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Biomedical subjects

J García

Publications and source records attributed to J García.

At least 217 records · Page 12Linked to original sources

Studies of the specificities of IgE antibodies found in sera from subjects with allergic reactions to penicillins.

Penicillins are immunogenic when administered to humans and in some instances they can also be allergenic, inducing specific IgE antibodies. Whilst the major haptenic group, the penicilloyl, is well characterised, less is known about the relative importance of the different parts of the structure for antibody binding and how this can influence the specificity of patients response. In order to investigate this further, sera from subjects who had suffered an IgE-mediated reaction to penicillins were studied using the radioallergosorbent test (RAST) and RAST inhibition. The assays employed reagents related to the penicillins causing the reaction. Using 173 sera, positive RAST results were only found with reagents based on benzyl penicillin (BP) and amoxicillin (AX). Fifty-three positive sera were selected for further studies and categorized into three groups: (A) sera only RAST positive to AX, (B) sera only positive to BP and (C) sera positive to both penicillins. RAST inhibition studies were then carried out using monomeric penicilloyl conjugates and compounds representing parts of the penicilloyl structures of BP and AX. For all three groups, monomeric penicilloyl conjugates were the most efficient inhibitors but there were differences for the other compounds. Group A sera were also inhibited by the side chain amoxicillin, whereas group B sera were poorly inhibited by all other inhibitors. Group C sera showed two patterns of inhibition, both consistent with their more cross-reactive profile.

Amoxicillin↗

[Dietary carotene intake and lung cancer among men from Santiago].

Aiming to assess the association between carotenes and vitamin A intake and lung cancer, a case control study was performed. Sixty one male subjects with lung cancer and 61 controls paired for age, sex and smoking habits from 6 hospitals in Santiago were analyzed. Based on a consumption tendency enquiry, the mean weekly intake of food groups, carotenes and vitamin A was calculated, considering seasonal variations. Results showed that cases consumed less "winter" vegetables than controls (chard, beet, chicory, spinach and cabbage) but no differences between groups in carotene and retinol consumption. It is concluded that patients with lung cancer have a lower consumption of carotene rich vegetables.

Carotenoids↗

Bowel inflammation in anterior uveitis and spondyloarthropathy.

OBJECTIVE: To assess the possible involvement of the bowel in the spectrum of anterior uveitis (AU). METHODS: Ileocolonoscopy and histology studies were performed on 27 patients with AU. RESULTS: Patients with AU had a higher incidence of chronic intestinal inflammation (CII) than controls. CII was present in AU regardless of HLA-B27 status, sacroiliitis or NSAID intake, and was more prevalent in uveitis with high recurrence. CONCLUSION: Bowl inflammation ia a component of the spectrum of AU. This finding supports a HIL-B27 independent common pathogenic link between AU and spondyloarthropathy.

Adolescent↗

Presentation of a PCR-nuclease protection strategy for minimal residual disease monitoring in B-ALL.

Methods for detecting residual malignant cells in patients suffering from lymphoid malignancies have neither been sufficiently sensitive nor easy to routinize, hampering a potential prediction of disease outcome. Taking advantage of clone-specific DNA sequences, generated during lymphocyte differentiation and the polymerase chain reaction, some strategies have been developed for several groups. Up to now the most specific and sensitive methodology, which consists of designing leukemia-specific oligonucleotides, requires sequencing of the complementary determining region III-DNA for each particular patient and is too laborious to be applied to each case for routine monitoring in most hospital laboratories. In an attempt to achieve an easy way to detect residual malignant cells in B lymphoproliferative diseases, we have used a new PCR-based approach, named here as PCR-nuclease protection assay, consisting of: (i) amplification of DNA segments corresponding to the complementarity determining region III of the immunoglobulin heavy chain genes from samples at disease diagnosis; (ii) isolation of the disease-specific single-stranded DNA; (iii) labeling of the single-stranded DNA to generate specific probes; (iv) hybridization to amplified DNA from samples corresponding to different disease phases; and (v) digestion with S1-nuclease. Using this approach, we could detect one malignant cell in a background of 10(5) healthy cells. The sensitivity and specificity of this approach compares with those of the above mentioned specific oligonucleotide strategy in detecting residual malignant B cells. Moreover, this strategy is much less tedious and could be used by most hospital laboratories.

