Biomedical subjects

L Lima

Publications and source records attributed to L Lima.

At least 37 records · Page 2Linked to original sources

PubMed2000

Unique frequency of known mutations in Brazilian MPS I patients.

The frequency of 10 known mutations in the IDUA gene-Q70X, A75T, H82P, R89Q, 678-7 g-->a, L218P, A327P, R383H, W402X, and P533R-was estimated in a group of 24 index cases with mucopolysaccharidosis type I. Three affected relatives were also analysed. Six of the 10 mutations screened were present in our patients (Q70X, R89Q, A327P, R383H, W402X, and P533R). These mutations account for 54% of the alleles; 37% of the genotypes were defined. Frequencies of these mutations are markedly different from those in the literature. A novel combination Q70X/A327P is described. This was the first time Brazilian MPS I patients were analysed with molecular techniques. The low frequency of common mutations indicates that a more comprehensive analysis of the IDUA gene should be done to delineate the mutation profile of MPS I better in our population.

Brazil↗

PubMed2000

Regulation of high affinity taurine transport in goldfish and rat retinal cells.

Adaptive regulation and modulation by phosphorylation are mechanisms by which some cells control taurine transport. Goldfish and rat retinal cells were incubated with the activator of protein kinase C, phorbol 12,13-dibutyrate (PDBu), or the inhibitor of protein phosphatases, okadaic acid (OKA). OKA, 1 nM, inhibited the uptake of taurine at short period of incubation in goldfish retinal cells, and at low concentrations in rat retinal cells incubated with the inhibitor for 1 h. PDBu treatment did not produce significant effects. Isolated Müller cells from the goldfish retina presented a clear adaptive regulation and a decrease of taurine uptake by increasing phosphorylation either by the stimulation of PKC with PDBu or the inhibition of phosphatases with OKA.

Animals↗

PubMed2000

Modulation of taurine uptake in the goldfish retina and axonal transport to the tectum. Effect of crushing the optic nerve or axotomy.

Although there are a great number of studies concerning the uptake of taurine in several tissues, the regulation of taurine transport has not been studied in the retina after lesioning the optic nerve. In the present study, isolated retinal cells of the goldfish retina were used either immediatly after cell suspension or in culture. The high-affinity transport system of [3H]taurine in these cells was sodium-, temperature- and energy-dependent, and was inhibited by hypotaurine and beta-alanine, but not by gamma-aminobutyric acid. There was a decrease in the maximal velocity (Vmax) without modifications in the substrate affinity (Km) after optic axotomy. These changes were mantained for up to 15 days after the lesion. The results might be the summation of mechanisms for providing extracellular taurine to be taken up by other retinal cells or eye structures, or regulation by the substrate taurine, which increases after lesioning the optic nerve. The in vivo accumulation of [3H]taurine in the retina after intraocular injection of [3H]taurine was affected by crushing the optic nerve or by axotomy. A progressive retinal decrease in taurine transport was observed after crushing the optic nerve, starting at 7 hours after surgery on the nerve. The uptake of [3H]taurine by the tectum was compensated in the animals that were subjected to crushing of the optic nerve, since the concentration of [3H]taurine was only different from the control value 24 hours after the lesion, indicating an efficient transport by the remaining axons. On the contrary, the low levels of [3H]taurine in the tectum after axotomy might be an index of the non-axonal origin of taurine in the tectum. Axonal transport was illustrated by the differential presence of [3H]taurine in the intact or crushed optic nerve. The uptake of [3H]taurine into retinal cells in culture in the absence or in the presence of taurine might indicate the existence of an adaptive regulation of taurine transport in this tissue, however taurine transport probably differentially occurs in specific populations of retinal cells. The use of a purified preparation of cells might be useful for future studies on the modulation of taurine transport by taurine in the retina and its role during regeneration.

Animals↗

PubMed2000

Modulation of outgrowth from goldfish retinal explants by a 5-HT2 receptor agonist and [3H]ketanserin binding sites in goldfish and rabbit retina.

