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1.
Rev. clín. esp. (Ed. impr.) ; 223(6): 331-339, jun.- jul. 2023.
Artigo em Espanhol | IBECS | ID: ibc-221348

RESUMO

Objetivos Este estudio tiene como objetivo la determinación de la incidencia de CPPD y la identificación de factores predisponentes en su aparición. Método Se lleva a cabo un estudio descriptivo, de carácter prospectivo en 57 pacientes a los que se les realiza una punción lumbar. Para ello, se han analizado variables relativas a factores de riesgo derivado del paciente, factores clínicos y del procedimiento con la presencia de CPPD. La incidencia de CPPD ha sido de 38,6% y entre los factores asociados a su aparición se ha identificado la edad joven y el antecedente de cefalea previa. Resultados La incidencia de CPPD ha sido mayor en mujeres, siendo de mayor intensidad en este grupo, si bien es necesaria la realización de estudios con mayor tamaño muestra. Conclusiones Debemos tener presente los factores asociados a la aparición de una CPPD como son: la edad joven, el antecedente de cefalea y la percepción de dificultad del proceso, para una mejor información a los pacientes y una optimización de la técnica empleada (AU)


Introduction Post-dural puncture headache (PDPH) is the most common complication following lumbar puncture. However, its incidence varies according to the series consulted. Different factors associated with its onset have been identified. Objectives The purpose of this study is to determine the incidence of PDPH and to identify predisposing factors for its appearance. Method Prospective, descriptive study in 57 patients who underwent lumbar puncture procedures. To this end, variables associated with patient-related risk factors, clinical and procedural factors with the presence of PDPH were analysed. The incidence of PDPH was 38.6% and factors associated with onset included young age and previous history of headache. Results The incidence of PDPH was higher in women and presented greater intensity in this group, though studies with a larger sample size would need to be conducted. Conclusions We must bear in mind the factors associated with the appearance of PDPH, which include: young age, history of headache, and the perception of procedural difficulty, to better inform patients and optimise the techniques used (AU)


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Cefaleia Pós-Punção Dural/etiologia , Punção Espinal/efeitos adversos , Estudos Longitudinais , Estudos Prospectivos , Fatores de Risco
2.
Rev Clin Esp (Barc) ; 223(6): 331-339, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37169081

RESUMO

INTRODUCTION: Post-dural puncture headache (PDPH) is the most common complication following lumbar puncture. However, its incidence varies according to the series consulted. Different factors associated with its onset have been identified. OBJECTIVES: The purpose of this study is to determine the incidence of PDPH and to identify predisposing factors for its appearance. METHOD: Prospective, descriptive study in 57 patients who underwent lumbar puncture procedures. To this end, variables associated with patient-related risk factors, clinical and procedural factors with the presence of PDPH were analysed. The incidence of PDPH was 38.6% and factors associated with onset included young age and previous history of headache. RESULTS: The incidence of PDPH was higher in women and presented greater intensity in this group, though studies with a larger sample size would need to be conducted. CONCLUSIONS: We must bear in mind the factors associated with the appearance of PDPH, which include: young age, history of headache, and the perception of procedural difficulty, to better inform patients and optimise the techniques used.


Assuntos
Cefaleia Pós-Punção Dural , Humanos , Feminino , Cefaleia Pós-Punção Dural/epidemiologia , Cefaleia Pós-Punção Dural/etiologia , Estudos Prospectivos , Cefaleia/complicações , Cefaleia/epidemiologia , Fatores de Risco , Punção Espinal/efeitos adversos
3.
Interv Neuroradiol ; 29(3): 301-306, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35285737

RESUMO

BACKGROUND AND PURPOSE: The carotid stent placement as a therapeutic option for carotid stenosis has been increasing among years; therefore, studies are required to evaluate the security and efficacy of its materials. The purpose of this study was to evaluate the distal filter and the proximal balloon-guided catheter with flow inversion as protection devices during carotid angioplasty and stenting. METHODS: This is a retrospective, observational study of patients diagnosed with carotid stenosis treated with angioplasty between January 1, 2014, and June 30, 2020; we analyzed a radiology service database to compare the distal filter and the proximal balloon-guided catheter as protection devices during angioplasty. RESULTS: One hundred seventy-five angioplasties were performed, the distal filter was the most prevalent embolic protection device used (66%), patients baseline characteristics did not differ between groups with different embolic protection devices, except for history of dyslipidemia (p < 0.000). As well, we did not find any significant differences between the groups in the device related complications, intervention time (p = 0.140), unrelated complications (p = 0.693) and functional independence at 90 days (p = 0.096). CONCLUSIONS: In our study the proximal balloon-guided catheter and the distal filter protection device as protection devices during the carotid stenting didn't show significant differences regarding complications related to the system.


