PubMed Health⌕ Search

Biomedical subjects

E Díaz

Publications and source records attributed to E Díaz.

At least 37 records · Page 2Linked to original sources
PubMed2000

Bacterial promoters triggering biodegradation of aromatic pollutants.

Unraveling the complex transcriptional regulation of bacterial catabolism of aromatic pollutants is a prerequisite for engineering efficient biological systems for many biotechnological applications. A first level of regulation relies on specific regulator-promoter pairs. There have been new insights into the molecular mechanisms that regulatory proteins use to sense a given signal and to activate transcription initiation from the cognate promoters. A second level of regulation allows adjustment of the expression of the particular catabolic operons in response to the global environmental conditions of the cells, and recent findings provide some clues about the mechanisms underlying such complex regulatory checkpoints.

Biodegradation, Environmental↗
PubMed2000

2D NMR analysis of highly restricted internal rotation in 2-methylthio-3H-4-p-bromophenyl)-7-[(ortho- and para-substituted)-phenylthio]-1, 5-benzodiazepines.

The complete assignments of twelve 4-aryl-7-thioaryl-1,5-benzodiazepines 1H and 13C spectra, performed with the use of high resolution variable solvent and temperature 1D and 2D techniques (e.g. HOMOCOSY, NOESY, HMQC and HMBC), lead to the determination of conformational equilibria between two rotamers having the aromatic ring of the thioaryl oriented in a perpendicular or helical orientation toward the benzodiazepine ring. The restricted rotation was evaluated from the population of these conformers.

Benzodiazepines↗
PubMed2000

13C and 1H NMR of 2,6-diaryl-1-hydroxy piperidin-4-one oximes; substituent effects on cis/trans ratio and conformational equilibria.

The reaction of substituted diarylidene acetones with hydroxylamine hydrochloride affords isomeric N-hydroxy diaryl piperidinone oximes as main products. The structures as well as conformational equilibria of these products were established by 1H and 13C NMR spectroscopy and further studied by variable temperature NMR. It was found that the cis/trans ratio of 2,6-substituted piperidine derivatives depended on the position of the substituent on the aromatic ring.

Acetone↗
PubMed2000

A gene containment strategy based on a restriction-modification system.

Engineering barriers to the spread of specific genes are of great interest both to increase the predictability of recombinant microorganisms used for environmental applications and to study the role of gene transfer in the adaptation of microbial communities to changing environments. We report here a new gene containment circuit based on a toxin-antidote pair that targets the cell DNA, i.e. the type II EcoRI restriction-modification system. The set-up involved linkage of the ecoRIR lethal gene encoding the EcoRI endonuclease (toxin) to the contained character in a plasmid and chromosomal insertion of the ecoRIM gene encoding the cognate EcoRI methylase (antidote) that protects the target DNA from restriction. Transfer of the contained character to a recipient cell lacking the antidote caused EcoRI-mediated chromosomal breaks, leading to cell death, thereby preventing gene spread. Using transformation and conjugation as mechanisms of DNA transfer and different environmentally relevant bacteria as recipients, we have shown that the potentially universal EcoRI-based containment system decreases gene transfer frequencies by more than four orders of magnitude. Analyses of the survivors escaping killing revealed a number of possible inactivation mechanisms.

Base Sequence↗
PubMed2000

Enhancing desulphurization by engineering a flavin reductase-encoding gene cassette in recombinant biocatalysts.

Biological desulphurization of petroleum feedstocks and products may offer an attractive alternative to reduce sulphur oxide emissions that cause serious environmental pollution. Dibenzothiophene (DBT) desulphurization via the Dsz pathway of Rhodococcus erythropolis IGTS8 is an energetically expensive process that consumes reducing equivalents. We have shown in this work that the HpaC oxidoreductase from Escherichia coli W is able to supply the required FMNH2 to the Dsz monooxygenases. The cloning and expression of the hpaC gene in Pseudomonas strains bearing the dszABC gene cluster significantly enhanced DBT desulphurization efficacy of the recombinant biocatalysts in a resting-cell process, thus indicating that overexpression of a heterologous flavin reductase in the host cell is critical for a high rate of sulphur removal in vivo. The hpaC and dszABC genes have been engineered as a single transcription unit under control of broadhost-range regulatory signals in a mobilizable DNA cassette that can be used to confer a DBT desulphurization phenotype to a wide variety of bacteria regardless of the expression of putative housekeeping flavin reductases within the host cells. This cassette will be very useful in exploring the biotechnological potential of novel biocatalysts for developing an efficient desulphurization process.

Air Pollutants↗
PubMed2000

Functional analysis of the small component of the 4-hydroxyphenylacetate 3-monooxygenase of Escherichia coli W: a prototype of a new Flavin:NAD(P)H reductase subfamily.