Base Sequence↗

[Intensive chemotherapy with support of peripheral hematopoietic progenitor cells, mobilized with high-dose cyclophosphamide and G-CSF: fast hematologic recovery].

BACKGROUND: Hematopoietic progenitor cells mobilized to peripheral blood by a chemotherapy combined or not with hematopoietic growth factors and harvested with cyto-apheresis (CTA) provide a rapid hematological recovery when infused as a support step after intensive chemotherapy (IC). METHODS: Cyclophosphamide (CI) 4 g/m2 and G-CSF 5 mcg/kg/d were administered to 19 patients with a solid tumor or lymphoma. Daily CTA were performed during hematological recovery to harvest > 2.5 x 10(6) cells CD34+/kg. Seventeen patients received IC with infusion of peripheral hematopoietic progenitor cells (PHPC) and G-CSF. RESULTS: A total of 52 CTA were performed, with a median (M) of 2 per patient. A M of 4.4 x 10(8) mononucleated cells/kg and 9.8 x 10(6) CD34+/kg were harvested per patient. Hematological recovery after IC with support of PHPC and G-CSF was rapid in all cases, but the aplastic period was shorter in the ten patients who received > 5 x 10(6) CD34+/kg cells than in the seven patients with < 5 x 10(6) kg: The median of recovery to neutrophils > 0.5 x 10(9)/l was 8 days compared with 9 days (p = 0.0005), to platelets > 20 x 10(9)/l of 8 days compared with 12 days (p = 0.001), and to platelets > 50 x 10(9)/l of 11 days compared with 14 days (p = 0.001). CONCLUSIONS: Toxicity of IC (4 g/m2) with G-CSF is moderate and allows the harvesting of an adequate number of PHPC. Its infusion after IC provides a rapid hematological recovery, which was more marked in patients receiving 5 x 10(6) CD34+/kg cells, than with the same IC schedules of IC with autologous bone marrow transplantation.

Adult↗

[Latex allergy in anesthesiology].

Allergy to latex, described 15 years ago, is increasing. It affects mainly certain risk groups, such as health-care workers and patients with spina bifida or genitourinary malformations. Symptoms vary in intensity from mild to life-threatening and can present during any medical procedure involving skin or mucosal contact, or inhalation, of the allergen in a previously sensitized patient. The most serious cases reported have generally involved patients undergoing surgery. Prevention is based mainly on tentative diagnosis made when the patient's history is taken, followed by laboratory tests and consultation with an allergist. Because prophylactic drug protocols have proven ineffective, the patient must be offered a latex-free environment in keeping with standard recommendations.

Allergens↗

Progression of natural immunity during one-year treatment of residual disease in neuroblastoma patients with high doses of interleukin-2 after autologous bone marrow transplantation.

The aim of this work was to monitor the functional and phenotypic variations of natural killer (NK) cells in seven children with stage IV neuroblastoma (NB) treated with recurrent 5-day cycles of interleukin-2 (IL-2) at a dose of 18 x 10(6) IU/m2/d by continuous intravenous infusion. All patients who entered the study had no detectable disease after hematologic recovery from intensive chemotherapy and autologous bone marrow transplantation (ABMT). To evaluate the effect of this treatment on tumor relapse, IL-2 immunotherapy was adjusted to maintain levels of NK activity above those of age-matched controls (threshold of 40 lytic units [LU]/10(9) mononuclear cells) during a 1-year period since hematologic recovery of ABMT. The levels of NK and endogenous lymphokine-activated killer (eLAK) cell cytotoxic activities, as well as phenotype-differentiated lymphocyte counts, were determined from patients' freshly isolated peripheral blood mononuclear cells (MNC). Data were analyzed at different points between each cycle of IL-2, and before and 36 hours after each infusion. NK and eLAK activities significantly increased in response to IL-2. Both cytotoxic parameters correlated with the serum levels of the soluble IL-2 receptor (sIL-2R). IL-2 increased the amounts of NK and T cell subsets but not of B cells. The effects of IL-2 were time-dependent. Early cycles of IL-2 preferentially increased cell numbers, especially of cells bearing a CD3-/CD16-/CD56+bright and CD8+dim phenotype. Conversely, late courses promoted higher cytotoxic effects but with a smaller increase in NK and T cell counts; the main NK subset became CD16+, and CD8+dim cells remained a minor subset. It is worthy to note that the patient who relapsed after completing immunotherapy showed only a slight increase of the NK subset in response to IL-2. These results show the feasibility of sustaining an increased NK activity during 1 year after ABMT in children with advanced neuroblastoma and suggest the occurrence of changes in the functional and phenotypic characteristics of the NK cells generated throughout the 1-year treatment.

