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Biomedical subjects

J Castro

Publications and source records attributed to J Castro.

At least 19 recordsLinked to original sources
PubMed2002

Analysis of endosulfan isomers and endosulfan sulfate in air and tomato leaves by gas chromatography with electron-capture detection and confirmation by gas chromatography-mass spectrometry.

Rapid analytical methods for the determination of endosulfan isomers and endosulfan-sulfate in air and plant samples were developed. The insecticides were trapped from air using a column containing Florisil and extracted with a low volume of ethyl acetate, assisted by sonication. Pesticide residues were determined by gas chromatography with electron-capture detection using a nonpolar capillary column. Residue identities were confirmed by gas chromatography coupled with mass spectrometry. Recoveries of these compounds from air samples were always higher than 78% with an RSD lower than 11% and the detection limits obtained were at least 0.3 ng/l air. Leaf samples were homogenised with ethyl acetate and extracts cleaned-up on an aluminium oxide column. Pesticides were eluted with a hexane-ethyl acetate (80:20, v/v) mixture. Recoveries obtained from plant samples were higher than 78% with an RSD lower than 14% and detection limits in leaves were 0.02 microg/g for each pesticide. These methods were applied to study the volatilisation of endosulfan from tomato leaves under laboratory conditions. A volatilisation rate near 1% of the initial amount of endosulfan per hour was obtained during the first 24 h at room temperature.

Air↗
PubMed2001

Effects of HsRad51 overexpression on cell proliferation, cell cycle progression, and apoptosis.

Expression of the DNA repair and recombination protein human Rad51 (HsRad51) is increased in transformed cells and in cancer cell lines. In order to study the effects of acute HsRad51 ectopic overexpression on cell proliferation, cell cycle progression, and apoptosis, we generated clones of the human fibrosarcoma cell line HT1080 carrying a HsRad51 transgene under a repressible promoter. The HsRad51-overexpressing cells showed decreased plating efficiency and growth rate in a dose-dependent manner with regard to the degree of overexpression. An accumulation of HsRad51-overexpressing cells in G(2) was observed following release of cells after synchronization with double thymidine block. Moreover, the fraction of apoptotic cells measured by annexin V-FACS increased with the time of HsRad51 overexpression. In the light of these observations, sustained increased levels of HsRad51 may contribute to tumor progression by causing a selection for cells tolerant to the growth-suppressive and apoptosis-inducing effects of acute HsRad51 overexpression.

Annexin A5↗
PubMed2001

Multiresidue analysis of insecticides in soil by gas chromatography with electron-capture detection and confirmation by gas chromatography-mass spectrometry.

A rapid multiresidue method has been developed for the analysis of nine insecticides (organochlorines, pyrethroids and organophosphorus) in soil. The method is based on the sonication extraction of residues from a certain amount of soil placed in a small column, using ethyl acetate. The effect of the residence time of insecticides in soil, the material of the columns used (glass or plastic columns) and the soil moisture content on the recovery of these compounds was also studied. Residues were determined by gas chromatography with electron-capture detection. The average recovery through the method obtained for these compounds varied from 90 to 108% with a relative standard deviation between 1 and 11%. The results of this study pointed out that the recoveries of insecticide residues obtained with plastic or glass columns at different soil moisture content were similar and that the residence of these compounds in soil during several days did not affect their recovery from soil. Confirmation of residue identity was performed by gas chromatography coupled with mass spectrometry.

Chromatography, Gas↗
PubMed2001

Hyperosmotic shock induces both activation and translocation of glucose transporters in mammalian cells.

The effect of osmotic stress on sugar transport was investigated in Clone 9 epithelial cells, which express the glucose uniporter GLUT1, and in 3T3-L1 adipocytes, which express both GLUT1 and GLUT4. An acute hyperosmotic shock increased the uptake of sugars in both cell types. In Clone 9 cells, this was followed by a regulatory volume increase (RVI) response. Stimulation of transport was rapid and reversible, with half-lives (t 1/2) for stimulation of 2-deoxy-D-glucose uptake of 5.6 +/- 0.9 (n=6) and 22.7 +/- 1.5 (n=4) min for Clone 9 cells and adipocytes respectively. The effect was dose dependent, reaching a maximum at 1.1 osM of 2.9 +/- 0.1-fold (n=3) for Clone 9 cells and 8.2 +/- 0.8-fold (n=3) for adipocytes. In the latter, this stimulation correlated with translocation of the glucose transporter isoform GLUT4 to the cell surface and was not significantly different from that elicited by 160 nM insulin (7.6 +/- 1.2-fold, n=3). The effect of osmotic shock was not, however, influenced by inhibitors of either phosphoinositide 3-kinase (PI 3-kinase) (wortmannin, 250 nM) or of p38 mitogen-activated protein kinase (p38 MAP kinase) (SB203580, 20 microM), which reportedly prevent GLUT4 translocation and/or activation by insulin respectively. These inhibitors also had no effect on the stimulation of transport by osmotic shock in Clone 9 cells. However, in contrast to adipocytes, the effect of osmotic shock on glucose transport in Clone 9 cells reflected primarily a change in the intrinsic activity of cell surface transporters and there was only a minor change in their subcellular distribution as assessed by cell immunostaining or no change as assessed by surface biotinylation. These results indicate that the response of cells to osmotic shock can involve changes both in transporter activity and location. The signal transduction pathways involved include neither PI 3-kinase nor the classical, osmotically-activated component, p38 MAP kinase.

