RESUMO
We assessed cerebral atrophy in mouse lemur primates (Microcebus murinus) by estimating CSF volume in their brains from 4.7 Tesla T2-weighted magnetic resonance images. Thirty animals aged from 1 to 10.3 years were imaged, 14 of them were followed for up to 2 years. Seven of these animals were examined for neuropathology. In 12 out of 17 animals older than 3.5 years, CSF volumes were increased. A subgroup of six animals had severe atrophy of the temporal lobe. Another subgroup of five animals displayed diffuse atrophy in addition to the temporal atrophy. One animal had a dilation of the external part of the temporal horn of the lateral ventricle in addition to the temporal atrophy. The three animals with diffuse atrophy that could be studied for neuropathology had diffuse cerebral amyloid deposits detected by immunocytochemistry. The other animals did not display amyloid deposits. Relations between the different types of atrophy as well as their causes will have to be assessed in future studies.
Assuntos
Envelhecimento/patologia , Encefalopatias/patologia , Encéfalo/patologia , Cheirogaleidae , Imageamento por Ressonância Magnética , Fatores Etários , Animais , Atrofia/patologia , Córtex Cerebral/patologia , Líquido Cefalorraquidiano , Feminino , Lobo Frontal/patologia , Hipocampo/patologia , Ventrículos Laterais/patologia , Masculino , Lobo Parietal/patologia , Placa Amiloide/patologia , Fatores Sexuais , Lobo Temporal/patologia , Tálamo/patologiaRESUMO
Controversy exists regarding the apolipoprotein E (ApoE) epsilon4 allele association with vascular dementia (VaD), ranging from increased epsilon4 frequency, similar to that found for Alzheimer's disease (AD), to no association between the epsilon4 allele and VaD. To clarify further the relationship between ApoE alleles polymorphism and cerebrovascular disease (CVD) in demented and cognitively impaired patients, we examined the ApoE phenotypes in a sample of 280 patients: 155 with AD, 21 with VaD, 32 with mixed dementia (MD), 45 with mild cognitive impairment (MCI) but without CVD, and 27 in which vascular disease was the most probable cause of cognitive decline [vascular mild cognitive impairment (VMCI)]. Our results show that the frequency of the ApoE epsilon4 allele in patients over 70 years old with clinically diagnosed VaD and VMCI does not differ significantly from that of controls. In contrast, ApoE epsilon4 allele-bearing individuals had greater risk of having late-onset AD (OR = 8.8; 95% CI 3.7-21.0), or non-vascular cognitive impairment (OR = 7.0; 95% CI 2.5-19.0).
Assuntos
Apolipoproteínas E/genética , Transtornos Cerebrovasculares/genética , Transtornos Cognitivos/genética , Demência/genética , Idoso , Alelos , Doença de Alzheimer/genética , Doença de Alzheimer/psicologia , Transtornos Cerebrovasculares/complicações , Transtornos Cognitivos/complicações , Demência/complicações , Demência Vascular/genética , Demência Vascular/psicologia , Feminino , Frequência do Gene , Humanos , Masculino , Fenótipo , Polimorfismo Genético/genéticaRESUMO
Evidence has indicated that circulating adrenal steroid quantitites were significantly changed in patients with Alzheimer's disease (AD). Aside of 3 beta-sulfatation and 3 beta-acylations, levels of dehydroepiandrosterone (DHEA) result from production and metabolic transformation yields. 7 alpha-Hydroxylation of DHEA has been described in humans, and 7 alpha-hydroxy-DHEA may be responsible for the known antiglucocorticoid effects of DHEA. Using a negative ion fragmentometry method with gas chromatography/mass spectrometry on trifluoroacetate derivatives, we measured levels of free 7 alpha-hydroxy-DHEA as well as its sulfated conjugate and its fatty acid esters in serum of 10 female patients with AD and of 8 age-matched healthy control women. Free 7 alpha-hydroxy-DHEA levels in AD and controls were not significantly different (240.2 +/- 37.2 pg/ml and 206.8 +/- 21.6 pg/ml, respectively), but sulfate conjugate levels were significantly increased in AD (p = .01) (262 +/- 28.4 and 145.4 +/- 27.6, respectively) as well as fatty acid esters (p = .041) (65.7 +/- 6.9 and 40.7 +/- 9.2, respectively). These results indicated that the total 7 alpha-hydroxy-DHEA produced was significantly increased in AD (p = .024) and may contribute to the disease-related disturbances of DHEA production and metabolism.