Adrenal Gland Neoplasms↗

Intestinal helminths of the red fox (Vulpes vulpes L.) in Galicia (Northwest Spain).

The intestinal helminth fauna of 201 specimens of Vulpes vulpes, captured in Galicia (northwest Spain), was investigated. Eight species were found: Toxocara canis (23% of host specimens), Toxocara cati (0.5%), Toxascaris leonina (1%), Uncinaria stenocephala (28%), Seuratascaris numidica (0.5%), Taenia crassiceps (23%), Mesocestoides litteratus (2.5%) and Dipylidium caninum (0.5%). We discuss possible relationships between human population density and the prevalence of infection of fox by intestinal helminths.

Animals↗

Overproduction and purification of biologically active native fungal alpha-sarcin in Escherichia coli.

An efficient system was developed to produce, in Escherichia coli, large amounts of native alpha-sarcin (alpha Sar), a cytotoxin from the mold Aspergillus giganteus. The protein has been purified to homogeneity with a yield of 1.5 micrograms/ml of original culture. The constructed expression vector (pINPG alpha S) is based on the synthesis of a fusion protein between alpha Sar and a modified version of the OmpA signal peptide. This peptide seems to favour the postranslational processing of the fusion protein. The purified recombinant alpha-sarcin (re-alpha Sar) is structurally identical to the mature fungal protein according to the following criteria: N-terminal amino acid (aa) sequence, aa composition, electrophoretic mobility, chromatographic behaviour, immunoreactivity and spectroscopic features. Indeed, the recombinant protein recovered is completely functional, since it cleaves, in vitro, eukaryotic rRNA and it is able to interact with phospholipid vesicles with the same specificity as the native fungal alpha Sar.

Amino Acid Sequence↗

Straw thermal stabilizer for embryo cryopreservation.

Embryo cryopreservation procedures have been highly developed to support in vitro fertilization techniques, but the clinical results have not met initial expectations. Many variables influence the outcome of the embryo cryopreservation procedure. Exposure to classical freezing media does not affect further embryo development. Therefore, the first crucial step seems to be embryo thermal behavior during the freezing/thawing procedure. With the aim of avoiding thermal oscillations to the embryos due to liquid nitrogen injection into the freezing chamber, we have developed a straw thermal stabilizer (STS) cryopreservation accessory that when adapted to a programmable biological freezer offers a more exact control of the straws temperature and also a greater reproducibility of the freezing process. The STS incorporates a seeding mechanism controlled by specially designed software. For the biological assays, 2-cell mouse embryos were cooled until seeding at -7 degrees C or frozen to -120 degrees C using 1.5 M 1,2-propanediol and 0.1 M sucrose. As control groups, embryos were either untreated or were exposed to cryoprotectants, developing to blastocyst at rates of 90.8 and 88.4%, respectively. The embryos cooled until seedling or subjected to the complete freezing procedure developed into blastocysts at rates of 95.7 and 82.8%, respectively, disregarding the effect of the cryoprotectant exposure. These results show a substantial improvement in cryopreservation with no loss in embryo development due to the seeding procedure.

Animals↗

Measurement of calcium transients and slow calcium current in myotubes.

