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R Sáez

Publications and source records attributed to R Sáez.

16 recordsLinked to original sources
PubMed2001

NMDA-induced phosphorylation of the microtubule-associated protein MAP-2 is mediated by activation of nitric oxide synthase and MAP kinase.

Microtubule-associated protein MAP-2 is a neuronal phosphoprotein which modulates microtubule stability and spatial organization of signal transduction pathways. The functions of MAP-2 are modulated by phosphorylation. We studied the modulation of MAP-2 phosphorylation using the N-methyl- D-aspartate (NMDA) type of glutamate receptors and the signal transduction pathways mediating this modulation in primary cultures of rat cerebellar neurons. NMDA induced a rapid increase (330% of basal at 5 min) in MAP-2 phosphorylation which was not prevented by KN-62, indicating that it is not mediated by activation of Ca-calmodulin-dependent protein kinase. NMDA-induced phosphorylation of MAP-2 was inhibited by the nitric oxide synthase inhibitors nitroarginine and 7-nitroindazole and by PD098059 (an inhibitor of MAP kinase kinase), but was only slightly reduced by calphostin C or U-73122, inhibitors of protein kinase C and of phospholipase C, respectively. This indicates that the main pathway mediating NMDA-induced phosphorylation of MAP-2 is activation of nitric oxide synthase and subsequent activation of MAP kinase. We show that activation of NMDA receptors induces an activation of MAP kinase which is prevented by nitroarginine. The nitric oxide-generating agent (+/-)-S-nitroso-N-acetylpenicillamine (SNAP) also induced activation of MAP kinase and increased phosphorylation of MAP-2. Other nitric oxide-generating agents (NOC-18 and NOR-3) also increased MAP-2 phosphorylation. The interplay between NMDA receptors-associated signal transduction pathways and MAP-2 may be involved in the modulation of neuronal responses to extracellular signals and in the regulation of neuronal function.

Animals↗
PubMed2001

Smoking-related attitudes, characteristics, and opinions in a group of young men with asthma.

UNLABELLED: The objective of this study was to examine the attitudes, characteristics, and opinions about smoking of a group of young asthmatic men. POPULATION AND METHODS: An anonymous, personal questionnaire was administered to 611 young male volunteers who had been diagnosed with asthma (according to the National Heart, Lung, and Blood Institute/World Health Organization Global Initiative for Asthma, 1995) in the respiratory disease and allergy clinics of the Burgos Military Hospital (Spain). This questionnaire contained items related to personal information, asthma characteristics, opinions about smoking, and information related to smoking habits. RESULTS: Six hundred patients with asthma completed the questionnaire. All were men, mean age 20.16 +/- 3.03 years; 189 (31.5%) were smokers and 16 (2.5%) were ex-smokers. Mean age at onset of regular smoking was 16.46 +/- 2 years. Sixty-five percent (65.07%) smoked fewer than 10 cigarettes per day. Most of the smoking asthmatics had mild asthma (58.9%). Eighty-eight percent (88.3%) had moderate dependence. Many of the smoking asthmatics were contemplating stopping smoking (54%), and 59% had tried before to stop. Concern about health was the main reason given for stopping smoking. Asthmatics who smoked had a higher percentage of smokers among family members, friends, and colleagues than nonsmoking asthmatics. Attitudes toward smoking were more permissive among smoking asthmatics. Only 36.64% of the total had received information about tobacco previously. In the sample group, 7% claimed that they did not smoke but their carbon monoxide concentration in exhaled air was 10 ppm or higher. CONCLUSIONS: There were no differences in the onset of the smoking habit between asthmatic and nonasthmatic young people. A large percentage of the smoking asthmatics were considering smoking cessation, motivated mainly by their asthma condition. The group as a whole had little previous information about tobacco.

Adult↗
PubMed1999

Prevention of glutamate neurotoxicity in cultured neurons by 3,4-dihydro-6-hydroxy-7-methoxy-2,2-dimethyl-1(2H)-benzopyran (CR-6), a scavenger of nitric oxide.