3T3 Cells↗
PubMed2001

Necrotic volume increase and the early physiology of necrosis.

Whether a lethally injured mammalian cell undergoes necrosis or apoptosis may be determined by the early activation of specific ion channels at the cell surface. Apoptosis requires K+ and Cl- efflux, which leads to cell shrinking, an active phenomenon termed apoptotic volume decrease (AVD). In contrast, necrosis has been shown to require Na+ influx through membrane carriers and more recently through stress-activated non-selective cation channels (NSCCs). These ubiquitous channels are kept dormant in viable cells but become activated upon exposure to free-radicals. The ensuing Na+ influx leads to cell swelling, an active response that may be termed necrotic volume increase (NVI). This review focuses on how AVD and NVI become conflicting forces at the beginning of cell injury, on the events that determine irreversibility and in particular, on the ion fluxes that decide whether a cell is to die by necrosis or by apoptosis.

Animals↗
PubMed2001

Localization of ribosomal genes in Pleuronectiformes using Ag-, CMA3-banding and in situ hybridization.

In this paper we present the analysis of nucleolar organizer regions (NORs) in five species of the order Pleuronectiformes (Scophthalmus maximus, Scophthalmus rhombus, Platichthys flesus, Solea solea and Solea lascaris), a group of fish poorly studied cytogenetically. In spite of the small chromosome sizes, which characterize the karyotypes of this group, we implement both classical (Ag- and CMA3-staining) and molecular (in situ hybridization with a major rDNA probe) techniques for NOR location. NORs were localized in a single chromosome pair in all species studied and showed size variation within each species. Evidence of a NOR-site polymorphism was detected in Platichthys flesus and Scophthalmus maximus. NOR location was apparently the same within families (S. rhombus vs. S maximus, and S. solea vs. S. lascaris) but clearly differed among families, and supports the phylogenetic relationships proposed for the order Pleuronectiformes.

Animals↗
PubMed2001

Population analysis of an unusual NOR-site polymorphism in brown trout (Salmo trutta L.).

A population analysis of an unusual NOR-site polymorphism previously detected in brown trout (Salmo trutta L.) from North-western Spain was carried out in 225 individuals from 20 native populations from this area. The analysis performed has permitted us to reveal: (i) the ubiquity of this phenomenon in most river basins from NW Spain, 13 extra-NORs not observed in standard trout being detected; (ii) the rDNA constitution of all extra-NORs, as confirmed by CMA3-staining and rDNA-FISH, and their capability to constitute their own nucleolus; (iii) the constant location of extra-NORs within individuals and their stable transmission across generations; and (iv) the telomeric location of Ag-NORs, which were randomly distributed in the karyotype of S. trutta, mostly in the heterozygous condition. Repetitive sequences in the IGS or scattered along rDNA units, rather than chromosome rearrangements, could play an important role in the dispersion of NORs. The polymorphism described seems to have a single geographical origin, since a positive correlation was demonstrated between the degree of polymorphism and geographical distance to a central point of the river basin where the phenomenon showed a higher intensity. The distribution of NOR-site variation as compared with allozyme variation in the populations studied, showed significant statistical differences. Selection against high copy number, or mutation due to changes in NOR location, could be acting on this polymorphism to explain the observed distribution. The high dispersion of NOR-site variants within and among populations suggests the transposition phenomenon responsible for NOR jumping could be still active.

Animals↗
PubMed2001

Allozymic evidence of parapatric differentiation of brown trout (Salmo trutta L.) within an Atlantic river basin of the Iberian Peninsula.