Assuntos
Doença de Alzheimer/sangue , Desidroepiandrosterona/análogos & derivados , Desidroepiandrosterona/sangue , Idoso , Idoso de 80 Anos ou mais , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
4.7 Tesla T2-weighted magnetic resonance images showed a highly significant signal decrease in the pallidum, substantia nigra, putamen, and a less significant decrease in the thalamus and the caudate of aging mouse lemurs (Microcebus murinus). We evaluated the contribution of iron deposits to the signal decrease comparing Perls' stained histological sections of six mouse lemurs brains aged 1 to 10 years to magnetic resonance images. In young animals, none of the brain structures was stained. A large number of iron deposits were visible in the pallidum and substantia nigra of aged animals and a moderate number in the middle aged ones. In the putamen, few iron deposits were visible in aged and middle-aged animals. The thalamus and the caudate appeared unstained with Perls' technique; iron was too low to be detected. The intensification of the reaction by diaminobenzidine revealed iron deposits in the thalamus of aging animals. This study suggests that in mouse lemurs, iron deposits are responsible for T2-weighted signal decrease in the central gray nuclei.
Assuntos
Envelhecimento/metabolismo , Química Encefálica/fisiologia , Encéfalo/fisiologia , Cheirogaleidae/metabolismo , Ferro/metabolismo , Animais , Encéfalo/anatomia & histologia , Cheirogaleidae/fisiologia , Feminino , Imageamento por Ressonância Magnética , MasculinoRESUMO
Previous histological and behavioral studies of aging mouse lemurs have demonstrated changes similar to those observed in elderly humans and in patients with Alzheimer's disease. We explored 18 animals of ages 6 months to 9 years. Axial T2-weighted images of the brain were performed on a 4.7 Tesla Bruker Biospec 47/30 system. We estimated cerebral atrophy by adding measures of high signal areas characteristic of cerebrospinal fluid (interlobular and sylvian fissures, lateral and third ventricles) of four contiguous cortical slices. We observed a significant increase of cerebral atrophy with aging and one case of an apathetic 8-year-old animal presenting a considerably higher cerebral atrophy. We also observed high correlations between decreased signal intensities and age for the pallidum, the substantia nigra, and the putamen. These results suggest that aging mouse lemurs present similar magnetic resonance images of cerebral alterations to those encountered in aging humans and that high-field T2-weighted magnetic resonance images can help in the early detection, in vivo, of animals suspected of pathological aging.
Assuntos
Envelhecimento/fisiologia , Encéfalo/anatomia & histologia , Encéfalo/crescimento & desenvolvimento , Cheirogaleidae/anatomia & histologia , Cheirogaleidae/crescimento & desenvolvimento , Animais , Líquido Cefalorraquidiano/fisiologia , Feminino , Imageamento por Ressonância Magnética , MasculinoRESUMO
Mouse lemurs (Microcebus murinus) are prosimian primates described to be convenient models of brain aging. We observed very high correlations between the T2-weighted magnetic resonance imaging (MRI) signal decrease and the natural logarithm of age in the basal ganglia. The correlation coefficient was higher for the pallidum (r = 0.95, P < 0.0001) than for other structures. We suggest that the ratio of the pallidum intensity divided by the amygdala and temporal lobe intensity should be a valuable non-invasive marker of age and of cerebral aging. It should be particularly useful for the non-invasive assessment of interventions and drugs that affect the aging process.
Assuntos
Envelhecimento/fisiologia , Encéfalo/fisiologia , Cheirogaleidae/fisiologia , Animais , Encéfalo/anatomia & histologia , Feminino , Imageamento por Ressonância Magnética , MasculinoRESUMO
The aim of the study was to assess the specificity of temporal amygdala (TA) atrophy with magnetic resonance imaging (MRI) by comparing a group of early impaired patients with Alzheimer's disease (AD) with 'other types of dementia' and controls. In this prospective case-control study, 41 patients were selected: 12 with probable AD according to NINCDS-ADRDA and CERAD inclusion and exclusion criteria, 14 with other types of dementia and 15 age-matched control subjects. Two radiologists blindly measured the TA volumes on coronal oblique contiguous slices with a 1.5-tesla MRI scanner. TA volume measurements obtained by the 2 observers and right-left TA values were not significantly different. A significant TA atrophy was found in the AD group as compared to the other groups, with 39.7% (p < 0.001) difference in TA volumes between AD and other types of dementia groups and 41.4% (p < 0.0005) difference between AD and control groups. There was no significant difference between other types of dementia and control groups. There was an overlap between the three groups for 4 patients. TA atrophy assessed with MRI could be of diagnostic value in AD, especially in the early stage of the disease.
