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L Alvarez

Publications and source records attributed to L Alvarez.

At least 127 records · Page 7Linked to original sources
PubMed1997

Antithrombotic map of the N-terminal domain of the GPIIIa subunit of the human platelet fibrinogen receptor (GPIIb-IIIa) determined in vivo by monoclonal, immunochemical inhibition of acute arterial thrombosis.

The inhibition of the platelet fibrinogen receptor, the glycoprotein IIb-IIIa (GPIIb-IIIa) or integrin alphaIIbbeta3, has recently became an accepted practice in clinical cardiology. The interest lies now in the improvement of the antithrombotic activity and the minimization of the secondary effects of the receptor inhibitors, by their evaluation in vivo in the different dynamic conditions and pathological states under which these inhibitors have to perform. In this paper, we functionally map in vivo the N-terminal domain of the GPIIIa subunit, using the antithrombotic activity of five murine monoclonal antibodies (mabs) (P37, P40, 95-1, P95-2 and P97), all of them inhibitors of platelet aggregation in vitro and directed to this ligand binding domain of the human fibrinogen receptor. Competition experiments have shown that these mabs bind with high affinity (5-7 nM) and compete very strongly among themselves for binding to human resting platelets, except P40, which neither binds nor competes. These antibodies were assayed in a dog model of acute thrombosis in the carotid artery, which were induced 15 min after their intravenous administration (0.8 mg/kg). The antithrombotic activity was quantified by the measurement of the [111In]oxine-labelled platelet deposition at the site of the arterial lesion and was expressed as the percentage of the total circulating platelets. Antibody P37, directed to the GPIIIa 101-109 sequence, decreased the platelet deposition 630-fold with respect to control animals. P95-2, P97 and P95-1 decreased the platelet deposition 160-, 32- and 25-fold, respectively, while P40, directed to the GPIIIa 260-302 sequence, did not show any antithrombotic activity. We conclude that all the mabs directed to the N-terminal domain of GPIIIa, which inhibit platelet aggregation in vitro and whose epitopes are very close to each other and exposed in resting platelets, have high antithrombotic activity in vivo, which varies depending on the actual location of the epitopes in the receptor topography. Among these antibodies, P37, the strongest receptor inhibitor in vivo and whose epitope is most probably the closest to the fibrinogen binding site(s), seems the best candidate for comparative studies in animal models with today's best GPIIb-IIIa inhibitors and for clinical trials in humans in order to arrest or prevent thrombosis, reocclusion and late restenosis.

Journal Article↗
PubMed1997

Glucocorticoid regulation of hepatic S-adenosylmethionine synthetase gene expression.

The effects of glucocorticoids on the regulation of rat liver S-adenosylmethionine synthetase were studied in vivo and in two culture systems. Livers from adrenalectomized animals were examined for enzyme activity, immunoreactive protein, and messenger RNA (mRNA) content. All three parameters showed a similar trend, i.e. they decreased 3-fold after adrenalectomy and increased over the control values upon triamcinolone replacement. These results suggested that glucocorticoid regulation of hepatic S-adenosylmethionine synthetase was mediated at the mRNA level. Triamcinolone and dexamethasone increased S-adenosylmethionine synthetase mRNA content in a time- and dose-dependent manner in both rat hepatoma H35 cells and primary cultures of adult rat hepatocytes. The kinetics of mRNA induction were identical in both culture systems, indicating that the hormone-mediated response is independent of the differentiated state of the cell. Insulin blocked the inducing effect of glucocorticoids on S-adenosylmethionine synthetase mRNA in a dose-dependent manner. On the other hand, the triamcinolone-dependent increase in mRNA levels was completely abolished by treatment with actinomycin D, whereas cycloheximide did not affect this response. The transcription rate of the gene, as measured by run-on assay, increased 3-fold after hormone addition. Transient transfections of H35 cells with 1.4 kilobases of the 5'-flanking region of the hepatic S-adenosylmethionine synthetase gene fused to a luciferase reporter gene showed that promoter activity is also increased 3-fold after triamcinolone treatment, suggesting that this promoter region contains the sequence elements necessary to confer glucocorticoid responsiveness. In addition to the transcriptional control of the hepatic S-adenosylmethionine synthetase gene, our results suggest that glucocorticoids may be acting at a posttranscriptional level.

Adrenalectomy↗
PubMed1997

Structural and ultrastructural study of the myocardium after 24-hour preservation in University of Wisconsin solution.