The genetic variation of brown trout from Duero, one of the main Atlantic Iberian river basins, was assessed at 34 enzymatic loci in 62 native populations. A strong intrabasin differentiation was detected (G(ST) = 0.46; range D: 0-0.066), mainly attributable to the existence of two divergent groups of populations within Duero: southern and northern groups. This divergence was mainly a consequence of the unequal distribution of *75 and *100 alleles at sMDH-B1,2* isoloci, which were correlated with substantial differences in genetic diversity among regions. The Lower Course region (nearly fixed for the *100 allele) and Pisuerga River (nearly fixed for the *75 allele) showed lower heterozygosities (H approximately 0.8%) in contrast with adjacent areas, which evidenced intermediate frequencies for both alleles and higher heterozygosities (H: 2.2-3.1%). Vicariance appeared as the more probable explanation for the significant positive correlation detected between genetic and geographical distances in Duero Basin. Genetic relationships with adjacent Iberian drainages indicate a close similarity between the southern group and Cantabric trout, whereas the northern group constitutes an ancient form from this basin. This study confirmed complex genetic relationships in brown trout from northwest Iberia, reasserting the existence of clines at several loci and for genetic diversity. The interaction between Cantabric and Duero trout, as well as the location of the limit of the anadromous form around the 42 degrees N parallel, are both required to understand the genetic characteristics of brown trout from this area.

Alleles↗
PubMed2001

Nonselective cation channels as effectors of free radical-induced rat liver cell necrosis.

Necrosis, as opposed to apoptosis, is recognized as a nonspecific cell death that induces tissue inflammation and is preceded by cell edema. In non-neuronal cells, the latter has been explained by defective outward pumping of Na(+) caused by metabolic depletion or by increased Na(+) influx via membrane transporters. Here we describe a novel mechanism of swelling and necrosis; namely the influx of Na(+) through oxidative stress-activated nonselective cation channels. Exposure of liver epithelial Clone 9 cells to the free-radical donors calphostin C or menadione induced the rapid activation of an approximately 16-pS nonselective cation channel (NSCC). Blockage of this conductance with flufenamic acid protected the cells against swelling, calcium overload, and necrosis. Protection was also achieved by Gd(3+), an inhibitor of stretch-activated cation channels, or by isosmotic replacement of extracellular Na(+) with N-methyl-D-glucamine. It is proposed that NSCCs, which are ubiquitous although largely inactive in healthy cells, become activated under severe oxidative stress. The ensuing influx of Na(+) initiates a positive feedback of metabolic and electrolytic disturbances leading cells to their necrotic demise.

Animals↗
PubMed2001

Enhanced magnification endoscopy: a new technique to identify specialized intestinal metaplasia in Barrett's esophagus.

BACKGROUND: Specialized intestinal metaplasia (SIM) in Barrett's esophagus (BE) is not identifiable by standard endoscopy. Acetic acid instillation enhances the ability to detect columnar epithelium at the squamocolumnar union. Enhanced magnification endoscopy involves the combined use of magnification endoscopy with acetic acid. This study assessed the value of enhanced magnification endoscopy in detecting SIM in patients with BE. METHODS: Patients undergoing endoscopic surveillance because of short segment BE without dysplasia underwent enhanced magnification endoscopy with 1.5% acetic acid instillation. Standard endoscopy was followed by magnification endoscopy and repeated after acetic acid spraying. Surface patterns were characterized prior to and after acetic acid spraying. The observed surface patterns were compared with histology results. RESULTS: Forty-nine patients, 9 women and 39 men, with a mean age of 50.5 years were studied. One was excluded because of unclear definition of the surface pattern. Enhanced magnification endoscopy detected 4 different mucosal surface patterns: I, round pits; II, reticular; III, villous; and IV, ridged. A total of 129 areas were examined. Standard endoscopy identified an endoscopic pattern in 1.5% of the areas, standard endoscopy and acetic acid in 8.5%, magnification endoscopy alone in 38%, and enhanced magnification endoscopy in all 129 endoscopic areas. The yields for detecting SIM according to endoscopic patterns were as follows: pattern I, 0%; II, 11% (odds ratio 0.5, p = 0.54); III, 87% (odds ratio 36, p = 0.001); and IV, 100% (odds ratio 14, p = 0.015). CONCLUSIONS: Enhanced magnification endoscopy is an accurate method of predicting SIM in BE. The simplicity of the technique and its ability to identify characteristic endoscopic patterns with outstanding clarity and resolution that correlate with histologic identification of specialized intestinal metaplasia make enhanced magnification endoscopy an excellent method for the evaluation of patients with BE.

