway

Hydroxylation of benzphetamine and other drugs by a solubilized form of cytochrome P-450 from liver microsomes: Lipid requirement for drug demethylation

A solubilized hepatic microsomal enzyme system previously shown to catalyze the [omega]-hydroxylation of fatty acids also catalyzes the hydroxylation of drugs. Benzphetamine, aminopyrine, ethylmorphine, hexobarbital, norcodeine, and -nitroanisole undergo aerobic demethylation in the presence of NADPH and the resolved enzyme system. The required submicrosomal components for benzphetamine demethylation, as determined either by formaldehyde liberation or by NADPH oxidation, are cytochrome P-450, NADPH-cytochrome P-450 reductase, and a heat-stable lipid fraction. Similar requirements were shown for the oxidation of aminopyrine, ethylmorphine, and hexobarbital. Laurate and benzphetamine were found to be mutually inhibitory, as would be expected if a common "methyl hydroxylase" were involved. The solubilized cytochrome P-450 preparation exhibits a difference spectrum in the presence of benzphetamine with a peak at 392 m[mu] and a trough at 427 m[mu] and difference spectra with aniline and hexobarbital typical of those obtained with the microsomal bound form of this hemoprotein.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/32916/1/0000296.pd

Hepatic and renal cytochrome p450 gene regulation during citrobacter rodentium infection in wild-type and toll-like receptor 4 mutant mice.

Citrobacter rodentium is the rodent equivalent of human enteropathogenic Escherichia coli infection. This study investigated regulation of hepatic and renal cytochrome P450 (P450) mRNAs, hepatic P450 proteins, cytokines, and acute phase proteins during C. rodentium infection. Female C3H/HeOuJ (HeOu) and C3H/HeJ (HeJ) mice [which lack functional toll-like receptor 4 (TLR4)] were infected with C. rodentium by oral gavage and sacrificed 6 days later. Hepatic CYP4A10 and 4A14 mRNAs were decreased in HeOu mice (\u3c4% of control). CYP3A11, 2C29, 4F14, and 4F15 mRNAs were reduced to 16 to 55% of control levels, whereas CYP2A5, 4F16, and 4F18 mRNAs were induced (180, 190, and 600% of control, respectively). The pattern of P450 regulation in HeJ mice was similar to that in HeOu mice for most P450s, with the exception of the TLR4 dependence of CYP4F15. Hepatic CYP2C, 3A, and 4A proteins in both groups were decreased, whereas CYP2E protein was not. Renal CYP4A10 and 4A14 mRNAs were significantly down-regulated in HeOu mice, whereas other P450s were unaffected. Most renal P450 mRNAs in infected HeJ mice were increased, notably CYP4A10, 4A14, 4F18, 2A5, and 3A13. Hepatic levels of interleukin (IL)-1beta, IL-6, and tumor necrosis factor alpha (TNFalpha) mRNAs were significantly increased in infected HeOu mice, whereas only TNFalpha mRNA was significantly increased in HeJ mice. Hepatic alpha1-acid glycoprotein was induced in both groups, whereas alpha-fibrinogen and angiotensinogen were unchanged. These data indicate that hepatic inflammation induced by C. rodentium infection is mainly TLR4-independent and suggest that hepatic P450 down-regulation in this model may be cytokine-mediated