The binding of [3H]ketanserin to goldfish and rabbit retinal membrane preparations and the possible role of 5-HT2A receptors in the in vitro outgrowth from goldfish retina were evaluated. Saturation experiments indicated a high-affinity binding site, and positive cooperativity for both tissues. The 5-HT2A/2C agonist (+/-)-2,5-dimetoxy-4-iodoamphetamine and serotonin inhibited outgrowth from goldfish retinal explants. These effects were blocked by the 5-HT2 antagonists ketanserin and 1-(1-naphthyl)piperazine and by the 5-HT2C antagonist mesulergine, respectively. Results make to suggest that [3H]ketanserin binds to 5-HT2A receptors in the rabbit and goldfish retina, but also to a monoamine transporter in the latter tissue. Subtypes of 5-HT2 receptors might mediate the 5-HT modulatory role on in vitro outgrowth of the goldfish retina.

Animals↗

PubMed2000

Differential taurine effect on outgrowth from goldfish retinal ganglion cells after optic crush or axotomy. Influence of the optic tectum.

The interaction between innervated tissues, targets and nerves is crucial in the maintenance of physiological conditions, and the disturbance of this harmony causes the production of morphological and biochemical changes. After lesion of the optic nerve, several modifications take place in the retina, the optic tectum and the optic nerve. The influence of the tectum on the outgrowth from the goldfish retina and the possible role of taurine was studied. Ganglion retinal cells were identified by retrolabeling with Dil. Crushing the optic nerve 10 days prior to plating retinal cells, as compared with optic axotomy, did not affect the survival of cultured retinal cells, as well as the length of the neurites. However, the number of neurites per cell and the branching of the longest fiber were higher after axotomy than after crushing. The addition of taurine to the medium did not modify this response at 5 days in culture. At early periods in culture, the stimulatory effect on isolated ganglion cell outgrowth produced by taurine was enhanced after axotomy respecting crushing of the optic nerve, but was not affected in retinal explants. The addition of medium from cultured optic tectum several days post-crush of the optic nerve to retinal explants from intact retinas or coming from post-crush retina modified the outgrowth, being inhibitory or stimulatory in a time-dependent manner. The co-culture of optic tectum and retina also affected the outgrowth from the retina with a byphasic shape. The results support the differential response of the retina facing partial or complete interruption with the target and limit the effect of taurine to early periods in culture. In addition, the production of inhibitory factors from the tectum, plus the stimulatory ones, are strongly supported by this work.

Animals↗

PubMed2000

Cellular expression and regulation of the Medicago truncatula cytosolic glutamine synthetase genes in root nodules.

In this paper we have studied the localisation of expression of the two functional cytosolic glutamine synthetase (GS) genes, MtGSa and MtGSb, in root nodules of the model legume Medicago truncatula. We have used a combination of different techniques, including immunocytochemistry, in situ hybridisation and promoter beta-glucuronidase (GUS) fusions in transgenic plants, to provide the means of correlating gene expression with protein localisation. These studies revealed that transcriptional regulation (mRNA synthesis) plays an important part in controlling GS protein levels in nodules of M. truncatula. The major locations of cytosolic GS mRNA and protein are the central tissue, the parenchyma and the pericycle of the vascular bundles. These findings indicate that in nodules, GS might be involved in other physiological processes in addition to the primary assimilation of ammonia released by the bacterial nitrogenase. The two genes show different but overlapping patterns of expression with MtGSa being the major gene expressed in the infected cells of the nodule. Promoter fragments of 2.6 kb and 3.1 kb of MtGSa and MtGSb, respectively, have been sequenced and primer extension revealed that the MtGSb promoter is expressed in nodules from an additional start site that is not used in roots. Generally these fragments in the homologous transgenic system were sufficient to drive GUS expression in almost all the tissues and cell types where GS proteins and transcripts are located except that the MtGSa promoter fragment did not express GUS highly in the nodule infected cells. These results indicate that the cis-acting regulatory elements responsible for infected-cell expression are missing from the MtGSa promoter fragment.