Glutamate neurotoxicity in cerebellar neurons in culture is mediated by excessive production of nitric oxide (NO). We anticipated that 3,4-dihydro-6-hydroxy-7-methoxy-2,2-dimethyl-1(2H)-benzopyran (CR-6) could act as a scavenger of NO since it contains a position (C-5) highly activated towards nitration reaction. The aim of this work was to assess whether CR-6 acts as an NO scavenger and prevents glutamate neurotoxicity in cultures of cerebellar neurons. It was shown that CR-6 reduced, in a dose-dependent manner, glutamate-induced formation of cGMP (EC50 approximately 15 microM) and prevented glutamate neurotoxicity. The protection was approximately 50% at 3-10 microM and nearly complete at 100 microM. CR-6 did not prevent glutamate-induced activation of NO synthase, but interfered with the glutamate-NO-cGMP pathway at a later step. CR-6 reduced the formation of cGMP induced by S-nitroso-N-acetylpenicillamine (SNAP), an NO-generating agent, indicating that CR-6 acts as a scavenger of NO in cultured neurons. This was further supported by experiments showing that in neurons treated with CR-6 and glutamate, the 5-nitro derivative of CR-6 was formed, as determined by GC-MS analyses. Moreover, in vitro incubation of CR-6 with SNAP also produced the 5-nitroderivative, thus confirming that CR-6 directly reacts with NO. The results reported indicate that CR-6 acts as an NO scavenger in neurons and prevents glutamate neurotoxicity.

Animals↗
PubMed1999

Chronic exposure to aluminium impairs the glutamate-nitric oxide-cyclic GMP pathway in the rat in vivo.

Aluminium is neurotoxic and is considered a possible etiologic factor in Alzheimer's disease, dialysis syndrome and other neurological disorders. The molecular mechanism of aluminium-induced impairment of neurological functions remains unclear. We showed that aluminium impairs the glutamate-nitric oxide-cGMP pathway in cultured neurons. The aim of this work was to assess by in vivo brain microdialysis whether chronic administration of aluminium in the drinking water (2.5% aluminium sulfate) also impairs the glutamate-nitric oxide-cGMP pathway in the cerebellum of rats in vivo. Chronic exposure to aluminium reduced NMDA-induced increase of extracellular cGMP by ca 50%. The increase in extracellular cGMP induced by the nitric oxide generating agent S-nitroso-N-acetylpenicillamine was higher (240%) in rats treated with aluminium than in controls. Immunoblotting experiments showed that aluminium reduced the cerebellar content of calmodulin and nitric oxide synthase by 34 and 15%, respectively. Basal activity of soluble guanylate cyclase was decreased by 66% in aluminium-treated rats, while the activity after stimulation with S-nitroso-N-acetylpenicillamine was similar to controls. Basal cGMP in the cerebellar extracellular space was decreased by 50% in aluminium-treated rats. These results indicate that chronic exposure to aluminium reduces the basal activity of guanylate cyclase and impairs the glutamate-nitric oxide-cGMP pathway in the animal in vivo.

Aluminum↗
PubMed1999

Chronic exposure to ammonia alters pathways modulating phosphorylation of microtubule-associated protein 2 in cerebellar neurons in culture.