Assuntos
Angioplastia com Balão , Estenose das Carótidas , Humanos , Estenose das Carótidas/diagnóstico por imagem , Estenose das Carótidas/cirurgia , Colômbia , Resultado do Tratamento , Catéteres , Stents
4.
Mar Policy ; 140: 105054, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35399704

RESUMO

The human response to the COVID-19 pandemic set in motion an unprecedented shift in human activity with unknown long-term effects. The impacts in marine systems are expected to be highly dynamic at local and global scales. However, in comparison to terrestrial ecosystems, we are not well-prepared to document these changes in marine and coastal environments. The problems are two-fold: 1) manual and siloed data collection and processing, and 2) reliance on marine professionals for observation and analysis. These problems are relevant beyond the pandemic and are a barrier to understanding rapidly evolving blue economies, the impacts of climate change, and the many other changes our modern-day oceans are undergoing. The "Our Ocean in COVID-19″ project, which aims to track human-ocean interactions throughout the pandemic, uses the new eOceans platform (eOceans.app) to overcome these barriers. Working at local scales, a global network of ocean scientists and citizen scientists are collaborating to monitor the ocean in near real-time. The purpose of this paper is to bring this project to the attention of the marine conservation community, researchers, and the public wanting to track changes in their area. As our team continues to grow, this project will provide important baselines and temporal patterns for ocean conservation, policy, and innovation as society transitions towards a new normal. It may also provide a proof-of-concept for real-time, collaborative ocean monitoring that breaks down silos between academia, government, and at-sea stakeholders to create a stronger and more democratic blue economy with communities more resilient to ocean and global change.

5.
Sci Rep ; 8(1): 15182, 2018 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-30315190

RESUMO

Intestinal fibrosis is thought to be a consequence of excessive tissue repair, and constitutes a common problem in patients with Crohn's disease (CD). While fibrosis seems to require inflammation as a prerequisite it is unclear whether the severity or persistence of inflammation influences the degree of fibrosis. Our aim was to investigate the role of sustained inflammation in fibrogenesis. For the initiation of fibrosis in vivo the models of Il10-/- spontaneous colitis, dextran sodium sulfate (DSS)-induced chronic colitis and heterotopic transplantation were used. In Il10-/- mice, we determined a positive correlation between expression of pro-inflammatory factors (Il1ß, Tnf, Ifnγ, Mcp1 and Il6). We also found a positive correlation between the expression of pro-fibrotic factors (Col3a1 Col1a1, Tgfß and αSma). In contrast, no significant correlation was determined between the expression of pro-inflammatory Tnf and pro-fibrotic αSma, Col1a1, Col3a1, collagen layer thickness and the hydroxyproline (HYP) content. Results from the DSS-induced chronic colitis model confirmed this finding. In the transplantation model for intestinal fibrosis a pronounced increase in Mcp1, inos and Il6 in Il10-/- as compared to WT grafts was observed, indicating more severe inflammation in Il10-/- grafts. However, the increase of collagen over time was virtually identical in both Il10-/- and WT grafts. Severity of inflammation during onset of fibrogenesis did not correlate with collagen deposition. Although inflammation might be a pre-requisite for the initiation of fibrosis our data suggest that it has a minor impact on the progression of fibrosis. Our results suggest that development of fibrosis and inflammation may be disconnected. This may be important for explaining the inefficacy of anti-inflammatory treatments agents in most cases of fibrotic inflammatory bowel diseases (IBD).


Assuntos
Fibrose/patologia , Inflamação/patologia , Intestinos/patologia , Animais , Colite/metabolismo , Colite/patologia , Colágeno/metabolismo , Colo/metabolismo , Colo/patologia , Doença de Crohn/metabolismo , Doença de Crohn/patologia , Modelos Animais de Doenças , Fibrose/metabolismo , Hidroxiprolina/metabolismo , Inflamação/metabolismo , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Interleucina-10/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL
6.
Clin Exp Immunol ; 193(3): 346-360, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29745420