Assuntos
Doença de Alzheimer/patologia , Tonsila do Cerebelo/patologia , Lobo Temporal/patologia , Idoso , Idoso de 80 Anos ou mais , Tonsila do Cerebelo/anatomia & histologia , Atrofia/patologia , Estudos de Casos e Controles , Demência/patologia , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Estudos Prospectivos , Valores de Referência , Lobo Temporal/anatomia & histologiaRESUMO
OBJECTIVE: To determine whether changes in phosphomonoester and phosphodiester levels could be detected in vivo with phosphorus magnetic resonance spectroscopy in the early stage of Alzheimer's disease (AD). DESIGN: Survey-type of case-control study using neuropsychological testing as criterion standard with blinded data analysis. SETTING: Patients were from a neurology clinic in Paris, France. The controls were from the community. Magnetic resonance measurements were performed in the prefrontal region of the brain with a clinical 1.5-T scanner. Blinded data analysis. PARTICIPANTS: Twenty-four patients with mild AD and 15 age-matched healthy volunteers. Subjects were separated into two groups, both composed of patients with AD and healthy volunteers. Two successive acquisition protocols were used in the two groups. RESULTS: A significant increase in the phosphomonoester-total phosphorus ratio was found in patients with AD compared with controls. In this series, use of a ratio above 11% as a threshold to test our sample yielded an 83.3% sensitivity and a 73.3% specificity test for AD. Other metabolite ratios (inorganic phosphate, phosphodiesters, phosphocreatine, and nucleotide phosphates to total phosphorus) were not significantly different between patients and controls. No metabolite ratio correlated with the neuropsychological status as assessed by the Mini-Mental State Examination. CONCLUSION: Changes in phospholipid metabolism can be detected in vivo in the early stage of AD. Discrepancies in the literature may be due to differences in technical setting or in subject population types.
Assuntos
Doença de Alzheimer/diagnóstico , Doença de Alzheimer/metabolismo , Espectroscopia de Ressonância Magnética , Fosfolipídeos/metabolismo , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , FósforoRESUMO
There is growing evidence that AD consists of different subtypes, and that language is a pertinent factor to identify a subgroup with a fast rate of cognitive decline. We report the first results of a longitudinal study in which we compared two groups of patients with probable AD. The main result showed that a subgroup with stable MMSE scores during a 1-year follow-up period had an impairment in language domains which are usually preserved until an advanced stage of the disease. It is proposed that this group may correspond to a variety of AD who, in addition to symptoms of AD, present characteristics of primary progressive aphasia. In other respects, we underscore that the high loading in language-mediated tasks of the MMSE makes it a poor index to accurately measure the rate of cognitive decline.
Assuntos
Doença de Alzheimer/psicologia , Transtornos Cognitivos/fisiopatologia , Transtornos da Linguagem/fisiopatologia , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/fisiopatologia , Feminino , Humanos , Masculino , Testes NeuropsicológicosRESUMO
OBJECTIVES: To study the ability of magnetic resonance imaging to measure the volume of the amygdala and detect amygdala atrophy in patients with early Alzheimer's disease. DESIGN: Prospective case-control study and "blind" measurements. SETTING: Subjects were ambulatory outpatients selected from an institutional practice in Paris, France. PATIENTS: We studied 11 patients with probable Alzheimer's disease according to National Institute of Neurologic and Communicative Disorders and Stroke/Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA) and Consortium to Establish a Registry for Alzheimer's Disease (CERAD) inclusion and exclusion criteria, as well as six age-matched control subjects. INTERVENTION: None. MAIN OUTCOME MEASURE: A 1.5-T magnetic resonance imager was used to acquire the images. Two neuroradiologists independently and blindly measured the volume of the right and left amygdalas on high-resolution contiguous slices. In addition, other cerebral structures, ie, the sylvian fissures, temporal lobes, lateral and third ventricles, corpus callosum, and hippocampal formation, were measured on a single slice. RESULTS: The values obtained by the two observers correlated highly (r = .90), and interrater variability was 13%. The Alzheimer's disease group showed significant (33%, P < .0001) atrophy of the amygdala when compared with the control group. The other structures showed less variation. CONCLUSION: Significant amygdala atrophy can be detected in vivo in patients with early Alzheimer's disease by means of standard magnetic resonance imaging. This technique may be useful in the early diagnosis of Alzheimer's disease.