This study deals with myocardial preservation after 24 hours of continuous perfusion with University of Wisconsin (UW) solution, focusing on the morphological changes produced by preservation and reperfusion, and their possible relationship to the composition of the solution and the immediate hemodynamic findings after orthotopic heart transplantation in dogs. Following preservation, the histological images of the myocardium were normal in almost every case. After revascularization, although 50% of the hearts recovered preoperative functional levels, there were relevant histological changes in all of them. These changes consisted of interstitial edema, damage to at least 50% of the mitochondria, presence of contraction bands, loss of intercellular junction structure and vasoconstriction. According to these results, the association of continuous perfusion and UW solution appears to be effective during the ischemic period of prolonged myocardial preservation, but not during reperfusion. Perhaps the proportion of viable hearts could be increased by preventing vasoconstriction.

Animals↗
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↗
PubMed1997

[Contribution of thalamic somatosensory evoked potentials to stereotaxic thalamometry].

INTRODUCTION: In order to use short latency somatosensory thalamic evoked potentials (PES) to locate therapeutic targets in functional surgery, thalamic PES were recorded during stereotactic thalamotomy in 25 patients with Parkinson's disease, using a concentric bipolar semi-micro-electrode, 4 mm in diameter. In the 72 trajectories planned. 628 registers were made, obtaining 314 PES in 55 trajectories. These recordings were divided into 5 groups, according to the electrical variables evaluated in each case (absolute latency, inter-peak latency, absolute amplitude and number of phases). MATERIAL AND METHODS: The electrophysiological characteristics of the PES groups obtained, and the spatial representation of these in a tridimensional system of coordinates, is shown. We analyze the sequence of the groups of potentials in each of the trajectories followed. CONCLUSIONS: We consider that the limit between adjacent nuclear edges, ventral intermediate (Vim)-ventro-caudal (Vc), may be represented by the transition of potentials in group 1 to potentials in group 4 and/or potentials of group 3 to those of group 2. This study shows that thalamic PES are useful for locating targets during stereotactic thalamotomy.

Adult↗
PubMed1997

Surgical treatment of Parkinson's disease.

At present, there are three major surgical approaches to Parkinson's disease (PD): (1) Ablative surgery (i.e. pallidotomy, thalamotomy); (2) deep brain stimulation (DBS) of the thalamus, internal globus pallidus (GPi) and subthalamic nucleus (STN); and (3) grafting fetal mesencephalic cells into the striatum. As a result of increasing understanding of the pathophysiology of the basal ganglia and the demonstration of surgical alleviation of experimental parkinsonism, surgery has regained a paramount importance in the management of PD. The aim of pallidotomy and DBS is to reduce the excessive inhibitory output from the GPi and substantia nigra reticulata (SNr). Pallidotomy and DBS of the STN or GPi aim to reverse the pathophysiological consequences of dopamine deficiency in PD, and should be considered entirely symptomatic treatments. The ideal candidates for pallidotomy are young patients in good general health in whom dyskinesias are the main reasons for disability. Patients with severe bilateral problems uncontrollable with present pharmacological tools are candidates for DBS. As yet, there are no formal data to help decide how to choose between GPi and STN stimulation. In our practice, patients are allocated to GPi stimulation when 'on' dyskinesias are extremely severe. In most other instances, we prefer to perform STN stimulation. At present there is almost no reason to decide for the thalamic stimulation since tremor is equally arrested by STN stimulation, which in addition improves all other features of PD. Equally the only indication for thalamotomy would be a patient with long-standing tremor as the main clinical manifestation, which can not be controlled with drugs. The proportion of patients in whom the thalamus will be the preferable target for either DBS or thalamotomy is small (less than 5%). Grafting aims to repair the nigrostriatal pathway and restore dopaminergic function in the striatum. In the future implants containing not only dopaminergic cells but also growth factors and a variety of other substances could become a method to not only functionally compensate the biochemical abnormalities of PD but also to arrest its progression. This technique is limited to a few centres around the world owing to the technical, logistical and ethical problems of obtaining and handling embryonic cells. At present, grafting of dopaminergic cells is perhaps best suited for patients with young-onset PD (less than 45 years old) who are at high risk of developing complications within a short time of beginning pharmacological treatment and in whom the idea of making lesions or implanting electrodes into the brain for decades seems less appealing. Consideration of surgery in any given patient should be weighed against the risks (about 1% mortality and 2-6% of severe morbidity-hemiplegia, cognitive deficit, speech problems, etc.) associated with these techniques. The development of better imaging methods and the growing expertise of multidisciplinary teams will undoubtedly make surgery for PD safer and more effective in the future.

Electric Stimulation Therapy↗
PubMed1996

Increased sensitivity to oxidative injury in chinese hamster ovary cells stably transfected with rat liver S-adenosylmethionine synthetase cDNA.

