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Biomedical subjects

F Moreno

Publications and source records attributed to F Moreno.

At least 109 records · Page 6Linked to original sources
PubMed1999

Interaction of acrylodan with human serum albumin. A fluorescence spectroscopic study.

The binding of the fluorescent probe acrylodan (AC) to human serum albumin (HSA) was studied by fluorescence spectroscopy. The binding isotherms could be fitted to two types of sites. Competition experiments using iodoacetamide suggested that AC binds tightly on HSA by the cysteine-34. Attempts were made to find the location of the second site using high concentrations of warfarin, phenylbutazone, diazepam, indomethacin, palmitic acid or bilirubin in order to displace the bound AC to the HSA. Bilirubin was the only ligand able to displace the bound AC. This result suggests that AC, which is a very hydrophobic molecule also capable of labeling lysine residues, should also bind the human albumin in the primary site of bilirubin, but with less affinity than to the cysteine-34.

2-Naphthylamine↗
PubMed1999

[Paternity analysis using four DNA markers amplified by polymerase chain reaction].

BACKGROUND: DNA typing in forensic analysis is a useful tool to analyze paternity due to its high discrimination power. AIM: To report the experience of Servicio Medico Legal in Santiago, resolving cases of dubious paternity. SUBJECTS AND METHODS: Four highly polymorphic loci, amplified by polymerase chain reactions, were analyzed in 153 cases of uncertain paternity. The paternity index was calculated for each case. RESULTS: The four genetic markers analyzed provided an exclusion probability of 0.933 for the general population in Santiago. Thirty-seven cases were excluded as parents. In 31 cases, the paternity index ranged from 19 to 100, considered as probable paternity and 77 cases had an index of over 100, considered as almost certain paternity. Eight cases had an index between 0.5 and 19, considered as inconclusive. All loci met Hardy-Weinberg expectations and their frequencies were similar to other data from people living in Santiago. CONCLUSIONS: The use of these genetic markers proved to be very useful, reliable and with a high exclusion power for paternity analysis.

DNA↗
PubMed1998

Analysis of cations in drainage water and soil solution by single-column ion chromatography.

Single-column ion chromatography (SCIC) for cation determination in drainage water and soil solution was tested. Using a 100-microliter sample loop, the SCIC detection limits were 0.04 mg l-1 for Na+, 0.02 mg l-1 for NH4-N, 0.06 mg l-1 for K+, 0.05 mg l-1 for Mg2+, and 0.085 mg l-1 for Ca2+. Results were highly reproducible in wide ranges of concentrations. Results obtained by the SCIC method were compared with those by atomic emission spectrometry (Na+ and K+) and atomic absorption spectrometry (Mg2+ and Ca2+). Student's t-test and regression analysis showed that corresponding methods agree closely.

Calcium↗
PubMed1998

Analysis of inorganic anions in drainage water and soil solution by single-column ion chromatography.

Single-column ion chromatography (SCIC) for anion determination in drainage water and soil solution was tested. The SCIC minimum detection limits (100-microliter sample loop) were 0.75 mg l-1 for Cl-, 0.2 mg l-1 for NO2-N, 0.02 mg l-1 for NO3-N, 1.25 mg l-1 for HPO4-P, and 0.5 mg l-1 for SO4-S. The results showed a high reproducibility. Results for Cl-, NO3-N and SO4-S obtained by the SCIC method were compared with those obtained by traditional methods; Student's t-test and regression analysis showed that the methods agree closely.

Anions↗
PubMed1998

The hexokinase 2 protein participates in regulatory DNA-protein complexes necessary for glucose repression of the SUC2 gene in Saccharomyces cerevisiae.

The HXK2 gene plays an important role in glucose repression in the yeast Saccharomyces cerevisiae. Recently we have described that the HXK2 gene product, isoenzyme 2 of hexokinase, is located both in the nucleus and in the cytoplasm of S. cerevisiae cells. In this work we used deletion analysis to identify the essential part of the protein-mediating nuclear localisation. Determinations of fructose-kinase activity and immunoblot analysis using anti-Hxk2 antibodies in isolated nuclei, together with observations of the fluorescence distribution of Hxk2-GFP fusion protein in cells transformed with an HXK2::gfp mutant gene, indicated that the decapeptide KKPQARKGSM, located between amino acid residues 7 and 16 of hexokinase 2, is important for nuclear localisation of the protein. Further experimental evidence, measuring invertase activity in wild-type and mutant cells expressing a truncated version of the Hxk2 protein unable to enter the nucleus, shows that a nuclear localisation of Hxk2 is necessary for glucose repression signalling of the SUC2 gene. Furthermore, we demonstrate using gel mobility shift analysis that Hxk2 participates in DNA-protein complexes with cis-acting regulatory elements of the SUC2 gene promoter.