Escherichia coli W uses the aromatic compound 4-hydroxyphenylacetate (4-HPA) as a sole source of carbon and energy for growth. The monooxygenase which converts 4-HPA into 3,4-dihydroxyphenylacetate, the first intermediate of the pathway, consists of two components, HpaB (58.7 kDa) and HpaC (18.6 kDa), encoded by the hpaB and hpaC genes, respectively, that form a single transcription unit. Overproduction of the small HpaC component in E. coli K-12 cells has facilitated the purification of the protein, which was revealed to be a homodimer that catalyzes the reduction of free flavins by NADH in preference to NADPH. Subsequently, the reduced flavins diffuse to the large HpaB component or to other electron acceptors such as cytochrome c and ferric ion. Amino acid sequence comparisons revealed that the HpaC reductase could be considered the prototype of a new subfamily of flavin:NAD(P)H reductases. The construction of a fusion protein between the large HpaB oxygenase component and the choline-binding domain of the major autolysin of Streptococcus pneumoniae allowed us to develop a rapid method to efficiently purify this highly unstable enzyme as a chimeric CH-HpaB protein, which exhibited a 4-HPA hydroxylating activity only when it was supplemented with the HpaC reductase. These results suggest the 4-HPA 3-monooxygenase of E. coli W as a representative member of a novel two-component flavin-diffusible monooxygenase (TC-FDM) family. Relevant features on the evolution and structure-function relationships of these TC-FDM proteins are discussed.

Amino Acid Sequence↗
PubMed2000

[Relationship between leptin and insulin blood levels in obese and lean Chilean women].

BACKGROUND: Leptin, a product of ob gene and insulin blood levels, are proportional to the amount of adipose tissue. Insulin could have an independent regulatory effect on leptin secretion. AIM: To assess the relationship between serum leptin and plasma insulin levels in obese and lean Chilean women. MATERIAL AND METHODS: One hundred forty five women, aged 20 to 60 years old, were studied. Weight, height, waist and hip circumference, fasting blood glucose, insulin and leptin levels were measured. Insulin resistance was assessed using the homeostasis model assessment. The relationship between different variables was determined using multiple linear regression, variance analysis and non parametric correlation. RESULTS: Leptin serum concentrations were positively correlated with body mass index, insulin plasma levels and degree of insulin resistance. The association of leptin with insulin was independent of body mass index and persisted after adjustments by body fat distribution and age. CONCLUSIONS: Insulin and insulin resistance are associated to high blood leptin levels and this association is independent of the degree of adiposity and body fat distribution.

Adult↗
PubMed2000

[Obesity and fatty acids in the etiology of insulin resistance].

Fatty acids, obesity and insulin resistance relationship are discussed. In the last decades fatty acids (FA) have been implicated in the etiology of insulin resistance. Initially, this process was related to FA inhibitory effects on glucose uptake mediated by the FA oxidation metabolites. This mechanism known as the Randle cycle has been presently discarded based on recent evidence for FA effects on glucose metabolism. Now is known that cytosolic lipid content and FA molecular structure determines higher or lower storage and oxidation capacity. Another factor is given by Tumor Necrosis Factor-alpha, which is overexpressed in animal and human obesity, producing insulin signaling and glucose uptake inhibition. This paper discuss the role played by FA and obesity on insulin resistance, mainly in relation to FA effects on glucose metabolism in the liver, muscle and adipose tissues. In the obesity condition adipose tissue releases higher levels of free FA which in turn stimulates hepatic glucose production. Adipose tissue also, increase TNF-alpha secretion impairing glucose utilization and insulin signaling. In muscle, cytosolic lipid content activate a Protein Kinase that inhibits the insulin signaling and reduce GLUT-4 translocation. The study of cellular and metabolic changes associated to weight gain and its relationship with insulin resistance etiology are encouraged.

Animals↗
PubMed1999

RegulonDB (version 2.0): a database on transcriptional regulation in Escherichia coli.

RegulonDB version 2.0, a database on transcriptional regulation and operon organization in Escherichia coli, is now available on the web at the following URL: http://www.cifn.unam. mx/Computational_Biology/regulondb/. In this paper we describe the main computational changes to the database, which include migrating the database to Sybase, providing graphical descriptions of the internal organization of operons and regulons, and direct links to MEDLINE references. The web interface offers searching either by mechanisms of regulation or by operon organization. The results of a search (operon organization, or site collection) are displayed as hypertext, and can also be displayed graphically. In terms of its contents, RegulonDB contains a large number of operons, as well as the absolute position in the completed genome sequence of sites, promoters, and individual genes of E.coli.

