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A Almeida

Publications and source records attributed to A Almeida.

At least 55 records · Page 3Linked to original sources
PubMed1998

A rapid method for the isolation of metabolically active mitochondria from rat neurons and astrocytes in primary culture.

A rapid method (about 1.5 h) for the isolation of intact functional mitochondria from neurons and astrocytes in primary culture is described. Mitochondria isolated by this method are metabolically active and tightly coupled as shown by respiratory control ratio values, which were about 4 with glutamate-malate as substrate. The activities of marker enzymes revealed the occurrence of a low degree of cytosolic (5%) or synaptosomal (5.5%) contamination in the mitochondrial fractions. In addition, the activity of citrate synthase was increased by 4 fold in both neuronal and astrocytic mitochondria with respect to values found in cell homogenates. These results confirm that the method affords mitochondrial preparations from cultured brain cells at suitable levels of purity and enrichment for the study of their mitochondrial function. Since mitochondrial damage has been associated with the pathogenesis of certain neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases (P. Chagnon, C. Betard, Y. Robitaille, A. Cholette, D. Gauvreau, Distribution of brain cytochrome oxidase activity in various neurodegenerative disease, Neuroreport 6 (1995) 711-715 [6]; S.J. Kish, C. Bergeron, A. Rajput, S. Dozic, F. Mastrogiacomo, L. Chang, J.M. Wilson, L.M. DiStefano, J.N. Nobrega, Brain cytochrome oxidase in Alzheimer's disease, J. Neurochem. 59 (1992) 776-779 [10]; A.H.V. Schapira, J.M. Cooper, D. Dexter, J.B. Clark, P. Jenner, C.D. Marsden, Mitochondrial complex I deficiency in Parkinson's disease, J. Neurochem. 54 (1990) 823-827 [15]), the method described here shed light on the possible susceptibility of neuronal or astrocytic mitochondria to deleterious effects of these diseases.

Animals↗
PubMed1998

[CHARGE association].

Posterior choanal atresia is a congenital malformation which can occur isolated or in combination to additional malformations. In CHARGE association the other anomalies are: coloboma, heart disease, retarded development/growth or central nervous system abnormalities, genital hypoplasia or hypogonadism and ear abnormalities or deafness. The authors present three cases of CHARGE association and they also review the clinical findings required for the diagnosis.

Abnormalities, Multiple↗
PubMed1998

[Iliac bone for secondary grafting in residual alveolar clefts].

Iliac cancellous bone has proven efficacy as a bone-graft donor. This study analyses the success of iliac bone autografts in secondary alveolar clefts. The study group was 30 patients with clefts with complete clinical charts and occlusal radiographs with surgery did in Cleft Unit Temuco Regional Hospital (Chile) between 1990-1996. The quality of graft "take" was measured radiologically and clinically. The results were named "excelent-good-regular-bad" and were studied by statistic methods. We did not find complications in donor site. We only had two partial dehiscences in recipient site. In 29 grafts, we feel "bone consistence" in clinical examination. We had one "nontake" graft. 80% of alveolar bone grafts showed similar bone density with respect normal bone. 80% of same grafts had similar height of interdental septum. We had 22 cases with "good result" and 3 cases with "regular result". All of them statistically significant. In our experience, iliac bone graft for alveolar clefts is a good technique for this difficult problem.

Adolescent↗
PubMed1998

Clinical, cytogenetic and toxicological studies in rural workers exposed to pesticides in Botucatu, São Paulo, Brazil.

Pesticides can cause gene mutations and chromosomal aberrations in exposed individuals. We have investigated 24 workers exposed to pesticides. Clinical examinations and cytogenetic and toxicological tests were performed. Ten non-exposed individuals were used as controls. Toxicological dosages of copper, zinc and manganese (metals found in some pesticides), hepatic enzyme dosage (GOT, GPT, AR) and acetylcholinesterase activity were performed in 16 workers and 8 controls. In the exposed workers, the most relevant clinical symptoms were poor digestion with fullness sensation after meals, irritated eyes, headache and fasciculations. The exposed group showed significantly lower manganese dosage and acetylcholinesterase activity, and significantly higher levels of alkaline phosphatase. Cytogenetic studies showed significantly higher chromosomal aberrations in the exposed group compared to the control group. Although the workers used protection against the pesticide's fog, the results revealed that the workers were contaminated with the pesticides. Therefore, the cytogenetic, toxicological studies with clinical examination are necessary for monitoring workers who are exposed to pesticides in any situation.

