Search PubMed⌕ Search

Biomedical subjects

J A Castro

Publications and source records attributed to J A Castro.

At least 37 records · Page 2Linked to original sources

Reduction of AOX in the bleach plant of a pulp mill.

The reconfiguration of an existing five-stage bleaching D0E1D1E2D2 sequence is proposed to ensure the minimal formation of organochlorine compounds, expressed as AOX, in the bleach plant of a kraft pulp mill processing Eucalyptus globulus wood. This reduction of the load of AOX in the effluents can be achieved without introducing new and expensive technologies in the bleaching process. In practice, this goal can be achieved by eliminating the washing step between the D0 and the E1 stages. With this strategy, the total AOX in the liquid effluent of the bleach plant can be reduced by almost 65% while maintaining a level of 90% ISO brightness of the pulp and even diminishing the degradation of its polysaccharides. With this process redesign, the total AOX discharge from the bleach plant can be decreased from 1.2 to 0.42 kg/t prior to any biological treatment. Furthermore, the proposed modified (D0E1)D1E2D2 sequence enables the reduction of the total flow rate of effluent to be treated and also the total consumption of water, which is quite critical in these types of plants. Moreover, with this strategy, one is able to move the first drum washer of a conventional bleaching sequence to the entrance of the bleach plant. This gives rise to a cleaner pulp and therefore to lower requirements of chlorine dioxide, which in turn will enable further reductions in the global AOX generation.

Adsorption↗

Cytosolic xanthine oxidoreductase mediated bioactivation of ethanol to acetaldehyde and free radicals in rat breast tissue. Its potential role in alcohol-promoted mammary cancer.

Epidemiological evidence links alcohol intake with increased risk in breast cancer. Not all the characteristics of the correlation can be explained in terms of changes in hormonal factors. In this work, we explore the possibility that alcohol were activated to acetaldehyde and free radicals in situ by xanthine dehydrogenase (XDh) and xanthine oxidase (XO) and/or aldehyde oxidase (AO). Incubation of cytosolic fraction with xanthine oxidoreductase (XDh+XO) (XOR) cosubstrates (e.g. NAD+, hypoxanthine, xanthine, caffeine, theobromine, theophylline or 1,7-dimethylxanthine) significantly enhanced the biotransformation of ethanol to acetaldehyde. The process was inhibited by allopurinol and not by pyrazole or benzoate or desferrioxamine and was not accompanied by detectable formation of 1HEt. However, hydroxylated aromatic derivatives of PBN were detected, suggesting either that hydroxyl free radicals might be formed or that XOR might catalyze aromatic hydroxylation of PBN. No bioactivation of ethanol to acetaldehyde was detectable when a cosubstrate of AO such as N-methylnicotinamide was included in cytosolic incubation mixtures. Results suggest that bioactivation of ethanol in situ to a carcinogen, such as acetaldehyde, and potentially to free radicals, might be involved in alcohol breast cancer induction. This might be the case, particularly also in cases of a high consumption of purine-rich food (e.g. meat) or beverages or soft drinks containing caffeine.

Acetaldehyde↗

Partial liquid ventilation with perfluorocarbons for treatment of ARDS in burns.

Pulmonary failure remains the major determinant of mortality and morbidity following burn injury. We hypothesized that intratracheal instillation of perfluorocarbon liquids could be a therapeutic measure in combination with conventional mechanical ventilation to improve pulmonary gas exchange in acute respiratory distress syndrome with thermal injury. Forty-five New Zealand rabbits were used for this prospective and randomized experimental study. The animals were burned by scald to reach full-thickness 40% burn surface area. After inducing respiratory distress by repeated lung lavage with saline, animals were divided randomly into three groups of 15 rabbits each. First group (control group) received conventional treatment (continuous positive-pressure ventilation) using a FiO(2) of 1.0, tidal volume of 12 ml/kg, respiratory frequency of 30 cycles/min and PEEP of 6 cm H(2)O. Second group was treated with 9 ml/kg of intratracheal perfluorocarbon. Third group was treated with 15 ml/kg of intratracheal perfluorocarbon. All groups were ventilated for 6 h. In the perfluorocarbon groups, PaO(2) increased significantly (P<0.05) from 46+/-4 to 439+/-10 mmHg compared to the control group in a dose-related manner. In pulmonary parameters we observed significant (P<0.05) decrease in mean airway pressures from the pre-treatment value of 11.44+/-0.15 cm H(2)O to the post treatment 10.22+/-0.12 cm H(2)O and increase (P<0.05) in respiratory system compliance from 1.8+/-0.02 to 2.46+/-0.07 ml/cm H(2)O with the perfluorocarbon. Perfluorocarbon instillation did not result in statistically significant changes in arterial pressure, heart rate and central venous pressure. In conclusion, partial liquid ventilation with perfluorocarbon is a new technique leading to a marked and sustained improvement in oxygenation and pulmonary function in an experimental model of ARDS in burns.

