NADPH oxidasa: rol en aterosclerosis y función plaquetaria / NADPH OXIDASE: role in atherosclerosis and platelet function

Reactive oxygen species have emerged as important molecules in cardiovascular function. Recent research has shown that the NADPH oxidases are important sources of superoxide in vascular cells and myocytes. The NADPH oxidases vascular share some, but not all, of the characteristics of the enzyme in neutrophils, both produce superoxide, which is metabolized to hydrogen peroxide, at the same time these reactive oxygen species serve as second messengers activate multiple intracellular signalling pathways. NADPH oxidases are essential in the physiological response of vascular cellsto pathological states such as atherosclerosis, and are functionally relevant in activation and recruitment of platelets. Recent studies suggest a key role for NADPH oxidase in the formation of a specific product from the oxidation of arachidonic acid, and a potential role in the process of recruitment of platelets. Taking into account these characteristics and evidence of the involvement of the NADPH oxidases in cardiovascular diseases as the thrombosis, inhibition of this enzymatic system appears as a promising therapy to treat and prevent these diseases.

Platelet enzyme abnormalities in leukemias.

Aim of the Study: The aim of this study was to evaluate platelet enzyme activity in cases of leukemia. Materials and Methods: Platelet enzymes glucose-6-phosphate dehydrogenase (G6PD), pyruvate kinase (PK) and hexokinase (HK) were studied in 47 patients of acute and chronic leukemia patients, 16 patients with acute myeloid leukemia (AML)(13 relapse, three in remission), 12 patients with acute lymphocytic leukemia (ALL) (five in relapse, seven in remission), 19 patients with chronic myeloid leukemia (CML). Results: The platelet G6PD activity was significantly low in cases of AML, ALL and also in CML. G6PD activity was normalized during AML remission. G6PD activity, although persistently low during ALL remission, increased significantly to near-normal during remission (P < 0.05) as compared with relapse (P < 0.01). Platelet PK activity was high during AML relapse (P < 0.05), which was normalized during remission. Platelet HK however was found to be decreased during all remission (P < 0.05). There was a significant positive correlation between G6PD and PK in cases of AML (P < 0.001) but not in ALL and CML. G6PD activity did not correlate with HK activity in any of the leukemic groups. A significant positive correlation was however seen between PK and HK activity in cases of ALL remission (P < 0.01) and CML (P < 0.05). Conclusions: Both red cell and platelet enzymes were studied in 36 leukemic patients and there was no statistically significant correlation between red cell and platelet enzymes. Platelet enzyme defect in leukemias suggests the inherent abnormality in megakaryopoiesis and would explain the functional platelet defects in leukemias.

Efeitos do tratamento periodontal não-cirúrgico na via L-arginina-óxido nítrico e no estresse oxidativo em plaquetas / Effects of nonsurgical periodontal treatment on L-arginine-nitric oxide pathway and oxidant status in platelets

Estudos publicados nas duas últimas décadas sugerem um aumento do risco de doença cardiovascular (DCV) em pacientes com periodontite, mas os mecanismos fisiopatológicos dessa associação ainda não estão completamente esclarecidos. Uma vez que foi demonstrado aumento da ativação plaquetária e do estresse oxidativo na periodontite, o objetivo do presente estudo foi investigar a via L-arginina-óxido nítrico (NO)- guanosina monofosfato cíclica (GMPc) e parâmetros de estresse oxidativo em plaquetas de pacientes com periodontite, bem como avaliar o efeito do tratamento periodontal não-cirúrgico nessas variáveis. Um total de 10 pacientes sem periodontite (periodontalmente saudáveis ou com gengivite) e 10 pacientes com periodontite participaram do estudo. A avaliação clínica, laboratorial e experimental foi realizada no início do estudo e 90 dias após realização da terapia periodontal básica (grupo periodontite). A avaliação clínica periodontal incluiu registros de: profundidade de bolsa à sondagem (PBS), nível de inserção (NIC), percentual de placa e percentual de sangramento à sondagem. Os seguintes experimentos foram realizados: influxo de L-arginina; atividade e expressão das enzimas óxido nítrico sintase e da arginase; expressão das enzimas guanilato ciclase solúvel e fosfodiesterase 5; determinação dos níveis intraplaquetários de GMPc; agregação plaquetária; avaliação do estresse oxidativo (atividade oxidante total, atividade das enzimas antioxidantes catalase e da superóxido dismutase - SOD); medição dos níveis de proteína C reativa (CRP) e de fibrinogênio. Os resultados obtidos no início do estudo demonstraram ativação do influxo de L-arginina em plaquetas via sistema y+L nos pacientes com periodontite, bem como concentrações intraplaquetárias de GMPc diminuídas e aumento sistêmico da CRP. Após o tratamento periodontal, observou-se redução do percentual de sítios com PBS ≥ 6 mm, NIC 4-5 mm e NIC ≥ 6 mm, aumento nos níveis de GMPc, para níveis ...

