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Biomedical subjects

C Ramos

Publications and source records attributed to C Ramos.

At least 55 records · Page 3Linked to original sources

Assessing the potential hazard of chemical substances for the terrestrial environment. Development of hazard classification criteria and quantitative environmental indicators.

Hazard assessment constitutes an essential tool in order to evaluate the potential effects of chemical substances on organisms and ecosystems. It includes as a first step, hazard identification, which must detect the potential dangers of the substance (i.e. the kind of effects that the substance may produce), and a second step to quantify each danger and to set the expected dose/response relationships. Hazard assessment plays a key role in the regulation of chemical substances, including pollution control and sustainable development. However, the aquatic environment has largely received more attention than terrestrial ecosystems. This paper presents the extrapolation of several basic concepts from the aquatic to the terrestrial compartment, and suggests possibilities for their regulatory use. Two specific proposals are discussed. The first focuses on the scientific basis of the hazard identification-classification criteria included in the EU regulations and their extrapolation to the terrestrial environment. The second focuses on the OECD programme for environmental indicators and the development of a soil pollution pressure indicator to quantify the potential hazards for the soil compartment and its associated terrestrial ecosystem related to the toxic chemicals applied deliberately (i.e. pesticides) or not (i.e. heavy metals in sludge-based fertilisers; industrial spills) to the soil.

Ecosystem↗

Ecological risk assessment of pesticides in the Mediterranean region. The need for crop-specific scenarios

The risk assessment procedures of pesticides in the European Union are mostly based on worst-case scenarios developed for central and northern European conditions. From the point of view of Mediterranean conditions, these assessment procedures are not always appropriate, and therefore the development of specific scenarios is required. There are differences not only in meteorological data (temperature, solar irradiation, or rainfall volume and annual distribution) between these countries, but also in farm distribution, crop characteristics, soil or sediment properties and surface or groundwater characteristics. These differences are more evident for the typical crops of the southern countries, such as olive groves, vineyards, citrus or rice fields, and in these cases more realistic estimates of exposure are required. Our proposal includes the development of new criteria to improve the ecological risk assessment process for the conditions of the Mediterranean region, and the justification of this need.

Journal Article↗

Tension-voltage relationship in membrane fusion and its implication in exocytosis.

In this study, new methods are used to control cellular membrane tension to evaluate the role it plays in electrofusion. The data show that membrane tension present during the application of an electric field facilitates electro-induced membrane fusion. No enhancement was detected if the strain was applied after the pulse. Analysis of the electromechanical process of fusion revealed a synergy between the two kinds of constraints in the membrane fusion. Both mechanical and electrical constraints apparently play a key role in membrane fusion between the granule membrane and the plasma membrane, i.e. the exocytosis process.

Animals↗

Rapid identification of a small dicentric supernumerary marker derived from chromosome 16 with a modified FISH technique on amniotic fluid.

Small supernumerary marker chromosomes are seldom found in prenatal diagnosis and the majority of them are difficult to identify. The only possibility to give a more precise prognosis is by establishing its origin. FISH is the best technique to identify the chromosomal origin, but in the majority of cases large amounts of chromosomal material are needed and this is time consuming. We have used a modification of the FISH technique that allows the hybridization of several probes on one slide. Using this method, we have identified the first de novo mosaic dicentric supernumerary marker derived from chromosome 16 (smaller than chromosome 21) in amniotic fluid. The gestation and the follow-up of the baby were normal.

Adult↗

Traumatic diaphragmatic injuries in infants and children: imaging findings.

OBJECTIVES: Traumatic diaphragmatic injuries (DI) in infants and children are uncommon and are often associated with multiple severe injuries. Delayed presentation can be life threatening due to organ herniation and strangulation. We present the imaging findings in a relatively large population of children who experienced this rare injury. METHODS: Medical records of all patients admitted to our Trauma Service from 1977 to 1998 with DI were retrospectively reviewed recording imaging, clinical and surgical or autopsy findings. RESULTS: Of sixteen patients with DI (7 females, 9 males; age 3 weeks to 15 years), 14 suffered from blunt trauma secondary to high-energy impact, and 2 from penetrating injuries. Unilateral DI occurred equally on each side, with one bilateral injury. Associated injuries, present in 81%, included severe head injuries, visceral, mesenteric and vascular injuries and multiple fractures. Six patients died from multiple organ failure (3), head injury (2), and shock (1). Findings in the initial chest X-ray suggested the diagnosis in 13 (81%) of 16 injuries, and CT demonstrated irregularity and thickening of the diaphragm in 4 out of 7. CONCLUSIONS: Plain film findings suggested the diagnosis in most; CT and MR were useful adjuncts. High index of suspicion and awareness of the mechanism of injury can lead to prompt diagnosis, early repair, and decreased morbidity and mortality.

