Biomedical subjects

A Machado

Publications and source records attributed to A Machado.

At least 37 records · Page 2Linked to original sources

PubMed2001

Enterococcus faecalis resistant to vancomycin and teicoplanin (VanA phenotype) isolated from a bone marrow transplanted patient in Brazil.

We report for the first time in Brazil, a patient from whom an Enterococcus faecalis VanA phenotype was isolated. Glycopeptide resistance is not commonly observed in Enterococcus faecalis, so this finding is of great concern since this species is responsible for 90% of enterococcal infections in Brazil. The isolate was recovered from a surveillance rectal swab culture from a patient with acute lymphocytic leukemia (ALL). Identification to the species level was performed by conventional biochemical tests and Vitek GPI cards. Antimicrobial susceptibility testing was evaluated by use of broth microdilution and Etest (AB BIODISK, Solna, Sweden) methods. The isolate was identified as E. faecalis and was considered resistant to both vancomycin (MIC, > 256 microg/mL) and teicoplanin (MIC, 256 microg/mL). The isolate also showed high level resistance to gentamicin and streptomycin (MICs, > 1024 microg/mL), but was considered susceptible to ampicillin (MIC, 4 microg/mL). Although the frequency of enterococcal infections is very low in most Latin America countries, the finding of glycopeptide (VanA) resistance in E. faecalis increases concern about apreading antimicrobial resistance in this region.

Adult↗

PubMed2000

Low selenium diet induces tyrosine hydroxylase enzyme in nigrostriatal system of the rat.

We have studied the effect of a selenium-deficient diet on the nigrostriatal dopaminergic system for 15 and 30 days. The neurochemical analysis demonstrated significant elevations in nigral DA levels after 15 and 30 days of selenium deficiency. The most significant change in striatum was an elevation in dopamine (DA) in 30-day-deficient animals. As a further step, we measured the levels of activity and mRNA expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT). There were significant elevations in all of these parameters in the nigrostriatal system following selenium deficiency at the two time-points studied except for the striatal DA uptake at day 30, which dropped to control levels. Altogether, our results suggest that the decrease in antioxidant capacity due to a selenium deficiency promotes an increase in DA synthesis and turnover, which are clearly associated to the induction of TH. This effect may trigger a positive feed-forward mechanism that could increase the oxidative stress considering the capacity of DA to generate free radicals.

Animals↗

PubMed2000

Neurotoxic relationship between dopamine and iron in the striatal dopaminergic nerve terminals.

The neurotoxic effect of dopamine (DA) and iron(III) on DAergic terminals in striatum has been studied by intracerebral microdialysis technique. Twenty-four hours after surgery (day 1), DA and/or iron(III) with and without DA reuptake inhibitor, nomifensine, were perfused for 1 h. Forty-eight hours after surgery (day 2), MPP(+) 1 mM was perfused for 15 min and the output of DA was measured, its amount being directly proportional to the remaining striatal DAergic terminals, supported by tyrosine hydroxylase immunohistochemistry technique. Perfusion of exogenous DA, as well as iron(III) 10 and 100 microM, did not produce any neurotoxic effect. However, perfusion of iron(III) (333 and 1000 microM) produced a concentration-dependent toxic effect. Co-perfusion of iron(III) at non-toxic concentration (100 microM) with DA (15 microM) produced a toxic effect. Elevation of the endogenous extracellular levels of DA by inhibiting its uptake with nomifensine increased the neurotoxic effect of iron(III) in a dose-dependent manner. The use of tetrodotoxin after elevation of DA with nomifensine partially prevented the neurotoxic effect of its co-perfusion with iron(III) (100 microM). These results suggest that DAergic system could be synergistically damaged by DA and iron(III). Thus, alterations in the clearance of DA from extracellular space along with an increase of iron may have significant consequences for DAergic system toxicity.

