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Biomedical subjects

J García-López

Publications and source records attributed to J García-López.

17 recordsLinked to original sources

Electromyostimulation and plyometric training effects on jumping and sprint time.

This study compared the effects of four-week training periods of electromyostimulation (EMS), plyometric training (P), or combined EMS and P training of the knee extensor muscles on 20 m sprint time (ST), jumping ability (Squat jump [SJ] and Countermovement jump [CMJ]), maximal isometric strength (MVC), and muscle cross-sectional area (CSA). Forty subjects were randomly assigned to one of the four treatment groups: electromyostimulation (EG), plyometric (PG), combined EMG, and P (EPG), that took place 4 times per week, and a control group (CG). Subjects were tested before and after the training program, as well as once more after 2 wk of detraining. A significant improvement (p < 0.05) in ST was observed after training (2.4 %) in EG while a significant slowing (p < 0.05) was observed (- 2.3 %) in EPG. Significant increases in EPG (p < 0.05) were observed in SJ (7.5 %) and CMJ (7.3 %) after training, while no significant changes in both jumps were observed after training and detraining for EG. A significant increase (p < 0.05) in MVC was observed after training (9.1 %) and after detraining (8.1 %) in EG. A significant increase (p < 0.05) in MVC was observed after training (16.3 %) in EPG. A significant increase (p < 0.01) in CSA was observed after training in EG (9.0 %) and in EPG (7.1 %). EMS combined with plyometric training increased the jumping height and sprint run in physically active men. In addition, EMS alone or EMS combined with plyometric training leads to increase maximal strength and to some hypertrophy of trained muscles. However, EMS training alone did not result in any improvement in jumping explosive strength development or even interfered in sprint run.

Adult↗

The validation of a new method that measures contact and flight times during vertical jump.

The aim of this study was to design and validate a vertical jump assessing system based on flight time measurements. Hence, the first phase of this study consisted of programming the software SportJumpv-1.0 and adapting a contact mat and a computer mouse in order to record the flight and contact times of a jump test. In the second phase, 9 subjects made 12 maximal and submaximal vertical single and rebound jumps. Thus 108 flight times and 36 contact times were simultaneously obtained with five different systems: ErgoJump Bosco System (PS), SportJump-v1.0 (SJ), Force Plate (FP), High-speed camera (HSC), and a Led (LED) filmed with a high-speed camera; FP was considered as the reference system. No significant differences were found either between SJ and LED, or between FP and HSC systems. SJ and LED overestimate flight time (9 ms and p < 0.001) and underestimate contact time in respect to FP values. PS underestimates flight time regarding FP values (4 ms and p < 0.01). In spite of the differences obtained between the systems, the errors found are constant. Those slightly depend on the subject's body mass for the systems that are connected to a contact mat. A high correlation between all of them was obtained (r approximately 1 and p < 0.001). The SJ system was validated for an accuracy of 500 Hz, and it was shown to be a useful tool in measuring flight and contact times during jumping. The differences and correlations found between the five systems show that it is necessary to take into account which system is being used when analyzing vertical jump performance.

Adolescent↗

Physical exercise and improvement of liver oxidative metabolism in the elderly.