Studies published over the last two decades have suggested an increase of cardiovascular disease (CVD) risk on periodontitis patients, but the physiopathological mechanisms involved in this association are not yet clear. Since it has been demonstrated an enhancement on both platelet activation and oxidative stress on periodontitis patients, the aim of this study was to investigate the L-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway on platelets from periodontitis patients, and the effect of non-surgical periodontal treatment in these variables. A total of 10 patients without periodontitis (periodontal healthy controls or gingivitis patients) and 10 periodontitis patients were included in this study. The clinical, laboratorial, and experimental evaluations were performed at the beginning of the study and 90 days after the basic periodontal therapy (periodontitis group). The clinical periodontal evaluation included the measurements of probing pocket depth (PPD), clinical attachment level (CAL), plaque percentage, and percentage of bleeding on probing. The following experiments were performed: L-arginine influx; nitric oxide synthase and arginase enzymes activity and expression; expression of guanylate cyclase and phosphodiesterase-5 enzymes; measurement of intraplatelet cGMP levels; platelet aggregation; oxidative stress evaluation (total oxidant activity and activity of both antioxidant enzymes catalase and superoxide dismutase – SOD); measurement of C reactive protein (CRP) and fibrinogen. The initial results demonstrated an activation of L-arginine influx in platelets from periodontitis patients via y+L system, reduced intraplatelet cGMP levels and increased CRP. After periodontal treatment, it was observed reduction on percentage of sites with PPD ≥ 6 mm, CAL 4-5 mm and CAL ≥ 6 mm, enhancement on cGMP levels, to levels comparables to patients without periodontitis, accompanied by a higher activity of both antioxidant ...

Efeitos do tratamento periodontal não-cirúrgico na via L-arginina-óxido nítrico e no estresse oxidativo em plaquetas / Effects of nonsurgical periodontal treatment on L-arginine-nitric oxide pathway and oxidant status in platelets

Estudos publicados nas duas últimas décadas sugerem um aumento do risco de doença cardiovascular (DCV) em pacientes com periodontite, mas os mecanismos fisiopatológicos dessa associação ainda não estão completamente esclarecidos. Uma vez que foi demonstrado aumento da ativação plaquetária e do estresse oxidativo na periodontite, o objetivo do presente estudo foi investigar a via L-arginina-óxido nítrico (NO)- guanosina monofosfato cíclica (GMPc) e parâmetros de estresse oxidativo em plaquetas de pacientes com periodontite, bem como avaliar o efeito do tratamento periodontal não-cirúrgico nessas variáveis. Um total de 10 pacientes sem periodontite (periodontalmente saudáveis ou com gengivite) e 10 pacientes com periodontite participaram do estudo. A avaliação clínica, laboratorial e experimental foi realizada no início do estudo e 90 dias após realização da terapia periodontal básica (grupo periodontite). A avaliação clínica periodontal incluiu registros de: profundidade de bolsa à sondagem (PBS), nível de inserção (NIC), percentual de placa e percentual de sangramento à sondagem. Os seguintes experimentos foram realizados: influxo de L-arginina; atividade e expressão das enzimas óxido nítrico sintase e da arginase; expressão das enzimas guanilato ciclase solúvel e fosfodiesterase 5; determinação dos níveis intraplaquetários de GMPc; agregação plaquetária; avaliação do estresse oxidativo (atividade oxidante total, atividade das enzimas antioxidantes catalase e da superóxido dismutase - SOD); medição dos níveis de proteína C reativa (CRP) e de fibrinogênio. Os resultados obtidos no início do estudo demonstraram ativação do influxo de L-arginina em plaquetas via sistema y+L nos pacientes com periodontite, bem como concentrações intraplaquetárias de GMPc diminuídas e aumento sistêmico da CRP. Após o tratamento periodontal, observou-se redução do percentual de sítios com PBS ≥ 6 mm, NIC 4-5 mm e NIC ≥ 6 mm, aumento nos níveis de GMPc, para níveis ...