Adolescent↗

A nuclear protein that binds specifically to several maize transposons is not essential for Ds1 excision.

Specific binding of plant nuclear proteins to GGTAAA-like motifs in the terminal regions of the transposable elements Ac and Mu1 has been detected in several laboratories. However, the role of these proteins in transposition remains unknown. To test the hypothesis that this binding activity is necessary for transposition, we identified and mutagenized all the binding motifs within the Ds1 element. This analysis enabled us to define more precisely the requirements for binding of the host protein. We then tested the ability of the mutated elements to excise from the maize streak virus (MSV) genome. We found that mutated Ds1 elements that do not bind the host proteins, as determined by gel-shift competition assay, are still capable of undergoing excision in maize, although for one of the maize lines the rate of excision was reduced. Excision of mutated Ds1 elements generated typical excision footprints. These data indicate that binding of host protein(s) to the GGTAAA-like motifs is not essential for Ds1 excision; however, it may contribute to the efficiency of the process.

DNA Transposable Elements↗

Electrofusion: a biophysical modification of cell membrane and a mechanism in exocytosis.

The molecular bases of the exocytosis process remain poorly known. Many proteins have been recognized to play key roles in the machinery. Their functions are well characterized in the specificity of the docking processes. Forces involved in the merging of the two partners must take into account the physics of membrane interfaces. The target membrane and the vesicle are both electrically charged interfaces. Strong electrostatic fields are triggered when they are brought in close neighborhood. These fields are high enough to induce an electropermeabilisation process. It is now well known that when applied on a cell, an external field induces a modulation of the transmembrane potential difference. When high enough the transmembrane potential may induce a membrane destabilisation. This results in a free exchange of polar molecules across well defined parts of the cell surface. Furthermore, when permeabilization is present on two cells, if those parts of the cell surfaces are brought in close contact, membrane merging occurs spontaneously. Cell fusion results from this membrane coalescence. The similarity with what is taking place in exocytosis is striking. The present review describes the state-of-the-art in the knowledge on electrofusion. It is emphasized that it results from electropermeabilisation and not from a direct effect of the external field. A local destabilisation of the vesicle membrane results from electrostatic interactions while keeping unaffected its viability. Such processes appear relevant for what takes place during exocytosis.

Animals↗

Antigen 43 facilitates formation of multispecies biofilms.

Antigen 43 (Ag43) is a surface-displayed autotransporter protein of Escherichia coli. By virtue of its self-association characteristics, this protein is able to mediate autoaggregation of E. coli cells in static cultures. Here, we show that Ag43 can be expressed in a functional form on the surface of Pseudomonas fluorescens. Ag43 expression dramatically enhances the biofilm-forming potential of both E. coli and P. fluorescens to abiotic surfaces in simple microtitre well assays and in flow chambers. Importantly, Ag43-expressing E. coli and P. fluorescens cells tagged with Gfp and Rfp were shown to form interwoven biofilms in flow chambers. The three-dimensional structures of the biofilms were analysed by laser-confocal microscopy. Heterogeneous expression of Ag43 induced interspecies cell-to-cell contact that generated multispecies biofilm formation. Our data indicate that this versatile molecular tool can be used for the rational design of multispecies biofilms. More specifically, this novel technology offers opportunities for the design of multispecies consortia in which the concerted action of several bacterial species is required, e.g. waste treatment and degradation of pollutants.

Adhesins, Bacterial↗

Bacterial activity in the rhizosphere analyzed at the single-cell level by monitoring ribosome contents and synthesis rates.

The growth activity of Pseudomonas putida cells colonizing the rhizosphere of barley seedlings was estimated at the single-cell level by monitoring ribosomal contents and synthesis rates. Ribosomal synthesis was monitored by using a system comprising a fusion of the ribosomal Escherichia coli rrnBP1 promoter to a gene encoding an unstable variant of the green fluorescent protein (Gfp). Gfp expression in a P. putida strain carrying this system inserted into the chromosome was strongly dependent on the growth phase and growth rate of the strain, and cells growing exponentially at rates of > or = 0.17 h(-1) emitted growth rate-dependent green fluorescence detectable at the single-cell level. The single-cell ribosomal contents were very heterogeneous, as determined by quantitative hybridization with fluorescently labeled rRNA probes in P. putida cells extracted from the rhizosphere of 1-day-old barley seedlings grown under sterile conditions. After this, cells extracted from the root system had ribosomal contents similar to those found in starved cells. There was a significant decrease in the ribosomal content of P. putida cells when bacteria were introduced into nonsterile bulk or rhizosphere soil, and the Gfp monitoring system was not induced in cells extracted from either of the two soil systems. The monitoring system used permitted nondestructive in situ detection of fast-growing bacterial microcolonies on the sloughing root sheath cells of 1- and 2-day-old barley seedlings grown under sterile conditions, which demonstrated that it may be possible to use the unstable Gfp marker for studies of transient gene expression in plant-microbe systems.