A Planet at 5 AU Around 55 Cancri

We report precise Doppler shift measurements of 55 Cancri (G8V) obtained from 1989 to 2002 at Lick Observatory. The velocities reveal evidence for an outer planetary companion to 55 Cancri orbiting at 5.5 AU. The velocities also confirm a second, inner planet at 0.11 AU. The outer planet is the first extrasolar planet found that orbits near or beyond the orbit of Jupiter. It was drawn from a sample of ~50 stars observed with sufficient duration and quality to detect a giant planet at 5 AU, implying that such planets are not rare. The properties of this jupiter analog may be compared directly to those of the Jovian planets in our Solar System. Its eccentricity is modest, e=0.16, compared with e=0.05 for both Jupiter and Saturn. Its mass is at least 4.0 jupiter masses (M sin i). The two planets do not perturb each other significantly. Moreover, a third planet of sub-Jupiter mass could easily survive in between these two known planets. Indeed a third periodicity remains in the velocity measurements with P = 44.3 d and a semi-amplitude of 13 m/s. This periodicity is caused either by a third planet at a=0.24 AU or by inhomogeneities on the stellar surface that rotates with period 42 d. The planet interpretation is more likely, as the stellar surface is quiet, exhibiting log(R'_{HK}) = -5.0 and brightness variations less than 1 millimag, and any hypothetical surface inhomogeneity would have to persist in longitude for 14 yr. Even with all three planets, an additional planet of terrestrial--mass could orbit stably at ~1 AU. The star 55 Cancri is apparently a normal, middle-aged main sequence star with a mass of 0.95 solar masses, rich in heavy elements ([Fe/H] = +0.27). This high metallicity raises the issue of the relationship between its age, rotation, and chromosphere.Comment: 47 pages, 4 tables, 12 figures, uses AASTE

Regulation of hepatic cytochrome P450 expression in mice with intestinal or systemic infections of citrobacter rodentium.

We reported previously that infection of C3H/HeOuJ (HeOu) mice with the murine intestinal pathogen Citrobacter rodentium caused a selective modulation of hepatic cytochrome P450 (P450) gene expression in the liver that was independent of the Toll-like receptor 4. However, HeOu mice are much more sensitive to the pathogenic effects of C. rodentium infection, and the P450 down-regulation was associated with significant morbidity in the animals. Here, we report that oral infection of C57BL/6 mice with C. rodentium, which produced only mild clinical signs and symptoms, produced very similar effects on hepatic P450 expression in this strain. As in HeOu mice, CYP4A mRNAs and proteins were among the most sensitive to down-regulation, whereas CYP4F18 was induced. CYP2D9 mRNA was also induced 8- to 9-fold in the C57BL/6 mice. The time course of P450 regulation followed that of colonic inflammation and bacterial colonization, peaking at 7 to 10 days after infection and returning to normal at 15 to 24 days as the infection resolved. These changes also correlated with the time course of significant elevations in the serum of the proinflammatory cytokines interleukin (IL)-6 and tumor necrosis factor-alpha, as well as of interferon-gamma and IL-2, with serum levels of IL-6 being markedly higher than those of the other cytokines. Intraperitoneal administration of C. rodentium produced a rapid down-regulation of P450 enzymes that was quantitatively and qualitatively different from that of oral infection, although CYP2D9 was induced in both models, suggesting that the effects of oral infection on the liver are not due to bacterial translocation

Kinematics, ages and metallicities for F and G type stars in the solar neighbourhood

A new metallicity distribution and an age-metallicity relation are presented for 437 nearby F and G turn-off and sub-giant stars selected from radial velocity data of Nidever et al. Photometric metallicities are derived from uvby-H\beta photometry, and the stellar ages from the isochrones of Bergbusch & VandenBerg as transformed to $uvby$ photometry using the methods of Clem et al. The X (stellar-population) criterion of Schuster et al., which combines both kinematic and metallicity information, provides 22 thick-disk stars. \sigma_{\rm W} = 32 \pm 5 km s^{-1}, = 154 \pm 6 km s^{-1} and = -0.55 \pm 0.03 dex for these thick-disk stars, which is in agreement with values from previous studies of the thick disk. \alpha -element abundances which are available for some of these thick-disk stars show the typical alpha-element signatures of the thick disk, supporting the classification procedure based on the $X$ criteria. Both the scatter in metallicity at a given age and the presence of old, metal-rich stars in the age-metallicity relation make it difficult to decide whether or not an age-metallicity relation exists for the older thin-disk stars. For ages greater than 3 Gyr, our results agree with the other recent studies that there is almost no correlation between age and metallicity, \Delta ([M/Fe])/\Delta(age) = -0.01 \pm 0.005 dex Gyr^{-1}. For the 22 thick-disk stars there is a range in ages of 7-8 Gyr, but again almost no correlation between age and metallicity.Comment: 11 pages, including 10 figures and 3 tables, accepted for publication in MNRA