Studies published over the last two decades have suggested an increase of cardiovascular disease (CVD) risk on periodontitis patients, but the physiopathological mechanisms involved in this association are not yet clear. Since it has been demonstrated an enhancement on both platelet activation and oxidative stress on periodontitis patients, the aim of this study was to investigate the L-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway on platelets from periodontitis patients, and the effect of non-surgical periodontal treatment in these variables. A total of 10 patients without periodontitis (periodontal healthy controls or gingivitis patients) and 10 periodontitis patients were included in this study. The clinical, laboratorial, and experimental evaluations were performed at the beginning of the study and 90 days after the basic periodontal therapy (periodontitis group). The clinical periodontal evaluation included the measurements of probing pocket depth (PPD), clinical attachment level (CAL), plaque percentage, and percentage of bleeding on probing. The following experiments were performed: L-arginine influx; nitric oxide synthase and arginase enzymes activity and expression; expression of guanylate cyclase and phosphodiesterase-5 enzymes; measurement of intraplatelet cGMP levels; platelet aggregation; oxidative stress evaluation (total oxidant activity and activity of both antioxidant enzymes catalase and superoxide dismutase – SOD); measurement of C reactive protein (CRP) and fibrinogen. The initial results demonstrated an activation of L-arginine influx in platelets from periodontitis patients via y+L system, reduced intraplatelet cGMP levels and increased CRP. After periodontal treatment, it was observed reduction on percentage of sites with PPD ≥ 6 mm, CAL 4-5 mm and CAL ≥ 6 mm, enhancement on cGMP levels, to levels comparables to patients without periodontitis, accompanied by a higher activity of both antioxidant ...(AU)

Regulating inflammation through the anti-inflammatory enzyme platelet-activating factor-acetylhydrolase

Platelet-activating factor (PAF) is one of the most potent lipid mediators involved in inflammatory events. The acetyl group at the sn-2 position of its glycerol backbone is essential for its biological activity. Deacetylation induces the formation of the inactive metabolite lyso-PAF. This deacetylation reaction is catalyzed by PAF-acetylhydrolase (PAF-AH), a calcium independent phospholipase A2 that also degrades a family of PAF-like oxidized phospholipids with short sn-2 residues. Biochemical and enzymological evaluations revealed that at least three types of PAF-AH exist in mammals, namely the intracellular types I and II and a plasma type. Many observations indicate that plasma PAF AH terminates signals by PAF and oxidized PAF-like lipids and thereby regulates inflammatory responses. In this review, we will focus on the potential of PAF-AH as a modulator of diseases of dysregulated inflammation.

Comparative study of platelet histamine and serotonin with their corresponding plasma oxidases in asthmatics with normals.

OBJECTIVE: Asthma is well controllable but non-curable disease. Exact pathophysiology involved is unresolved till today. Role of allergic hypersensitivity reaction in asthmatic on-set is well established. Present work is an effort to elucidate some basic points of unresolved pathophysiology of asthma taking platelets as marker. MATERIAL AND METHODS: A group of 52 normal human subjects in the age group of 20-60 years were studied for platelet histamine and serotonin levels and also for their plasma metabolising enzymes diamine oxidase (DAO) and monoamine oxidase (MAO). The data was collected for 79 asthmatic patients at different stages of asthma and accordingly were studied as four different groups of seventy nine asthmatics those were on regular treatment and were comfortable with drugs and were free from symptomatic attack formed gr. I; these (79) patients were followed-up during their symptomatic phase (gr. II) and same (79) patients immediately after their recovery from symptomatic stage studied as gr. III members. All the 79 asthmatic patients fall in gr. I, II and III in a serial manner i.e. all (n = 79) in each group. A separate group of thirty seven patients with known history of asthma but were symptom free and also off drugs for last 2-4 years formed gr. IV. RESULTS: Results showed mean platelet count in asthmatics at all four stages were in the normal range but were slightly low in comparison with normals. Both the enzymatic levels (DAO and MAO) in gr. I, II and III were significantly higher than normals but were same in the case of gr. IV patients. Low levels of platelet biogenic amines were observed in asthmatics (gr. I to gr. IV) than normals. CONCLUSIONS: Thus, study parameters showed significant difference in asthmatics and normals. Findings of the study have been utilized to understand unanswered hypersensitivity shown by the asthmatics over normal individuals (non-asthmatics).