Assuntos
Doença de Alzheimer/patologia , Tonsila do Cerebelo/patologia , Imageamento por Ressonância Magnética , Idoso , Doença de Alzheimer/epidemiologia , Doença de Alzheimer/psicologia , Atrofia , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Estudos Prospectivos , Escalas de Graduação PsiquiátricaRESUMO
Using a combination of in situ mapping and DNA analysis with recombinant DNA probes specific for the Sxr region of the mouse Y chromosome, we show that both the gene(s) controlling sex determination and the expression of the male-specific antigen H-Y (Tdy and Hya, respectively) are located on the minute short arm of the mouse Y chromosome. We demonstrate that the H-Y- variant of Sxr (Sxr') arose by a partial deletion within the Sxr region. Also, we show that intrachromosomal recombination between the Y short arm and Sxr' can sometimes occur during male meiosis, restoring the deleted DNA sequences and resulting in an H-Y+ mouse (male 719 in this paper). Based on these results, we propose a model for the generation of the original Sxr region and the Sxr' and Sxr719 variants.
Assuntos
Camundongos/genética , Análise para Determinação do Sexo , Cromossomo Y , Animais , Mapeamento Cromossômico , DNA/genética , DNA/isolamento & purificação , DNA Recombinante/metabolismo , Fígado/fisiologia , Masculino , Hibridização de Ácido NucleicoRESUMO
We present genetic and molecular data demonstrating linkage of the gene for steroid sulfatase (Sts) to the mutation sex reversed (Sxr) definitively showing the existance of a functional allele for Sts mapping to the pseudoautosomal region of the mouse Y chromosome. Thus, in mouse, functional Sts genes are present in the pseudoautosomal region of both the X and Y chromosomes. This is in contrast to man where Sts has been mapped to the short arm of the X just centromeric to the pseudoautosomal region. Only a single recombinant separating Sts and Sxr was found out of 103 male meioses analyzed; double recombinants were not found between sex (Tdy), Sts and Sxr. If the rate of recombination in the pseudoautosomal region in male mice is equivalent to that in man and thus 7-10X higher than normal, then our data suggest that the distance between Sts and Sxr (or the telomere of the Y) is approximately 100-200 kb in length. Our data is in contrast to a recent report of a recombination frequency separating Sts and Sxr of as high as 6.2-9.8%.
Assuntos
Transtornos do Desenvolvimento Sexual , Genes , Ligação Genética , Mutação , Sulfatases/genética , Animais , DNA Recombinante/metabolismo , Feminino , Genótipo , Masculino , Meiose , Camundongos , Camundongos Endogâmicos , Hibridização de Ácido Nucleico , Fenótipo , Esteril-SulfataseRESUMO
The mouse Y chromosome plays a fundamental role in the control of primary sex determination and fertility. Both genetic and molecular biological evidence has shown that much of the necessary information is contained in a minute piece of the Y (the Sxr region) which has arisen by a duplication of the pericentric region of the normal Y and the transposition of one copy to the distal pseudoautosomal region. The present article describes the isolation of random Y-chromosome probes and their use to investigate this Sxr region at the molecular level. Total mouse Y-chromosome libraries were constructed from flow-sorted material and a Sxr regional library after specific microdissection and cloning. Transcription has been detected in the testis using both Sxr-specific and non Sxr-located genomic probes taken from these libraries. In addition, we have been able to confirm the presence of an active steroid sulphatase gene on the mouse Y. This gene is located in the distal portion of the pseudoautosomal region and is tightly linked to Sxr. Finally, using an Sxr-specific probe we can define multiple Y-chromosome haplotypes in the mouse showing that the region is evolving very rapidly.
