Older donors in kidney transplantation.
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Biomedical subjects
Publications and source records attributed to A Díaz.
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Pachydysostosis of the fibula is an isolated deformity of the fibula with bowing and lengthening. Only one report has appeared in the literature, describing four lesions which underwent spontaneous regression during early childhood; no therapy was recommended. We report a 12-year-old boy with this type of deformity, who developed progressive angulation and deformation of the ankle joint. An osteotomy was necessary to realign the fibula. One year later the patient was free of symptoms.
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Cytochrome c6 from the cyanobacterium Synechocystis 6803 has been isolated and purified to electrophoretic homogeneity. The gene coding for such a heme protein (petJ) has been cloned and properly expressed in E. coli. This procedure yields a protein preparation completely identical to that obtained from the cyanobacterial cells. The N-terminal amino acid sequences of cytochrome c6 synthesized in both organisms are the same, thus allowing us to conclude that the petJ gene product is correctly processed in E. coli. To the best of our knowledge, this is the first time that any cytochrome c6 is produced in the enterobacterium. The identical physicochemical and kinetic properties of the proteins isolated from both sources confirm that expression of the petJ gene in E. coli is an adequate tool to address the study of Synechocystis cytochrome c6 by site-directed mutagenesis in a parallel way to that carried out with plastocyanin from the same organism.
Plastocyanin and cytochrome c6 from the green alga Monoraphidium braunii reduce the photo-oxidized algal photosystem I (PSI) reaction center chlorophyll (P700) with similar kinetics, as expected from their functional equivalence. The observed P700+ reduction rate constants show a non-linear dependence on metalloprotein concentration, which indicates a (minimal) two-step kinetic mechanism involving complex formation prior to electron transfer. The dependence of the observed rate constants on NaCl concentration suggests that the electrostatic interaction forces between the negatively charged donor proteins and PSI are repulsive at neutral pH and relatively low ionic strength (I), although attractive dipole-dipole interactions may play a role at higher ionic strengths. Activation parameters for P700+ reduction by cytochrome c6 and plastocyanin have been determined by studying the temperature dependence of the respective rate constants at varying ionic strength and pH. Changes in NaCl concentration and pH induce significant changes in the activation free energy of the overall reaction, even though the corresponding values for activation enthalpy and entropy undergo changes in opposite directions. Such a compensation effect between enthalpy and entropy is observed with both cytochrome c6 and plastocyanin. Protein concentration dependencies of the observed rate constants at different temperatures has allowed an estimate of the free energy change during complex association, as well as the activation parameters for electron transfer, according to a two-step kinetic model.
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The appearance and distribution of 5-HT1A receptors in the human cerebellum during the ontogenetic development was studied in samples from a group of fetal, neonatal and adult cases, by means of quantitative autoradiography and membrane binding. [3H]8-OH-DPAT was used as a ligand. High densities of 5-HT1A receptors were found during the fetal and neonatal stages over the whole cerebellar cortex, with a slight predominance in the external part, in a band which included the molecular layer. In contrast, the adult cerebellum was nearly devoid of such sites. This finding supports a role for 5-HT1A receptors in the regulation of development in the human cerebellum.
The polymerase activity of DNA polymerase I is important for the establishment of the pLS1 replicon by reconstitutive assembly in Streptococcus pneumoniae after uptake of exogenous pLS1 plasmid DNA. In polA mutants lacking the polymerase domain, such establishment was reduced at least 10-fold in frequency. Chromosomally facilitated establishment of pLS1-based plasmids carrying DNA homologous to the host chromosome was not so affected. However, both types of plasmid transfer gave mostly small colonies on initial selection, which was indicative of a defect in replication and filling of the plasmid pool. Once established, the pLS1-based plasmids replicated in polA mutants, but they showed segregational instability. This defect was not observed in strains with the wild-type enzyme or in an S. pneumoniae strain that encodes the polymerase and exonuclease domains of the enzyme on separate fragments. The role of DNA polymerase I in stably maintaining the plasmids depends on its polymerizing function in three separate steps of rolling-circle replication, as indicated by the accumulation of different replication intermediate forms in polA mutants. Furthermore, examination of the segregational stability of the pLS1 replicon in an Escherichia coli mutant system indicated that both the polymerase and the 5'-to-3' exonuclease activities of DNA polymerase I function in plasmid replication.