RESUMO

In inflammatory bowel disease (IBD), inflammation is sustained by an exaggerated response of lymphocytes. This results from enhanced expression of anti-apoptotic B cell lymphoma (BCL-2) and BCL-XL associated with a diminished turnover. Azathioprine (AZA) directly targets BCL-2 family-mediated apoptosis. We investigated whether the BCL-2 family expression pattern could be used to predict treatment response to AZA and determined whether BCL-2 inhibitor A-1211212 effectively diminishes lymphocytes and ameliorates inflammation in a model of colitis. BCL-2 family expression pattern was determined by next-generation sequencing (NGS). BCL-2 inhibitor was administered orally to Il10-/- mice. Haematological analyses were performed with an ADVIA 2120 and changes in immune cells were investigated using quantitative polymerase chain reaction (qPCR) and fluorescence activated cell sorter (FACS). We determined similar expression levels of BCL-2 family members in patients with remission and patients refractory to treatment, showing that BCL-2 family expression can not predict AZA treatment response. Expression was not correlated with the modified Truelove and Witts activity index (MTWAI). BCL-2 inhibitor initiated cell death in T cells from patients refractory to AZA and reduced lymphocyte count in Il10-/- mice. FACS revealed diminished CD8+ T cells upon BCL-2 inhibitor in Il10-/- mice without influencing platelets. Tnf, Il1ß, IfnƔ and Mcp-1 were decreased upon BCL-2 inhibitor. A-1211212 positively altered the colonic mucosa and ameliorated inflammation in mice. Pro-apoptotic BCL-2 inhibitor A-1211212 diminishes lymphocytes and ameliorates colitis in Il10-/- mice without inducing thrombocytopenia. BCL-2 inhibition could be a new therapy option for patients refractory to AZA.


Assuntos
Azatioprina/uso terapêutico , Colite/tratamento farmacológico , Doenças Inflamatórias Intestinais/tratamento farmacológico , Linfócitos/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Animais , Apoptose , Células Cultivadas , Colite/diagnóstico , Colite/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Humanos , Mediadores da Inflamação/metabolismo , Doenças Inflamatórias Intestinais/diagnóstico , Doenças Inflamatórias Intestinais/genética , Interleucina-10/genética , Linfócitos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Prognóstico , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Resultado do Tratamento
7.
Biochim Biophys Acta ; 1804(7): 1467-75, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20215052

RESUMO

Under aerobic or anaerobic conditions, tyrosinase undergoes a process of irreversible inactivation induced by its physiological substrate L-dopa. Under aerobic conditions, this inactivation occurs through a process of suicide inactivation involving the form oxy-tyrosinase. Under anaerobic conditions, both the met- and deoxy-tyrosinase forms undergo irreversible inactivation. Suicide inactivation in aerobic conditions is slower than the irreversible inactivation under anaerobic conditions. The enzyme has less affinity for the isomer D-dopa than for L-dopa but the velocity of inactivation is the same. We propose mechanisms to explain these processes.


Assuntos
Di-Hidroxifenilalanina/química , Monofenol Mono-Oxigenase/química , Agaricales/enzimologia , Catálise , Domínio Catalítico , Catecol Oxidase/química , Cinética , Espectroscopia de Ressonância Magnética , Modelos Biológicos , Modelos Químicos , Oxigênio/química , Ligação Proteica , Espectrofotometria/métodos , Fatores de Tempo
8.
Biochim Biophys Acta ; 1794(12): 1766-74, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19695351

RESUMO

There is controversy in the literature concerning the action of tetrahydropterines on the enzyme tyrosinase and on melanogenesis in general. In this study, we demonstrate that tetrahydropterines can inhibit melanogenesis in several ways: i) by non-enzymatic inhibition involving purely chemical reactions reducing o-dopaquinone to L-dopa, ii) by acting as substrates which compete with L-tyr and L-dopa, since they are substrates of tyrosinase; and iii) by irreversibly inhibiting the enzymatic forms met-tyrosinase and deoxy-tyrosinase in anaerobic conditions. Three tetrahydropterines have been kinetically characterised as tyrosinase substrates: 6-R-L-erythro-5,6,7,8-tetrahydrobiopterin, 6-methyl-5,6,7,8-tetrahydropterine and 6,7-(R,S)-dimethyl-5,6,7,8-tetrahydropterine. A kinetic reaction mechanism is proposed to explain the oxidation of these compounds by tyrosinase.