1-Methyl-4-phenylpyridinium↗

PubMed2000

Upregulation of BDNF mRNA and trkB mRNA in the nigrostriatal system and in the lesion site following unilateral transection of the medial forebrain bundle.

We have performed unilateral transection of the medial forebrain bundle (MFB) and studied BDNF mRNA and trkB mRNA levels at different postlesion times in the nigrostriatal system by means of in situ hybridization. BDNF mRNA levels were transiently induced in the substantia nigra pars compacta at 1 day postaxotomy. The disposition of BDNF mRNA expressing cells at this postlesion time in substantia nigra mimicked that of the dopaminergic neurons expressing the mRNA for the dopamine transporter. TrkB mRNA levels remained unaltered in the ventral mesencephalon at the different postlesion times examined-1 to 14 days. In contrast, trkB mRNA levels were significantly induced in the striatum at the longer postlesion time examined-14 days-when all neurodegenerative events are completed. It is becoming apparent that nigral BDNF mRNA levels are anterogradely transported to its target tissue in striatum. However, following axotomy, the lesion site represents a second potential target for BDNF action. Consequently, we also analyzed the pattern of mRNA expression for BDNF and trkB at the lesion site where dopaminergic axons are disconnected. There, we found notable inductions of both BDNF mRNA and trkB mRNA levels at 4 days postaxotomy. BDNF mRNA expressing cells were confined at the site of axotomy, which coincided precisely to that showing induction of trkB mRNA. Altogether, our results anticipate promising trophic roles of BNDF in the injured nigrostriatal system.

Animals↗

PubMed2000

The single intranigral injection of LPS as a new model for studying the selective effects of inflammatory reactions on dopaminergic system.

We have injected lipopolysaccharide (LPS) into the nigrostriatal pathway of rats in order to address the role of inflammation in Parkinson's disease (PD). LPS induced a strong macrophage/microglial reaction in Substantia nigra (SN), with a characteristic clustering of macrophage cells around blood-vessels. The SN was far more sensitive than the striatum to the inflammatory stimulus. Moreover, only the dopaminergic neurons of the SN were affected, with no detectable damage to either the GABAergic or the serotoninergic neurons. The damage to the DA neurons in the SN was permanent, as observed 1 year postinjection. Unlike the direct death of dopaminergic neurons caused by agents as MPP(+) or 6-OHDA, LPS seems to cause indirect death due to inflammatory reaction. Therefore, we suggest that the injection of a single dose of LPS within the SN is an interesting model for studying the selective effects of inflammatory reaction on dopaminergic system and also potentially useful for studying PD.

Animals↗

PubMed2000

Decreased messenger RNA expression of key markers of the nigrostriatal dopaminergic system following vitamin E deficiency in the rat.

We have evaluated the effect of a vitamin E-deficient diet on the rat nigrostriatal dopaminergic system. After 15 days of deficient diet, the amount and activity of striatal and nigral tyrosine hydroxylase increased, which contrasted with a decreased messenger RNA expression for tyrosine hydroxylase and the dopamine transporter in the ventral mesencephalon. When we prolonged the deficiency of vitamin E for 30 days, dopamine levels did not differ in both areas. In contrast, messenger RNA levels for tyrosine hydroxylase and the dopamine transporter were markedly reduced in 30-day deficient rats. In addition, the number of oxidatively modified proteins significantly increased in the striatal and nigral areas studied. Overall, we propose that these changes suggest an important role of vitamin E in maintaining the normal equilibrium of the dopaminergic nigrostriatal system.

Animals↗

PubMed2000

Protective effect of melatonin against the 1-methyl-4-phenylpyridinium-induced inhibition of complex I of the mitochondrial respiratory chain.