Hyperammonemia is considered the main cause for the neurological alterations found in hepatic failure. However, the mechanisms by which high ammonia levels impair cerebral function are not well understood. It has been shown that chronic hyperammonemia impairs signal transduction pathways associated with NMDA receptors and also alters phosphorylation of some neuronal proteins. The aim of the present work was to analyze the effects of chronic exposure to ammonia on phosphorylation of microtubule-associated protein 2 (MAP-2) in intact neurons in culture and to assess whether modulation of MAP-2 phosphorylation by glutamate receptor-associated transduction pathways is altered in neurons chronically exposed to ammonia. It is shown that chronic exposure to ammonia increases basal phosphorylation of MAP-2 by approximately 70%. This effect seems to be due to a decreased tonic activation of NMDA receptors and of calcineurin. Chronic exposure to ammonia also alters the modulation of MAP-2 phosphorylation by NMDA receptors and metabotropic glutamate receptors. In neurons exposed to ammonia, treatment with NMDA for 30 min induced a significant decrease in phosphorylation of MAP-2. Activation of metabotropic glutamate receptors with (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid significantly increased phosphorylation of MAP-2 in control neurons, whereas in neurons exposed to ammonia the response was the opposite, with 1-aminocyclopentane-1,3-dicarboxylic acid inducing a dephosphorylation of MAP-2. These results indicate that ammonia alters significantly signal transduction pathways associated with different types of glutamate receptors. This would lead therefore to significant alterations in glutamatergic neurotransmission, which would contribute to the neurological alterations found in hyperammonemia and in hepatic encephalopathy.

Ammonia↗
PubMed1998

Chronic exposure to aluminum impairs neuronal glutamate-nitric oxide-cyclic GMP pathway.

Humans are exposed to aluminum from environmental sources and therapeutic treatments. However, aluminum is neurotoxic and is considered a possible etiologic factor in Alzheimer's disease and other neurological disorders. The molecular mechanism of aluminum neurotoxicity is not understood. We tested the effects of aluminum on the glutamate-nitric oxide-cyclic GMP pathway in cultured neurons. Neurons were exposed to 50 microM aluminum in culture medium for short-term (4 h) or long-term (8-14 days) periods, or rats were prenatally exposed, i.e., 3.7% aluminum sulfate in the drinking water, during gestation. Chronic (but not short-term) exposure of neurons to aluminum decreased glutamate-induced activation of nitric oxide synthase by 38% and the formation of cyclic GMP by 77%. The formation of cyclic GMP induced by the nitric oxide-generating agent S-nitroso-N-acetylpenicillamine was reduced by 33%. In neurons from rats prenatally exposed to aluminum but not exposed to it during culture, glutamate-induced formation of cyclic GMP was inhibited by 81%, and activation of nitric oxide synthase was decreased by 85%. The formation of cyclic GMP induced by S-nitroso-N-acetylpenicillamine was not affected. These results indicate that chronic exposure to aluminum impairs glutamate-induced activation of nitric oxide synthase and nitric oxide-induced activation of guanylate cyclase. Impairment of the glutamate-nitric oxide-cyclic GMP pathway in neurons may contribute to aluminum neurotoxicity.

Aluminum↗
PubMed1991

Evolution of several cytoskeletal proteins of astrocytes in primary culture: effect of prenatal alcohol exposure.

In the present work we have analyzed, using immunoblotting and immunofluorescence techniques, the evolution of several cytoskeletal proteins during the development of astrocytes in primary culture. The effect of prenatal exposure to alcohol on these proteins was also evaluated. Microtubular protein alpha-tubulin decreased approximately 47% from 4 to 7 days after which its content remained practically constant. Immunofluorescence studies showed also that the content of alpha-tubulin was greater at day 4 of culture. This increase in fluorescence was coincident with the presence of globular particles which were found in interphase astrocytes and stained with both anti alpha- and anti-beta tubulin. These structures appeared only in proliferating cells. Glial fibrillary acidic protein (GFAP) and vimentin were analyzed as intermediate filament (IF) proteins. GFAP, in cytoskeletal preparations, increased regularly for 14 days followed by a decrease to day 21. In contrast, vimentin showed a progressive increase throughout the entire culture period. Fluorescence studies revealed some differences between the IF distribution patterns of GFAP and vimentin. In astrocytes obtained from rats prenatally exposed to ethanol, decreases in the amounts of all the cytoskeletal proteins studied were found during the entire culture period. In these cells a striking disorganization of cytoskeleton was also observed. The alcohol-induced decrease of GFAP in cultured astrocytes was also found when this protein was studied in preparations from whole brain developed "in vivo".