Drug metabolizing capacity is generally reduced in the elderly, and physical exercise has been reported to increase drug oxidative metabolism. The purpose of this investigation was to study the effects of engagement in a program of regular physical exercise on the clearance and metabolite excretion of antipyrine, a marker of oxidative metabolism, in elderly subjects. The saliva clearance of antipyrine and the production clearances of antipyrine metabolites were studied in 37 elderly women (mean age 66 years). Subjects attended 60-min sessions three times a week for 12 weeks. Each session consisted of both aerobic (training of cardiorespiratory capacity) and nonaerobic (training of muscular strength/endurance and flexibility/coordination) exercises performed at 50-75% of maximum oxygen uptake. Antipyrine was administered orally and pharmacokinetic parameters were obtained from saliva and urine samples. After 3 months of participation in the exercise program, salivary antipyrine clearance was significantly increased by 17% mean (SEM) 0.42 (0.02) vs 0.36 (0.02) ml/min/kg; P < 0.05) and the half-life of antipyrine was significantly reduced by 18% (17.9 (1.1) vs 22.3 (1.3) h; P < 0.05). No significant change with exercise was observed in the renal clearance of antipyrine or in the norantipyrine formation clearance, but significant increases were found for hydroxymethylantipyrine [42 (5) vs 32 (4) microl/kg/min; P < 0.05; +31%] and 4-hydroxyantipyrine [243 (18) vs 194 (17) microl/kg/min; P < 0.05; +25%] formation clearances. These findings indicate that regular exercise leads to increased disposition of antipyrine in the elderly and that the main metabolic pathways of the compound are changed differentially.

Aged↗

Genomic typing of minor histocompatibility antigen HA-1 by reference strand mediated conformation analysis (RSCA).

Disparities in minor histocompatibility antigen (mHAg) HA-1 are involved in the development of acute graft-versus-host disease (GvHD) in adult recipient after HLA-identical sibling donor hematopoietic stem cell transplantation. The mHAg HA-1 is an HLA-A*0201-restricted nonapeptide, which derives from the cleavage of a protein encoded at chromosome 19. The sequence analysis of HA-1 cDNA identified two alleles, termed HA-1H and HA-1R, which differ in only two nucleotides at 3' end of exon A, at positions 500 and 504. DNA-based methods for HA-1 typing were developed in 1998, using polymerase chain reaction with sequence-specific primers (PCR-SSP) and restriction fragment length polymorphism (PCR-RFLP). Here, we report the usefulness of reference strand mediated conformation analysis (RSCA), which was developed for mutation detection and typing of polymorphic loci, to discriminate between the two HA-1 alleles. We performed genomic typing of HA-1 locus in 203 HLA-A*0201-positive samples using RSCA and we confirmed these results by PCR-SSP. The results demonstrate the high reproducibility of this method and their strong correlation with the results obtained by PCR-SSP (99%). Only two samples showed disparity between the RSCA typing and the PCR-SSP. Direct sequencing of these samples confirmed that the correct allele assignment was that obtained by the RSCA typing. Furthermore, HA-1- RSCA-based typing provides additional information about the intronic structure of both alleles. With this approach, we describe the almost constant presence (99.2%) of a 5-bp deletion at intronic position 214-218 associated to the HA-1H allele, previously unidentified. We conclude that HA-1 genomic typing by RSCA is easy to perform and that could be used as a routine typing method.

Alleles↗

HLA-B44 subtyping in the Catalan population using reference strand mediated conformation analysis. Implications for the selection of unrelated bone marrow donors.

Cytotoxic T lymphocytes (CTLs) reactive against the disparity between HLA-B*4402 and HLA-B*4403 have been reported after unrelated donor bone marrow transplantation. These CTLs have been associated with acute graft-versus-host disease and graft rejection. This study describes the HLA-B44-subtyping in the Catalan population using reference-strand mediated conformation analysis. It has been performed on 297 unrelated HLA-B44+ cord blood units from the Barcelona Cord Blood Bank (Barcelona, Spain). We have found a predominance of HLA-B*4403 (66.04%) over HLA-B*4402 (33.02%), whereas the predominant HLA-B44 allele in Northern Europe and the United States is HLA-B*4402. This inverted proportion between HLA-B44 subtypes in Mediterranean populations compared with other Caucasian populations suggests that HLA-B44 subtyping should be performed when an HLA-B44+ unrelated donor marrow is identified.

Alleles↗

Follow-up of chimerism status after allogeneic HLA-mismatched stem cell transplantation by detection of non-shared HLA alleles.