In the present study, a novel property of melatonin is shown: a protective effect of melatonin on the respiratory chain in isolated rat liver mitochondria and in striatal synaptosomes treated with 1-methyl-4-phenylpyridinium ion (MPP+). The cellular damage caused by MPP+, a compound that produces a Parkinsonian-like syndrome in humans, is the result of the mitochondrial respiration inhibition at the Complex I level and oxidative stress induction. Treatment of mitochondria with MPP+ inhibits the respiration rate. This effect was prevented by the inclusion of melatonin in the incubation mixture. This preventive effect, which is not related to the antioxidative properties of melatonin, seems to be due to the fact that melatonin prevents MPP+ interaction with Complex I. These results suggest that melatonin may protect against the effect of several Parkinsonogenic compounds that are associated with progressive impairment of mitochondrial function and increased oxidative damage.

1-Methyl-4-phenylpyridinium↗

PubMed2000

The endogenous amine 1-methyl-1,2,3,4- tetrahydroisoquinoline prevents the inhibition of complex I of the respiratory chain produced by MPP(+).

The endogenous monoamine 1-methyl-1,2,3,4-tetrahydroisoquinoline has been shown to prevent the neurotoxic effect of MPP(+) and other endogenous neurotoxins, which produce a parkinsonian-like syndrome in humans. We have tested its potential protective effect in vivo by measuring the protection of 1-methyl-1,2,3,4-tetrahydroisoquinoline in the neurotoxicity elicited by MPP(+) in rat striatum by tyrosine hydroxylase immunocytochemistry. Because we know that cellular damage caused by MPP(+) is primarily the result of mitochondrial respiratory inhibition at the complex I level, we have extended the study further to understand this protective mechanism. We found that the inhibitory effect on the mitochondrial respiration rate induced by MPP(+) in isolated rat liver mitochondria and striatal synaptosomes was prevented by addition of 1-methyl-1,2,3,4-tetrahydroisoquinoline. This compound has no antioxidant capacity; therefore, this property is not involved in its protective effect. Thus, we postulate that the preventive effect that 1-methyl-1,2,3,4-tetrahydroisoquinoline has on mitochondrial inhibition for MPP(+) could be due to a "shielding effect," protecting the energetic machinery, thus preventing energetic failure. These results suggest that this endogenous amine may protect against the effect of several parkinsonism-inducing compounds that are associated with progressive impairment of the mitochondrial function.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed2000

Histamine infusion induces a selective dopaminergic neuronal death along with an inflammatory reaction in rat substantia nigra.

We have evaluated the effects of a direct infusion of histamine, as mediator of inflammatory response, in substantia nigra, striatum, medial septum, and medial lemniscus. Injection of 100 and 250 nmol of histamine in substantia nigra produced a selective damage in dopaminergic neurons evidenced by the loss of tyrosine hydroxylase mRNA-expressing cells, tyrosine hydroxylase-immunolabeled-positive cell bodies, and dopamine and 3,4-dihydroxyphenylacetic acid levels. In parallel we found an acute inflammatory response manifested by a loss of glial fibrillary acidic protein-immunolabeled astrocytes and, at precisely the same area, an activation of microglia. In the striatum, only high doses (500 nmol) produced an evident terminal degeneration. The selective neurotoxicity of histamine for dopaminergic cells was demonstrated by the unaltered transcription of glutamic acid decarboxylase mRNA in substantia nigra. Moreover, intraseptal injection of 100 nmol of histamine failed to alter the pattern of choline acetyltransferase mRNA-expressing cells, and intraparenchymal injection of histamine in medial lemniscus failed to alter the pattern of serotonin-immunolabeled cells. We conclude that the substantia nigra is highly sensitive to histamine-derived neurotoxicity, where inflammatory processes mediated by histamine could be important in the pathological changes that lead to dopaminergic neuronal damage after histamine infusion.

3,4-Dihydroxyphenylacetic Acid↗

PubMed2000

Laser peel: facial rejuvenation with a superficial erbium:YAG laser treatment.