Animals↗

Genetic analyses in five western Mediterranean populations: variation at five tetrameric short tandem repeat loci.

A population study of the Balearic (Majorca, Minorca, Ibiza, Chueta) and Valencia populations was carried out using the short tandem repeat loci HUMTHO1, D4S243, HUMF13A1, D18S535, and D12S391. All the populations sampled were found to be in Hardy-Weinberg equilibrium for the five markers analyzed. Several statistical analyses showed a clear displacement of the Chueta and Ibiza populations from the other populations sampled. These results are in agreement with those obtained from the analysis of classical markers and mitochondrial DNA restriction fragment length polymorphisms, as well as with the history of these populations. A comparative study performed with other European populations using three of the five markers selected for this study (HUMTHO1, HUMF13A1, and D12S391) revealed a clear differentiation only of the Chueta population. We detected a tendency for a west-east clinal distribution in the frequency of the HUMTH01*6 allele in the European and Mediterranean area. This distribution could reflect some of the migratory events that have happened throughout that area's history. The forensic usefulness of these markers can be judged by their highly combined power of discrimination (0.999997).

DNA Fingerprinting↗

Rat ventral prostate xanthine oxidase bioactivation of ethanol to acetaldehyde and 1-hydroxyethyl free radicals: analysis of its potential role in heavy alcohol drinking tumor-promoting effects.

The ability of the ventral prostate cytosolic fractions to biotransform ethanol to acetaldehyde and 1-hydroxyethyl (1HEt) radicals was tested. Acetaldehyde formation was determined by GC-FID analysis in the head space of incubation mixtures. 1HEt was determined by spin trapping with PBN followed by extraction, silylation of the adduct and GC-MS of the product. Prostate cytosol was able to biotransform ethanol to acetaldehyde in the presence of NADH, hypoxanthine, xanthine, caffeine, theobromine, theophylline, and 1,7-dimethylxanthine but not in the presence of N-methylnicotinamide. All these biotransformations were inhibited by allopurinol and were sensitive to heating for 5 min at 100 degrees C. The biotransformation of ethanol to acetaldehyde in the presence of purines as cosubstrates was accompanied by the formation of hydroxyl and 1HEt radicals as detected by GC-MS, and the process was inhibited by allopurinol. Results suggest that prostate cytosolic xanthine oxidase is able to bioactivate ethanol to acetaldehyde and free radicals. The potential of these processes to be involved in tumor-promoting effects of heavy alcohol drinking in conjunction with high meat and/or purines consumption is analyzed. Multifactorial epidemiological studies considering that possibility might be convenient. Teratogenesis Carcinog. Mutagen. 21:109-119, 2001.

Acetaldehyde↗

[Ultrastructural alterations in colonic mucosa of nifurtimox treated rats].

Nifurtimox (Nfx) is a chemotherapeutic agent used in the treatment of acute Chagas' disease. Clinical and experimental studies with this nitroheterocyclic compound evidenced serious undesirable side effects. These were correlated with Nfx nitroreduction to a nitroanion radical followed by superoxide anion generation through a redox cycling process. The aim of this study was to verify whether the oral administration of Nfx to Sprague Dawley male rats (100 mg.kg-1, p.o.) produced any observable ultrastructural alteration in the cells of the colonic mucosa. Results showed that 24 h after Nfx administration there were observable alterations in this type of cells. They essentially consisted of moderate dilatation of their endoplasmic reticulum and intense dilatation of their Golgi complex. Already 1 and 3 h after Nfx administration, the original compound reached a concentration of 9.7 +/- 2.9 and 7.0 +/- 1.7 nmol.g-1 respectively in the colonic tissue. Studies on Nfx nitroreductase activity of colonic mucosa as determined spectrophotometrically and by HPLC methods showed that the microsomal fraction (from 0.72 +/- 0.29 to 0.26 +/- 0.04 nmol Nfx.min-1.mg-1 protein) but not the cytosol had the ability to nitroreduce Nfx. The results obtained show a correlation between the ultrastructural localization of injury and that of nitroreductase activity. The intense deleterious effects of Nfx in the Golgi apparatus suggest the potential occurrence of alterations in the synthesis/storage of secretory products of the colonic mucosa.