Assuntos
Melaninas/biossíntese , Monofenol Mono-Oxigenase/antagonistas & inibidores , Pterinas/farmacologia , Agaricales/enzimologia , Ligação Competitiva , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Cinética , Levodopa/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Oxirredução , Pterinas/química , Especificidade por Substrato , Tirosina/metabolismo
9.
J Agric Food Chem ; 57(4): 1143-50, 2009 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-19170505

RESUMO

Captopril and mesna are molecules with a free thiol group, used as active ingredients due to their hypotensor and mucolytic properties, respectively. These compounds cross the hematoencephalic barrier and, due to the reactivity of their thiol group, can form adducts with the o-quinones formed during the oxidation of mono- and o-diphenols. Polyphenol oxidase from plants and fungi can be used as a tool for generating o-quinones in their action on o-diphenols and facilitate the formation of adducts in the presence of captopril or mesna. The spectrophotometric characterization of these adducts is useful from several points of view. Here, using the end-point method, which involves the exhaustion of oxygen in the medium, we determined the molar absorptivity of the adducts of different o-diphenols with captopril and mesna. Besides the analytical interest of this approach, we also use it to make a kinetic characterization of polyphenol oxidase as it acts on o-diphenolic substrates that produce unstable o-quinones.


Assuntos
Captopril/química , Mesna/química , Fenóis/química , Catecol Oxidase/metabolismo , Flavonoides/química , Flavonoides/metabolismo , Neurotransmissores/química , Oxirredução , Fenóis/metabolismo , Polifenóis , Quinonas/química , Espectrofotometria , Compostos de Sulfidrila/química
10.
J Agric Food Chem ; 55(3): 920-8, 2007 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-17263494

RESUMO

Chlorogenic acid is the major diphenol of many fruits, where it is oxidized enzymatically by polyphenol oxidase (PPO) or peroxidase (POD) to its o-quinone. In spectrophotometric studies of chlorogenic acid oxidation with a periodate ratio of [CGA]0/[IO4-]0 < 1 and [CGA]0/[IO4-]0 > 1, the o-quinone was characterized as follows: lambda(max) at 400 nm and epsilon = 2000 and 2200 M-1 cm-1 at pH 4.5 and 7.0, respectively. In studies of o-quinone generated by the oxidation of chlorogenic acid using a periodate at ratio of [CGA]0/[IO4-]0 > 1, a reaction with the remaining substrate was detected, showing rate constants of k = 2.73 +/- 0.17 M-1 s-1 and k' = 0.05 +/- 0.01 M-1 s-1 at the above pH values. A chronometric spectrophotometric method is proposed to kinetically characterize the action of the PPO or POD on the basis of measuring the time it takes for a given amount of ascorbic acid to be consumed in the reaction with the o-quinone. The kinetic constants of mushroom PPO and horseradish POD are determined.


Assuntos
Catecol Oxidase/metabolismo , Ácido Clorogênico/metabolismo , Peroxidase/metabolismo , Quinonas/metabolismo , Agaricales/enzimologia , Armoracia/enzimologia , Concentração de Íons de Hidrogênio , Cinética , Oxirredução , Ácido Periódico/metabolismo
11.
J Clin Pathol ; 58(6): 605-10, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15917411

RESUMO

BACKGROUND: Ciliated cells in gastrectomies from patients dwelling in the Pacific and Atlantic basins have been reported previously. AIM: To compare all the results in an attempt to explain the findings. METHODS: Sections from 3406 gastrectomies were reviewed: 1966 and 1440 from the Atlantic and Pacific basins, respectively. Ciliated cells and intestinal metaplasia (IM) were recorded; IM was classified into focal or extensive IM. The total number of sections/gastrectomy was noted. RESULTS: In the Atlantic basin, 5% of specimens had ciliated metaplasia (CM); it was more frequent in intestinal carcinoma (IC; 9%) than diffuse carcinoma (DC; 3%) or miscellaneous gastric diseases (MGD; 3%). In the Pacific basin, the frequency of specimens with CM was 29%: it was more frequent in IC (43%) than in DC (16%) or MGD (10%). The difference between the frequency of CM in specimens with IC or with DC/MGD in the Atlantic and the Pacific basins was significant (p < or = 0.05). The presence of CM was influenced by age and the extent of IM in both basins, but not by sex or the number of sections investigated. CONCLUSIONS: CM-apparently an independent microscopic marker-was significantly higher in the Pacific than in the Atlantic basin. Environmental carcinogens involved in the evolution of IM and IC seem to be implicated in gastric ciliogenesis. Carcinogens that differ in nature and/or in strength in both basins might activate the latent natural genes encoding ciliated processes in gastric cells in patients subsequently developing gastric carcinoma, more notably of intestinal type.