BACKGROUND: Facial rejuvenation is a popular procedure to temporarily mask the effects of aging. Most patients desiring this treatment are younger and want improvement without any down time. This study was conducted to evaluate the use of Er:YAG laser as a facial rejuvenation tool. METHODS: The full faces of 18 volunteers were treated with an Er:YAG laser using a fluence of either 5 or 10 J/cm2. All volunteers applied EMLA cream (lidocaine 2.5% and prilocaine 2.5%) two hours before the procedure and were treated with a single pass using a pulse duration of about 300 microseconds. Follow-up visits were made in order to evaluate the degree of discomfort, erythema, swelling and improvement in skin aging. Skin biopsy was performed in one volunteer before and two hours after EMLA application, although preceding laser treatment. RESULTS: Most volunteers experienced moderate discomfort during the treatment. There was mild to moderate erythema and mild swelling. The improvement in general skin appearance, actinic bronzing and photo-damage was mild to moderate. The microscopic evaluation of pre-laser treated skin two hours after EMLA application was suggestive of increased water content in the dermis. CONCLUSION: The Er:YAG laser is an effective and safe tool for facial rejuvenation. With a superficial treatment, resolution of intense erythema is fairly rapid, averaging two to three days. The improvement, however, is mild compared to full laser skin resurfacing (LSR).

Adult↗

PubMed2000

Shifts in the psychometric function and their implications for models of timing.

This study examined how two models of timing, scalar expectancy theory (SET) and learning to time (LeT), conceptualize the learning process in temporal tasks, and then reports two experiments to test these conceptualizations. Pigeons responded on a two-alternative free-operant psychophysical procedure in which responses on the left key were reinforceable during the first two, but not the last two, quarters of a 60-s trial, and responses on the right key were reinforceable during the last two, but not the first two, quarters of the trial. In Experiment 1 three groups of birds experienced a difference in reinforcement rates between the two keys only at the end segments of the trial (i.e., between the first and fourth quarters), only around the middle segments of the trial (i.e., between the second and third quarters), or in both end and middle segments. In Condition 1 the difference in reinforcement rate favored the left key; in Condition 2 it favored the right key. When the reinforcement rates differed in the end segments of the trial, the psychometric function--the proportion of right responses across the trial--did not shift across conditions; when it occurred around the middle of the trial or in both end and middle segments, the psychometric function shifted across conditions. Experiment 2 showed that the psychometric function shifts even when the overall reinforcement rate for the two keys is equal, provided the rates differ around the middle of the trial. This pattern of shifts of the psychometric function is inconsistent with SET. In contrast, LeT provided a good quantitative fit to the data.

Animals↗

PubMed1999

Differential upregulation of aquaporin-4 mRNA expression in reactive astrocytes after brain injury: potential role in brain edema.

Astrocytes and aquaporin-4 (AQP4) play a significant role in brain ion homeostasis. Consequently the regulation of AQP4 mRNA in the CNS after different neurological insults was of interest. A single intrastriatal injection of ringer or quinolinic acid strongly induced AQP4 mRNA in the striatum, specially at the core of the lesion. Colocalization studies demonstrated that AQP4 mRNA induction was restricted to hypertrophic astrocytes. The extent of striatal AQP4 mRNA induction did not correlate with neuronal degeneration, but it did with extravasation of Evans blue dye as a marker of BBB disruption. Distant lesions were additionally induced by either 6-OHDA or a knife cut in the medial forebrain bundle (MFB). The former, but not the latter, induced a high AQP4 mRNA expression in the lesioned substantia nigra. However, axotomy of the MFB induced a high AQP4 mRNA expression at the lesion site. We conclude that the induction of AQP4 mRNA expression is related to disruption of the blood-brain barrier and under brain edema conditions this water channel plays a key role in the reestablishment of the brain osmotic equilibrium.

Animals↗

PubMed1999

Serotonin hyperinnervation in the adult rat ventral mesencephalon following unilateral transection of the medial forebrain bundle. Correlation with reactive microglial and astroglial populations.