Aging↗
PubMed1990

Ethanol alters astrocyte development: a study of critical periods using primary cultures.

Using astrocytes obtained from 21-day-old rat fetuses, in primary culture, we have analyzed the effect of prenatal alcohol consumption on DNA and protein synthesis of astrocytes during their development. The variation in sensitivity of astrocytes to ethanol "in vitro" during the proliferation and maturation periods was also assessed. Control astrocytes showed peaks of DNA and protein synthesis at 8 and 15 days, respectively. A significant decrease in both DNA and protein synthesis was found in astrocytes from fetuses prenatally exposed to ethanol. This effect on DNA synthesis was also observed when control astrocytes were exposed to ethanol (100 mM) "in vitro" during the entire culture period. The effects on astrocytes of short term (48h) exposure to ethanol during the proliferation or differentiation periods on the above mentioned parameters and on the cell cycle as well as the possible recovery from these effects were also evaluated. Decreases in DNA and protein synthesis were found in both periods. However, DNA synthesis and content were more affected in astrocytes exposed to ethanol during the proliferation period. This effect correlates with an accumulation of cells in the G0/G1 phase of the cell cycle. On the other hand, when cells exposed to ethanol were cultured in alcohol-free medium to assess recovery, only cells exposed to ethanol during days 4 to 6 still showed DNA ethanol-induced effects at 21 days. In conclusion, our results show that ethanol consumption during gestation induces serious damage to cortical astrocyte progenitor cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1982

Increase in size of sonicated phospholipid vesicles in the presence of detergents.

Triton X-100, sodium dodecylsulphate, sodium cholate, and beta-octylglucoside increase the size of sonicated, but not of unsonicated, phospholipid vesicles above the Tc gel to liquid-crystalline transition temperature. Lysophosphatidylcholine or glyceryl monooleate do not increase liposome size under these circumstances. The observed phenomenon of vesicle growth is virtually unaffected by phospholipid composition, surface potential, calcium ions, EDTA, or albumin. The presence of cholesterol makes the vesicles more susceptible to detergent-induced growth. The presence of detergents, even at concentrations much lower than required to observe any increase in vesicle size, favors the liberation of vesicle contents. These results may have implications concerning membrane fusion as well as the reconstitution of membrane proteins in the presence of detergents.

Calcium↗
PubMed1981

Constancy of liver lipid composition in two genera of toads after a short-term temperature acclimation.

The lipid and fatty acid composition of Bufo calamytes and Alytes obstetricans livers has been studied. Data for both species are similar, and resemble closely those published for Rana sculenta. Total lipids constitute 20-25% of the total liver dry weight; about one fifth of these are phospholipids; cholesterol makes up 1.7-2.2% of the total liver dry weight. The most abundant phospholipids are phosphatidylcholine and phosphatidylethanolamine. The various lipid cases differ in their fatty acid composition: neutral lipids contain high proportions of palmitoleic and linoleic acids; phosphatidylcholine is characterized by its contents in palmitic acid, whereas cardiolipin contains polyunsaturated fatty acids. Thermal acclimation of toads for 96 h produces but few changes in liver lipid composition.

Acclimatization↗
PubMedDate not supplied

Catecholaminergic polymorphic ventricular tachycardia detected by an insertable loop recorder in a pediatric patient with exercise syncopal episodes.

Syncope is a common clinical problem that occurs at all ages and is particularly prevalent in childhood and adolescence. The evaluation of syncope is best approached using the history and physical examination, electrocardiogram and risk stratification to guide further diagnostic tests. Syncope associated with exercise or exertion must be considered dangerous, it can be a predictor of sudden cardiac death and therefore it must be taken seriously. The insertable loop recorder has been proven to be useful in the diagnosis of recurrent syncope in adult patients in whom the cause of the syncope cannot be found by other noninvasive means. We report the first known case of cathecolaminergic polymorphic ventricular tachycardia detected by an insertable loop recorder in a 5-year-old boy with a syncopal history and no structural heart disease.

Child↗