Assuntos
Cílios/patologia , Lesões Pré-Cancerosas/etnologia , Gastropatias/etnologia , Estômago/patologia , Adulto , Fatores Etários , Idoso , América/epidemiologia , Europa (Continente)/epidemiologia , Feminino , Gastrectomia , Mucosa Gástrica/patologia , Humanos , Masculino , Metaplasia/etnologia , Metaplasia/patologia , Pessoa de Meia-Idade , Ilhas do Pacífico/epidemiologia , Lesões Pré-Cancerosas/patologia , Antro Pilórico/patologia , Fatores Sexuais , Gastropatias/patologia , Neoplasias Gástricas/etnologia , Neoplasias Gástricas/patologia
12.
Biochem J ; 380(Pt 3): 643-50, 2004 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-15025557

RESUMO

A solvent deuterium isotope effect on the catalytic affinity (km) and catalytic constant (kcat) of tyrosinase in its action on different monophenols and o-diphenols was observed. The catalytic constant decreased in all substrates as the molar fraction of deuterated water in the medium increased, while the catalytic affinity only decreased for the o-diphenols with an R group in C-1 [-H, -CH3 and -CH(CH3)2]. In a proton inventory study of the oxidation of o-diphenols, the representation of kcat fn/kcat f0 against n (atom fractions of deuterium), where kcat fn is the catalytic constant for a molar fraction of deuterium (n) and kcat f0 is the corresponding kinetic parameter in a water solution, was linear for all substrates, indicating that only one of the four protons transferred from the hydroxy groups of the two molecules of substrate, which are oxidized in one turnover, is responsible for the isotope effects, the proton transferred from the hydroxy group of C-4 to the peroxide of the oxytyrosinase form (Eox). However, in the representation of Km fn/Km f0 against n, where Km fn represents the catalytic affinity for a molar fraction of deuterium (n) and Km f0 is the corresponding kinetic parameter in a water solution, a linear decrease was observed as n increased in the case of o-diphenols with the R group [-H, -CH3 and -CH(CH3)2], and a parabolic increase with other R groups, indicating that more than one proton is responsible for the isotope effects on substrate binding. In the case of monophenols with six protons transferred in the catalytic cycle, the isotope effect occurs in the same way as for o-diphenols. In the present paper, the fractionation factors of different monophenols and o-diphenols are described and possible mechanistic implications are discussed.


Assuntos
Deutério/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Fenóis/metabolismo , Agaricales/enzimologia , Proteínas Fúngicas/metabolismo , Isótopos/metabolismo , Oxirredução
13.
Biochim Biophys Acta ; 1650(1-2): 128-35, 2003 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-12922176

RESUMO

A solvent deuterium isotope effect on the catalytic affinity (K(m)) and rate constant (k(cat)) of tyrosinase in its action on 4-tert-butylcatechol (TBC) was observed. Both parameters decreased as the molar fraction of deuterated water in the medium increased, while the k(cat)/K(m) ratio remained constant. In a proton inventory study, the representation of k(cat)(f(n))/k(cat)(f(0)) and K(m)(f(n))/K(m)(f(0)) vs. n (atom fractions of deuterium) was linear, indicating that, of the four protons transferred from the two molecules of substrate and which are oxidized in one turnover, only one is responsible for the isotope effects. The fractionation factor of 0.64+/-0.02 contributed to identifying the possible proton acceptor. Possible mechanistic implications are discussed.


Assuntos
Catecóis/metabolismo , Deutério/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Agaricales/enzimologia , Cinética , Oxirredução
14.
Biochim Biophys Acta ; 1548(2): 238-56, 2001 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-11513969