Acetic Acid↗
PubMed2001

High-LET radiation induces apoptosis in lymphoblastoid cell lines derived from atazia-telangiectasia patients.

PURPOSE: To investigate and compare the propensity of Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCL), derived from ataxia-telangiectasia (A-T) patients and from unaffected healthy individuals (controls), to undergo apoptosis after exposure to high-linear energy transfer (LET) radiation. MATERIALS AND METHODS: Four A-T (ARO, BMA, CSA and RJO) and two control (JAC and KKB3) LCL were exposed to doses of up to 4Gy of accelerated nitrogen ions (32-45 MeV/u, 8-12Gy/min). For comparative purposes X-ray irradiation (1.36 Gy/min) was also performed. The induction of apoptosis was studied 0-48 h after irradiation with the use of two methods: (1) monitoring of high molecular weight (HMW) DNA fragments by field inversion pulse gel electrophoresis (FIGE); and (2) morphological characterization ofapoptotic cells after fluorescent staining. In parallel, cell-cycle distribution, monitored by DNA flow cytometry, as well as measurements of p53/p21(WAF1) protein levels by Western blots, were investigated in these cells. RESULTS: High-LET radiation-induced apoptosis and G2/M-arrest in both A-T and control LCL. No significant increase in the amount of p53/p21(WAF1) proteins preceded apoptosis in control or in A-T LCL after high-LET irradiation. However, low-LET radiation did induce significant enhanced levels of p53 proteins in control but not in A-T LCL. CONCLUSIONS: LCL from both A-T homozygous and unaffected healthy individuals undergo apoptosis without accumulation of p53/p21(WAF1) proteins after exposure to high-LET radiation. In contrast, low-LET radiation induces apoptosis and significantly increases levels of p53 protein in control but not in A-T LCL.

Apoptosis↗
PubMed2001

Adolescent anorexia nervosa: the catch-up effect in bone mineral density after recovery.

OBJECTIVE: To determine whether bone mineral density (BMD) loss can be reversed in adolescent anorexic patients. METHOD: A prospective study with 108 anorexia nervosa patients (DSM-IV) from 12 to 17 years of age at the Eating Disorders Unit in the Hospital Clinic of Barcelona (Spain). They were first evaluated by dual-energy x-ray absorptiometry in lumbar spine and femoral neck consecutively from 1997 until 1999 and reexamined after 6 to 30 months. Results were compared with normative values of bone mass. RESULTS: Patients with poor outcome (n = 44) had a bone mass loss. Patients with good short-term outcome were divided in two groups. The group with normal BMD at first evaluation (n = 41) had a bone mass gain per year of 3.0% at lumbar spine and 0.5% at femoral neck. The group with low BMD at first evaluation (n = 23) had an increase per year of 9.1% at lumbar spine and 4.5% at femoral neck. In a multiple linear regression analysis with the variables body mass index, age, months with menstruation, and BMD zscore at first evaluation, the only predictor of BMD increase was the first z score both at the lumbar spine (coefficient R = 0.64; p < .001) and at the femoral neck (coefficient R = 0.5; p < .001). CONCLUSIONS: There is a catch-up effect in adolescent patients with low BMD but good short-term outcome.

Adolescent↗
PubMed2001

Clinical isolation and resistance patterns of and superinfection with 10 nosocomial pathogens after treatment with ceftriaxone versus ampicillin-sulbactam.

Isolation of pathogens from clinical cultures and their resistance patterns may be altered by antecedent antibiotic treatment. The objective of this study was to assess the influence of treatment with ceftriaxone versus that with ampicillin-sulbactam on recovery and superinfections with 10 nosocomial pathogens. The study was designed as a historical cohort study, using a propensity score to adjust for confounding by indication and multivariate survival analyses to adjust for other confounding. Two thousand four hundred forty-five patients were treated with ampicillin-sulbactam, and 1, 308 were treated with ceftriaxone. The study analyzed two outcomes: (i) recovery of pathogens from clinical cultures and (ii) microbiologically documented infections. Data were obtained from administrative, pharmacy, clinical, and laboratory databases and by chart extraction. Following treatment, new isolation of at least 1 of the 10 target pathogens occurred for 244 patients. After adjustment, more infections occurred in the ampicillin-sulbactam group (hazard ratio [HR], 1.55; P = 0.009). This was observed with all gram-negative rods combined (HR, 3.6; P < 0.001) and with each genus of the family Enterobacteriaceae. No differences in isolation of gram-positive bacteria were evident (P = 0.33). Microbiologically documented superinfections occurred in 172 patients and were less frequent in the ceftriaxone group (3.8% versus 5%; HR, 1.6; P = 0. 015). All the Escherichia coli and Klebsiella spp. isolates were susceptible to ceftriaxone, but half were resistant to ampicillin-sulbactam. The prevalence of oxacillin resistance among Staphylococcus aureus isolates was higher in the ceftriaxone group (63% versus 31%; odds ratio, 3.8; P = 0.08). Differences in the rates of superinfections and the likely causative organisms following treatment with ceftriaxone or ampicillin-sulbactam were evident. This may guide clinicians in empirical choices of antibiotics to treat superinfection.

