Partial recovery from rabies in a nine-year-old boy.
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Biomedical subjects
Publications and source records attributed to L Alvarez.
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In March 1993, a patient with idiopathic Parkinson disease who had received an adrenal brain autograft 6 years before underwent bilateral stereotactic transplantation of a mesencephalic cell suspension into the striatum, in order to prevent further deterioration in his neurological condition. During the CT-guided surgical planning and the transplantation procedures, electrophysiological evidence of the adrenal graft viability was assessed. Based on these findings, we drew the preliminary conclusion that the chromaffin tissue implanted in April 1987 is still functional, which explains the clinical evolution of this patient during the postoperative long-term follow-up.
A further trial of CT-oriented microrecording guided stereotactic selective thalamotomy was conducted at the Centro Internacional de Restauración Neurológica, Havana City as treatment of resting tremor in 11 patients with idiopathic Parkinson's disease (PD), and in 3 other patients with intentional tremor associated with multisystemic atrophy and cerebral palsy. Three of the parkinsonian patients had undergone fetal mesencephalic tissue transplantation with significant improvement of the most debilitating symptoms of PD and stabilization of the motor state, but predominantly unilateral tremor had impaired them progressively despite increased levodopa doses. A Leksell frame was used with a novel surgical planning system and electrophysiological recordings to identify the optimal target point inside the ventralis intermedius. In all but 1 case, the tremor was totally arrested. No persistent complications were observed.
This paper summarizes the results of three controlled clinical trials related to the transplantation of embryonic ventral mesencephalic tissue into the striatum of 46 idiopathic parkinsonian patients exhibiting motor complications on standard levodopa therapy. From January 1988 to April 1990, 30 subjects with fluctuating Parkinson's disease received fetal dopaminergic tissue implants by the open microsurgical technique. In March 1992 the stereotactic approach was adopted for successive fetal mesencephalic cell suspension transplants (7 unilateral and 9 bilateral) into the caudate and putamen of parkinsonian patients with levodopa-induced complex fluctuations and dyskinesias. The neurological assessment performed 12 months before and 3-18 months after transplantation demonstrated a reduction of both the daily time spent in the 'off' condition and the number of daily 'off' periods, and a significant improvement of the motor scale. The stereotactic selective thalamotomy with microelectrode recording was introduced in January 1993, in order to provide a further potential treatment strategy; i.e., the combination of the two surgical trends in Parkinson's disease, the restorative neurotransplantation technique, and the selective lesional approach. In addition to that, microelectrode recording is also used for implantation site selection and functional characterization.
From March 1991 to September 1993, 26 patients (aged 4-78 years) with brain tumors (4 glioblastoma multiforme, 10 nonglioblastoma multiforme, 1 mixed oligoastrocytoma, 2 carniopharyngiomas, 2 meningiomas and 7 metastases) were treated with stereotactic techniques at the Centro Internacional de Restauración Neurológica, La Habana, Cuba. A total of 28 stereotactic surgical procedures were performed with no operative mortality; they included biopsies in all cases, 1 stereotactic microsurgical resection and 12 permanent implants of 192Ir, followed by external beam fractionated radiation therapy (40-60 Gy). The present paper shows that the combined use of a stereotactic approach, a comprehensive and reliable stereotactic dosimetric planning system, stereotactic brachytherapy with 192Ir and complementary percutaneous radiation treatment constitutes a promising strategy for brain tumor management.
In the present study we quantified the contractile protein troponin-T at the cellular and subcellular level in chick embryo cardiomyocytes to investigate the modulation of cardiac development by catecholamines. We analyzed the effects of these drugs on cultures of chick cardiomyocytes obtained from Hamburger and Hamilton's (HH) stage 21, HH stage 29 and HH stage 40 embryos; cardiomyocytes are considered to be mature at HH stage 40. We analyzed the modifications these drugs induced in the transcription of the gene for chick cardiac troponin-T. Sodium dodecyl sulfate-gel electrophoresis and immunobloting showed that cytoplasmic and cytoskeletal concentrations of troponin-T are dependent on the stage of embryonic development analyzed, and on the type of catecholamine added to the culture. The most significant finding was the increase in troponin-T mRNA in the chick heart at HH stage 40, accompanied by an increase in the increase in the expression of this protein in the cytoskeletal compartment after treatment with norepinephrine. At HH stage 21, norepinephrine induced less marked changes in the accumulation of troponin-T in comparison with untreated cardiomyocytes.