The purpose of this study was to characterize excitation-contraction (e-c) coupling in myotubes for comparison with e-c coupling of adult skeletal muscle. The whole cell configuration of the patch clamp technique was used in conjunction with the calcium indicator dye Fluo-3 to study the calcium transients and slow calcium currents elicited by voltage clamp pulses in cultured myotubes obtained from neonatal mice. Cells were held at -80 mV and stimulated with 15-20 ms test depolarizations preceded and followed by voltage steps designed to isolate the slow calcium current. The slow calcium current had a threshold for activation of about 0 mV; the peak amplitude of the current reached a maximum at 30 to 40 mV a and then declined for still stronger depolarizations. The calcium transient had a threshold of about -10 mV, and its amplitude increased as a sigmoidal function of test potential and did not decrease again even for test depolarizations sufficiently strong (> or = 50 mV) that the amplitude of the slow calcium current became very small. Thus, the slow calcium current in myotubes appears to have a negligible role in the process of depolarization-induced release of intracellular calcium and this process in myotubes is essentially like that in adult skeletal muscle. After repolarization, however, the decay of the calcium transient in myotubes was very slow (hundreds of ms) compared to adult muscle, particularly after strong depolarizations that triggered larger calcium transients. Moreover, when cells were repolarized after strong depolarizations, the transient typically continued to increase slowly for up to several tens of ms before the onset of decay. This continued increase after repolarization was abolished by the addition of 5 mM BAPTA to the patch pipette although the rapid depolarization-induced release was not, suggesting that the slow increase might be a regenerative response triggered by the depolarization-induced release of calcium. The addition of either 0.5 mM Cd2+ + 0.1 mM La3+ or the dihydropyridine (+)-PN 200-110 (1 microM) reduced the amplitude of the calcium transient by mechanisms that appeared to be unrelated to the block of current that these agents produce. In the majority of cells, the decay of the transient was accelerated by the addition of the heavy metals or the dihydropyridine, consistent with the idea that the removal system becomes saturated for large calcium releases and becomes more efficient when the size of the release is reduced.

Action Potentials↗

Relationship of calcium transients to calcium currents and charge movements in myotubes expressing skeletal and cardiac dihydropyridine receptors.

In both skeletal and cardiac muscle, the dihydropyridine (DHP) receptor is a critical element in excitation-contraction (e-c) coupling. However, the mechanism for calcium release is completely different in these muscles. In cardiac muscle the DHP receptor functions as a rapidly-activated calcium channel and the influx of calcium through this channel induces calcium release from the sarcoplasmic reticulum (SR). In contrast, in skeletal muscle the DHP receptor functions as a voltage sensor and as a slowly-activating calcium channel; in this case, the voltage sensor controls SR calcium release. It has been previously demonstrated that injection of dysgenic myotubes with cDNA (pCAC6) encoding the skeletal muscle DHP receptor restores the slow calcium current and skeletal type e-c coupling that does not require entry of external calcium (Tanabe, Beam, Powell, and Numa. 1988. Nature. 336:134-139). Furthermore, injection of cDNA (pCARD1) encoding the cardiac DHP receptor produces rapidly activating calcium current and cardiac type e-c coupling that does require calcium entry (Tanabe, Mikami, Numa, and Beam. 1990. Nature. 344:451-453). In this paper, we have studied the voltage dependence of, and the relationship between, charge movement, calcium transients, and calcium current in normal skeletal muscle cells in culture. In addition, we injected pCAC6 or pCARD1 into the nuclei of dysgenic myotubes and studied the relationship between the restored events and compared them with those of the normal cells. Charge movement and calcium currents were recorded with the whole cell patch-clamp technique. Calcium transients were measured with Fluo-3 introduced through the patch pipette. The kinetics and voltage dependence of the charge movement, calcium transients, and calcium current in dysgenic myotubes expressing pCAC6 were qualitatively similar to the ones elicited in normal myotubes: the calcium transient displayed a sigmoidal dependence on voltage and was still present after the addition of 0.5 mM Cd2+ + 0.1 mM La3+. In contrast, the calcium transient in dysgenic myotubes expressing pCARD1 followed the amplitude of the calcium current and thus showed a bell shaped dependence on voltage. In addition, the transient had a slower rate of rise than in pCAC6-injected myotubes and was abolished completely by the addition of Cd2+ + La3+.