BACKGROUND AND OBJECTIVES: Chimerism studies after allogeneic transplantation are usually performed using cytogenetic analysis, PCR-VNTR or PCR-STR. Here, we report an alternative method for following the chimerism status after an HLA-mismatched stem cell transplantation (SCT), detecting the presence of non-shared HLA alleles by reference-strand mediated conformation analysis (RSCA). DESIGN AND METHODS: We tested this new approach on allogeneic related haploidentical SCT, unrelated cord blood transplantation, and HLA-mismatched unrelated donor SCT. The quantification of the chimerism was performed by laser detection of fluorescent-labeled primers on an automated DNA sequencer. RESULTS: In all cases this technique was able to detect mixed chimeras. The technique detected above 5% of residual cells when the analysis was based on HLA-class I and above 3% for HLA-class II. This sensitivity is similar to that of the PCR-VNTR analysis. INTERPRETATION AND CONCLUSIONS: This method avoids the need to search for an informative locus (which is essential for PCR-VNTR or -STR). Moreover, we did not find the phenomenon of preferential amplification that is observed with most VNTR, thus avoiding the need for construction of standard curves to quantify mixed chimeras. We conclude that the detection of the non-shared HLA alleles by RSCA is a useful approach for chimerism follow-up after HLA-mismatched SCT.

Alleles↗

Metastatic endometrial cancer in lung and liver: complete and prolonged response to hormonal therapy with progestins.

A double complete and prolonged response of metastatic endometrial carcinoma to medroxyprogesterone is reported. A 61-year-old woman with metastatic endometrial carcinoma in lung and liver achieved a complete clinical response with medroxyprogesterone lasting for 2 years. She discontinued the therapy by herself and developed a pulmonary relapse, which disappeared after retreatment with the same hormonal therapy. At present, she is alive without evidence of disease 6 years after starting progestins for metastatic disease and 14 years after treatment of the primary tumor. Progestin therapy in metastatic endometrial carcinoma is discussed, emphasizing the factors predicting response.

Carcinoma↗

Granulocyte colony-stimulating factor administered as a single intraperitoneal injection modifies the lethal dose95/30 in irradiated B6D2F1 mice.

Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor that stimulates the proliferation of progenitor myeloid cells. We have previously demonstrated that recombinant human G-CSF (rhG-CSF) significantly improves survival of lethally irradiated B6D2F1 mice when administered as a single intraperitoneal dose of 1 mg/kg 2 hours after a lethal dose (LD)95/30 irradiation. In our model, rhG-CSF is also able to modify the LD95/30 in irradiated animals and 1.1 has been found to be the dose modification factor (the ratio of LD95/30 for mice treated with rhG-CSF to that for control animals).

Animals↗

Low-dose donor CD8+ cells in the CD4-depleted graft prevent allogeneic marrow graft rejection and severe graft-versus-host disease for chronic myeloid leukemia patients in first chronic phase.

Based on previous experiences in animals and humans, low doses of CD8+ lymphocytes infused together with the marrow graft seem to enhance engraftment after allogeneic T cell-depleted marrow transplantation. From April 1994 to February 1997, 12 patients with chronic myelogenous leukemia in first chronic phase receiving a bone marrow transplant (BMT) from an HLA-identical sibling were included in a pilot study of T cell subset depletion. Total depletion of CD4+ cells of the marrow graft and partial depletion of CD8+ cells was performed by immunomagnetic separation. In order to improve the engraftment rate, we infused a low fixed number of CD8+ lymphocytes (0.25 x 10(6)/kg). All the patients were at high risk of developing acute graft-versus-host disease (GVHD), with a recipient age of >30 years, and/or donor sensitized by previous pregnancies or transfusions. All of them received cyclosporin A and methotrexate post-BMT. No graft failure was observed. The grade III-IV GVHD rate was 16.6%, and the actuarial survival at 3 years is 81.8%. Immunological recovery showed persistent CD8+ HLA-DR+ lymphocytosis 8 months after transplant. Relapses were not observed. This experience shows the importance of CD8+ cells to ensure correct engraftment, decreasing the GVHD rate.