Adult↗
PubMed1997

Isolation and characterization of tightly coupled mitochondria from neurons and astrocytes in primary culture.

This work provides a rapid method for isolation of intact functional mitochondria from neurons and astrocytes in primary culture. By using this method, it was found that the respiratory control ratio was 1.5-fold greater in neuronal than in astrocytic mitochondria using both NAD-linked (glutamate/malate) and FAD-linked (succinate) substrates. The difference observed in RCR values was due to the lower rate of respiration in state 4 found in neurons as compared to that found in astrocytes, because both cell types showed the same rate of respiration in state 3. The P/O ratio was also higher in neurons than in astrocytes. Our results suggest that the coupling between the mitochondrial respiratory chain and oxidative phosphorylation is stronger in neurons than in astrocytes. These results may be of relevance for the understanding of the differential susceptibility of brain cells to impairments of energy metabolism observed in certain neurodegenerative diseases.

Animals↗
PubMed1997

DNA hypomethylation in breast cancer: an independent parameter of tumor progression?

The global DNA methylation status was investigated on a series of 59 breast cancers by Southern blotting, using methylation sensitive restriction enzymes. By comparison to control DNA, almost all tumor DNAs were found globally hypomethylated. However, the demethylation was variable from tumor to tumor. Compared to other biological parameters, the methylation did not correlate with chromosome alterations, steroid hormone receptor status, or histopathological grading. Tumors which appeared to be the most evolved for other parameters were only mildly hypomethylated, whereas tumors with strongly hypomethylated DNA corresponded to those with slight alterations of the other parameters. Thus, DNA hypomethylation is a consistent characteristic of breast cancer, but its variations may not correlate with tumor progression of most breast cancers.

Age Factors↗
PubMed1997

Interrelationships between astrocyte function, oxidative stress and antioxidant status within the central nervous system.

Astrocytes have, until recently, been thought of as the passive supporting elements of the central nervous system. However, recent developments suggest that these cells actually play a crucial and vital role in the overall physiology of the brain. Astrocytes selectively express a host of cell membrane and nuclear receptors that are responsive to various neuroactive compounds. In addition, the cell membrane has a number of important transporters for these compounds. Direct evidence for the selective co-expression of neurotransmitters, transporters on both neurons and astrocytes, provides additional evidence for metabolic compartmentation within the central nervous system. Oxidative stress as defined by the excessive production of free radicals can alter dramatically the function of the cell. The free radical nitric oxide has attracted a considerable amount of attention recently, due to its role as a physiological second messenger but also because of its neurotoxic potential when produced in excess. We provide, therefore, an in-depth discussion on how this free radical and its metabolites affect the intra and intercellular physiology of the astrocyte(s) and surrounding neurons. Finally, we look at the ways in which astrocytes can counteract the production of free radicals in general by using their antioxidant pathways. The glutathione antioxidant system will be the focus of attention, since astrocytes have an enormous capacity for, and efficiency built into this particular system.

Animals↗
PubMed1997

Lesions of the caudal ventrolateral medulla block the hypertension-induced inhibition of noxious-evoked c-fos expression in the rat spinal cord.

The effect of lesioning the lateral portion of the caudal ventrolateral medullary reticular formation (VLMIat) on the noxious-evoked expression of the c-fos proto-oncogene in spinal neurons, was studied in short-term hypertensive rats. Occlusion of the renal artery for 96 h in unlesioned animals induced a 52% increase in blood pressure (BP) and a 66% decrease in the number of Fos-immunoreactive (Fos-IR) spinal cells following noxious cutaneous stimulation, as compared to values in normotensive controls. Lesioning the VLMIat in hypertensive rats by unilateral quinolinic acid (QA) injection (0.3 microl of a 180 nmol/microl solution) 24 h before noxious stimulation, prevented the Fos-IR cell decrease. In normotensive rats, lesioning the VLMIat produced no changes in c-fos expression. To investigate the role played by the VLMIat in cardiovascular control, BP and heart rate (HR) were measured during local injections of QA or glutamate (0.5 microl of a 100 nmol/microl solution) to normotensive animals. Injections of QA produced an immediate rise in BP and HR which reached maximal values (18 and 14% increase, respectively) 5 min after the administration onset, then returning gradually to baseline levels. Glutamate injections resulted in an immediate decrease of the same values, which reached 29 and 39%, respectively, 4 min after the beginning of injection, after which they decreased to baseline levels. These results suggest that VLMIat neurons inhibit nociceptive spinal neurons in response to rises in blood pressure, while exerting negative control of cardiovascular parameters. It is suggested that the VLMIat is involved in the genesis of hypoalgesia during hypertension.

