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Biomedical subjects

M Ruiz

Publications and source records attributed to M Ruiz.

At least 19 recordsLinked to original sources

A fully automated method for mononuclear bone marrow cell concentration.

We describe our experience in processing 40 bone marrow aspirates harvested for autotransplantation from patients with several hematological diseases using the CS-3000 blood cell separator. The bone marrow of the first 30 patients was processed by a semiautomated method, and a fully automated procedure was used for the remaining 10 cases. Both procedures were developed in our laboratory and yielded a similar average mononuclear cell recovery of 87.78% and 86.98%, respectively, and similar nucleated cell recovery (27.39% and 27.11%). The cloning efficiency of hematopoietic progenitor cells, measured as the total CFU-GM colony recovery in the in vitro cultures, did not differ between processed and recovered mononuclear cells. On the other hand, all the patients with transplants showed complete hematologic recovery, and the time to engraftment was similar to that described for other procedures. The automated procedure resulted in an average red cell removal of 97.81%, similar to the semiautomated procedure (94.19%), though with a narrower range (96.31-98.6% vs. 80.34-98.34%). The time taken to process a similar amount of bone marrow cell suspension was very different for each method: 1 hour for the fully automated vs. 2 1/2 hours for the semiautomated method to process 1,000 ml. Furthermore, the semiautomated procedure required the addition of homologous or irradiated plasma in a laminar air flow chamber, while the automated method is performed in a closed sterile system. We conclude that our procedure using the CS-3000 processor is an efficient method for fully automated large-scale processing of human bone marrow cells.

Adolescent

Isoform-specific subcellular targeting of glucose transporters in mouse fibroblasts.

GLUT1, the erythrocyte glucose transporter, and GLUT4, the adipose/muscle transporter, were each expressed in NIH-3T3 cells by retrovirus-mediated gene transfer. In fibroblasts overexpressing GLUT1, basal as well as insulin-stimulated deoxyglucose uptake was increased. Expression of GLUT4 was without affect on either basal or hormone stimulated hexose uptake. Localization of each of the transporters by indirect immunofluorescence revealed that, whereas GLUT1 was found primarily on the cell surface, GLUT4 was directed to vesicles in a perinuclear distribution and throughout the cytoplasm. The GLUT4-containing compartment represented neither Golgi complex nor lysosomes, as evidenced by the failure of lgp110 or Golgi mannosidase to co-localize. However, there was substantial overlap between the distribution of GLUT4 and the transferrin receptor, and some colocalization of the transporter isoform with the manose-6-phosphate receptor. In addition, when FITC-wheat germ agglutinin bound to the cell surface was allowed to internalize at 37 degrees C, it concentrated in vesicular structures coincident with GLUT4 immunoreactivity. These data establish that GLUT1 and GLUT4 contain within their amino acid sequences information which dictates targeting to distinct cellular compartments. Moreover, GLUT4 can be recognized by those cellular factors which direct membrane proteins to the endosomal pathway.

3T3 Cells

Influence of subendocardial ischemia on transmural myocardial function.

The relationship between regional myocardial perfusion and function under ischemic conditions was examined by using a nontraumatic single-crystal pulsed Doppler system that permits complete transmural assessment of myocardial thickening. Sixteen open-chest dogs underwent either 50 (n = 4) or 180 (n = 12) min of partial coronary artery occlusion. Simultaneous measurements of myocardial thickening fraction (TF) and microsphere-determined blood flow (BF) were taken in the subepicardial, midwall, and subendocardial thirds of the left ventricular wall. During ischemia, there was an excellent correlation between BF and TF in the subendocardium. Mean subendocardial BF was reduced to 0.45 +/- 0.3 ml.min-1.g-1, resulting in a subendocardial TF of 0.8 +/- 19%. Although subepicardial BF was relatively preserved at 1.03 +/- 0.4 ml.min-1.g-1, subepicardial TF was diminished markedly and not significantly different from subendocardial TF. Subepicardial and midwall TF were highly dependent on subendocardial flow rather than on the actual flow in these areas. Hence these studies show a marked dependence of transmural myocardial function on subendocardial blood flow. Outer wall function is more dependent on subendocardial than subepicardial blood flow.