Amino Acid Sequence↗
PubMed1998

Hexokinase PII has a double cytosolic-nuclear localisation in Saccharomyces cerevisiae.

We describe here that the HXK2 gene product, isoenzyme PII of hexokinase, is located in both the nucleus and the cytoplasm of Saccharomyces cerevisiae cells. This conclusion is supported by assays of hexokinase-specific activity in isolated nuclei from wild-type and hxk1lhxk2 double mutant strains, by immunoblot experiments using anti-Hxk2 antibodies and by observation of the fluorescence distribution of a Hxk2-GFP fusion protein in cells transformed with the HXK2::gfp gene.

Cell Fractionation↗
PubMed1998

A 27 kDa protein binds to a positive and a negative regulatory sequence in the promoter of the ICL1 gene from Saccharomyces cerevisiae.

IsocitrateICL1, is one of the key enzymes of the glyoxylate pathway, which operates as an anaplerotic route for replenishing the tricarboxylic acid cycle; it is required for growth of Saccharomyces cerevisiae on carbon sources such as ethanol, but is dispensable when fermentable carbon sources are available. The positive regulation of the ICL1 gene by an upstream activating sequence (UAS) element located between -397 and -388 has been previously reported. In this paper we show that the ICL1 promoter sequence 5'-AGTCCGGACTAGCATCCCAG-3' located between -261 and -242 contains an upstream repressing sequence (URS) element. We have identified and partially purified a 27 kDa protein that binds specifically to both the UAS and URS sequences of the ICL1 promoter. For both UAS and URS, binding requires the protein Snf1 (Cat1), a protein kinase essential for the derepression of genes repressed by glucose. Binding does not take place with extracts from glucose-grown strains, unless they lack Mig1, a negative regulatory protein involved in glucose repression.

Chromatography, Affinity↗
PubMed1998

Effect of a necrogenic dose of diethylnitrosamine on vitamin E-deficient and vitamin E-supplemented rats.

In order to evaluate the effects of a necrogenic dose of diethylnitrosamine (DEN) on vitamin E-deficient and vitamin E-supplemented rats, a single dose of the drug (200 mg/kg body weight) was injected intraperitoneally at the end of 10 weeks of treatment with the diets. The hepatic necrosis and lipoperoxidation provoked by DEN were evaluated 24, 48, 72 and 120 hours after the injection and were found to be more intense in the deficient group (thiobarbituric acid reactive substances (TBARS): 5.20 +/- 1.48 nmol/mg protein; necrosis volume: 68.99 +/- 8.36%; P < 0.05) during the second period. Also, in the same group and during the same period, mean plasma and hepatic vitamin E concentrations and mean liver glutathione concentration were the lowest detected, suggesting the occurrence of antioxidant consumption due to the toxic action of DEN. In contrast to vitamin E deficiency, which permitted the drug to exert stronger toxic effects, 20-fold supplementation with vitamin E did not provide additional protection against the lipoperoxidation and necrosis provoked by DEN (P < 0.05). The results suggest that other mechanisms in addition to lipoperoxidation provoked by free radicals originating from the metabolism of nitrosamines by the cytochrome P-450-dependent enzymatic system may be involved in the hepatotoxic action of these substances.

Animals↗
PubMed1998

Changes in quality of life after renal transplantation.

The objective of this study was to evaluate the modifications that renal transplantation produces on the quality of life (QOL) of patients with chronic renal failure (CRF) previously undergoing hemodialysis (HD) and to analyze the possible factors implicated. A multicenter study of QOL was performed on 1,023 patients undergoing dialysis, using as QOL indicators the Karnofsky Scale (KS) and the Sickness Impact Profile (SIP). Among this group, 93 patients received a renal transplant and QOL was re-studied in them; each subject, therefore, was his own control. In the 88 patients with a functioning graft, an improvement in QOL indices was globally observed; this improvement was much more marked in men than in women, for unclear reasons. Older age and greater prior comorbidity diminished the beneficial effects of transplantation.

Age Factors↗
PubMed1998

The post-larval development of lateral musculature in gilthead sea bream Sparus aurata (L.) and sea bass Dicentrarchus labrax (L.).