We produced and characterized a specific monoclonal antibody (mAB) designated GR-ICOR-2. This mAb recognizes sarcomeric actin molecules (43 kDa) and was used in an immunohistochemical analysis of staining patterns in Hamburger and Hamilton's stages 18, 22 and 25 (HH 18, 22 and 25) embryonic chick hearts. Staining showed a mainly cytoplasmic distrubition in three regions: the atrioventricular (AV) canal cushion tissue, the primitive ventricle, and conal crests. In addition, this mAb-cross-reacted with rabbit and human cardiac and skeletal muscle tissue; but not with smooth muscle tissue.
The emergence of drug-resistant tumor cells remains a major problem in cancer chemotherapy. Resistance to multiple unrelated antineoplastic drugs may be related, in part, to expression of the P-glycoprotein. The cell line RD, derived from an embryonic rhabdomyosarcoma tumor, was used as an in vitro model to examine the development of drug resistance. A cell line resistant to actinomycin D (RD-DAC) was developed by growing RD in increasing concentrations of the drug. The ID50 (concentration of drug needed to induce a 50% reduction in cell growth) of the resultant line to actinomycin D was more than 15 times that of the parental line. The resistant line was cross-resistant to vincristine and doxorubicin. Resistance to actinomycin D resulted in increased P-glycoprotein expression, which was associated with a change in desmin and vimentin expression. These results suggest that exposure to chemotherapeutic drugs can induce not only classical multidrug resistance, but also a process of cellular differentiation in rhabdomyosarcoma cells.
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The epidermal growth factor receptor is related with processes of cell replication and differentiation. We used the intermediate filament protein desmin as a marker to study the relation between muscle cell differentiation and modifications in the expression of this receptor during heart development in the chick embryo. Epidermal growth factor receptor was expressed as early as Hamburger and Hamilton's stage 17, when myocardiocytes are still poorly differentiated and desmin-negative. Expression became steadily weaker as the heart matured, and decreased after Hamburger and Hamilton's stage 25, a key stage in heart maturation characterized by a sharp increase in desmin expression. Our findings suggest that in the chick embryo, the expression of epidermal growth factor receptor becomes steadily weaker as myocardiocyte differentiation progresses.
The sequence of a full-length cDNA coding for human liver S-adenosylmethionine synthetase has been determined. It spans 3217 nucleotides and encodes a protein of 395 amino acid residues, with a calculated molecular mass of 43,647 Da. The structural features deduced from the amino acid sequence show a close similarity to those of the rat liver enzyme. The liver-specific S-adenosylmethionine synthetase gene appears to be present as a single copy in the genome, as revealed by Southern analysis. The occurrence of a single mRNA species for this enzyme has been determined by primer extension and Northern analysis. Among several human tissues examined, this gene is expressed only in the liver. Similar S-adenosylmethionine synthetase mRNA levels have been detected in biopsies from normal human liver and from patients with alcoholic cirrhosis and hepatocellular carcinoma. Based on these results, a possible mechanism of regulation of human liver S-adenosylmethionine synthetase is discussed.
Telomeric DNA is composed of highly conserved sequences which are present at the termini of chromosomes as well as at intrachromosomal locations. Here, we studied a Chinese hamster ovary (CHO) cell line, BL-10, with highly stable amplified telomeric DNA at the termini as well as at intrachromosomal locations. We show that intrachromosomal or interstitial telomeric sites in this cell line and in another CHO cell line, HA-I, are radiosensitive in that they are more prone to breakage than would be expected based on the percentage of the genome composed of telomeric sequences. The frequency of breakage at interstitial telomeric sites is 4.3 to 8.3 times higher than that in the CHO genome overall. These conclusions are reached by both conventional cytogenetic analysis of two CHO cell lines which have the same survival rates after exposure to ionizing radiation, and by use of double fluorescence in situ hybridization (FISH) with a pan-telomere-specific probe and a CHO chromosome-specific library in the same metaphase cells after irradiation.
The developmental toxicity of trans-1,2-dichloroethylene (t-DCE), a component of certain Freon cleaning agents, was examined in pregnant rats. t-DCE was administered by inhalation 6 hr daily on Days 7-16 of gestation (the day copulation was confirmed was termed Day 1 of gestation) at exposure levels of 0, 2000, 6000, or 12,000 ppm. The offspring were then examined on Day 22 of gestation. Overt maternal toxicity was expressed as a significant reduction in weight gain at 12,000 ppm and in feed consumption at 6000 and 12,000 ppm. During the exposure period, lacrimation and stained periocular hair, and signs of ocular irritation, were observed in all groups. In addition, increased incidences of alopecia, lethargy, and salivation were observed in the high-dose dams. Significant increases in the mean number of resorptions per litter were seen in the litters of dams exposed to 6000 and 12,000 ppm of t-DCE; however, these values are within the range of historical controls and not considered to be treatment related. The mean combined and female fetal weights were significantly reduced in the litters of dams exposed to the highest concentration (12,000 ppm) of t-DCE. Marginal effects on feed consumption, unaccompanied by other changes and reflective of the pattern seen at higher doses, were seen at 2000 ppm. Thus, marginal maternal toxicity was seen at 2000 ppm and exposures to 6000 ppm t-DCE or higher caused frank maternal toxicity while the fetus was affected only at 12,000 ppm. Therefore, t-DCE is not considered to be uniquely toxic to the rat conceptus.