Animals↗

Cytochrome P450 reductase-mediated anaerobic biotransformation of ethanol to 1-hydroxyethyl-free radicals and acetaldehyde.

The ability of cytochrome P450 reductase to metabolize ethanol (EtOH) to acetaldehyde (AC) and 1-hydroxyethyl free radicals (1HEt) in anaerobic media was studied. Determination of AC was made by GC-FID analysis of the head space of incubation mixtures. The formation of 1HEt was established by GC-MS analysis of the adduct formed between the radical and the spin trap PBN. Results showed that pure human P450 reductase is able to biotransform EtOH to AC and 1HEt in a NADPH-dependent process under an oxygen-free nitrogen atmosphere. Pure FAD in the presence of NADPH was also able to generate AC and 1HEt from the alcohol. Anaerobic incubation mixtures containing either rat liver microsomes or pure nuclei were also able to biotransform EtOH to AC and 1HEt in the presence of NADPH. These processes were inhibited by antibody against rat liver microsomal P450 reductase. Results suggest that semiquinone forms of the flavin in P450 reductase may biotransform EtOH. These reactions might be of some significance in tissues where the P450 reductase is present in the absence of specific forms of cytochrome P450 known to be involved in EtOH metabolism (e.g. CYP2E1). However the toxicological significance of this enzymatic process remains to be established.

Acetaldehyde↗

Antagonism of direct alloreactivity of an HLA-B27-specific CTL clone by altered peptide ligands of its natural epitope.

Antagonism of allospecific CTL by altered MHC ligands is a potential approach to specific immunomodulation of allogeneic T cell responses in acute graft rejection and graft-vs-host disease. In this study we have analyzed the capacity of peptide analogs of a natural HLA-B27-allospecific CTL epitope to antagonize direct alloreactivity. Alanine scanning demonstrated that positions 4, 5, and 7 of the peptide epitope were critical for allorecognition. A number of relatively conservative substitutions at each of these positions were then tested for their effect on allorecognition and antagonism. All substitutions at position 5 abrogated cytotoxicity. In contrast, a few changes at positions 4 and 7 were tolerated, indicating a limited flexibility of the allospecific CTL in recognition of peptide epitope variants. Most of the substitutions impairing cytotoxicity actually induced antagonism. However, whereas epitope variants with changes at positions 4 and 7 behaved as weak or intermediate antagonists, some of the variants with changes at position 5 antagonized CTL alloreactivity almost completely. The results in this study demonstrate for the first time that antagonism of direct class I-mediated alloreactivity can be achieved by variants of a natural allospecific peptide epitope.

Amino Acid Substitution↗

Genetic variability at nine STR loci in the Chueta (Majorcan Jews) and the Balearic populations investigated by a single multiplex reaction.

A study of the genetic variability in the Chueta (Majorcan Jews) and the Balearic (Majorca and Minorca Islands) populations was carried out using a multiplex system containing the nine tetrameric STRs D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820. The Chueta population has remained isolated because intermarriage with non-Jews did not take place until the middle of this century, which has resulted in it being a small inbred community. The results indicate the existence of HW equilibrium for the Chueta and Balearic populations. No pair-wise correlation was observed between the nine markers. Consequently, they seem to comprise a suitable group of markers for population genetics purposes and for paternity and forensic testing.

DNA↗

Prevalence of the C282Y mutation for haemochromatosis on the Island of Majorca.