RESUMO

Using gas chromatography-mass spectrometry, the direct enzymatic release of o-diphenol (4-tert-butylcatechol) during the action of tyrosinase on a monophenol (4-tert-butylphenol) has been demonstrated for the first time in the literature. The findings confirm the previously proposed mechanism to explain the action of tyrosinase on monophenols (J.N. Rodríguez-López, J. Tudela, R. Varón, F. García-Carmona, F. García-Cánovas, J. Biol. Chem. 267 (1992)). Oxytyrosinase, the oxidized form of the enzyme with a peroxide group, is the only form capable of catalysing the transformation of monophenols into diphenols, giving rise to an enzyme-substrate complex in the process. The o-diphenol formed is then released from the enzyme-substrate complex or oxidized to the corresponding o-quinone. In order to detect the enzymatic release of o-diphenol, the non-enzymatic evolution of the o-quinone to generate o-diphenol by weak nucleophilic attack reactions and subsequent oxidation-reduction was blocked by the nucleophilic attack of an excess of cysteine. Furthermore, the addition of catalytic quantities of an auxiliary o-diphenol (e.g. catechol) considerably increases the accumulation of 4-tert-butylcatechol. The enzyme acting on 4-tert-butylphenol generates the enzyme-4-tert-butylcatechol complex and 4-tert-butylcatechol is then released (with k(-2)) generating mettyrosinase. The auxiliary o-diphenol added (catechol) and the 4-tert-butylcatechol generated by the enzyme then enter into competition. When [catechol] >> [4-tert-butylcatechol], the enzyme preferentially binds with the catechol to close the catalytic cycle, while 4-tert-butylcatechol is accumulated in the medium. In conclusion, we demonstrate that the enzyme produces 4-tert-butylcatechol from 4-tert-butylphenol, the concentration of which increases considerably in the presence of an auxiliary o-diphenol such as catechol.


Assuntos
Catecóis/química , Monofenol Mono-Oxigenase/química , Fenóis/química , Agaricales , Benzoquinonas/química , Catecóis/farmacologia , Cisteína/química , Cisteína/farmacologia , Cromatografia Gasosa-Espectrometria de Massas/métodos , Cinética , Melanossomas/metabolismo , Modelos Químicos , Modelos Teóricos , Monofenol Mono-Oxigenase/metabolismo , Oxirredução
15.
J Cancer Educ ; 16(2): 75-9, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11440066

RESUMO

BACKGROUND: In spite of an early cancer detection program (CCSP), Mexico has a mortality rate for cervical cancer of 16.5 per 100,000 women. METHOD: A cross-sectional study of 330 physicians at the Mexico City General Hospital evaluated their knowledge of the CCSP, etiology, diagnostic alternatives, and treatment guidelines. Variance analysis was the statistical procedure used. Replies to a questionnaire about cervical cancer prevention awareness were scored on a scale from 1 to 9. RESULTS: According to the awareness scale, the global average classification was 4.4, with 50% of the physicians scoring 4 or less. There was no difference in the CCSP knowledge scores of gynecologists (mean 4.92, 95% CI 4.2-5.3), oncologists (mean 4.85, 95% CI 4.3-5.5), pathologists (mean 5.23, 95% CI 4.9-5.6), and those in other specialties (mean 4.29, 95% CI 4.2-5.0), p > 0.05. Many respondents attributed CCSP's lack of effectiveness to public apathy (68.12%). CONCLUSIONS: The effectiveness of the CCSP can be improved by educating health professionals if this education is combined with elimination of obstacles to its use. More information is needed to justify revising how doctors are educated in terms of not only quality of the training but also the contents of pre- and postgraduate training programs.


Assuntos
Competência Clínica , Medicina , Especialização , Neoplasias do Colo do Útero/prevenção & controle , Análise de Variância , Estudos Transversais , Feminino , Humanos , Entrevistas como Assunto , México , Neoplasias do Colo do Útero/diagnóstico
16.
Biochim Biophys Acta ; 1548(1): 1-22, 2001 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-11451433

RESUMO

Tyrosinase can act on monophenols because of the mixture of met- (E(m)) and oxy-tyrosinase (E(ox)) which exists in the native form of the enzyme. The latter form is active on monophenols, while the former is not. However, the kinetics are complicated because monophenols can bind to both enzyme forms. This situation becomes even more complex since the products of the enzymatic reaction, the o-quinones, are unstable and continue evolving to generate o-diphenols in the medium. In the case of substrates such as L-tyrosine, tyrosinase generates very unstable o-quinones, in which a process of cyclation and subsequent oxidation-reduction generates o-diphenol through non-enzymatic reactions. However, the release of o-diphenol through the action of the enzyme on the monophenol contributes to the concentration of o-diphenol in the first pseudo-steady-state [D(0)](ss). Hence, the system reaches an initial pseudo-steady state when t-->0 and undergoes a transition phase (lag period) until a final steady state is reached when the concentration of o-diphenol in the medium reaches the concentration of the final steady state [D(f)](ss). These results can be explained by taking into account the kinetic and structural mechanism of the enzyme. In this, tyrosinase hydroxylates the monophenols to o-diphenols, generating an intermediate, E(m)D, which may oxidise the o-diphenol or release it directly to the medium. We surmise that the intermediate generated during the action of E(ox) on monophenols, E(m)D, has axial and equatorial bonds between the o-diphenol and copper atoms of the active site. Since the orbitals are not coplanar, the concerted oxidation-reduction reaction cannot occur. Instead, a bond, probably that of C-4, is broken to achieve coplanarity, producing a more labile intermediate that will then release the o-diphenol to the medium or reunite it diaxially, involving oxidation to o-quinone. The non-enzymatic evolution of the o-quinone would generate the o-diphenol ([D(f)](ss)) necessary for the final steady state to be reached after the lag period.