Assuntos
Sondas de DNA , Mutação , Cromossomo Y , Animais , Arilsulfatases/genética , Separação Celular , Clonagem Molecular , Transtornos do Desenvolvimento Sexual , Feminino , Ligação Genética , Masculino , Camundongos , Polimorfismo Genético , Análise para Determinação do Sexo , Esteril-Sulfatase , Testículo/análise , Transcrição GênicaRESUMO
Hog thyroid peroxidase (TPO) was highly purified in order to study the spectral properties and catalytic specificities of its H2O2 compounds in iodothyronine biosynthesis. Purified TPO exhibited a Soret spectrum with an absorption maximum at 410 nm and had an A410/A280 value of 0.55. Protein iodination was only catalyzed under conditions which allowed formation of the transient TPO compound I (Fe(IV)-pi o+). On addition of an equimolar amount of H2O2, TPO formed a stable compound with an absorption maximum at 417 nm. This compound efficiently catalyzed the coupling reaction, but was unable to iodinate proteins. It catalyzed the formation of 1 mol iodothyronines/mol TPO, and therefore retained two oxidizing equivalents per molecule. It is proposed that this compound constitutes a second form of compound I whose structure might be Fe(IV)-Ro, analogous to that of cytochrome c peroxidase compound I. In the presence of an excess of H2O2, it formed TPO-compound III with an absorption maximum at 420 nm. TPO-compound III catalyzed neither the iodination nor the coupling reaction.
Assuntos
Peróxido de Hidrogênio/metabolismo , Iodeto Peroxidase/metabolismo , Iodo/metabolismo , Animais , Citocromo-c Peroxidase/metabolismo , Monoiodotirosina/metabolismo , Proteínas/metabolismo , Espectrofotometria , Suínos , Tireoglobulina/análiseRESUMO
In thyroid gland, iodination takes place on the apical plasma membrane and requires the presence of the thyroid peroxidase and H2O2 generating system. H2O2 generation and NBT (nitro blue tetrazolium) reductase activity (both of which are NADPH-dependent) as well as peroxidase activity were compared for their respective orientations in membrane vesicles. The possible role of NADPH-NBT reductase activity in H2O2 generation was also examined. Results favor the conclusion that thyroid peroxidase is oriented towards the luminal side of the vesicles, whereas the NADPH site of NADPH oxidase-dependent H2O2 generation is located on the external side of the same or of different vesicles. Furthermore, it is shown that different NADPH-NBT reductase activities are present on both the outer and inner surfaces of the membrane vesicles, and that none of these activities is able to produce either H2O2 or O-2. The idea that a multi-component complex is involved in H2O2 generation is discussed, and a model is proposed which takes into account the possible spatial separation of the thyroid peroxidase site from the NADPH site of this H2O2 generation system on the apical membrane of the thyrocyte.
Assuntos
Peróxido de Hidrogênio/biossíntese , NADP/metabolismo , Oxirredutases/metabolismo , Peroxidases/metabolismo , Glândula Tireoide/metabolismo , Animais , Digitonina , Complexos Multienzimáticos/metabolismo , NADH Desidrogenase/metabolismo , Nitroazul de Tetrazólio/metabolismo , Fosfolipases A , Frações Subcelulares/metabolismo , Suínos , TripsinaRESUMO
We have examined the respective roles played by guaiacol and scopoletin in NADPH oxidation catalyzed by the peroxidase/H2O2 system. It was shown that NADPH was not oxidized by either the horseradish or lactoperoxidase/H2O2 systems alone; oxidation occurred immediately after the addition of guaiacol or scopoletin. In both cases, the oxidation product was enzymatically active NADP+. Differences were observed in the NADPH oxidation mechanism depending on whether guaiacol or scopoletin was the mediator molecule. In guaiacol-mediated NADPH oxidation, the stoichiometry between H2O2 and oxidized NADPH was about 1; superoxide dismutase did not affect the oxidation rate. In scopoletin-mediated oxidation, the stoichiometry was much higher (1:14 in the present experiments); superoxide dismutase considerably increased the oxidation rate. It is concluded that catalysis of NADPH oxidation by the horse radish peroxidase/H2O2 system requires the presence of a mediator molecule. The NADPH oxidation mechanism depends on the intermediary oxidation state of this molecule.