The C282Y mutation of the HFE gene has been reported to be present in most of the patients with hereditary haemochromatosis (HH) of Northern European ancestry. HH affects approximately 1/300 individuals, but it is not evenly distributed in the different European countries. In the present study, polymerase chain reaction (PCR) and restriction-enzyme digestion were used to analyse the frequency of the most important mutation in haemochromatosis (C282Y) in subjects from Majorca (Balearic Islands, Spain) and patients with haemochromatosis. The results were compared with other studies from Spain and Europe. A total of 420 Majorcan chromosomes were analysed and the C282Y mutation was observed at a frequency of 2.62%+/-0.8 (11 heterozygotes: eight men and three women). In the group of hereditary haemochromatosis probands, 13 out of 14 were homozygous for the C282Y mutation. In the distribution of the C282Y mutation, a north-west to south-east cline was detected, supporting the Celtic origin of this mutation.

Adult↗

Benznidazole-induced ultrastructural and biochemical alterations in rat colon.

AIM: To study the effects of benznidazole (Bz), a drug used in the chemotherapy of the acute and the intermediate phase of Chagas' disease, on the colon of rats. METHODS: Sprague Dawley male rats received Bz 100 mg/kg ig. After 24 h colons were examined by electron microscopy. Concentrations of Bz in colonic tissue were measured by HPLC. Bz nitroreduction was followed by the decrease in the drug concentration using spectrophotometry and HPLC or by covalent binding to proteins of reactive products formed under in vivo and in vitro conditions. RESULTS: Colon mucosa of Bz-treated rats showed intense ultrastructural alterations: abundant mucus secretion at the level of the Goblet cells and dilatation of the endoplasmic reticulum and the Golgi apparatus in epithelial cells. The concentration of Bz in tissue was (59 +/- 18) and (93 +/- 14) nmol/g (protein) 1 and 3 h after oral administration to rats, respectively. Colonic microsomes anaerobically activated Bz in the presence of NADPH. This activating nitroreductive pathway only involved a minor part of the total Bz and could not be detected spectrophotometrically or by HPLC analysis of the Bz consumed. Reactive metabolites that bound covalently to microsomal proteins were formed in this process. The covalent binding was also observed in vivo 1, 3, 6, and 24 h after administration of the labeled drug to rats. CONCLUSION: Reactive Bz metabolites produced during nitroreductive bioactivation of the drug in the colonic mucosa could interact with proteins and other cellular constituents to cause injury.

Animals↗

A liver nuclear ethanol metabolizing system. Formation of metabolites that bind covalently to macromolecules and lipids.

Recent studies from the laboratory reported the presence in highly purified liver nuclear preparations free of endoplasmic reticulum, mitochondria or cytosol, of an ethanol metabolizing group of enzymes (NEMS) leading to acetaldehyde and to hydroxyl and 1-hydroxyethyl (1HEt) free radicals. In the present study it is reported that when NEMS metabolize [14C]ethanol using NADPH as cofactor, its reactive metabolites bind covalently to nuclear proteins and lipids. No covalent binding to DNA was detected with presently used procedures. The covalent binding to nuclear proteins was acid labile and is mostly attributable to acetaldehyde. Additional evidence was attempted through studies where the acetaldehyde was identified as its 2,4-dinitrophenylhydrazone or as its pentafluorphenylhydrazone and gas chromatography (GC) analysis using electron capture detection. Values obtained were close to detection limit and of variable nature. The covalent binding to nuclear lipids involved phospholipids, fatty acids and esters and cholesterol free and esterified and it was only partially labile to acid treatment. Production of ethanol reactive metabolites such as acetaldehyde and free radicals, nearby liver nuclear DNA and nuclear proteins or lipids, might have significant toxicological consequences.

Acetaldehyde↗

The genetic structure of Drosophila subobscura populations from the Islands of Majorca and Minorca (Balearic Islands, Spain) based on allozymes and mitochondrial DNA.

The genetic structure of seven populations of Drosophila subobscura from different locations on Majorca and Minorca (Balearic Islands, Spain) was studied using two types of markers: allozyme and mitochondrial DNA restriction analyses. Both markers showed congruent results. In the allozyme data, when the Acph-1 locus was excluded from the joint FST statistics, only three out of 21 comparisons were statistically significant, lending support to the hypothesis of low genetic differentiation. The mtDNA restriction analyses showed two haplotypes at a high frequency (more than 40% each), irrespective of the location considered, and a number of endemic haplotypes at very low frequencies (not higher than 2% each). The analyses of the genetic structure yielded a pattern similar to the allozymes. The cytonuclear disequilibrium analyses showed the difficulty of detecting cytonuclear associations in natural populations because they are mainly transient.