Animals↗

Calcium transients associated with the T type calcium current in myotubes.

Immature skeletal muscle cells, both in vivo and in vitro, express a high density of T type calcium current and a relatively low density of the dihydropyridine receptor, the protein thought to function as the Islow calcium channel and as the voltage sensor for excitation-contraction coupling. Although the role of the voltage sensor in eliciting elevations of myoplasmic, free calcium (calcium transients) has been examined, the role of the T type current has not. In this study we examined calcium transients associated with the T type current in cultured myotubes from normal and dysgenic mice, using the whole cell configuration of the patch clamp technique in conjunction with the calcium indicator dye Fluo-3. In both normal and dysgenic myotubes, the T type current was activated by weak depolarizations and was maximal for test pulses to approximately -20 mV. In normal myotubes that displayed T type calcium current, the calcium transient followed the amplitude and the integral of the current at low membrane potentials (-40 to -20 mV) but not at high potentials, where the calcium transient is caused by SR calcium release. The amplitude of the calcium transient for a pulse to -20 mV measured at 15 ms after depolarization represented, on average, 4.26 +/- 0.68% (n = 19) of the maximum amplitude of the calcium transient elicited by strong, 15-ms test depolarizations. In dysgenic myotubes, the calcium transient followed the integral of the calcium current at all test potentials, in cells expressing only T type current as well as in cells possessing both T type current and the L type current Idys. Moreover, the calcium transient also followed the amplitude and time course of current in dysgenic myotubes expressing the cardiac, DHP-sensitive calcium channel. Thus, in those cases where the transient appears to be a consequence of calcium entry, it has the same time course as the integral of the calcium current. Inactivation of the T type calcium current with 1-s prepulses, or block of the current by the addition of amiloride (0.3-1.0 mM) caused a reduction in the calcium transient which was similar in normal and dysgenic myotubes. To allow calculation of expected changes of intracellular calcium in response to influx, myotubes were converted to a roughly spherical shape (myoballs) by adding 0.5 microM colchicine to culture dishes of normal cells. Calcium currents and calcium transients recorded from myoballs were similar to those in normal myotubes.(ABSTRACT TRUNCATED AT 250 WORDS)

Aniline Compounds↗

Bone marrow purging in acute lymphoblastic leukemia: biological and clinical features.

A group of 56 patients with immunophenotyped acute lymphoblastic leukemia had their bone marrow purged using monoclonal antibody (MAb) and complement. Mononuclear marrow cells were treated with panels of MAb directed against T cells (n = 19), CD10-CALLA (n = 16), and B cells (n = 21), resulting in median specific cell depletions of 96.50, 97.59, and 96.97%, respectively. The resulting CFU-GM recoveries were 60.15, 110.6, and 72.9%. Long-term bone marrow cultures showed normal formation of adherent cell layers in 17 of 23 cases, and failure to form an adherent layer correlated with poor hematological reconstitution. A total of 32 patients (aged 4-40 years) received purged marrow grafts containing a median of 1.32 x 10(7) mononuclear cells/kg and 1.64 x 10(4) CFU-GM/kg; 7 patients required backup marrow. Time to engraftment of 500 granulocytes, 1000 leukocytes, 20,000 platelets, and 50,000 platelets was 37, 38, 39, and 57 days, respectively. A total of 16 patients (50%) remain alive and in complete remission 313-913 days posttransplant (median 565 days); 13 patients (41%) relapsed between 50 and 365 days posttransplant (median 100 days), and 3 patients (9%) died from transplant-associated complications.

Adolescent↗

[Characterization of Neisseria meningitidis isolated fron systemic infections. Chile, 1992-1993].