Fibre-type differentiation of lateral musculature has been studied in gilthead sea bream Sparus aurata (L.) and sea bass Dicentrarchus labrax (L.) during post-larval development using ultrastructural, histochemical and morphometric techniques. The study showed three muscle layers: red, intermediate (or pink) and white. Initially, most of the red muscle showed low myosin ATPase (m-ATPase) activity fibres, whereas near the transverse septum some small high m-ATPase activity fibres appeared and later acquired a rosette aspect. Afterwards, during adult growth the red muscle showed a histochemical mosaic appearance. The pink muscle in sea bass was observed at the beginning of juvenile development by the oxidative technique (NADH-RT reaction) whereas in gilthead sea bream it was also observed at the end of larval development. The pink layer consists of high m-ATPase activity fibres. However, along the muscle development other low and moderate m-ATPase activity fibres were observed close to the red and white muscles, respectively. The white muscle of juvenile fish showed a histochemical mosaic appearance near the pink muscle. In adult specimens the mosaic white muscle spread out occupying the whole of the myotome. Morphometric analysis shows a significant increase in mean fibre diameter during post-larval development, as shown by the Student's t-test (hypertrophic growth). Skewness and kurtosis values of fibre diameters point to the generation of a new fibres from the myosatellite cells (hyperplastic growth).

Animals↗
PubMed1998

Muscle development in gilthead sea bream (Sparus aurata, L.) and sea bass (Dicentrarchus labrax, L.): further histochemical and ultrastructural aspects.

The histochemical profiles--mATPase and NADH-TR reactions--of the red and white muscle fibres of gilthead sea bream and sea bass were determined from the first week after hatching. Modifications of the mATPase technique by combinations of pH/time/molarity were carried out in order to compare the sensitivity of the myosin ATPase of each muscle fibre type of the lateral muscle. Results showed that the staining of muscle fibres was independent of small modifications in the technique. The intermediate 'pink' muscle was histochemically defined towards the end of the larval life and is considered to be implicated in the growth of the myotome. A layer of external cells was observed, by electron microscopical examination, between the connective tissue of the skin and the superficial red muscle fibres of larvae and postlarvae. It is suggested that the external cells are unlikely to be a source of red muscle fibres and implicated on the growth of the myotome, but rather a part of the dermatome. The timing, areas and mechanisms of hyperplastic growth of the myotome were defined and discussed.

Animals↗
PubMed1998

Lens injuries induced by occupational exposure in non-optimized interventional radiology laboratories.

Several cases of ophthalmologically confirmed lens injuries, caused by occupational radiation exposure, have occurred in two X-ray rooms devoted to vascular and visceral interventional radiology procedures. Both laboratories were equipped with overcouch X-ray systems not designed for interventional radiology and without specific tools for radiation protection of the eyes. Typical workloads ranged from between two and five procedures per day. For the two radiologists affected, estimates for the dose to eye lens ranged from 450 to 900 mSv per year, over several years. Once the incidents had been detected, the X-ray systems in both rooms were removed and new equipment specifically designed for interventional radiology was installed, including suspended shielding screens. Since these lens injuries were only detected accidentally, measures to avoid similar occurrences in the future are discussed.

Cataract↗
PubMed1997

Isocitrate lyase localisation in Saccharomyces cerevisiae cells.

The isocitrate lyase from Saccharomyces cerevisiae was only located in the cell cytoplasm. This protein was found not to be associated with cell organelles, even under growth conditions that induce peroxisome proliferation. This conclusion is supported by experiments carried out by damaging the protoplast plasma membrane with DEAE-dextran, by differential centrifugation of osmotically lysed protoplast and by using the green fluorescent protein (GFP) of Aequorea victoria as a reporter fusion tag to localise the subcellular compartment to which isocitrate lyase is targeted.

Cell Compartmentation↗
PubMed1997

Determination of IgE antibodies to the benzylpenicilloyl determinant: a comparison of the sensitivity and specificity of three radio allergo sorbent test methods.

The quantitation of in vitro IgE antibodies to the benzylpenicilloyl determinant (BPO) is a useful tool for evaluating suspected penicillin allergic subjects. Although many different methods have been employed, few studies have compared their diagnostic specificity and sensitivity. In this study, the sensitivity and specificity of three different radio allergo sorbent test (RAST) methods for quantitating specific IgE antibodies to the BPO determinant were compared. Thirty positive control sera (serum samples from penicillin allergic subjects with a positive clinical history and a positive penicillin skin test) and 30 negative control sera (sera from subjects with no history of penicillin allergy and negative skin tests) were tested for BPO-specific IgE antibodies by RAST using three different conjugates coupled to the solid phase: benzylpenicillin conjugated to polylysine (BPO-PLL), benzylpenicillin conjugated to human serum albumin (BPO-HSA), and benzylpenicillin conjugated to an aminospacer (BPO-SP). Receiver operator control curves (ROC analysis) were carried out by determining different cut-off points between positive and negative values. Contingence tables were constructed and sensitivity, specificity, negative predictive values (PV-), and positive predictive values (PV+) were calculated. Pearson correlation coefficients (r) and intraclass correlation coefficients (ICC) were determined and the differences between methods were compared by chi 2 analysis. Analysis of the areas defined by the ROC curves showed statistical differences among the three methods. When cut-off points for optimal sensitivity and specificity were chosen, the BPO-HSA assay was less sensitive and less specific and had a lower PV- and PV+ than the BPO-PLL and BPO-SP assays. Assessment of r and ICC indicated that the correlation was very high, but the concordance between the PLL and SP methods was higher than between the PLL and HSA or SP and HSA methods. We conclude that for quantitating IgE antibodies by RAST to the BPO determinant, BPO-SP or BPO-PLL conjugates offer advantages in sensitivity and specificity compared with BPO-HSA. These results support and extend previous in vitro studies by our group and highlight the importance of the carrier for RAST assays.