We used Western blot, a highly sensitive technique that detects amounts of protein as low as 0.1 to 1.0 ng, to investigate the possible presence in the blood stream of the contractile protein alpha-actin in 29 patients diagnosed with angina pectoris (Braunwald's classification). Circulating protein was identified with a monoclonal antibody specific for cardiac alpha-actin. Of the 20 control samples of blood, the immunoblot results were negative for alpha-actin in 19. Of the 30 patients with skeletal muscle damage caused by surgery, 27 were negative for circulating alpha-actin. Of the 29 patients with angina pectoris, circulating alpha-actin was found in 19 as a 43 kDa band in immunoblots. Of the four patients with anterior acute myocardial infarction, mean concentration of circulating alpha-actin was 58 mg/l. Among the patients with angina pectoris, the highest circulating concentrations (mean 40 mg/l) was found in those with prolonged angina (class III B, according to Braunwald's classification). In the entire group of individuals with angina pectoris alpha-actin was detectable in serum for up to 175 h after the onset of pain, and showed two peaks, one at 1 h (112 mg/l) and one at 50 h (82 mg/l) after the onset of pain. These findings reinforce the notion that unstable angina should be considered a serious condition.
We report our preliminary results related to CT-guided stereotactic transplantation of foetal ventral mesencephalic cell suspension into the striatum of five patients with idiopathic Parkinson's disease. The mean age was 51 years, the evolution time of the disease ranged from 7 to 14 years, and all of them had motor complications associated with chronic L-dopa therapy. The patients were evaluated according to the Core Assessment Program for Intracerebral Transplantations (CAPIT) for one year before and three months after surgery. The postoperative clinical assessment demonstrated significant improvement of neurological symptoms and reduction of daily L-dopa dosage.
We used Western-blot analysis to investigate the possible presence in the bloodstream of the contractile protein alpha-actin in 70 patients diagnosed with acute myocardial infarction on the basis of clinical, electrocardiographic and laboratory (creatine kinase and lactate dehydrogenase) criteria. Circulating protein was identified with a monoclonal antibody specific for cardiac alpha-actin. Of the 70 control samples of blood, the immunoblot results were negative for alpha-actin in 98% of the cases. Of the 30 patients with skeletal muscle damage caused by surgery, 26 were negative for circulating alpha-actin. Of the 70 patients with acute myocardial infarction, circulating alpha-actin was found in 67 (95%) as a 43 kDa band in immunoblots; the highest circulating concentrations (0.0580 micrograms/microliters) were found in those with anterior acute myocardial infarction. Circulating alpha-actin was detected in samples taken between 1 and 180 h after the onset of pain, and showed a biphasic pattern of appearance. Our findings for serum alpha-actin, together with the relationship between serum concentrations of this protein and sex (p = 0.001), tobacco use (p = 0.007) and postepisode complications (p = 0.002), should make it possible to gain a deeper understanding of acute myocardial infarction as a clinical entity.
We analyzed the influence of 6 and 24 h of treatment with the fibric acid derivatives bezafibrate (10 micrograms/ml), gemfibrozil (23 micrograms/ml), and fenofibrate (30 micrograms/ml) on alpha-actinin, troponin-T, and tropomyosin proteins in the cytoplasmic and cytoskeletal fractions of cultured chick myocardiocytes. The findings with sodium dodecyl sulfate-gel electrophoresis and immunoblotting showed that all three drugs modified cellular and subcellular protein levels in different ways: bezafibrate and fenofibrate produced the most significant alterations in both fractions, modifying alpha-actinin, troponin T, and tropomyosin compartmentalization in myocardiocytes, whereas gemfibrozil altered these proteins less notably. Given the role of these proteins in heart muscle contraction, fibric acid derivative-induced changes may be related with the secondary effects of these drugs on heart rhythmicity.
The right ventricle was studied in 75 anatomically normal swine hearts. Nine parameters in the papillo-tendino valvular system and three corresponding to the tricuspid orifice, pulmonary orifice and length of the inflow tract were measured. Correlations were established between the parameters and heart weight in grams, between the different parameters themselves, and between heart weight and body weight. The results were compared with similar data from human hearts, and were considered of use to researchers planning to use the swine heart as an experimental model to study congenital or induced heart diseases, or as a reference for the clinical interpretation of spontaneous cardiac, anomalies in swine.