Assuntos
Agaricales/enzimologia , Monofenol Mono-Oxigenase/química , Fenóis/química , Quinonas/química , Catecol Oxidase/química , Relação Dose-Resposta a Droga , Cinética , Levodopa/química , Modelos Químicos , Modelos Teóricos , Monofenol Mono-Oxigenase/farmacologia , Espectrofotometria , Tirosina/química
17.
Eur J Biochem ; 267(19): 5865-78, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-10998046

RESUMO

Tyrosinase can act on monophenols because of the mixture of mettyrosinase (Em) and oxytyrosinase (Eox) that exists in the native form of the enzyme. The latter form is active on monophenols although the former is not. However, the kinetics are complicated because monophenols can bind to both enzyme forms. This situation becomes even more complex as the products of the enzymatic reaction, the o-quinones, are unstable and continue evolving to generate o-diphenols in the medium. In the case of substrates such as 4-methoxyphenol, 4-ethoxyphenol and 4-tert-butylphenol, tyrosinase generates o-quinones which become unstable with small constants of approximately < 10-3 s-1. The system evolves from an initial steady state, reached when t-->0, through a transition state towards a final steady state, which is never reached because the substrate is largely consumed. The mechanisms proposed to explain the enzyme's action can be differentiated by the kinetics of the first steady state. The results suggest that tyrosinase hydroxylates monophenols to o-diphenols, generating an intermediate Em-diphenol in the process, which may oxidize the o-diphenol or release it directly into the medium. In the case of o-quinone formation, its slow instability generates o-diphenol which activates the enzymatic system yielding parabolic time recordings.


Assuntos
Monofenol Mono-Oxigenase/metabolismo , Fenóis/metabolismo , Proteínas de Plantas/metabolismo , Isoformas de Proteínas/metabolismo , Quinonas/metabolismo , Agaricales/enzimologia , Simulação por Computador , Cromatografia Gasosa-Espectrometria de Massas , Cinética , Espectroscopia de Ressonância Magnética , Modelos Químicos , Oxirredução
18.
Biochemistry ; 39(34): 10497-506, 2000 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-10956040

RESUMO

The reaction of mushroom (Agaricus bisporus) tyrosinase with dioxygen in the presence of several o-diphenolic substrates has been studied by steady-state and transient-phase kinetics in order to elucidate the rate-limiting step and to provide new insights into the mechanism of oxidation of these substrates. A kinetic analysis has allowed for the first time the determination of individual rate constants for several of the partial reactions that comprise the catalytic cycle. Mushroom tyrosinase rapidly reacts with dioxygen with a second-order rate constant k(+8) = 2.3 x 10(7) M(-)(1) s(-)(1), which is similar to that reported for hemocyanins [(1.3 x 10(6))-(5.7 x 10(7)) M(-)(1) s(-)(1)]. Deoxytyrosinase binds dioxygen reversibly at the binuclear Cu(I) site with a dissociation constant K(D)(O)()2 = 46.6 microM, which is similar to the value (K(D)(O)()2 = 90 microM) reported for the binding of dioxygen to Octopus vulgaris deoxyhemocyanin [Salvato et al. (1998) Biochemistry 37, 14065-14077]. Transient and steady-state kinetics showed that o-diphenols such as 4-tert-butylcatechol react significantly faster with mettyrosinase (k(+2) = 9.02 x 10(6) M(-)(1) s(-)(1)) than with oxytyrosinase (k(+6) = 5.4 x 10(5) M(-)(1) s(-)(1)). This difference is interpreted in terms of differential steric and polar effects that modulate the access of o-diphenols to the active site for these two forms of the enzyme. The values of k(cat) for several o-diphenols are also consistent with steric and polar factors controlling the mobility, orientation, and thence the reactivity of substrates at the active site of tyrosinase.