Base Sequence↗

PubMed1999

[3H]Paroxetine binding to human peripheral lymphocyte membranes of patients with major depression before and after treatment with fluoxetine.

Serotonin has been involved in major depression and is also related to central and peripheral mechanisms of neuroimmunomodulation. Recently, the uptake of [3H]serotonin into human peripheral blood lymphocytes has been reported. We determined the density of serotonin uptake sites by the binding of [3H]paroxetine to blood peripheral lymphocyte membrane preparations of controls and of patients with major depression before and after treatment with fluoxetine for six weeks. The severity of depression was assessed by the use of Hamilton Rating Scale for Depression and of Beck Depression Inventory. There was a reduction in the number of sites for [3H]paroxetine in patients before administration of the antidepressant respecting controls, and a recovery after the treatment. Affinity was unchanged. No correlation was obtained between the severity of symptoms determined by Hamilton Rating Scale for Depression or by Beck Depression Inventory, and the number of binding sites for [3H]paroxetine. The levels of the plasma serotonin metabolite, 5-hydroxyindoleacetic acid, did not differ between the three groups of subjects, but serotonin was lower in patients after treatment respecting controls and in patients before treatment. The significant correlation between specific binding of [3H]paroxetine and plasma serotonin levels in controls was not present in the patients. Lymphocyte serotonin transporter is decreased in major depressed patients and is modulated by antidepressant treatment. In addition of counting with a peripheral marker in depression, the study of serotonin system in lymphocytes might contribute to understand the bi-directional interaction between the nervous and the immune systems.

Adult↗

PubMed1999

Taurine and its trophic effects in the retina.

The sulphur amino acid taurine possesses variable functions during development and regeneration of the central nervous system. The retina synthesize and uptake taurine, which is the amino acid present in the highest concentration in this tissue. Deficiency of taurine alters the structure and the function of the cerebral and cerebellar cortex, as well as the retina. Taurine increases outgrowth of postcrush goldfish retina in culture, partially by elevating calcium influx, and also by the modulation of protein phosphorylation. Its concentration increases in the retina after the lesion of the optic nerve, and the intraocular injection of it, between the crush and the explantation, stimulates the outgrowth of neurites. Taken together, although there are a great number of unresolved questions on the mechanisms of action of this amino acid as a trophic substance, the results support the role of taurine during regeneration of the optic nerve.

Animals↗

PubMed1999

Perioperative management in minimally invasive coronary surgery.

OBJECTIVE: It is important to apply the same rules used for classical coronary revascularization to beating heart coronary surgery. The surgeons must have a strategy, and be prepared for adverse events, and complications. METHODS: A careful analysis of the coronarography is essential to predict eventual contraindications or causes of possible operative difficulties. All the team must be involved in the surgical protocol. Three main problems need to be solved: the prevention of ischemia during the procedure, good stabilization of the anastomotic site and arterial occlusion. The first goal is reached by using a pre-conditioning technique or an intracoronary shunt. Vessel stabilization may be obtained by several methods: pressure adhesion devices, patch technique or suction devices. The final problem is to have a blood free field, this requires arterial occlusion which may be achieved in several ways: silastic Snares or sutures, aclan clamps, coronary occluder. RESULTS: We have developed at la Pitié a protocol to apply all these principles. From February 1997 to November 1998, multiple revascularization was performed on 167 patients. A total of 344 coronary anastomosis were carried out (mean: 2.05 graft/patient). CONCLUSION: Some simple rules had to be applied to reduce the rate of complications to the minimum: a good clinical case selection, use of a routine protocol, and recognize the need to adapt the indications to the learning curve of the operator.

Coronary Angiography↗

PubMed1998

Taurine might be acting as a trophic factor in the retina by modulating phosphorylation of cellular proteins.