Animals

[A molar pregnancy coexistent with a viable fetus in a twin pregnancy].

A case of molar gestation associated with an only fetus, both product of an twin gestation is presenting. We analyzed the clinical picture, the diagnostic method, the after birth resolution and evolution of this gestation. The result of a live newborn of 1,300 g and verify coexisting of complete hydatidiform mole.

Adolescent

[Isolation and identification of Gardnerella vaginalis in women with symptoms of bacterial vaginosis].

70 samples of vaginal secretions were collected from women with bacterial vaginosis syndrome, that were attended in Nuevo Veranillo' Health Center. All samples were tested in order to determinate the presence of amine and to Gram'methods stain to observe the morphologic characteristics of the "Clue" cells (epithelial cells with adhered bacteria). Samples were cultured in a selective media of blood agar in Columbia base with colistine and nalidixic acid, and incubated in an environment of CO2 at 37 degrees C. Tests of oxidase, catalase, glucid fermentation, hippurate hydrolysis and starch hydrolysis were made on the grown colonies. 27 out of 70 samples resulted in positive cultures for Gardnerella vaginalis, 11 to pregnant women and 16 to nonpregnant women; among these, 14 women used some kind of contraceptive methods as DIU or pill.

Adolescent

Cytogenetic analysis of human oocytes from fertile women.

Several reports have shown a relatively high incidence of chromosome anomalies in human inseminated-unfertilized oocytes from infertile women. In the present study, cytogenetic analysis was attempted in 73 human oocytes from 17 fertile women in order to establish the incidence of chromosome anomalies in the fertile population and to compare this with the incidence in inseminated-unfertilized oocytes from infertile women. Of 56 oocytes that could be analysed, 42 oocytes were haploid, 12 were hypohaploid and two exhibited fragmented chromosomes. The low rate of chromosome anomalies (3.6%) found in this population suggests that there is natural selection at fertilization against diploid oocytes and oocytes with fragmented chromosomes. This result also questions the high incidence of chromosome anomalies found in previous reports using inseminated-unfertilized oocytes.

Chromosome Aberrations

Comparison of implantation and early development of human embryos fertilized in vitro versus in vivo using transvaginal ultrasound.

Several reports in the literature suggest delayed implantation of in vitro-fertilized human embryos compared to in vivo-fertilized eggs. The use of high-frequency transvaginal transducers for early detection of pregnancy has allowed the identification of the gestational sac with very low serum human chorionic gonadotropin (beta-hCG) levels. Thus, the present study evaluated whether retarded implantation can be identified using this novel technology. We studied 13 single pregnancies after in vitro fertilization (IVF) and 14 pregnancies after artificial insemination either by husband (n = 6) or donor (n = 8). In the IVF patients, oocytes were retrieved 35 hours after hCG administration. Embryo transfer occurred approximately 48 hours after retrieval. Artificial insemination was performed 24 and 48 hours after hCG administration. Transvaginal ultrasound scans and serum beta-hCG levels were evaluated every 3 days starting day 12 post-hCG administration. Serum beta-hCG levels rose in parallel when in vitro- and in vivo-fertilized embryos were compared. Similarly, there was no difference between groups in the mean time needed to detect early embryonic structures, such as the embryonic sac, yolk sac, and heartbeats, or the growth rate of the gestational sac. In conclusion, there was no difference in detecting implantation and early embryonic development of human embryos fertilized in vivo versus in vitro as ascertained by ultrasound scans and serum beta-hCG levels. An embryonic sac is detected 23-24 days after hCG administration in pregnancies achieved by assisted reproductive techniques.

Amnion

Renin-angiotensin system in thyroidectomized rats at different periods of development.

The relationship between the renal function and some components of the renin-angiotensin system has been studied in hypothyroid rats thyroidectomized surgically at different periods of their life. Changes in plasma renin concentration (PRC) depending on the period hypothyroidism were induced. Results showed that the renin release control could result from an equilibrium between the reduced beta-adrenergic activity and the marked natriuresis observed in hypothyroidism. A reduction in plasma angiotensinogen concentration (PAC), due to a decrease in its hepatic production, was observed in thyroidectomized animals. PAC reduction was independent of the hypothyroidism induction period. Alterations in plasma renin activity (PRA) were a consequence of PRC and PAC changes in thyroidectomized animals, as an increase in fractional sodium excretion (FENa) time course dependent, was found in these rats.