Databases, Factual↗
PubMed1999

Persistent neuronal density changes related to the establishment of a motor memory.

Rats were trained in a lateralized reaching motor task during either an 'early' (22-31 days old) or a 'late' (62-71 days old) postnatal period. The 'late' group showed significant neuronal density reduction in cortical layers II-III of the contralateral motor forelimb representation. The 'early' group evidenced a similar localized contralateral effect that persisted after a subsequent period without training. Furthermore, in this group, a bilateral overall decrease in neuronal density was found throughout the motor cortex. This bilateral experience and age-dependent effect is conceivably related to a critical period of motor cortical development. The localized reduction of neuronal density strongly indicates a morphological expression of the motor engram. Our present study supports the concept that the acquisition and retention of motor learning involves the persistence of structural changes in the brain.

Animals↗
PubMed1999

Short-term effects of marathon running in master runners: no evidence of myocardial injury.

The purpose of this investigation was to analyze the short-term effects of marathon running on serum levels of cardiac markers in a group of master runners (> 60 yrs). Ten marathoners (9 male and 1 female; 63+/-4 yrs) were enrolled in the study. All of them completed the Madrid Marathon (1998). Venous blood was drawn from each subject three times during the study (48 h before the race, immediately after the race, and 24 h post-competition) for the determination of the several biochemical markers, such as total creatine kinase catalytic activity (total CK), mass concentration of creatine kinase isoenzyme MB (CK-MB mass), and cardiac troponin I (Tnl-c). The most important finding was that in each sample (pre- or post-race) serum Tnl-c was below the commonly accepted level of 0.1 ng x m(-1) indicative of myocardial injury. Although further research is needed using more complete methodology, our results suggest that marathon running does not have an acute deleterious effect on the hearts of the elderly.

Aged↗
PubMed1999

Effect of exogenous melatonin on neuroendocrine-reproductive function of middle-aged female rats.

The possible role of melatonin in the regulation of the reproductive system of female rats during ageing was investigated in middle-aged female rats showing irregular duration of the oestrous cycle (n = 30). Blood samples were obtained by jugular venepuncture during the oestrous cycle in control rats. After this experiment was completed, the female rats were treated with melatonin for 2 months and blood samples were obtained at different stages of the oestrous cycle. Plasma LH, FSH and prolactin concentrations were significantly increased in the afternoon of the day of pro-oestrus after melatonin treatment compared with control rats. Moreover, FSH concentrations too were significantly increased on the morning of pro-oestrus and oestrus in melatonin treated rats compared with control rats. Similarly, oestradiol concentrations were significantly higher on the morning of pro-oestrus in melatonin treated rats compared with controls. Another group of rats showing irregular duration of the oestrous cycle was used to study the possible effect of melatonin treatment on the timing of pro-oestrous surges of LH and FSH. The results showed that LH and FSH peak values occurred at 5 h after melatonin treatment. Pituitary responsiveness to LHRH in a 90 min test was also studied in middle-aged rats showing irregular duration of the oestrous cycle that had been injected for 1 month with either melatonin or saline. Prolactin response was unaffected by exogenous melatonin, but a stimulatory effect of melatonin on LH and FSH pituitary responsiveness to LHRH was observed. The results indicate an improved function of the neuroendocrine-reproductive axis in middle-aged rats after melatonin treatment.

Aging↗
PubMed1999

[Metabolic and dietary factors involved in the etiology of obesity in Chilean women].

This review discusses the factors involved in the aetiology of obesity. The effect of dietary macronutrient composition on the metabolic fate of fat (oxidation or storage) is emphasised. Available information on dietary intake in adults from food balance sheets or dietary surveys, show that females eat a relatively low amount of fat (50-70 g/d on average), although this seems to be rising. The increasing prevalence of obesity in our population could be related to the glycemic index of meals, their fatty acid composition and the time interval between meals. This paper compares the oxidation rates of different fatty acids, particularly polyunsaturated fatty acids (PUFA), and among these, the high oxidation rates found for alpha-linolenic and linoleic acids. Medium chain saturated fatty acids (C8 to C12) are more likely to be oxidised in comparison with PUFA and longer chain saturated fatty acids. The latter are more likely to be directed to fat storage, particularly when they are combined with highly glycemic carbohydrates (CHO) in the same meal. Given the large proportion of CHO in the usual diet, the relevance of de-novo lipogenesis from CHO is questioned. It is concluded that this route is not readily used in humans consuming normal diets. Therefore, this mechanism can not be responsible for the rising prevalence of obesity.

Energy Metabolism↗
PubMed1999

Construction of plasmid vectors bearing a NotI-expression cassette based on the lac promoter.