Adult↗

Flow cytometric analysis of P-glycoprotein function using rhodamine 123.

The MDR1 gene product, P-glycoprotein (P-gp), works as a transmembrane efflux pump for several cytotoxic products, representing a major cause for cancer treatment failure. Rhodamine 123 (Rh123), a low toxic fluorescent probe commonly used to assess mitochondrial bioenergetics in living cells, has also been used to measure the efflux activity of P-gp in both normal and malignant cells. Analysis of variation in cellular fluorescence by measuring the rates of Rh123 influx and efflux, together with the effect of mdr reversing agents, allows the investigation of drug-resistant phenotypes in cancer samples. We have studied the functional activity of P-gp in human leukemic cell lines using flow cytometry, taking into consideration that variables such as Rh123 cytotoxicity, culture conditions, cell membrane integrity, as well as the effect of specific P-gp modulators, can impair the resolution of the Rh123-efflux measurements. The studies show that: (1)optimal non-cytotoxic concentrations of Rh123 which allow appropriate color compensation are in the range of 50-200 ng/ml; (2) life-gating allows accurate measurement on the 50% average rate of Rh123 efflux; (3) relative efficiency of P-gp inhibitors was PSC-833 > cyclosporin A > verapamil; and (4) the presence or absence of fetal calf serum had no effect on the bioavailability of chemosensitizer agents, with the exception of serum-free experiments, which showed a significant decrease in P-gp activity under the presence of PSC-833 (P = 0.05). Hence, we recommend this experimental strategy for clinical practice better to study the cellular drug resistance phenotype.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Is rhodamine 123 an appropriate fluorescent probe to assess P-glycoprotein mediated multidrug resistance in vinblastine-resistant CHO cells?

Cellular drug resistance, which involves several mechanisms such as P-glycoprotein (P-gp) overexpression, kinetic and metabolic quiescence, or the increase in the intracellular levels of glutathione, limits the effectiveness of cancer treatment. It has been reported that functional assessment of the cationic dye rhodamin 123 (Rho123) efflux reveals accurately the drug-resistant phenotype. To study cellular drug resistance, we have obtained a CHO-K1 derived cell line resistant to vinblastine by means of multistep selection. This cell line (CHOVBR) displays high reactivity with a monoclonal antibody (MAb) (C219) directed against an internal domain of P-gp, and an active Rho123 efflux, as shown by parallel flow cytometric and fluorometric assays. However, under similar experimental conditions, the drug-sensitive parental cell line CHO-K1 (as well as the myeloblastic KG1 and KG1a cell lines), was also able to pump Rho123 out. These parental CHO-K1 cells had a very low reactivity against the C219 Mab, as confirmed by Western blot analysis. Both vinblastine and verapamil inhibited Rho123 efflux in CHO-K1 cells, but had no effect on CHOVBR cultures. Also, deprivation of vinblastine for one month did not affect Rho123 efflux in these cells. Our results suggest that the activity of P-gp appears to be essential, but not sufficient to confer drug resistance, and that Rho123-based functional assays of drug resistance should be evaluated for each cellular experimental model.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Comparative effect of verapamil, cyclosporin A and SDZ PSC 833 on rhodamine 123 transport and cell cycle in vinblastine-resistant Chinese hamster ovary cells overexpressing P-glycoprotein.