Aging

[Spontaneous bacterial pleuritis in 3 patients with liver cirrhosis].

We report 4 episodes of spontaneous bacterial pleuritis observed in 3 patients with liver cirrhosis complicated by ascites and pleural effusion. This infection mimics spontaneous bacterial peritonitis. Three episodes were successfully treated. Proposed pathogenesis, diagnostic methods and therapy are discussed.

Ascites

[Portal venous flow (ultrasonography-Doppler) in patients with alcoholic liver cirrhosis].

To evaluate the clinical usefulness of the ultrasonographic (echo-Doppler) measurements of portal blood flow their results were compared with several clinical and biochemical parameters in alcoholic cirrhotic patients. The technique was standardized and its reproducibility was checked in 30 cirrhotics and in 20 control subjects. In controls, portal area was greater when measured on its transversal axis and in deep inspiration. In cirrhotics the area did not change neither according to the axis nor the respiratory movements. The estimated velocity of blood flow was dependent on the angle of insonation. Measurements performed on longitudinal axis, at 50 degrees in expiratory apnea showed, in the same subject, an interday variability of 7%. In cirrhotic patients portal blood flow was higher than in controls (0.93 +/- 0.32 L/min vs 0.64 +/- 0.12, p < 0.001) being the difference due to a greater area. Considering controls and Child's A and B cirrhotic patients, portal blood flow correlated with the severity of disease (r = 0.738, p < 0.001). In Child C, portal blood flow was decreased, compared to Child's B patients. The presence and size of esophageal varices was also correlated to portal blood flow (r = 0.461, p < 0.05). However no differences were observed between the groups with or without previous variceal bleeding. It is concluded that echo-doppler measurement of portal blood flow is a reproducible technic in standardized conditions, detecting changes related to global liver function and the presence and size of esophageal varices.

Blood Flow Velocity

[Cystic fibrosis. Prenatal diagnosis].

The clinical case of heterozygote parents, both bearers of cystic fibrosis, whose three newborn infants died of this disease is presented. The authors describe the prevention of and the progress in its early prenatal diagnosis and management.

Carrier State

Cell wall structures which may be important for successful immunization with Salmonella-Shigella hybrid vaccines.

Three separate lots of S. typhi/S. sonnei hybrid (Ty/Shig) live oral vaccine strain 5076-1C were tested for efficacy in human volunteers challenged with virulent S. sonnei. Two lots (2 and 5) protected volunteers, a third lot (8) did not. The three lots were evaluated by immunological tests and electron microscopy. Lots 2 and 5, which protected, contained bacteria that reacted with anti-flagellar serum and had observable attached flagella and pili. Lot 8, which failed to protect, did not react with anti-flagellar serum, and had no observable pili. There was no correlation between vaccine efficacy and the reaction of IgG in patient's sera in western blot analysis. Surface structures on the Ty-Shig hybrid may be important for generating a protective immune response.

Animals

An enzyme-linked antiglobulin test for assessing anti-D immunoglobulin preparations.

Two enzyme-linked antiglobulin tests (ELAT) for assessing anti-D IgG preparations are described; one is performed in tubes and the other in microtitre plates. An anti-human IgG alkaline phosphatase conjugate and the substrate p-nitrophenyl phosphate are used. Both methods were sensitive and reproducible, with variations coefficients of 7.8 and 8.6% for enzyme immunoassay in tubes and microplates, respectively. The linear relationship between the amount of red cell-bound anti-D and the optical density shows that the method is suitable for quantitative studies. Results obtained by the two methods show a very good correlation (r = 0.99) in 12 of the 14 samples assayed, and both give good agreement with results obtained in automated haemagglutination. Since microtitre plate ELAT has numerous advantages over the tube method, it could provide an alternative method for assessing anti-D activity of specific IgG preparations in control laboratories.

Antibodies, Anti-Idiotypic