Assuntos
Guaiacol/metabolismo , Peroxidase do Rábano Silvestre/farmacologia , Peróxido de Hidrogênio/farmacologia , NADP/metabolismo , Peroxidases/farmacologia , Escopoletina/metabolismo , Umbeliferonas/metabolismo , Catálise , Cinética , NADP/isolamento & purificação , Oxirredução , Superóxido Dismutase/farmacologiaRESUMO
Oxidation of NADPH catalyzed by the peroxidase/H2O2 system is known to require the presence of mediating molecules. Using either lactoperoxidase or horseradish peroxidase, we demonstrated that in the peroxidase/H2O2 system, NADPH oxidation was mediated by iodide. The oxidation product was the iodinated NADP. This product was shown to possess spectral characteristics different from those of NADP+ and NADPH, since for iodinated NADP, increased absorbance was observed in the 280-nm region and was directly proportional to the rate of iodination. It is suggested that oxidation and iodination of NADPH proceed via a single reaction between the intermediary iodide oxidation species and NADPH. Experiments with different molecules of NADPH analogues indicated that iodination occurred in the nicotinamide part of the NADPH molecule.
Assuntos
Iodetos , NADP , Peroxidases/metabolismo , Catálise , Fenômenos Químicos , Química , Peroxidase do Rábano Silvestre/metabolismo , Lactoperoxidase/metabolismo , Oxirredução , Espectrofotometria UltravioletaRESUMO
Cytochrome c peroxidase (CcP) and horseradish peroxidase (HRP), when combined with a stoichiometric amount of H2O2, form stable compounds I which are known as FeIV Ro and FeIV o pi + structures, respectively. These compounds were assayed in the catalysis of thyroid hormone synthesis and the iodination reaction. As previously shown for the lactoperoxidase FeIV Ro compound, the CcP FeIV Ro compound was involved in the coupling and not in the iodination reactions. In contrast, the HRP FeIV o pi + compound catalyzed both iodination and hormone formation. The possible role of the different peroxidase-H2O2 compounds in the two sequential reactions, thyroglobulin iodination and thyroid hormone formation, is discussed.
Assuntos
Citocromo-c Peroxidase/metabolismo , Peroxidase do Rábano Silvestre/metabolismo , Iodo/metabolismo , Peroxidases/metabolismo , Tireoglobulina/metabolismo , Hormônios Tireóideos/biossíntese , Catálise , Fenômenos Químicos , Química , Peróxido de Hidrogênio/análise , Concentração de Íons de Hidrogênio , Cinética , Lactoperoxidase/metabolismo , OxirreduçãoRESUMO
The titration by ferrocyanide and the localization of the oxidizing equivalents of lactoperoxidase "compound II" were studied as a function of pH. It was demonstrated that 1) whatever the pH, the structure of lactoperoxidase "compound II" was compatible with a Fe IV R degree state, 2) at acidic pH, ferrocyanide preferentially reduced the oxidizing equivalent localized on the heme iron to give an Fe III R degree compound, 3) at pH 4.2 only the Fe III R degree form was obtained after reduction of lactoperoxidase "compound II" with one mole of ferrocyanide and whereas at pH greater than 4.2, a mixture of both Fe III R degree and Fe IV R forms was present, 4) lowering the pH from 7.2 to 4.0 induced a transition of Fe IV R state to Fe III R degree state, but increasing the pH from 4.0 to 7.2 did not permit the formation of Fe IV R compound from Fe III R degree compound.
Assuntos
Apoenzimas , Apoproteínas , Ferrocianetos , Peróxido de Hidrogênio , Lactoperoxidase , Peroxidases , Sítios de Ligação , Concentração de Íons de Hidrogênio , Técnicas de Diluição do Indicador , Cinética , Modelos Químicos , OxirreduçãoRESUMO
A NADPH-dependent H2O2 generating system associated with a thyroid particular fraction is described. H2O2 is measured by two different methods: iodination of NADPH itself when the system is supplemented with lactoperoxidase and [125I]iodide, and by the scopoletin method. It is shown that: H2O2 generation is inhibited by catalase and is dependent on NADPH or particulate protein concentration; radical scavengers of OH and of singlet oxygen have no effect while superoxide dismutase has only a marginal effect; disruption of the particular fraction by phospholipase A2 or digitonin treatment completely abolished H2O2 generation activity while thyroid peroxidase activity appears, suggesting different sites for the two activities in the membrane vesicles.