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Biomedical subjects

P Vera

Publications and source records attributed to P Vera.

At least 37 records · Page 2Linked to original sources

Adsorption of a Corticoid on Colloidal Hematite Particles of Different Geometries

An electrokinetic study of the adsorption of a glucocorticoid drug, betamethasone disodium phosphate (BMP), on hematite (alpha-Fe2O3) particles is presented in this work. We analyze the electrophoretic mobility of both spherical and elongated hematite as a function of the concentration of BMP in the dispersion medium; it is found that, even for the lowest BMP concentration, the drug molecules adsorb onto the particles, changing their surface charge from positive to negative. When the drug concentration is increased between 10(-5) and 5 x 10(-3) M, the mobility of spherical hematite shows a monotonous increase from approximately -1.5 to approximately -3.5 &mgr;ms-1/Vcm-1; in the case of elongated particles, the mobility increases with BMP concentration up to 10(-4) M, larger concentrations of the corticoid having little effect on the electrokinetic properties of the particles. The effect of pH on the electrophoretic mobility of hematite is also investigated for two constant drug concentrations (0.05 and 0.5 mM); the isoelectric point of the particles is shifted to a pH below 3, another indication of adsorption of the negatively charged phosphate groups of BMP. These qualitative reasonings are confirmed by direct spectrophotometric determinations of the amount of BMP adsorbed on both hematite samples: spherical and ellipsoidal. The effect of pH on the adsorption density, Gamma, suggests that electrostatic interactions are not the only forces responsible for adsorption. In fact, although Gamma decreases with pH for both types of particles, it remains measurable up to pH 10, when strong electric repulsion is to be expected between hematite and phosphate groups. The effect of initial BMP concentration on Gamma is also studied. It is shown that a plateau region is reached in both samples, this indicating that their surfaces get saturated with BMP: the saturation begins when the coverage is of the order of approximately 1/8 (spherical particles) and approximately 1/40 (elongated particles) of a drug monolayer. Such difference might be due to the ellipsoidal hematite particles having a lower positive surface charge, since they are synthesized in a different dispersion medium.

Journal Article↗

Complementarity of lung scintigraphy and D-dimer test in pulmonary embolism.

D-dimer assay (DDA), measuring fibrin degradation products, was compared with lung scintigraphy (LS) in a prospective unselected series of 83 consecutive patients referred owing to suspicion of pulmonary embolism (PE). This patient series was also used to compare several methods of performing and interpreting LS images. The final diagnosis was established independently by a separate panel with all available information except for the result of DDA. D-dimer was determined by ELISA (threshold value 500 ng/ml). LS, including perfusion (.Q) and pseudo-ventilation (Technegas) (.V), was classified according to PIOPED, (1) immediately by the physician on duty, and (2) retrospectively by a blinded panel. A positive (19) or negative (61) diagnosis of PE was achieved in 80 patients, the prevalence of PE being 24%. Only one false-negative was noted on DDA (sensitivity=95%) but there were 42 false-positives (specificity=31%), resulting in a positive predictive value of 30% and a negative predictive value of 95%. Emergency and retrospective interpretations of LS were close (kappa=0.4). In a minority of patients, PE may be excluded with reasonable certainty if DDA is normal, resulting in a significant saving in terms of time and money.

Enzyme-Linked Immunosorbent Assay↗

Two PR-1 genes from tomato are differentially regulated and reveal a novel mode of expression for a pathogenesis-related gene during the hypersensitive response and development.

Pathogenesis-related (PR) proteins form a heterogeneous family of plant proteins that are likely to be involved in defense and are inducible by pathogen attacks. One group of PRs, represented by the subfamily PR-1, are low-molecular-weight proteins of unknown biochemical function. Here we describe the cloning and characterization of two closely related genes encoding a basic and an acidic PR-1 protein (PR1b1 and PR1a2) from tomato (Lycopersicon esculentum). We present a comparative study of the mode of transcriptional regulation of these two genes in transgenic tobacco plants using a series of promoter-GUS fusions. Unexpectedly, the chimeric PR1a2/GUS gene is not induced by pathogenic signals but instead shows constitutive expression with a reproducible developmental expression pattern. It is expressed in shoot meristems, trichomes, and cortical cells as well as in vascular and nearby tissues of the mature stem. This constitutive expression pattern may represent preemption of plant defenses against potential pathogens. Conversely, the chimeric PR1b1/GUS gene does not show any constitutive expression in the plant, but it is transcriptionally activated following pathogen attack. Upon infection by tobacco mosaic virus, the PR1b1 gene is strongly activated locally in tissues undergoing the hypersensitive response but not systemically in uninoculated tissues. Furthermore, its expression is induced by both salicylic acid and ethylene precursors, two signals that coexist and apparently mediate the activation of local defenses during the hypersensitive response. We speculate that the different mode of expression of the two genes presented here, together with that reported previously for the induction of other PR-1 genes in systemic, uninoculated tissues, may all be complementary and necessary for the plant to acquire an efficient refractory state to resist pathogen attacks.