Journal Article↗
PubMed1997

Nitric oxide-mediated mitochondrial damage in the brain: mechanisms and implications for neurodegenerative diseases.

Within the CNS and under normal conditions, nitric oxide (.NO) appears to be an important physiological signalling molecule. Its ability to increase cyclic GMP concentration suggests that .NO is implicated in the regulation of important metabolic pathways in the brain. Under certain circumstances .NO synthesis may be excessive and .NO may become neurotoxic. Excessive glutamate-receptor stimulation may lead to neuronal death through a mechanism implicating synthesis of both .NO and superoxide (O2.-) and hence peroxynitrite (ONOO-) formation. In response to lipopolysaccharide and cytokines, glial cells may also be induced to synthesize large amounts of .NO, which may be deleterious to the neighbouring neurones and oligodendrocytes. The precise mechanism of .NO neurotoxicity is not fully understood. One possibility is that it may involve neuronal energy deficiency. This may occur by ONOO- interfering with key enzymes of the tricarboxylic acid cycle, the mitochondrial respiratory chain, mitochondrial calcium metabolism, or DNA damage with subsequent activation of the energy-consuming pathway involving poly(ADP-ribose) synthetase. Possible mechanisms whereby ONOO- impairs the mitochondrial respiratory chain and the relevance for neurotoxicity are discussed. The intracellular content of reduced glutathione also appears important in determining the sensitivity of cells to ONOO- production. It is concluded that neurotoxicity elicited by excessive .NO production may be mediated by mitochondrial dysfunction leading to an energy deficiency state.

Animals↗
PubMed1997

Activation by cutaneous or visceral noxious stimulation of spinal neurons projecting to the medullary dorsal reticular nucleus in the rat: a c-fos study.

The involvement of spinal neurons in the transmission of cutaneous and visceral nociceptive input to the medullary dorsal reticular nucleus was studied. Rats were injected with cholera toxin subunit B in the left dorsal reticular nucleus and subjected 4 days later to noxious mechanical, thermal or chemical stimulation of the proximal internal aspect of the left thigh, or to chemical stimulation of the urinary bladder. Sections of spinal segments T13-L3 were processed immunocytochemically for cholera toxin subunit B and Fos protein. The percentage of double-labelled cells in the population of Fos-positive cells was higher in lamina I (1-4%) than in deeper laminae (0-0.7%) following all stimuli. The percentage of double-labelled cells in the population of retrogradely labelled cells was 30-53% in lamina I and 0-5% in laminae III-X. Visceral stimulation activated more retrogradely labelled lamina I cells than any kind of cutaneous stimulation. Pyramidal cells were activated in higher numbers than multipolar and flattened cells after thermal cutaneous or visceral stimulation, and in lower numbers than multipolar cells after mechanical stimulation. These results suggest that, in the experimental conditions used, spinal cord cells conveying noxious input to the dorsal reticular nucleus are concentrated in lamina I. They further indicate that the spinal-dorsal reticular nucleus pathway plays a major role in the transmission of nociceptive visceral input, and point to the preferential involvement of pyramidal cells in cutaneous thermal and visceral processing.

Animals↗
PubMed1997

Thyroid hormones regulate the onset of osmotic activity of rat liver mitochondria after birth.

The effect of thyroid hormone deprivation on the osmotic activity of liver mitochondria from early newborn rats was studied. Experimentally induced hypothyroidism prevented the increase in the osmotic activity of mitochondria observed immediately after birth. Osmotic activity was restored by T4 and T3 treatment to hypothyroid newborns but not when this treatment was supplemented with cycloheximide. Under the same circumstances, streptomycin had no effect. Hypothyroidism abolished the change in the slope of the osmotic curve (plot of inverse absorbance of mitochondrial suspensions incubated in sucrose solutions vs. inverse sucrose concentration) observed in mitochondria from euthyroid newborns at 110-120 mOsm sucrose, suggesting that hypothyroidism prevents the formation of tight physical connections between mitochondrial outer and inner membranes. Thyroid hormone deprivation increased the passive permeability of the mitochondrial inner membrane to protons, resulting in a decreased respiratory control ratio. Hypothyroidism prevented the sharp decrease in the affinity of mitochondria for ATP observed in euthyroid newborns immediately after birth. These results corroborate our previous suggestion (Endocrinology, 1995, 136:4448) that, during the early neonatal period, thyroid hormones control the synthesis of some nucleus-coded protein(s) involved in the assembly of F0,F1-ATPase.