Protein phosphorylation is involved in the regeneration of the nervous system. Taurine modulates the phosphorylation of specific proteins in the retina, and also increases outgrowth from ganglion cells. In order to test the possible role of protein phosphorylation on the outgrowth from the retina and on the trophic effect of taurine, in vitro studies were performed in the presence of phorbol and nonphorbol protein kinase C activators, the protein kinase C inhibitor tamoxifen, and phosphatase inhibitors. After crush of the optic nerve, explants of the goldfish retina were cultured and the outgrowth was evaluated by measuring the length and the density of neurites. The activation of protein kinase C decreased the outgrowth from the explants and impaired the stimulatory effect of taurine. Phosphatase inhibitors produced a similar effect on basal and taurine-modulated outgrowth. In certain concentrations, some of these drugs did not affect the emission of neurites in the absence of taurine, but decreased the effect of the amino acid. Tamoxifen also reduced the outgrowth, probably acting at other cellular levels or indicating that the regulation of outgrowth by phosphorylation is a complex and dual process. The response to the drugs, evaluated by length or density of fibers, was not the same, since rate of outgrowth was more affected than density, which suggests that both parameters are modulated at differential stages or sensitivities to the tested agents.

Animals↗

PubMed1998

Taurine-stimulated outgrowth from the retina is impaired by protein kinase C activators and phosphatase inhibitors.

Taurine increases neurite elongation of post-crush goldfish retinal explants, as well as the number of outgrowing isolated cells from goldfish and rat retina in culture. The trophic effect of taurine is related to an elevation in calcium flux rather than increased cell proliferation. Since taurine regulates phosphorylation in rat retina, we investigated if this process could be involved in the mechanism of taurine action on outgrowth. Control and taurine-supplemented post-crush goldfish retinal explants were cultured in the presence of protein kinase C activators or phosphatase inhibitors, and the length of neurites was measured after five days in culture. In some cases, there was an inhibition of the stimulatory effect of taurine without a modification in basal outgrowth. In others, outgrowth of control explants was also reduced. A certain level of protein phosphorylation seems to be critical for the trophic effect of taurine in the retina.

Animals↗

PubMed1998

Plasma concentration of taurine is higher in malnourished than control children: differences between kwashiorkor and marasmus.

Plasma free amino acids were determined in the plasma of severely malnourished children under two years of age. A total of thirty-one patients and eleven controls were evaluated: seventeen cases of kwashiorkor, eight cases of marasmus, and six cases of marasmic-kwashiorkor. Fasting plasma samples were taken in the morning on the day of admission. Fasting plasma samples were also taken from nine patients at discharge after two months in the hospital where they received a balanced diet as treatment. A partial reversal of the signs of malnutrition was observed at discharge. In the whole group of patients ad admission, lower concentrations of tyrosine, methionine, tryptophan, and leucine and higher concentrations of aspartate, glutamate, and taurine were observed compared to controls. Taurine continued to be elevated in the malnourished group at the time of discharge. Marasmic children, as compared to controls, had high aspartate and low tryptophan levels, but taurine levels were not significantly different from controls. Kwashiorkor patients had low tyrosine, methionine, tryptophan, and lysine, and significantly higher taurine plasma levels. The elevated concentration of taurine might be the result of a redistribution of this amino acid to provide specific tissues with the required amount for development.

Amino Acids↗

PubMed1998

Taurine, glutamate and GABA modulate the outgrowth from goldfish retinal explants and its concentrations are affected by the crush of the optic nerve.