Amino Acid Sequence↗

Primary structure and expression of a pathogen-induced protease (PR-P69) in tomato plants: Similarity of functional domains to subtilisin-like endoproteases.

A 69-kDa proteinase (P69), a member of the pathogenesis-related proteins, is induced and accumulates in tomato (Lycopersicon esculentum) plants as a consequence of pathogen attack. We have used the polymerase chain reaction to identify and clone a cDNA from tomato plants that represent the pathogenesis-related P69 proteinase. The nucleotide sequence analysis revealed that P69 is synthesized in a preproenzyme form, a 745-amino acid polypeptide with a 22-amino acid signal peptide, a 92-amino acid propolypeptide, and a 631-amino acid mature polypeptide. Within the mature region the most salient feature was the presence of domains homologous to the subtilisin serine protease family. The amino acid sequences surrounding Asp-146, His-203, and Ser-532 of P69 are closely related to the catalytic sites (catalytic triad) of the subtilisin-like proteases. Northern blot analysis revealed that the 2.4-kb P69 mRNA accumulates abundantly in leaves and stem tissues from viroid-infected plants, whereas the mRNA levels in tissues from healthy plants were undetectable. Our results indicate that P69, a secreted calcium-activated endopeptidase, is a plant pathogenesis-related subtilisin-like proteinase that may collaborate with other defensive proteins in a general mechanism of active defense against attacking pathogens.

Amino Acid Sequence↗

Adsorption of betamethasone disodium phosphate on ethylcellulose latex: an electrokinetic study.

The use of polymer latexes as vehicles for drug transport and controlled delivery in the body is a very active research field in applied colloid science. In this work, the polymer chosen is a commercially available ethylcellulose-based material, Aquacoat. We report electrokinetic (electrophoresis) data on this polymer in the presence of the soluble glucocorticoid betamethasone disodium phosphate (BMP). The electrophoretic mobility of the particles becomes more negative as the concentration of BMP in the medium is increased between 10(-5) and 10(-3) M, this demonstrating that the negative species produced upon dissolution of BMP adsorb on the particles; this trend is reversed for higher concentrations due to the phenomenon of double layer thickness. When pH is increased at constant BMP concentration, the negative mobility increases in absolute value, but it reaches lower values than attained in the absence of BMP. This is interpreted as another consequence of adsorption, since the ionization constants of the phosphate and sulfate groups are different. Such adsorption is confirmed and measured by spectrophotometric analysis of the supernatants after centrifugation of the suspensions; it is found that the adsorption density increases with the initial concentration of BMP in the medium, an almost vertical isotherm being obtained for concentrations above 5 x 10(-3) M, possibly due to bidimensional aggregation of the adsorbed molecules and multilayer adsorption at higher concentrations. It is found that the adsorbed amount increases with the ionic strength of the medium as a consequence of screening of the particle and BMP charges, whereas these electrostatic contributions to adsorption manifest also in the pH dependence of adsorption. IR analysis of the particles after equilibration with BMP solutions of increasing concentration agrees well with adsorption data.

Adsorption↗

Characterization of LRP, a leucine-rich repeat (LRR) protein from tomato plants that is processed during pathogenesis.

This paper describes the isolation and characterization of LRP, a new gene from tomato plants. The deduced amino acid sequence showed that the encoded protein is enriched in leucine, and contains interesting structural motifs. LRP contains four tandem repeats of a canonical 24 amino acid leucine-rich repeat (LRR) sequence present in different proteins that mediates molecular recognition and/or interaction processes. Genomic organization and intron-exon arrangement of LRP favor the hypothesis that the LRR domains present in LRP evolved by exon duplication and shuffling. LRP expression analysis and immunohistochemical localization studies of the encoded protein indicate that the gene is under developmental regulation exhibiting tissue-specificity, particularly in certain cell types of the stele, like phloem fibers, parenchyma cells of the protoxylem, and in the cell files that constitute the rays of the secondary xylem. It is shown that this gene is upregulated in diseased tomato plants infected with citrus exocortis viroid. However, in this pathogenic context, LRP is processed proteolytically to a lower molecular weight form by a host-induced extracellular protease. The structural characteristics of LRP, its spatio-temporal pattern of expression, and its post-translational processing during pathogenesis, suggest this protein as a candidate molecule that may mediate recognition and interaction events taking place in the plant extracellular matrix under normal and/or pathogenesis-related conditions.

Amino Acid Sequence↗

Phloem-specific expression of a plant homeobox gene during secondary phases of vascular development.