Chinese hamster ovary cells were stably transfected with rat liver S-adenosylmethionine synthetase cDNA. As a result, S-adenosylmethionine synthetase activity increased 2.3-fold, an effect that was accompanied by increased S-adenosylmethionine, a depletion of ATP and NAD levels, elevation of the S-adenosylmethionine/S-adenosylhomocysteine ratio (the methylation ratio), increased DNA methylation and polyamine levels (spermidine and spermine), and normal GSH levels. By contrast, the transfected cells showed normal growth curves and morphology. Exposure to an oxidative stress by the addition of H2O2 resulted in a greater consumption of ATP and NAD in the transfected cells than in the wild-type cells. In turn, cell killing by H2O2 was greater in the transfected cells than in the wild-type cells. This killing of Chinese hamster ovary cells by H2O2 involved the activation of poly(ADP-ribose) polymerase with the resultant loss of NAD and ATP. 3-Aminobenzamide, an inhibitor of poly(ADP-ribose) polymerse, but not the antioxidant N,N'-diphenylphenylenediamine, prevented the killing of Chinese hamster ovary cells by H2O2 and maintained the contents of NAD and ATP. The results of this study indicate that a moderate activation of the synthesis of S-adenosylmethionine leads to ATP and NAD depletion and to a greater sensitivity to cell killing by oxidative stress.

Adenosine Triphosphate↗
PubMed1996

Site-directed mutagenesis of rat liver S-adenosylmethionine synthetase. Identification of a cysteine residue critical for the oligomeric state.

We have examined the functional importance of the cysteine residues of rat liver S-adenosylmethionine synthetase. For this purpose the ten cysteine residues of the molecule were changed to serines by site-directed mutagenesis. Ten recombinant enzyme mutants were obtained by using a bacterial expression system. The same level of expression was obtained for the wild type and mutants, but the ratio of S-adenosylmethionine synthetase between soluble and insoluble fractions differed for some of the mutant forms. The immunoreactivity against an anti-(rat liver S-adenosylmethionine synthetase) antibody was equivalent in all the cases. Effects on S-adenosylmethionine synthetase activities were also measured. Mutants C57S, C69S, C105S and C121S showed decreased relative specific activity of 68, 85, 63 and 29%, respectively, compared with wild-type, whereas C312S resulted in an increase of 1.6-fold. Separation of tetramer and dimer forms for wild type and mutants was carried out by using phenyl-Sepharose columns. The dimer/tetramer ratio was calculated based on the activity and on the protein level estimated by immunoblotting. No monomeric forms of the enzyme were detected in any case. Comparison of dimer/tetramer ratios indicates the importance of cysteine-69 (dimer/tetramer protein ratio of 88 versus 10.2 in the wild type) in maintaining the oligomeric state of rat liver S-adenosylmethionine synthetase. Moreover, all the mutations carried out of cysteine residues between cysteine-35 and cysteine-105 altered the ratio between oligomeric forms.

Amino Acid Sequence↗
PubMed1996

Inverse expression of mdr 1 and c-myc genes in a rhabdomyosarcoma cell line resistant to actinomycin d.

Cytotoxic agents used in cancer therapy may induce differentiation in tumour cells with no proliferative potential. However, chemotherapy can also induce multidrug resistance, a formidable obstacle to the successful treatment of tumours. Both events were recently shown to occur in a rhabdomyosarcoma cell line (RD-DAC) resistant to actinomycin D, a drug of choice in the treatment of these tumours. To analyse this connection, cell line RD cultures were investigated with progressive concentrations of actinomycin D and it was shown that a minimum dose (1.2 x 10(-6) mM) of the drug was necessary to increase mdr 1 mRNA in RD-DAC. The mechanism of mdr 1 overexpression was an increase in the number of copies of the mdr 1 gene, although the mRNA levels were not correlated with mdr 1 amplification. Drug resistance mediated by mdr 1 overexpression coincided with the development of myogenic differentiation in RD-DAC and with a decrease in c-myc mRNA levels, whereas levels of N-myc mRNA showed no modulation. These findings suggest that factors implicated in cell proliferation and differentiation, such as c-myc, may be responsible for the control of genes related to the development of multidrug resistance in rhabdomyosarcomas. Modulation of these factors may determine the sensitivity of rhabdomyosarcoma cells to drugs and may play an important role in triggering the differentiation programme found in these resistant rhabdomyosarcoma cells.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

Experimental study of the antithrombogenic behavior of Dacron vascular grafts coated with hydrophilic acrylic copolymers bearing salicylic acid residues.