The amino acid taurine plays an important trophic role during development and regeneration of the central nervous system. Other amino acid systems, such as those for glutamate and gamma-aminobutyric acid (GABA), are modified during the same physiological and pathological processes. After crushing the optic nerve, goldfish retinal explants were plated in the absence and in the presence of different amino acids and amino acid receptor agonists. The length and the density of the neurites were measured at 5 days in culture. Taurine increased the length and the density of neurites. Glutamate and glycine increased them at low concentration, but were inhibitors at higher concentration. The combination of N-methyl-D-aspartate (NMDA) and glycine produced a greater inhibitory effect than NMDA alone. NMDA or alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) added simultaneously with taurine impaired the stimulatory effect of the latter. GABA stimulated the emission of neurites in a concentration dependent manner. Hypotaurine also elevated the length of neurites, but cysteinsesulfinic acid did not produce a significant effect. The concentrations of taurine, glutamate and GABA were determined by HPLC with fluorescent detection in the retina of goldfish at various days post-crushing the optic nerve. The levels of taurine were significantly increased at 48h after the crush, and were elevated up to 20 days. Glutamate level decreased after the lesion of the optic nerve and was still low at 20 days. GABA concentration was not significantly different from the control. The interaction of these amino acids during the regenerative period, especially the balance between taurine and glutamate, may be a determinant in restoring vision after the crush.

Animals↗

PubMed1998

Serotonergic projections to the retina of rat and goldfish.

Serotonin amacrine cells have been described in the retina of vertebrates, except mouse and rat. Moreover, serotonin immunoreactive fibers have been reported in the optic nerve of rodents, frog and stingray. The purpose of this work was to study serotonin dorsal raphe nucleus projections to the retina, and to determine whether this pathway occurs in teleosts. The existence of specific connections was investigated in the rat and in the goldfish by the use of the neurotoxic substance 5,7-dihydroxytryptamine followed by monoamines determination in the retina by HPLC. The administration of 5,7-dihydroxytryptamine into the central or the lateral area of the rat dorsal raphe nucleus decreased serotonin levels in the raphe area and in the hippocampus, but only the bilateral injection into the dorsal raphe nucleus decreased it in the retina. In the goldfish, the injection of 5,7-dihydroxytryptamine into the optic tectum decreased serotonin concentration in situ and in the retina. The binding of [3H]paroxetine, a marker of serotonin transporter, was reduced in the retina of both species after the central treatment with the neurotoxic substance. In addition, the administration of the serotonin precursor 5-hydroxytryptophan into the optic tectum increased serotonin levels in the site of the injection and in the retina. The intraocular administration of 5,7-dihydroxytryptamine produced a big decrease in the content of retinal serotonin. This indoleamine and 5-hydroxyindoleacetic acid were detected in the optic nerves of rat and goldfish. The results indicate the existence of serotonergic retinopetal fibers in the rat and in the goldfish, a pathway that was not specifically demonstrated for the rat and was not previously proposed for the fish. The study of these serotonergic projections from the brain to the retina could be of interest in the understanding of the functional role of serotonin in the retina.

5,7-Dihydroxytryptamine↗

PubMed1998

[Amino acids as biochemical markers in epidemic and endemic optic neuropathies].

It was studied the amino acids pattern in serum and cerebrospinal fluid of 12 and 8 patients, respectively, suffering from optic epidemic neuropathy diagnosed between 1995 and 1997 (endemic period). The cerebrospinal fluid of 16 patients diagnosed during the epidemic (1992) was also studied. The analysis of amino acids in serum and cerebrospinal fluid was made by high resolution liquid chromatography with fluorescent detection, previous derivation with orthofthallic aldehyde. As in the previous study, no important deficiencies of the essential amino acids were observed in the serum of the studied patients. Lower concentrations of threonine, aspartic acid and taurine were found in the serum of patients with epidemic optic neuropathy diagnosed in the endemic period. Taurine plays an important role in the structure and function of photoreceptors of the retina. It is synthesized in the necessary amounts in man, so it must be supplied in the diet. As it is only present in food of animal origin, it is considered that the deficiency of taurine may contribute to the development of optic epidemic neuritis in our patients. Samples of cerebrospinal fluid were analyzed during the epidemic and endemic periods and it was observed an increase of the glutamic and aspartic acid in the epidemic period and of glutamic acid during the endemic. The excess of excitotoxic amino acids in the cerebrospinal fluid supports the previous neurocognitive studies that suggested the affection of the central nervous system in these patients.