Adenosine Triphosphate↗
PubMed1997

[Clinico-pathological correlation in the main types of dementia].

INTRODUCTION: Dementia has became a serious health problem in developed countries. The objective of this study was to establish the possible correlation between the initial clinical diagnosis and the anatomopathological criteria. Pathological confirmation of the cases clinically diagnosed as Alzheimer disease/senile dementia Alzheimer type (AD/SDAT) and multi-infarct dementia (MID) was carried out. MATERIAL AND METHODS: Twelve brains from demented patients were studied. Brains were removed at post-mortem intervals of 1-3 hours to guarantee an adequate conservation of the tissue. The brains were weighed, fixed for 4 weeks in 10% buffered neutral formalin and coronally sectioned at intervals of approximately 1 cm. Bilateral sections of neocortex from frontal, temporal, parietal lobes, cingulate gyrus, amygdala, hippocampus, thalamus, cerebellum and unilateral sections of locus ceruleus and substantia nigra were taken. Five micrometer sections of the paraffin embedded material were stained by the following methods: hematoxylin-floxine, Congo red and Bielschowsky silver impregnation. RESULTS: Our neuropathological results showed a high correlation with the initial clinical classification and confirmed the diagnosis of AD/ SDAT in 6 cases, MID in 3 cases and mixed dementia in 1 case. Two cases did not exhibited morphological evidence of dementia. CONCLUSIONS: We concluded that the methodology applied for the morphologic diagnosis of dementia was feasible, useful and reproducible. Further studies will be necessary using a larger number of sample.

Aged↗
PubMed1996

Cross-talk between topoisomerase I and HU in Escherichia coli.

In Escherichia coli about one half of the negative supercoiling of DNA is constrained by proteins, in contrast to the situation in eukaryotic cells where most of the DNA is constrained by histones. The level of supercoiling in the unrestrained portion is controlled by a balance between the supercoiling activity of gyrase and the relaxing activity of DNA topoisomerase I. In the present work we show, by disrupting one or both genes encoding the heterodimeric protein HU, that an interplay exists in bacteria between HU and topoisomerase I activity: a decrease in the intracellular concentration of HU was accompanied by an increase in relaxing activity as measured in cell extracts. Conversely, a topA10 mutant of topoisomerase I, which has low levels of relaxing activity, was unable to accept an HU deficiency introduced by transduction. Thus it appears that the ability to increase relaxing activity, or to decrease an excess of supercoiling, is important for cells to survive in the absence of HU. These data can be explained in terms of HU constraining supercoiling in vivo as it does in vitro: the absence of HU would generate more unconstrained supercoiling, which in turn would require an increase in relaxing activity to maintain physiological levels.

Bacterial Proteins↗
PubMed1996

The medullary dorsal reticular nucleus facilitates acute nociception in the rat.

The influence on pain processing caused by destruction or stimulation of the dorsal reticular nucleus (DRt) was studied using the tail-flick and the increasing temperature hot-plate tests. Lesions of the DRt were obtained by injecting quinolinic acid (180 nmol/microliters) unilaterally or bilaterally, and nociceptive responses were evaluated by both tests. Following unilateral lesions, the tail-flick latencies and the hot-plate response temperatures were increased, values differing statistically from controls in the latter test. Bilateral lesions resulted in statistically significant increases of both tail-flick latency and hot-plate response temperature. Stimulation of the DRt was performed by injecting glutamate (100 nmol/microliters) unilaterally, which was followed 1 min later by a significant decrease in the tail-flick latency compared to saline injected controls. These results suggest that the DRt is involved in the facilitation of nociception after acute thermal noxious stimulation. This effect may be mediated through a spino-DRt-spinal loop causing a rebound of excitation in lamina I cells receiving noxious input from their own receptive field.

Animals↗