The objective of the present work was study of the behavior of active coatings of hydrophilic acrylic polymers bearing salicylic acid residues linked covalently to the macromolecular chains, after their application to woven and knitted Dacron vascular grafts. In vitro tests were carried out under dynamic flow conditions using equipment especially designed to reproduce physiologic conditions, to determine the retention of the coating using a saline solution. Ex vivo tests were carried out in an extracorporeal circuit using the dog as an animal model. The study of the deposition of platelets was followed by labeling of autologous platelets with 111In-oxine, as well as by analysis of the surfaces of the prostheses by scanning electron microscopy. An application of thin coatings of hydrophilic acrylic copolymers improves the antithrombogenicity of the vascular grafts with respect to the uncoated prosthesis. The presence of relatively small amounts of units bearing salicylic acid residues in the copolymer chains (5-20 wt %) gives good results when they are applied to woven and knitten Dacron meshes which have been quantified by analysis of the percentage of radiotracer on the surface of the vascular grafts tested in ex vivo experiments. The salicylic acid residues are released slowly to the medium by hydrolysis of the reversible covalent bonds of this compound to the acrylic macromolecular chains, which provides an additional antiaggregating effect for platelets. The polymeric coating forms a thin active film which improves the antithrombogenic properties of the surface of woven or knitted Dacron vascular grafts in ex vivo experiments.

Animals↗
PubMed1996

Language cortex representation: effects of developmental versus acquired pathology.

Relatively little is known about language cortex representation in patients with developmental pathology and epilepsy. We report the results of mapping language by electrical stimulation of chronically implanted subdural electrodes in 34 patients (mean age, 12.2 years) evaluated for epilepsy surgery, 28 of whom had magnetic resonance imaging or histological evidence of developmental tumors or cortical dysplasia. Language cortex was identified in the temporal or frontal lobe of 19 patients (left hemisphere in 17, right hemisphere in 2), and overlapped or bordered the epileptogenic region in 12. Language cortex was not found in the frontal or temporal lobe of 15 patients (left hemisphere in 4, right hemisphere in 11) and was presumed to be contralateral to grid placement. Three patients with left-hemisphere perinatal or postnatal cerebral insults before the age of 5 years had no language in the left hemisphere, while 3 patients with insults between the ages of 6 and 16 years had preserved left-hemisphere language. Developmental lesions and early-onset seizures do not displace language cortex from prenatally determined sites, whereas lesions acquired before the age of 5 years may cause language to relocate to the opposite hemisphere, but only when language cortex is destroyed.

Adolescent↗
PubMed1996

Effect of venovenous bypass on perioperative renal function in liver transplantation: results of a randomized, controlled trial.

Although venovenous bypass (VVBP) has been suggested to protect the kidneys during liver transplantation and its systematic use has therefore been recommended, this beneficial effect of VVBP has not been clearly demonstrated. In a prospective, randomized, controlled trial, 77 patients receiving liver transplants for chronic liver disease were allocated to be supported with VVBP (group 1, 38 patients) or not (group 2, 39 patients). Both groups were similar in relation to preoperative clinical and laboratory data and operative transfusion requirements. Inulin clearance and urinary beta(2)-microglobulin and N-acetyl-beta-D-glucosaminidase (NAG) excretion (to determine glomerular filtration rate and tubular damage, respectively) were measured at different perioperative periods (anesthesia induction, hepatectomy, anhepatic phase, biliary anastomosis, and 24 hours after surgery). A significant decrease in inulin clearance and increase in tubular damage markers were observed in the anhepatic phase, which only partly improved in the subsequent phases. No significant differences were observed between groups 1 and 2 at any perioperative phase, except during the anhepatic phase, in which a more marked renal function impairment occurred in group 2 patients. However, renal function on the 7th postoperative day and the need for hemodialysis/ hemofiltration during the 1st week were similar in both groups. Among 40 variables analyzed, only low mean arterial pressure at anesthesia induction was identified as an independent predictor for early postoperative severe renal failure (inulin clearance < 10 mL/min/1.73 m(2) at the 24th postoperative hour), with no significant relationship between this complication and the use of venovenous bypass. Renal function markedly deteriorates during liver transplantation, and renal impairment persists during the early postoperative period. Because VVBP support is not associated with any clear benefit in renal function, its systematic use does not seem to be justified.

Acute Kidney Injury↗
PubMed1996

Differential expression pattern of S-adenosylmethionine synthetase isoenzymes during rat liver development.