We have previously studied changes in the serotoninergic and dopaminergic nigrostriatal systems following transection of the medial forebrain bundle and found a long-term axotomy-induced increase in the levels of serotonin and its main metabolite, 5-hydroxyindolacetic acid in substantia nigra [Venero et al. (1997) J. Neurochem. 68, 2458-2468]. In an attempt to find a rationale for this effect, we have performed an immunohistochemical study. Transection of the medial forebrain bundle of the rat interrupted most of the ascending serotoninergic pathways from the raphe nuclei as revealed by serotonin immunoreactivity. While serotonin immunostaining was almost absent in striatum, it doubled in the ventral mesencephalon at 21 days postlesion. This axotomy-induced increase was accompanied by an increased density of the serotonin nerve terminal network in the ipsilateral substantia nigra and ventral tegmental area. The increase in serotonin immunoreactivity was in line with the measured levels of serotonin and 5-hydroxyindolacetic acid in substantia nigra. In addition, the distribution pattern of glial fibrillary acidic protein-immunoreactive astrocytes and OX42-immunoreactive microglia correlated highly with the location of increased serotonin fibre density in the ventral mesencephalon, especially in ventral tegmental area and in the most medial part of substantia nigra. We suggest that a pruning effect may underly the axotomy-induced increase in serotonin immunoreactivity in the ventral mesencephalon, and further, that activated astroglia and microglia may play a role in directing serotoninergic axonal regeneration following axotomy.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1999

Detailed localization of aquaporin-4 messenger RNA in the CNS: preferential expression in periventricular organs.

We have performed a detailed in situ hybridization study of the distribution of aquaporin-4 messenger RNA in the CNS. Contrary to expectation, we demonstrate that aquaporin-4 is ubiquitously expressed in the CNS. Strong hybridization labeling was detected in multiple olfactory areas, cortical cells, medial habenular nucleus, bed nucleus of the stria terminalis, tenia tecta, pial surface, pontine nucleus, hippocampal formation and multiple thalamic and hypothalamic areas. A low but significant hybridization signal was found, among others, in the choroid plexus of the lateral ventricles, ependymal cells, dorsal raphe and cerebellum. Overall, a preferential distribution of aquaporin-4 messenger RNA-expressing cells was evident in numerous periventricular organs. From the distribution study, the presence of aquaporin-4 messenger RNA-expressing cells in neuronal layers was evident in neuronal layers including the CA1 -CA3 hippocampal pyramidal cells, granular dentate cells and cortical cells. Further evidence of neuronal expression comes from the semicircular arrangement of aquaporin-4 messenger RNA-expressing cells in the bed nucleus of the stria terminalis and medial habenular nucleus exhibiting Nissl-stained morphological features typical of neurons. Combined glial fibrillary acidic protein immunohistochemistry and aquaporin-4 messenger RNA in situ hybridization demonstrated that aquaporin-4 messenger RNA is expressed by glial fibrillary acidic protein-lacking cells. We conclude that aquaporin-4 messenger RNA is present in a collection of structures typically involved in the regulation of water and sodium intake and that aquaporin-4 water channels could be the osmosensor mechanism responsible for detecting changes in cell volume by these cells.

Animals↗

PubMed1999

Delayed apoptotic pyramidal cell death in CA4 and CA1 hippocampal subfields after a single intraseptal injection of kainate.