Aged↗
PubMed2001

Susceptibility genes for asthma and allergy: hits and questions.

Allergy and asthma are closely related conditions which result from a complex interaction between several genetic and environmental factors. On the basis of familial linkage analysis data from different populations, some chromosomal regions containing a series of susceptibility loci have been identified. To date, genome-wide search studies have reported a group of chromosomal regions which include most likely several candidate genes for asthma and allergy. Some of these genes are excellent candidates, as they are known to modulate the immune response and airways inflammation processes. So, by focusing efforts on the analysis of those chromosomal regions, the characterization of the potentially important genes can be achieved. This will undoubtedly provide a clue for a better understanding of those pathologic components involved in the onset and development of asthma and allergic disorders.

Asthma↗
PubMed2000

Skin and wound infection by rapidly growing mycobacteria: an unexpected complication of liposuction and liposculpture. The Venezuelan Collaborative Infectious and Tropical Diseases Study Group.

OBJECTIVE: To describe 10 patients with skin and soft tissue infection caused by rapidly growing mycobacteria after cosmetic liposuction and liposculpture. DESIGN: Case series. SETTINGS: Eight private geographically separate surgical facilities from a single metropolitan area. PATIENTS: Eight patients with definite and 2 with presumptive cases of skin and soft tissue infection caused by rapidly growing mycobacteria after cosmetic surgery procedures during a 24-month period. Microorganisms were isolated from the purulent drainage obtained from wounds or fistulas in 8 cases and were identified as Mycobacterium fortuitum (3 cases) and Mycobacterium abscessus (5 cases) by routine microbiologic techniques. Acid-fast bacilli were observed on Ziehl-Neelsen-stained smears in the 2 remaining cases, but these ultimately failed to grow. In 2 of the surgical units, no apparent environmental predisposing factors were identified after thorough microbiologic environmental investigation. Clinically, all patients exhibited signs of inflammation, microabscesses, and purulent wound drainage within 24 months of abdominal and/or thigh liposuction or homologous fat tissue injection. INTERVENTION: A combined therapeutic approach including surgical drainage, debridment, and prolonged (>3 months) treatment with combined antimicrobial agents including clarithromycin was used in all cases. RESULTS: Nine of 10 patients responded to the combined therapeutic approach, and no evidence of infection was present during at least 12 months of follow-up. CONCLUSION: To our knowledge, this is the first series of patients with rapidly growing mycobacterial infections to be described after liposuction and liposculpture. Rapidly growing mycobacteria should be included in the differential diagnosis of skin and soft tissue infection after cosmetic surgery.

Abdomen↗
PubMed2000

Quality of rearing practices as predictor of short-term outcome in adolescent anorexia nervosa.

BACKGROUND: Studies of family relationships in anorexia nervosa have produced conflicting results. Some authors claim that family factors are related to short-term outcomes. METHODS: Perceived rearing practices, as measured by the EMBU (Egna Minnen Betraffande Uppfostran: 'My memories of Upbringing') were examined in a sample (N = 158) of adolescents with anorexia nervosa and compared with the perceptions of adolescents (N = 159) from the general population. A further comparison was made between the groups of patients with good and bad short-term outcomes. Logistic regression analysis was performed to evaluate the predictive value of different variables on short-term outcome. RESULTS: Overall, small differences were observed in the perceptions of rearing practices as expressed by the controls and the anorexic patients. Patients with bad short-term outcome perceived more rejection and control-overprotection from both parents than those with good outcome. In the logistic regression analysis only Rejection from father and the EAT (Eating Attitudes Test) total score gave independent prediction of treatment response. CONCLUSIONS: Taken as a whole, these results do not support the idea of altered rearing practices in anorexic patients, at least in young patients with a short evolution of the disease. Perceived rearing practices, especially 'rejection', appear to have an appreciable effect on the short-term outcome.

Adolescent↗