Drug Metabolism in Human Brain: High Levels of Cytochrome P4503A43 in Brain and Metabolism of Anti-Anxiety Drug Alprazolam to Its Active Metabolite

Cytochrome P450 (P450) is a super-family of drug metabolizing enzymes. P450 enzymes have dual function; they can metabolize drugs to pharmacologically inactive metabolites facilitating their excretion or biotransform them to pharmacologically active metabolites which may have longer half-life than the parent drug. The variable pharmacological response to psychoactive drugs typically seen in population groups is often not accountable by considering dissimilarities in hepatic metabolism. Metabolism in brain specific nuclei may play a role in pharmacological modulation of drugs acting on the CNS and help explain some of the diverse response to these drugs seen in patient population. P450 enzymes are also present in brain where drug metabolism can take place and modify therapeutic action of drugs at the site of action. We have earlier demonstrated an intrinsic difference in the biotransformation of alprazolam (ALP) in brain and liver, relatively more α-hydroxy alprazolam (α-OHALP) is formed in brain as compared to liver. In the present study we show that recombinant CYP3A43 metabolizes ALP to both α-OHALP and 4-hydroxy alprazolam (4-OHALP) while CYP3A4 metabolizes ALP predominantly to its inactive metabolite, 4-OHALP. The expression of CYP3A43 mRNA in human brain samples correlates with formation of relatively higher levels of α-OH ALP indicating that individuals who express higher levels of CYP3A43 in the brain would generate larger amounts of α-OHALP. Further, the expression of CYP3A43 was relatively higher in brain as compared to liver across different ethnic populations. Since CYP3A enzymes play a prominent role in the metabolism of drugs, the higher expression of CYP3A43 would generate metabolite profile of drugs differentially in human brain and thus impact the pharmacodynamics of psychoactive drugs at the site of action

Micronutrient status in lactating mothers before and after introduction of fortified flour: cross-sectional surveys in Maela refugee camp

Background Deficiency of micronutrients is common in refugee populations. Objectives Identify deficiencies and whether provided supplements and wheat flour fortified with 10 micronutrients impacts upon status among breast-feeding women from Maela refugee camp. Methods Two sequential cross-sectional studies were conducted in different groups of lactating mothers at 12 weeks postpartum. The first survey was before and the second 4-5 months after micronutrient fortified flour (MFF) had been provided to the camp (in addition to the regular food basket). Iron status and micronutrients were measured in serum, whole blood, and in breast milk samples. Results Iron and zinc deficiency and anemia were highly prevalent while low serum retinol and thiamine deficiency were rarely detected. Iron and zinc deficiency were associated with anemia, and their proportions were significantly lower after the introduction of MFF (21 vs. 35% with soluble transferrin receptor (sTfR)>8.5 mg/L, P = 0.042, and 50 vs. 73% with serum zinc<0.66 mg/L, P = 0.001). Serum sTfR, whole-blood thiamine diphosphate (TDP) and serum β-carotene were significant predictors (P<0.001) of milk iron, thiamine and β-carotene, respectively. Lower prevalence of iron deficiency in the MFF group was associated with significantly higher iron and thiamine in breast milk. Conclusions High whole-blood TDP and breast milk thiamine reflected good compliance to provided thiamine; high prevalence of iron deficiency suggested insufficient dietary iron and low acceptance to ferrous sulfate supplements. MFF as an additional food ration in Maela refugee camp seemed to have an effect in reducing both iron and zinc deficiency postpartum. © Springer-Verlag 2012