Adult↗
PubMed1997

Identification of de novo deletions at the NF1 gene: no preferential paternal origin and phenotypic analysis of patients.

Neurofibromatosis type 1 (NF1) is a common autosomal dominant disorder. To date, a relatively small number of NF1 mutations have been characterized, thus precluding genotype-phenotype correlations. By genotyping 75 NF1 families, we have detected six hemizygous patients (two of whom are members of the same family). The five presumed deletions were confirmed by two quantitative methods of analysis of NF1 copy number: Southern hybridization with cDNA probes and a single-strand conformation polymorphism analysis that discriminates between the NF1 gene and the pseudogene sequences. The five deletions remove most of the NF1 gene, at least 225 kb, from exon 9 to the 3' end of the coding sequence. The origin of de novo mutations in the NF1 gene has been reported to be mainly paternal but we have determined that four of the de novo deletions involved the maternal chromosome and one the paternal chromosome. The six patients with deletions exhibited precocious, multiple clinical features of the disease. The incidence of tumor complications, particularly plexiform neurofibromas and intracranial tumors, among this group of patients is higher than the observed incidence in our NF1 population, suggesting that NF1 haploinsufficiency may cause a more severe phenotype with regard to tumor development. In contrast to other reports that associated large deletions with mildly dysmorphic facies, mental retardation and a large number of cutaneous neurofibromas, only one out of our six patients presented this phenotype.

Adolescent↗
PubMed1997

Novel stop and frameshifting mutations in the autosomal dominant polycystic kidney disease 2 (PKD2) gene.

Autosomal dominant polycystic kidney disease (ADPKD) is one of the most frequent inherited disorders. The majority of cases are due to mutation of the PKD1 gene, on 16p13.3, while in most of the remainder the disease maps to the PKD2 locus, at chromosome 4q21-q23. Recently, the PKD2 gene has been positionally cloned and three nonsense mutations within the coding sequence of the gene identified. Here we report a systematic mutation screening of all 15 exons of the PKD2 gene in chromosome 4-linked ADPKD families, using heteroduplex and SSCP analyses. We have identified and characterized seven novel mutations, with a detection rate of approximately 90% in the population studied. All of the mutations result in the premature stop of translation: four nonsense changes and three deletions. The deletions are all frameshifting, of four T nucleotides in one case and one G nucleotide in the other two. All mutations are unique and are distributed throughout the gene without evidence of clustering. Comparison of specific mutations with the clinical profile in ADPKD2 families shows no clear correlation.

Chromosomes, Human, Pair 4↗
PubMed1997

The Escherichia coli K-12 sheA gene encodes a 34-kDa secreted haemolysin.

Haemolytic toxins belong to one of several classes of virulence factors that contribute to bacterial pathogenicity. The non-pathogenic Escherichia coli K-12 laboratory strain was considered for years to be non-haemolytic. However, overproduction of several transcriptional regulators induced the appearance of a haemolytic activity that is absent under usual laboratory growth conditions. In this work, we have cloned and characterized an E. coli K-12 gene, sheA, whose overexpression results in a haemolytic phenotype. It maps to min 27 on the genetic map, and codes for a 34-kDa polypeptide with at least one putative transmembrane segment. This polypeptide, which has neither signal peptide nor other known secretory motifs, is secreted to the medium during the exponential growth phase. In vitro coupled transcription/translation assays, using a plasmid carrying only the sheA gene as template, resulted in the production of a polypeptide with haemolytic activity per se. Our results demonstrate that the sheA gene actually encodes the E. coli K-12 chromosomal haemolysin. The SheA haemolysin does not resemble other known cytolytic toxins, and it may represent the prototype of a novel family, as suggested by the presence of homologues in several E. coli pathogenic strains and in Shigella flexneri.

Amino Acid Sequence↗