Colony Count, Microbial↗

Proline catabolism by Pseudomonas putida: cloning, characterization, and expression of the put genes in the presence of root exudates.

Pseudomonas putida KT2442 is a root-colonizing strain which can use proline, one of the major components in root exudates, as its sole carbon and nitrogen source. A P. putida mutant unable to grow with proline as the sole carbon and nitrogen source was isolated after random mini-Tn5-Km mutagenesis. The mini-Tn5 insertion was located at the putA gene, which is adjacent to and divergent from the putP gene. The putA gene codes for a protein of 1,315 amino acid residues which is homologous to the PutA protein of Escherichia coli, Salmonella enterica serovar Typhimurium, Rhodobacter capsulatus, and several Rhizobium strains. The central part of P. putida PutA showed homology to the proline dehydrogenase of Saccharomyces cerevisiae and Drosophila melanogaster, whereas the C-terminal end was homologous to the pyrroline-5-carboxylate dehydrogenase of S. cerevisiae and a number of aldehyde dehydrogenases. This suggests that in P. putida, both enzymatic steps for proline conversion to glutamic acid are catalyzed by a single polypeptide. The putP gene was homologous to the putP genes of several prokaryotic microorganisms, and its gene product is an integral inner-membrane protein involved in the uptake of proline. The expression of both genes was induced by proline added in the culture medium and was regulated by PutA. In a P. putida putA-deficient background, expression of both putA and putP genes was maximal and proline independent. Corn root exudates collected during 7 days also strongly induced the P. putida put genes, as determined by using fusions of the put promoters to 'lacZ. The induction ratio for the putA promoter (about 20-fold) was 6-fold higher than the induction ratio for the putP promoter.

Amino Acid Sequence↗

Surfactant components modulate fibroblast apoptosis and type I collagen and collagenase-1 expression.

During lung injury, fibroblasts migrate into the alveolar spaces where they can be exposed to pulmonary surfactant. We examined the effects of Survanta and surfactant protein A (SP-A) on fibroblast growth and apoptosis and on type I collagen, collagenase-1, and tissue inhibitor of metalloproteinase (TIMP)-1 expression. Lung fibroblasts were treated with 100, 500, and 1,000 microg/ml of Survanta; 10, 50, and 100 microg/ml of SP-A; and 500 microg/ml of Survanta plus 50 microg/ml of SP-A. Growth rate was evaluated by a formazan-based chromogenic assay, apoptosis was evaluated by DNA end labeling and ELISA, and collagen, collagenase-1, and TIMP-1 were evaluated by Northern blotting. Survanta provoked fibroblast apoptosis, induced collagenase-1 expression, and decreased type I collagen affecting mRNA stability approximately 10-fold as assessed with the use of actinomycin D. Collagen synthesis and collagenase activity paralleled the gene expression results. SP-A increased collagen expression approximately 2-fold and had no effect on collagenase-1, TIMP-1, or growth rate. When fibroblasts were exposed to a combination of Survanta plus SP-A, the effects of Survanta were partially reversed. These findings suggest that surfactant lipids may protect against intraluminal fibrogenesis by inducing fibroblast apoptosis and decreasing collagen accumulation.

Apoptosis↗

Choroideremia, sensorineural deafness, and primary ovarian failure in a woman with a balanced X-4 translocation.

We present clinical and cytogenetic studies of a female patient affected with choroideremia, mild sensorineural deafness, and primary amenorrhea showing a balanced translocation between chromosomes X and 4. The breakpoint was precisely defined applying FISH techniques: 46,X,t(X;4)(q21.2;p16.3).ish t(X;4)(D4S96+, D4F26+; wcpX+). The X-chromosomal breakpoint was located within a region where both the choroideremia locus and a deafness locus (DFN3/POU3F4) have been mapped. The presence of X-linked disorders in this balanced carrier of X-autosomal translocations (XAT) can be explained either by the disruption of the structural coding or regulatory sequences of the gene(s) or by the submicroscopic deletion of this region leading to a contiguous gene deletion syndrome. The primary ovarian failure (POF) found in the present case has been already observed in XAT when the breakpoint is within a previously defined critical region (Xq13-26). A position effect is postulated as a possible explanation.

Adult↗

A MELAS phenotype and a paternal inherited inversion of chromosome 10 in a female patient.