The product of the mdr1 gene, P-glycoprotein (P-gp), represents a common mechanism of cellular resistance to a wide variety of structurally and functionally unrelated drugs. A range of structurally different P-gp inhibitors, such as verapamil, cyclosporin A and SDZ PSC 833, have been shown to modify multidrug resistance (MDR). We used flow cytometry to investigate in vitro modulation of P-gp-dependent efflux of rhodamine 123 (Rh123). The capacity to modulate the MDR phenotype of vinblastine-resistant Chinese hamster ovary (CHO) cells was assessed by analyzing the concentration of modulator needed to decrease the Rh123 mean fluorescence intensity by 50%. We found that the cyclosporin derivative SDZ PSC 833 was significantly more effective than cyclosporin A and verapamil, either in the presence or absence of fetal calf serum-supplemented media. This study indicates that analysis of Rh123 efflux modulation can be used to determine the optimal doses of MDR inhibitors in vitro and suggests that more than one modulator is needed to measure P-gp function, since verapamil had no effect on Rh123 modulation when MDR cells were used.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Relevance of forward scatter and side scatter in aneuploidy detection by flow cytometry.

The study of the nuclear DNA content by flow cytometry (FCM) has been classically accomplished by selecting the nuclear population on the biparametric forward scatter (FS)-DNA fluorescence or FS-DNA fluorescence peak histograms to determine ploidy and DNA index (DI). Different cellular factors such as nuclear morphological heterogeneity of the neoplastic cells, intratumoral variability, histological origin, displasia grade, necrosis, and size of the tumoral piece analyzed constitute important problems in ploidy studies and, consequently, residual or underrepresented clones with different ploidy levels can be masked by populations with a large cell number. In the present report, an alternative methodology is proposed for aneuploidy detection, since populations coinciding with DNA content may be different with respect to morphological criteria. The discrimination of aggregates and background noise by using peak or logarithmic fluorescence signal, and backgating in side scatter (SS)/FS histograms, permits the establishment of specific bounds through complete scatterplot mapping and to distinguish between scarce or minor populations in association with small or abnormal DNA peaks. Moreover, variations in the DNA modal channel value and the peak coefficient of variation value remained unmodified, also maintaining the quality of cytometric measurement data.

Aneuploidy↗

Peripheral blood CD34+ cell immunomagnetic selection in breast cancer patients: effect on hematopoietic progenitor content and hematologic recovery after high-dose chemotherapy and autotransplantation.

BACKGROUND: Tumor cells have been detected in mobilized peripheral blood of breast cancer patients, and they may contribute to tumor recurrence after the transplantation of peripheral blood progenitor cells. One of the most widespread technologies for tumor purging of the graft is immunomagnetic hematopoietic progenitor cell selection. STUDY DESIGN AND METHODS: The study assessed the effectiveness of a magnetic cell-separation system in selecting functional subpopulations of hematopoietic progenitors from 14 blood-derived harvests of 11 patients with high-risk breast cancer after mobilization following cytotoxic chemotherapy supported by granulocyte--colony-stimulating factor, as well as the feasibility of transplanting these selected subpopulations. RESULTS: CD34(+)-enriched cell fractions had a median purity of 93.0 percent (72.7-98.5%). The procedure yielded 52.6 percent of the CD34+ cell input (39.4-116.8%). Median recoveries of colony-forming units (CFUs) (36.87%) and cobblestone area-forming cells (CAFCs) (152.5%) were, respectively, 0.70 and 2.87 times those of CD34+ cells (52.6%). Moreover, CAFC efficiency in the positive cell fraction was 2.57 times that in the starting cell fraction. Peripheral blood neutrophil counts of 0.5 x 10(9) per L and platelet counts of 20 x 10(9) per L were reached after median times of 9 and 11 days, respectively. The number of transfused CAFCs per kg, CD34+ cells per kg, and postthaw CFU-granulocyte-macrophage per kg was correlated, respectively, with the speed of engraftment of neutrophils, platelets, or both. Tumor cells detected in one patient's peripheral blood were not found after CD34+ cell selection. CONCLUSION: Transplantation of immunomagnetically purified peripheral blood CD34+ cells does not increase transplantation-related morbidity. It induces a selective enrichment of more immature hematopoietic progenitors, which makes it suitable for use in cell expansion and gene therapy protocols.

Adult↗