We have constructed two plasmid vectors for cloning and expression of DNA fragments controlled by the lac promoter as a NotI-expression cassette. Whereas plasmid pSJ33 allows mobilization of the expression cassette into a wide variety of Gram-negative bacteria by RP4-mediated conjugation, the low-copy-number plasmid pSJP18Not facilitates cloning and expression in Escherichia coli when high gene dosage may be detrimental. In addition to their suitable cloning features (e.g. multiple cloning site, lacZ alpha fragment, compatible with ColE1-derived vectors), these plasmids are particularly useful as auxiliary vectors for cloning of the expression cassettes at the NotI site of mini-transposon elements [1, 2] and their eventual stable insertion into the host chromosome.

Cloning, Molecular↗
PubMed1998

Catabolism of phenylacetic acid in Escherichia coli. Characterization of a new aerobic hybrid pathway.

The paa cluster of Escherichia coli W involved in the aerobic catabolism of phenylacetic acid (PA) has been cloned and sequenced. It was shown to map at min 31.0 of the chromosome at the right end of the mao region responsible for the transformation of 2-phenylethylamine into PA. The 14 paa genes are organized in three transcription units: paaZ and paaABCDEFGHIJK, encoding catabolic genes; and paaXY, containing the paaX regulatory gene. The paaK gene codes for a phenylacetyl-CoA ligase that catalyzes the activation of PA to phenylacetyl-CoA (PA-CoA). The paaABCDE gene products, which may constitute a multicomponent oxygenase, are involved in PA-CoA hydroxylation. The PaaZ protein appears to catalyze the third enzymatic step, with the paaFGHIJ gene products, which show significant similarity to fatty acid beta-oxidation enzymes, likely involved in further mineralization to Krebs cycle intermediates. Three promoters, Pz, Pa, and Px, driven the expression of genes paaZ, paaABCDEFGHIJK, and paaX, respectively, have been identified. The Pa promoter is negatively controlled by the paaX gene product. As PA-CoA is the true inducer, PaaX becomes the first regulator of an aromatic catabolic pathway that responds to a CoA derivative. The aerobic catabolism of PA in E. coli represents a novel hybrid pathway that could be a widespread way of PA catabolism in bacteria.

Acetyl Coenzyme A↗
PubMed1998

Characterization of a 76 kDa endosomal, multispanning membrane protein that is highly conserved throughout evolution.

We report here the identification and characterization of a human 76kDa membrane protein that is found predominantly in endosomes. This protein is related to the Saccharomyces cerevisiae EMP70 gene product, a precursor protein whose 24kDa cleavage product (p24a) was found in yeast endosome-enriched membrane fractions (Singer-Krüger et al., 1993. J. Biol. Chem. 268, 14376-14386). Northern blot analysis indicated that p76 mRNA is highly expressed in human pancreas but could be detected in all tissues examined. p76 is highly conserved throughout evolution, as related proteins have also been detected in Caenorhabditis elegans and Arabidopsis thaliana. This family of proteins has a relatively divergent, hydrophilic N-terminal domain and a well-conserved, highly hydrophobic C-terminal domain which contains nine potential membrane-spanning domains. Transiently expressed, myc-tagged human p76 appears to be localized to endosomes by virtue of its apparent colocalization with transferrin receptors and some mannose 6-phosphate receptors. Furthermore, p76 adopts a type-I topology within the membrane, with its hydrophilic N-terminus facing the lumen of cytoplasmic membranes. The structural features of p76 suggest that it may function as a channel or small molecule transporter in intracellular compartments throughout phylogeny. 1998 Elsevier Science B.V.

Amino Acid Sequence↗
PubMed1998

Interleukin-2 stimulates a late increase in phosphatidic acid production in the absence of phospholipase D activation.

The signal transduction pathways involving phospholipid metabolism during T-cell proliferation remain partly undefined. Herein we show that interleukin-2 caused a late (> 12 h) rise in the intracellular phosphatidic acid content of CTLL-2 cells which was a consequence of the activation of the enzyme diacylglycerol kinase. No activation of phospholipase D was observed at similar times. Incubation of the cells with a recognized diacylglycerol kinase a isoform inhibitor, R59499, prior to interleukin-2 stimulation was able to block cell cycle entry, diacyglycerol kinase activation and phosphatidic acid accumulation. In contrast, when R59499 was added 3 h after interleukin-2, few or no observable effects on the above three parameters were noticed. These results suggest that the early signaling employed by IL-2 involving the alpha isoform of diacylglycerol kinase is sufficient to control the late increase in phosphatidic acid and that phosphatidic acid is a mitogenic agent in T-cells.

3T3 Cells↗