Amino Acids↗

PubMed1997

Atypical in vitro and in vivo binding of [3H]S-14506 to brain 5-HT1A receptors.

The tritiated derivative of the potent 5-HT1A receptor agonist S-14506 ¿1[2-(4-fluorobenzoylamino)ethyl]-4-(7-methoxynaphtyl)pipera zine¿ was tested for its capacity to selectively label the serotonin 5-HT1A receptors both in vitro in the rat and the mouse brain, and in vivo in the mouse. In vitro studies showed that the pharmacological profile and the distribution of [3H]S-14506 specific binding sites (Kd = 0.15 nM) in different brain regions matched perfectly those of the prototypical 5-HT1A receptor ligand [3H]8-OH-DPAT. However, in the three regions examined (hippocampus, septum, cerebral cortex), the density of [3H]S-14506 specific binding sites was significantly higher (+66-90%) than that found with [3H]8-OH-DPAT. Whereas the specific binding of [3H]8-OH-DPAT was markedly reduced by GTP and Gpp(NH)p and increased by Mn2+, that of [3H]S-14506 was essentially unaffected by these compounds. In addition, the alkylating agent N-ethylmaleimide was much less potent to inhibit the specific binding of [3H]S-14506 than that of [3H]8-OH-DPAT. Measurement of in vivo accumulation of tritium one hour after i.v. injection of [3H]S-14506 to mice revealed marked regional differences, with about 2.5 times more radioactivity in the hippocampus than in the cerebellum. Pretreatment with 5-HT1A receptor ligands prevented tritium accumulation in the hippocampus but not in the cerebellum. Autoradiograms from brain sections of injected mice confirmed the specific in vivo labeling of 5-HT1A receptors by [3H]S-14506, therefore suggesting further developments with derivatives of this molecule for positron emission tomography in vivo in man.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

PubMed1997

Nonpenetrating clips for coronary anastomosis.

BACKGROUND: A nonsuture clip technique (nonpenetrating titanium clips applied to everted tissue edges at high compressive forces) was used to perform coronary anastomoses in a clinical setting. METHODS: Clipped coronary anastomoses were performed in 10 patients. The anastomoses incorporated the left internal mammary artery to the left anterior descending artery (n = 1) and the saphenous vein to the right coronary artery (n = 5), the posterior descending artery (n = 2), the diagonal artery (n = 2), and one vein-to-vein proximal anastomosis (n = 1). RESULTS: The mean duration for completion of the anastomoses was 15 minutes (range, 7 to 20 minutes). This time was reduced from 20 minutes at the beginning of the clinical experience to 7 minutes for the last 3 patients. No technical complication was related to clip application and all patients had uneventful outcomes. Three anastomoses studied by coronary angiography were patent without stenosis. CONCLUSION: The clipped anastomotic technique has a rapid learning curve, the same safety as suture methods, and the potential for facilitating endoscopic vascular reconstructions.

Anastomosis, Surgical↗

PubMed1997

Thoracoscopic internal mammary artery harvesting: technical considerations.

BACKGROUND: Technical details of thoracoscopic harvesting of the internal mammary artery (IMA) are reported. This procedure allows a complete dissection of the left IMA from its origin at the subclavian artery to the sixth intercostal branches with transection of all collateral branches. METHODS: Between September 1995 and September 1996, thoracoscopic harvesting of the left IMA was performed on 32 patients who had undergone a minimally invasive coronary artery bypass grafting procedure. RESULTS: There were no conversions to a standard approach because of an injury to the graft and no reoperations for bleeding. The mean duration of the IMA harvesting procedure was 58.7 minutes (range, 20 to 130 minutes). CONCLUSIONS: This procedure enlarges the field of minimally invasive coronary artery bypass grafting techniques. The thoracoscopic harvest of the full length of the IMA allows the procedure to more closely replicate the open approach.

Endoscopy↗