The ecto-ATPases of endoparasites and of blood cells and vessels

The cells of blood vessel walls and the external surface of all blood cells have an ecto-ATPase which hydrolyzes ATP to ADP and also ADP to AMP. This enzyme has also been called apyrase or ATP-diphosphohydrolase. The enzyme hydrolyzes a broad range of tri-and diphosphate nucleosides such as UTP and UDP, GTP and GDP in additon to the adenine nucleotides and because of that it has also been called a nucleoside triphosphate hydrolase. The possible physiological roles for this ecto-ATPase involve the control of vascular tone by modulation of the levels of ATP and ADP binding to purino-receptors of the vasculature, the modulation of thrombogenesis by controlling the extracellular level of ADP which is known to activate platelet aggregation, and the protection from cytolytic effects of extracellular ATP. An ATP-diphosphohydrolase activity has been characterized on the external surface of Schistosoma mansoni, a parasite that lives in the circulation of the human host, and on the outer surface of Entamoeba histolytica, a parasite that may enter the circulation of the host through ulceration in the intestinal mucosa. The endoparasite Toxoplasma gondii also exhibits a nucleoside triphosphate hydrolase of high activity, although in this case the ecto-localization is still not documented. We raise the possibility that the endoparasites have evolved in a way to possibly mimic some of the conditions on the surface of cells normally present in the host circulation, thus escaping hemostatic defense responses of the host which require extracellular ADP or ATP.

El aumento del calcio ionizado intracelular depende de algún factor presente en el suero de las pacientes preeclámpticas / The intracellular increase of ionized calcium depend on some factor present in pre-eclamptic patient sera

En trabajos previos nuestro grupo ha demostrado que la concentración intracelular de calcio ionizado es mayor en pacientes con preeclampsia que en embarazadas normotensas y que estas cifras correlacionan con la tensión arterial. Ambos indicadores se normalizan 6 semanas después del parto. Con el fin de investigar la posibile participación de factores solubles en el plasma, en el aumento del calcio libre intracelular, se estudiaron 8 pacientes preeclámpticas diagnosticadad por los criterios del Colegio Americano de Ginecólogos y Obstetras. Como grupo testigo se incluyeron 8 embarazadas normotensas pareadas por edad cronológica y gestacional. Para medir flujos de calcio transmembrana se utilizaron plaquetas de varones sanos. Las plaquetas fueron incubadas durante 0, 15, 30 y 60 minutos en suero de pacientes preeclámpticas o de embarazadas normotensas marcado con Ca. Las diferencias en el transporte de calcio se evaluaron con análisis de varianza de Kruskal Wallis. El transporte de calcio fue mayor cuando las plaquetas se incubaron en suero de pacientes preeclámpticas, Md = 1.475 ñ 0.311 nanomolas que cuando fueron incubadas en suero de embarazadas normotensas, Md = 0.9725 ñ 0.58 nanomolas, p < 0.02. Este hallazgo sugiere que en el suero de las pacientes preeclámpticas existe algún factor que facilicita la entrada de calcio a la célula, que provoca un aumento en la concentración de calcio libre y participa en la hipertensión gestacional.