Assuntos
Agaricus/enzimologia , Monofenol Mono-Oxigenase/metabolismo , Animais , Hemocianinas/análogos & derivados , Hemocianinas/metabolismo , Técnicas In Vitro , Cinética , Modelos Químicos , Oxirredução , Oxigênio/metabolismo , Fenóis/química , Fenóis/metabolismo , Especificidade por Substrato
19.
Biol Chem ; 381(4): 313-20, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10839460

RESUMO

The relationship between the structure and activity of meta- and para-hydroxylated monophenols was studied during their tyrosinase-catalysed hydroxylation and the rate-limiting steps of the reaction mechanism were identified. The para-hydroxylated substrates permit us to study the effect of a substituent (R) in the carbon-1 position (C-1) of the benzene ring on the nucleophilic attack step, while the meta group permits a similar study of the effect on the electrophilic attack step. Substrates with a -OCH3 group on C-1, as p-hydroxyanisol (4HA) and m-hydroxyanisol (3HA), or with a -CH2OH group, as p-hydroxybenzylalcohol (4HBA) and m-hydroxybenzylalcohol (3HBA), were used because the effect of the substituent (R) size was assumed to be similar. However, the electron-donating effect of the -OCH3 group means that the carbon-4 position (C-4) is favoured for nucleophilic attack (para-hydroxylated substrates) or for electrophilic attack (meta-hydroxylated substrates). The electron-attracting effect of the -CH2OH group has the opposite effect, hindering nucleophilic (para) or electrophilic (meta) attack of C-4. The experimental data point to differences between the maximum steady-state rate (V(M)Max) of the different substrates, the value of this parameter depends on the nucleophilic and electrophilic attack. However, differences are greatest in the Michaelis constants (K(M)m), with the meta-hydroxylated substrates having very large values. The catalytic efficiency k(M)cat/K(M)m is much greater for thepara-hydroxylated substrates although it varies greatly between one substrate and the other. However, it varies much less in the meta-hydroxylated substrates since this parameter describes the power of the nucleophilic attack, which is weaker in the meta OH. The large increase in the K(M)m of the meta-hydroxylated substrates might suggest that the phenolic OH takes part in substrate binding. Since this is a weaker nucleophil than the para-hydroxylated substrates, the binding constant decreases, leading to an increase in K(M)m. The catalytic efficiency of tyrosinase on a monophenol (para or meta) is directly related to the nucleophilic power of the oxygen of the phenolic OH. The oxidation step is not limiting since if this were the case, the para and meta substrates would have the same V(M)max. The small difference between the absolute values of V(M)max suggests that the rate constants of the nucleophilic and electrophilic attacks are on the same order of magnitude.


Assuntos
Monofenol Mono-Oxigenase/metabolismo , Fenóis/metabolismo , Agaricales/enzimologia , Anisóis/química , Anisóis/metabolismo , Álcoois Benzílicos/química , Álcoois Benzílicos/metabolismo , Elétrons , Concentração de Íons de Hidrogênio , Hidroxilação , Cinética , Monofenol Mono-Oxigenase/química , Monofenol Mono-Oxigenase/farmacologia , Oxirredução , Fenóis/química , Espectrofotometria
20.
Biophys Chem ; 84(1): 65-76, 2000 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-10723545

RESUMO

Tyrosinase hydroxylates 3-hydroxyanisole in the 4-position. The reaction product accumulates in the reaction medium with a lag time (tau) which diminishes with increasing concentrations of enzyme and lengthens with increasing concentrations of substrate, thus fulfilling all the predictions of the mechanism proposed by us for 4-hydroxyphenols. The kinetic constants obtained, kcatM = (46.87 +/- 2.06) s-1 and KmM = (5.40 +/- 0.60) mM, are different from those obtained with 4-hydroxyanisole, kcatM = (184.20 +/- 6.1) s-1 and KmM = (0.08 +/- 0.004) mM. The catalytic efficiency, kcatM/KmM is, therefore, 265.3 times greater with 4-hydroxyanisole. The possible rate-determining steps for the reaction mechanism of tyrosinase on 3- and 4-hydroxyanisole, based on the NMR spectra of both monophenols, are discussed. These possible rate-determining steps are the nucleophilic attack of hydroxyl's oxygen on the copper and the electrophilic attack of the peroxide on the aromatic ring. Both steps may be of similar magnitude, i.e. take place in the same time scale.


Assuntos
Anisóis/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Agaricales/enzimologia , Benzotiazóis , Catálise , Hidrazonas , Hidroxilação , Isomerismo , Cinética , Ressonância Magnética Nuclear Biomolecular , Oxirredução , Tiazóis
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