Laser-flash absorption spectroscopy has been used to investigate the kinetics of electron transfer from reduced cytochrome c6 and plastocyanin, isolated from Anabaena PCC 7119, to oxidized P700 in photosystem-I particles isolated from the same cyanobacterium and from spinach. For all metalloproteins and photosystems, the observed rate constant has a non-linear protein-concentration dependence, thus suggesting complex formation preceding electron transfer. Plastocyanin and cytochrome c6 have similar association constants for complex formation with spinach photosystem I, but the copper protein exhibits a higher intracomplex-electron-transfer rate constant (twofold). With Anabaena photosystem I, the two redox proteins are more effective with respect to both complex formation (5-10-fold) and electron transfer (1.5-4-fold) than with the spinach photosystem. In all cases, the observed rate constants for electron-transfer monotonically decrease with increasing NaCl or MgCl2 concentration. This is interpreted in terms of the involvement of attractive electrostatic interactions, which result in the initial collision complex having the most productive orientation for the electron transfer process, without a requirement for further reorientation. The magnitude of the response to MgCl2 suggests the occurrence of specific ion effects as well. In the absence of added salts, the reduction rate of oxidized P700 increases with pH from approximately 6 to 8, but decreases slightly at pH 8.5.
Native plastocyanin from Synechocystis 6803 has been isolated and purified to electrophoretic homogeneity. The corresponding gene (petE) has been cloned and expressed in E. coli, thus leading to a protein completely identical to plastocyanin purified from the cyanobacterial cells. The petE gene product is correctly processed in E. coli as deduced from the N-terminal amino acid sequences. These results, along with the identical physicochemical and kinetic properties of the two protein preparations, confirm that expression of petE in E. coli is an adequate tool to address the study of Synechocystis plastocyanin by site-directed mutagenesis.
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1. Specific antibodies against a whole preparation of glycophorins from rat erythrocyte membrane and against its most prominent component (32 kDa) were prepared. 2. Both antisera and the respective affinity-purified antibodies recognize the 74, 32 and 25 kDa components of rat glycophorins; therefore, a close antigenic relationship between them can be inferred. 3. On the other hand, a cross-reaction with human glycophorin A (both MM- and NN-type) is observed for both antisera. 4. This suggests the existence of epitopes common to human and rat glycophorins.
The release of dopamine (DA) from corpus striatum is affected by the endocrine state of the animal being progesterone suggested as a potential hormonal modulatory signal. Most of its actions have been described on endogenous DA release induced by amphetamine. However, the release of DA and the mechanism of the drug effect have been shown to be highly complexes. Considering that DA recently incorporated and/or synthetized is preferentially used we have characterized the effect of progesterone in vitro on the K(+)-induced release of 3H-dopamine (3H-DA) from rat corpus striatum slices. These were obtained during the estrous cycle or under conditions of high or low levels of endogenous progesterone (pregnant and ovariectomized rats). The release of 3H-DA was independent of the cycle. However, progesterone in vitro modified the induced release in a cycle-dependent way. Low concentrations of the hormone (100-200 mM) reduced the K+ (30 mM) effect while higher doses (300-500 mM) were facilitatories. After 7 days of ovariectomy, the induced release of 3H-DA was unchanged while in pregnant rats it was found decreased. In both cases the inhibitory effect of the hormone disappeared. Both progesterone (200 nM) and omission of Ca++ from the superfusion medium did not modified tyramine (20 microM) or K+ induced release, respectively. Data suggest that the pool of DA, related to exocytotic mechanism of release, could be specifically affected by progesterone, in a bimodal way, probably through independent genomic and non-genomic influences.
We observed that Sendai virus preinduction of peripheral blood mononuclear cells and subsequent mitogenic stimulation resulted in: (i) Superproduction of interferon-gamma, (IFN-gamma) (ii) an increase in interleukin-2 (IL-2) synthesis that correlates with DNA synthesis when stimulated with phytohemagglutinin (PHA) or pokeweed mitogen (PWM) after treatment with the Sendai virus, while stimulation with Protein A from Staphylococcus aureus was not affected, and (iii) enhanced tumor necrosis factor-alpha (TNF-alpha) production in response to bacterial lipopolysaccharide (LPS). Treatment of monocyte cultures with LPS and cycloheximide or actinomycin-D inhibited the superinduction phenomenon. When cycloheximide was added at the viral induction time, the inhibition of TNF-alpha superproduction and DNA synthesis was still observed. These results suggest that Sendai virus lymphocyte superinduction is specific for a particular stimulatory pathway, not dependent on mRNA accumulation, and probably mediated by induction of an activating protein.