Cambios de algunos parámetros bioquímicos en linfocitos y plaquetas de pacientes con demencia de tipo Alzheimer / Changes of some biochemical parameters in lymphocytes and platelets of patients with Alzheimer type dementia

La demencia tipo Alzheimer (DTA) es una de las más frecuentes entre los adultos mayores de 65 años de edad. En el sistema nervioso central de los pacientes con enfermedad de Alzheimer se observa tempranamente una marcada alteración del sistema colinérgico, lo que se correlaciona con una disminución de la actividad enzimática de la acetilcolinesterasa (AChE) y un aumento de la butirilcolinesterasa (BuChE). Es probable que alguno de estos cambios bioquímicos pueda tener su reflejo también a nivel periférico, considerando que tanto la AChE como la BuChE se expresan en células no-neuronales, incluyendo a células sanguíneas. En el presente trabajo se evaluaron las actividades de la AchE y de la BuChE en linfocitos y plaquetas tanto en individuos normales como en pacientes con DTA, encontrándose que la actividad de la AChE está disminuida (60 por ciento ) en linfocitos obtenidos de pacientes con DTA, sin observarse cambios aparentes en la actividad de la BuChE. En plaquetas no se observaron diferencias en las actividades de la AChE y de la BuChE entre individuos con DTA y controles. Se evaluó también la captación de 14 C-serotonina por las plaquetas. No se observaron diferencias en la velocidad máxima de captación de 14 C-serotonina; sin embargo, la Km para la captación fue menor para los pacientes con DTA que para los controles. Finalmente el recuento de plaquetas y leucocitos evidenció un aumento en el número total de células en los pacientes con DTA

Effect of retinoic acid on adenosine diphosphate and collagen-induced alterations in enzymes of GSH-linked antioxidant defence system of human blood platelets in vitro.

Collagen stimulation of blood platelets resulted in significant increases in malondialdehyde (MDA) formation and activity of glucose-6-phosphate dehydrogenase (G6PDH) and a decrease in catalase and glutathione peroxidase (GPx). Retinoic acid (RA) pretreatment did not show any appreciable changes except for a decrease in G6PDH activity as compared with collagen alone. RA pretreatment of human blood platelets resulted in an increase in the activities of catalase and GPx, two important radical scavenging enzymes, with significant decrease in MDA formation when compared with ADP alone. It is suggested that RA has a significant effect on the antioxidant defence system in ADP stimulated platelets but not in the collagen stimulated platelets.

Actividad de la monoamino-oxidasa plaquetaria en pacientes con depresión mayor con y sin melancolia: resultados preliminares / Activity of platelet monoamino-oxidase in major depressive illness: melancholic and non-melancholic patients: preliminary data

Se analizaron muestras sanguíneas de 99 pacientes libres de tratamiento (80 mujeres y 19 hombres) con diagnóstico de depresión mayor (DSM-III), 41 con melancolía y 58 sin melancolía. Se encontró que la actividad de la MAO plaquetaria fue significativamente mayor en las mujere que en los hombres, y en los pacientes con depresión mayor sin melancolía, que en aquellos con melancolía. No se encontraron correlaciones significativas de los valores de actividad de la enzima con la edad , ni con la severidad de la sintomatología depresiva medida por la EDH. Estos hallazgos coinciden con los de investigaciones previas al confirmar el hecho de que la mujer tiende a presentar una mayor actividad de la MAO plaquetária que el hombre, y apoyan la hipótesis de la presencia de una actividad enzimática elevada en las depresiones no endógenas. Sin embargo, para ubicar adecuadamente este posible marcador, deverán desarrolarse estudios cuidadosos, con un enfoque fenomenológico y controles adecuados, de las variables orgánicas y metodológicas que sabemos afectan la actividad de la MAO

Platelet monoamineoxidase activity in chronic schizophrenics

Se dosó la actividad de monoamino oxidasa (MAO) plaquetaria en 17 pacientes esquizofrénicos crónicos y en 13 sujetos-control, utilizándose kinuramina como substrato. La actividad de MAO fue aproximadamente 10 veces mayor en los sujetos del grupo control que en los esquizofrénicos (controles: 31,7 + ou - 4,5 nmoles.mg proteína-1.h-1, n=13; esquizofrénicos: 3,3 + ou - 0,6 nmoles.mg proteína-1.h-1, n=17). Los valores de Vmáx y Km para kinuramina fueron significativamente más bajos en os pacientes que en los controles. La actividad de MAO fue aun menor en el subgrupo de pacientes cuya forma clínica previa actual estado residual fue paranoide. Estos resultados permitirían utilizar la determinación de MAO plaquetaria como una variable en la investigación, sobre esquizofrenía