The pattern of expression of liver-specific and extrahepatic S-adenosylmethionine (SAM) synthetase in developing rat liver was established by determining steady-state levels of the respective messenger RNAs (mRNAs) and protein content. Levels of liver-specific SAM synthetase mRNA increased progressively from day 20 of gestation, increased 10-fold immediately after birth, and reached a peak at 10 days of age, decreasing slightly by adulthood. Conversely, mRNA levels of extrahepatic isoenzyme decreased toward birth, increased threefold in the newborn, and decreased further in the postnatal life, reaching a minimum in the adult. Similar expression profiles were observed in isolated hepatocytes, indicating that both mRNAs are differentially regulated in the same cell type. Western blot analysis showed that levels of immunoreactive liver-specific isoenzyme followed a trend similar to the mRNA, indicating that developmental regulation of this enzyme is mediated at the mRNA level. Developmental patterns of expression of albumin and alpha-fetoprotein (AFP) mRNAs were closely related to those for liver-specific and extrahepatic isoenzymes, respectively. Therefore, it is suggested that liver-specific SAM synthetase may be a marker for hepatocyte differentiation. Incubation of primary cultures of hepatocytes from 21-day-old fetuses with permeant cyclic adenosine monophosphate (cAMP) analogues elicited an up-regulation of the mRNA for the liver-specific isoenzyme with a concomitant down-regulation of the extrahepatic message, suggesting a physiological role for the increased postnatal glucagonemia in the control of this isoenzyme switching. In contrast with the isoenzyme expression profiles, the levels of SAM, the product of SAM synthetase reaction, were determined to be greater during gestation than in immediate postnatal periods. These results indicate that synthesis and utilization of SAM may be regulated differentially in fetal and adult hepatocytes.

Albumins↗
PubMed1996

Subchronic toxicity and teratogenicity of 2-chloro-1,1,1,2-tetrafluoroethane (HCFC-124).

Inhalation studies were conducted to determine the potential toxicity of HCFC-124. Groups of rats and mice were exposed to HCFC-124 6 hr/day, 5 days/week for 13 weeks at 0, 5000, 15,000, and 50,000 ppm. Subgroups of rats and mice were held for a 1-month recovery period. A functional observational battery (FOB) was conducted on rats at 0, 4, 13, and 16 weeks. Clinical pathology evaluations were conducted at 7, 13, and 17 weeks. Thirteen or 17 weeks after study initiation, rats and mice underwent gross and microscopic evaluation, and livers were evaluated for hepatic beta-oxidation activity. In addition, groups of female rats and rabbits were exposed to HCFC-124 by inhalation during gestation to 0, 5000, 15,000, or 50,000 ppm. Exposure of rats and mice to HCFC-124 caused minimal compound-related effects. Compound-related changes occurred in several clinical pathology parameters in rats and mice. Hepatic beta-oxidation activity was significantly higher in 5000, 15,000, and 50,000 ppm male mice; however, there were no compound-related effects on beta-oxidation activity in rats. During the daily exposures, rats, mice, and rabbits exposed to 50,000 ppm were less responsive to auditory stimuli or less active compared to controls. At the 13-week FOB, male rats exposed to 15,000 or 50,000 ppm had decreased arousal. There were no compound-related effects on mortality, clinical signs, ocular tissues, hematology parameters, organ weights, and tissue morphology at any concentration in rats or mice. Maternal toxicity in rats was evident by a significant decrease in weight gain and food consumption at 50,000 ppm. Similarly, 50,000 ppm pregnant rabbits had lower food consumption. However, for both rats and rabbits, there was no evidence of fetal toxicity at any concentration.

Administration, Inhalation↗
PubMed1996

Low concentrations of actinomycin D potentially cause therapeutic differentiation in human rhabdomyosarcoma cell line RD.

Neoplastic transformation may be an alteration in the process of cell maturation that leads to an infinite capacity for proliferation. Because the cytodestruction caused by most drugs available for cancer chemotherapy is often accompanied by significant morbidity and poor response, the induction of differentiation has been proposed as an alternative approach to conventional anticancer therapy. We used human rhabdomyosarcoma cell line RD to analyze the differentiation process induced by actinomycin D, a drug of choice in the conventional treatment of rhabdomyosarcomas. Low concentrations of actinomycin D induced a terminal process of morphological and ultrastructural myogenic differentiation in rhabdomyosarcoma cells, which concluded with cell death. However, this potential therapeutic effect cannot be considered complete because of the presence of tumoral cells that are heterogeneous with respect to actinomycin D chemosensitivity. This heterogeneity led to the appearance of foci of resistant cells which, despite their greater degree of differentiation in comparison with the parental cell line, escaped from terminal myogenic differentiation. This subgroup of tumoral cells may be responsible for the failure of cytotoxic treatment.

Cell Differentiation↗