We have performed a detailed time-course analysis of cell death in the hippocampal formation, basal forebrain and amygdala following a single intraseptal injection of kainate in adult rats. Acetylcholinesterase histochemistry revealed a profound loss of staining in the medial septum but not in the diagonal band, and cholinergic fiber density was highly reduced in the hippocampus and amygdala at 10 days postinjection. Terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphatebiotin nick end labeling (TUNEL) histochemistry was performed for precise location of apoptotic cells. Both the medial septum and amygdala exhibited numerous TUNEL-positive nuclei after the intraseptal injection of kainate, while the lateral septum exhibited a lower but significant incidence in terms of apoptotic cells. In the medial septum, the presence of apoptotic cells was at a location displaying acetylcholinesterase staining. TUNEL histochemistry revealed a time-dependent sequential apoptotic cell death in hippocampal pyramidal cells. During the first two days postinjection, apoptosis in the hippocampus was only evident in the CA3 region. At five days postinjection, the entire CA4 region became apoptotic. At 10 days postinjection, the whole extent of the CA1 pyramidal cell layer exhibited numerous TUNEL-positive nuclei. The time-course of kainate-induced apoptosis in Ammons's horn correlated with the disappearance of hippocampal pyramidal neurons as detected by Nissl staining, which is suggestive of a prominent apoptotic death for these cells. The temporal delayed distant damage to CA4 and CA1 hippocampal subfields after a single intraseptal kainate injection is not seen in other models employing kainate and may be a valuable tool for exploring the cellular mechanisms leading to cell death in conditions of status epilepticus.

Acetylcholinesterase↗

PubMed1999

Effects of aging on the various steps of protein synthesis: fragmentation of elongation factor 2.

The possible mechanism responsible for the in vivo protein synthesis decline during aging was studied. In order to determine the effect of aging on the various steps of protein synthesis, we determined the ribosomal state of aggregation and the time of assembly and release of polypeptide chains in the process of protein synthesis in rat liver. The results suggest that elongation is the most sensitive step to aging. A molecular study of the Elongation Factor 2 (EF-2), the main protein involved in the elongation step, shows that this protein has a higher content of carbonyl groups and is less active in old rats. In addition, the molecular mass analysis of EF-2 shows that this protein becomes fragmented in old rats. A similar pattern of fragmentation is found in 3-month-old rats suffering oxidative stress, in that the decline in protein synthesis is similar to that found in old rats. These data suggest that: i) oxidative stress seems to be involved in the modifications of EF-2 observed during aging, and ii) the observed modifications (oxidation and fragmentation) of EF-2 could account for the decline in protein synthesis in old animals.

Aging↗

PubMed1999

Induced mono-(ADP)-ribosylation of rat liver cytosolic proteins by lipid peroxidant agents.

We have studied the effect of free radical generating agents on the mono-(ADP)-ribosylation of rat liver cytosolic proteins. Our results show that this post-translational modification, whose physiological significance is still unclear, is activated by lipid peroxidant agents via activation of cytoplasmatic mono-(ADP)-ribosyltransferases. The implication of free radicals in this process is demonstrated by the fact that mono-(ADP)-ribosylation can be prevented by melatonin, N-tert-butyl-alpha-phenylnitrone and dithiothreitol. On the basis of our results, we discuss the modification of proteins caused by free radicals as a possible mechanism by which they damage the cell.

ADP Ribose Transferases↗

PubMed1999

A signalling theory of excessive technological adoption.

Technology adoption has been identified as one of the main elements behind the growth of health care expenditures. It has been argued that the health insurance arrangements in the US justify, to a certain extent, the technology-driven rise in costs. Moreover, it eases the adoption of less cost-effective procedures and devices. This paper presents an additional argument by which excessive technology investments may occur: providers of care invest in technology as a way to "signal" their intrinsic (and unobservable) quality. Providers face the option of adopting a new technology. The decision of adoption in itself may convey information about his/her quality: for example, patients conjecture that providers who display newer technology are of higher quality. Providers, being aware of this, may invest in technology to reveal themselves as high quality. Thus, technology adoption could result only from the desire to attract patients. The investment is self-defeating in the sense that if all providers invest, no information about quality is transmitted to patients. We evaluate the argument in a context of demand for health care services where patients have initially no information about the quality of different providers. We show that an incentive to invest as a way to signal quality may or may not lead to overinvestment. It is also possible that only some providers invest. They reveal themselves as high quality providers. The analysis suggests that the argument is more important for some services than for others. Overall, an additional argument for overinvestment in technology in some circumstances is provided.

Attitude of Health Personnel↗