BACKGROUND AND OBJECTIVES: Exercise Doppler echocardiography allows measure valvular gradient during exercise, that could be useful in the prescription of physical activity in patients with a prosthetic aortic valve. METHODS: Exercise Doppler echocardiography was performed in 50 patients (mean age 53 +/- 12 years) with normally-functioning aortic prosthesis. Maximal exercise was performed by supine bicycle ergometry in 35 and modified Bruce protocol in 15. Continuous wave Doppler examinations were obtained at rest, during and at maximal exercise with a nonimaging transducer. RESULTS: The mean value of workload was 6.9 +/- 2 METS and there were no complications. Gradients at peak exercise were available in 43/50 patients. Exercise induced a statistic significant increase in the heart rate (76 +/- 14 to 136 +/- 26), systolic blood pressure (128 +/- 19 to 182 +/- 19 mmHg), peak gradient (29 +/- 9 to 61 +/- 17 mmHg) and mean gradient (16 +/- 5 to 32 +/- 9 mmHg). The gradient was greater than 59 mmHg in 18/43 patients, greater than 69 mmHg in 12/43 and greater than 79 mmHg in 5/43. CONCLUSIONS: Prescription of exercise in patients with aortic prosthesis requires perform an exercise Doppler echocardiography to measure the increase of valvular gradient with the exercise in each patient, since the stress testing could be normal, it can advise a rate of physical activity that determine very high prosthetic gradients.
Three different mutations were introduced in the polA gene of Streptococcus pneumoniae by chromosomal transformation. One mutant gene encodes a truncated protein that possesses 5' to 3' exonuclease but has lost polymerase activity. This mutation does not affect cell viability. Other mutated forms of polA that encode proteins with only polymerase activity or with no enzymatic activity could not substitute for the wild-type polA gene in the chromosome unless the 5' to 3' exonuclease domain was encoded elsewhere in the chromosome. Thus, it appears that the 5' to 3' exonuclease activity of the DNA polymerase I is essential for cell viability in S. pneumoniae. Absence of the polymerase domain of DNA polymerase I slightly diminished the ability of S. pneumoniae to repair DNA lesions after ultraviolet irradiation. However, the polymerase domain of the pneumococcal DNA polymerase I gave almost complete complementation of the polA5 mutation in Escherichia coli with respect to resistance to ultraviolet irradiation.
Twenty-eight allogeneic BMT patients (16 with acute leukemia, 12 with chronic myeloid leukemia) were included in a single center, prospective, randomized, controlled trial to assess the value of recombinant human erythropoietin (rh-Epo) in this setting. rh-Epo was administered through a central venous catheter as a single bolus injection (days 0-7: 100 U/kg/d; days 7-30: 150 U/kg/d). No secondary effects to rh-Epo treatment were detected. An earlier appearance of reticulocytes and a diminished need of red blood cells (RBCs) transfusions were observed in patients who were treated with rh-Epo (4 units vs 12 units; p < 0.05). The time to unsupported platelets above 25 x 10(9)/l was less in patients treated with rh-Epo than in control patients (19 days vs 31; p < 0.05), and they received significantly fewer platelet transfusions (36 units vs 138.5; p < 0.05). Our results show that rh-Epo treatment is capable of accelerating the erythroid reconstitution and decreasing the need for RBC transfusions. A beneficial effect on platelet reconstitution is also suggested, but further studies are necessary to confirm this point.
Four consecutive years (1987-1990) outpatients attendance at a cardiology clinic was recorded in a district hospital with a catchment area of 200,000 inhabitants. The aim being to identify and evaluate the demand for cardiology consultations. A system for recording diagnoses was developed, based on the WONCA. The total number of consultations was 26,273, 27% being new patients, 78% of whom were referred by family doctors and health centers. Of the total, 57% were male. The most common age groups being from 60 to 69 (25%) and from 50 to 59 (23%); 22% were over 69. In 13.7% of the consultations, the find diagnosis was of "healthy hearts", in 16.7% valvular heart disease, in 31.5% ischaemic heart disease, in 15.1% arrhythmias, in 8.4% hypertension, and the remainder were other diseases. We carried out 4,406 echocardiograms, 1,408 exercise tests and 511 Holter monitoring traces. There were 322 direct hospital admissions, and 6,094 discharges in the four years period. We consider this type of records to be useful organizing cardiology outpatients clinics, analyzing the demand, and planning the use of health care resources, as well as estimating future needs and assessing quality of care.