BACKGROUND: in Chile, all systemic infections caused by Neisseria meningitidis must be reported and the bacterial strain must be sent to a Reference Laboratory at the Instituto de Salud Pública de Chile (ISP). AIM: to report the characterization of strains of N. meningitidis isolated during systemic infections in Chile during the years 1992 and 1993. METHODS: the serogroup, serotype, subtype and antimicrobial susceptibility of every strain of N. meningitidis received at the ISP during 1992 and 1993 was studied. RESULTS: six hundred twenty eight strains of N. meningitidis were confirmed during 1992 and 1993. B serogroup was responsible of 91.1% and 94.7% of confirmed cases during 1992 and 1993 respectively. Serotypes and subtypes most frequently associated to B serogroup were B: 15: P1.3 (63.2%) in 1992 and 51.8% in 1993) and B:NT:P1.3 (11.7% in 1992 and 21.3% in 1993). In 1992, all strains were susceptible to penicillin, chloramphenicol, ceftriaxone and rifampicin. During 1993, 7 (2%) strains were found, for the first time in Chile, moderately susceptible to penicillin and rifampicin MIC90 increased fourfold in respect of 1992, although all strains continued to be susceptible to this antimicrobial. CONCLUSIONS: the increasing frequency of NT (non typified strains) isolation will demand the use of molecular biology techniques for their identification. The appearance of penicillin resistant strains in our country is worrisome.

Chile↗

[Evaluation of three new culture media: brilliant-glycerol-lactose novobiocin-green agar, modified iron lysin agar and Rambach agar for the isolation of enteropathogenic E. coli, Salmonella sp. in acute gastroenteritis].

BACKGROUND: Acute gastroenteritis (AGE) is one of the most common diseases in children, particularly in those under the age of 5 years in whom a severe picture of hydroelectrolyte imbalance may be triggered accompanied, in most cases, by leukocyte response. The aim of this study was to evaluate three new culture media for the isolation of enteropathogens in infantile diarrhea. MATERIAL AND METHODS: From April to September, 1993, 170 samples of diarrhea stools from children up to 7 years of age attending pediatric hospitals in Bogota were studied. Three new culture media were used in the isolation of the enteropathogenic bacteria: BGLN, MILA agar, Rambach agar and a conventional agar medium S.S. Biochemical tests were performed to identify the isolated bacteria. RESULTS: A total of 98.5% of enteropathogenic E. coli was isolated in Rambach agar, which also presented excellent recovery rates of Salmonella sp. (100%) versus S.S> (64.3%) and BGLN (77.2%), respectively. The best selectivity for Shigella sp. was observed with BGLN with a 100% recovery rate. Out of the 170 samples 105 showed a leukocyte count of 70-75% and positive isolation for enteropathogenic bacteria. Six samples with the same leukocyte count did not present enteropathogenic bacteria, with the 59 remaining samples with a 20-25% PMN count being negative for enteropathogenic bacteria. CONCLUSIONS: The results suggest that the new culture media used in this study may have better recovery rates for enteropathogenic bacteria in acute gastroenteritis. Likewise, a correlation was observed between leukocyte count and isolation enteropathogenic bacteria.

Acute Disease↗

[Colostomy closure: is it an intervention without risk?].

OBJECTIVE: To examine whether colostomy closure is an operation with a high risk of complications. DESIGN: Retrospective study of colostomy closures in a 14-year period. PARTICIPANTS: 60 patients, averaging 54 years, males in 63%. The main indication for colostomy was colorectal neoplasm (47%), followed by trauma (23%). RESULTS: There were 15 Hartman's procedure reconstructions, 4 reconstructions of colostomy and mucous fistula, and 41 "simple" colostomy closures. The closure was extraperitoneal in 40%, suturing only the anterior colonic wall in 42%. We had 27 postoperative complications in 33% of the patients, without mortality. The average hospital stay was 8 days longer in the group with complications. The complication rate was 29% in the "simple" closure versus 42% in the reconstruction group (this difference was not statistically significant). The statistic analysis (chi-square and Mann-Whitney tests) showed no influence on morbidity of factors such as age, sex, previous disease, age of colostomy, and type and closure technique of colostomy. CONCLUSIONS: Colostomy closure has a high rate of postoperative complications (33%). These are minor in almost all cases, without mortality.

Adult↗