A MELAS phenotype and a paternal inherited inversion of chromosome 10 in a female patient: We describe a patient suffering from encephalomyopathy with overlapping symptoms, including MELAS and Kearn-Sayre syndrome features. Mutations in tRNA LEU (UUR) were not found in mtDNA of blood cells, suggesting a different genetic defect. Cytogenetic studies revealed a paternal inherited pericentric inversion of chromosome 10 (p13;q22) pat. Although the presence of the same inversion in the father and in the apparently asymptomatic sister does rather suggest that the concurrence of the mitochondrial disease in the patient was due to chance, some alternative explanations to associate both events might be proposed.

Adult↗

Ser186Pro mutation of RHO gene in a Spanish autosomal dominant retinitis pigmentosa (ADRP) family.

A Spanish family affected with autosomal dominant retinitis pigmentosa (ADRP) with a diffuse phenotype showed a mutation in the rhodopsin gene. The mutation was the transition T-->C in codon 186, which has been reported once before in an American patient (Dryja et al., Proc Natl Acad Sci USA 1991;88:9370-9374). This change replaces a serine by a proline in the second intradiscal loop of the protein, generating a molecule that is probably folding- and transport-defective.

Adult↗

Genomic cloning and characterization of the human homeobox gene SIX6 reveals a cluster of SIX genes in chromosome 14 and associates SIX6 hemizygosity with bilateral anophthalmia and pituitary anomalies.

The Drosophila gene sine oculis (so), a nuclear homeoprotein that is required for eye development, has several homologues in vertebrates (the SIX gene family). Among them, SIX3 is considered to be the functional orthologue of so because it is strongly expressed in the developing eye. However, embryonic SIX3 expression is not limited to the eye field, and SIX3 has been found to be mutated in some patients with holoprosencephaly type 2 (HPE2), suggesting that SIX3 has wide implications in head development. We report here the cloning and characterization of SIX6, a novel human SIX gene that is the homologue of the chick Six6(Optx2) gene. SIX6 is closely related to SIX3 and is expressed in the developing and adult human retina. Data from chick and mouse suggest that the human SIX6 gene is also expressed in the hypothalamic and the pituitary regions. SIX6 spans 2567 bp of genomic DNA and is split in two exons that are transcribed into a 1393-nucleotide-long mRNA. Chromosomal mapping of SIX6 revealed that it is closely linked to SIX1 and SIX4 in human chromosome 14q22.3-q23, which provides clues about the origin and evolution of the vertebrate SIX family. Recently three independent reports have associated interstitial deletions at 14q22.3-q23 with bilateral anophthalmia and pituitary anomalies. Genomic analyses of one of these cases demonstrated SIX6 hemizygosity, strongly suggesting that SIX6 haploinsufficiency is responsible for these developmental disorders.

Amino Acid Sequence↗

A comparison of the bronchodilatory effect of 50 and 100 microg salbutamol via Turbuhaler and 100 microg salbutamol via pressurized metered dose inhaler in children with stable asthma.

AIM: The aim of the study was to compare the efficacy of single doses of salbutamol Turbuhaler (50 and 100 microg), salbutamol pressurized metered dose inhaler (pMDI) (100 microg) and placebo in children with stable chronic reversible airway obstruction. Primary efficacy variable (FEV1-av) was calculated as the area under the curve of forced expiratory volume in one second (FEV1) (AUC, 0-4 h) and divided by the observed time. DESIGN: The study was of a randomized, single-dose, crossover and double-blind design. Seven centres participated. FEV1 was measured pre-dose and at 15 min, 0.5, 1, 1.5, 2, 3 and 4 h post study dose. PATIENTS: Forty asthmatic children (9 girls) with a mean age of 9 years (range: 6-12), mean FEV1 of 1.6 l (range: 0.9-2.4) and a mean FEV1 in percentage of predicted normal value of 80% (range: 61-109) were randomized into the study. The mean reversibility 30 min after inhaling 2x100 microg salbutamol from pMDI was 20% (range: 9-45) or 15% (range: 8-27) in percentage of predicted normal value. RESULTS: The mean FEV1-av was 1.63 l for placebo, 1.71 l for 50 microg salbutamol Turbuhaler, 1.76 l for 100 microg salbutamol Turbuhaler and 1.76 for 100 microg salbutamol pMDI. Corresponding values for maximum FEV1 were 1.76, 1. 85, 1.87 and 1.87 l, respectively. There were no statistically significant differences between the active treatments in FEV1-av or maximum FEV1. All active treatments were significantly better than placebo. CONCLUSION: No significant differences in bronchodilating effect between 50, 100 microg salbutamol Turbuhaler and 100 microg salbutamol pMDI in children, aged 6-12 years, with stable asthma could be demonstrated. All active treatments were significantly better than placebo.

Albuterol↗