Alleles↗

Liver nuclear ethanol metabolizing systems (NEMS) producing acetaldehyde and 1-hydroxyethyl free radicals.

Biotransformation of ethanol by liver nuclei was studied. The formation of acetaldehyde was determined by GC/FID. The 1-hydroxyethyl (1HEt) formation was established by spin trapping of the radical with N-t-butyl-alpha-phenylnitrone (PBN) followed by GC/MS. Liver nuclei, free of endoplasmic reticulum, cytosol or mitochondria, were able to biotransform ethanol to acetaldehyde in the presence of NADPH under air. Only 22% activity was observed in the absence of the cofactor. Twenty-six percent of the NADPH-dependent activity and 47% of the NADPH-independent activity were observable under nitrogen. Aerobic biotransformation was inhibited by CO, SKF 525A, 4-methylpyrazole and by diethyldithiocarbamate. This suggests that CYP2E1 is involved in the process. However, the formation of acetaldehyde was able to proceed under a pure CO atmosphere. The lack of inhibitory effects of 2-mercapto-1-methylimidazol and thiobenzamide excludes the potential participation of the NADPH flavin monooxigenase system. The formation of hydroxyl radicals in the process is suggested by the partial inhibitory effect of 5 mM mannitol and 5 mM sodium benzoate and by the fact that the 1HEt was detected. The NADPH-dependent anaerobic ethanol biotransformation pathway was stimulated by FAD and inhibited to some extent by iron chelators. The relevance of a liver nuclear ethanol biotransformation, generating reactive metabolites, such as acetaldehyde and free radicals, nearby DNA, nuclear proteins and lipids is discussed.

Acetaldehyde↗

Determination of immuno-cross-reactivity between Cryptosporidium parvum and Eimeria spp.

Immuno-cross-reactivity between Cryptosporidium parvum and Eimeria spp. was studied by the indirect fluorescent antibody test (IFAT) and Western blot procedure. Thirty-seven sera from asymptomatic (non-diarrheic) cattle, with known coprological (presence-absence of coccidia) and serological data respecting C. parvum, were tested by IFAT using Eimeria oocysts as antigen. Most sera (54%) displayed immunofluorescence around the surface of the Eimeria oocysts. Simultaneously, serum samples from rabbits naturally infected with Eimeria spp. (E. magna, E. intestinalis and E. residua), but free of C. parvum infection, were used to investigate the recognition of C. parvum oocyst antigens by the Western blot procedure. Fractions in the 11.5-94 kDa range, as well as others with molecular masses over 94 kDa, were recognized by sera from rabbits. Sera collected during patency period showed low or moderate reaction with antigenic fractions in the 11.5-25 kDa range. However, 29, 58 and 71 to 75 kDa proteic fractions were moderately or strongly recognized even after rabbits finished oocyst excretion.

Animals↗

Mitochondrial DNA haplotype frequencies in natural and experimental populations of Drosophila subobscura.

The evolution of Drosophila subobscura mitochondrial DNA has been studied in experimental populations, founded with flies from a natural population from Esporles (Majorca, Balearic Islands, Spain). This population, like other European ones, is characterized by the presence of two very common (>96%) mitochondrial haplotypes (called I and II) and rare and endemic haplotypes that appear at very low frequencies. There is no statistical evidence of positive Darwinian selection acting on the mitochondrial DNA variants according to Tajima's neutrality test. Two experimental populations, with one replicate each, were established with flies having a heterogeneous nuclear genetic background, which was representative of the composition of the natural population. Both populations were started with the two most frequent mitochondrial haplotypes, but at different initial frequencies. After 13 to 16 generations, haplotype II reached fixation in three cages and its frequency was 0.89 by generation 25 in the fourth cage. Random drift can be rejected as the force responsible for the observed changes in haplotype frequencies. There is not only statistical evidence of a linear trend favoring a mtDNA (haploid) fitness effect, but also of a significant nonlinear deviation that could be due to a nuclear component.

Animals↗