This paper reports the isolation and characterization of a homeobox gene (VAHOX1) from Lycopersicon esculentum encoding a homeodomain protein that contains a leucine zipper motif. The bipartite homeodomain-leucine zipper (HD-Zip) motif has only been found in homeobox genes from dicotyledonous plants, indicating that this type of transcription factor regulates particular developmental processes in these plant species. Here, the genomic organization, sequence comparison and expression analysis of VAHOX1 are described. Transcriptional fusion of the 5' promoter region of VAHOX1 with the reporter GUS gene (VAHOX1-GUS) and expression analysis of this construct in transgenic plants indicates that VAHOX1 is specifically expressed in the phloem during phases of secondary growth. Unlike other plant homeobox genes, VAHOX1 does not appear to be expressed in meristems, and the function of VAHOX1 is considered a likely candidate molecule that may participate in the regulation of the identity and/or activity of phloem tissues during secondary phases of vascular development.

Amino Acid Sequence↗

Characterization of defense-related genes ectopically expressed in viroid-infected tomato plants.

Differential hybridization was used to isolate genes induced by viroid infection in tomato plants. Four new cDNA clones encoding a peroxidase, a desaturase-like enzyme, a lipoxygenase, and a proteinase inhibitor, were selected and characterized. All of these genes display a characteristic expression pattern, showing constitutive expression in roots of healthy plants and being ectopically activated in aerial tissues upon viroid infection and ethephon treatment. Possible functions for these genes in the viroid-tomato interaction are proposed. The existence of an integrated program that compiles developmental and defense-related responses is also suggested to explain the characteristic expression pattern detected for these genes as well as for other defense-related genes.

Amino Acid Sequence↗

Effect of amino acids on the stability properties of nitrofurantoin suspensions. Flocculation and redispersion compared with interaction energy curves.

The influence of the addition of amino acids, lysine, alanine and glutamic acid on the properties of nitrofurantoin suspensions was investigated. Flocculation and redispersability was investigated, taking into account the electrical properties of the nitrofurantoin/solution interface. We conclude that lysine and glutamic acid, but not alanine, would be suitable amino acids for control of the stability of nitrofurantoin suspensions.

Alanine↗

Effects of different amino acids on the electrical characteristics of pharmaceutical polymers.

The effect of variations in pH and in concentration of five different amino acids (alanine, glycine, lysine, threonine, and glutamic acid) on the electrical properties of the Aquacoat polymer/solution interface was investigated by means of electrophoretic mobility measurements. The results are explained on the basis of determinations of adsorption of the amino acids on the polymer particles. The pH appears to be a determining factor in the way amino acids interact with the negatively charged polymer particles. The net, pH-dependent charge on the amino acid molecules may significantly affect their interaction with Aquacoat; thus, electrostatic effects seem to be most important in the case of lysine and glutamic acid. However, specific effects related to the structure (polarizability, size, charge) of the side group of the amino acid species appear to play an important role in the interaction and adsorption phenomena involved.

Alanine↗

5HT2 receptors in cerebral cortex of migraineurs studied using PET and 18F-fluorosetoperone.

Since the brain 5HT2 receptors might be implicated in migraine pathogenesis, we have used positron emission tomography and 18F-fluorosetoperone, a 5HT2 specific radioligand, to investigate in vivo the cortical 5HT2 receptors in migraine subjects. Nine migraineurs who had either migraine with and without aura (n = 5) or only migraine without aura (n = 4) were studied between attacks. Twelve unmedicated healthy subjects of similar mean age were used as controls. Brain radioactivity was measured after 18F-setoperone IV injection for 90 min. A decrease of the regional specific distribution volumes (SDV) of the ligand was observed both in migraineurs and in controls. The age adjusted group means of SDV did not differ between patients and controls for the whole and for the right or left frontal, temporal, parietal and occipital cortex. These results suggest that cortical 5HT2 receptors may be unaltered between attacks in migraine sufferers.

Adult↗

Properties of shoe insert materials related to shock wave transmission during gait.

The influence of the mechanical characteristics of certain insole materials in the generation and transmission of heel strike impacts while walking was studied. Three insole materials were selected according to their mechanical characteristics under heel strike impacts. The selection of materials has made it possible to distinguish the effect of rigidity and loss tangent in the transmission of heel strike impacts. A lower rigidity and a high loss tangent have been shown to reduce the transmission of impacts to the tibia. A low rigidity was seen to significantly increase the transmission of impacts from tibia to forehead.

Adult↗

[Neurological applications of single photon emission tomoscintigraphy].

Brain single photon emission tomography is a functional imaging modality in constant evolution. Tracers belong to 4 categories; blood pool tracers, perfusion tracers, ligands of receptors, "metabolic" tracers. Examination conditions must be accurately described since they may cause specific cerebral activation. The interpretation must be done in comparison with a morphological examination (CT or NMR). Cerebrovascular diseases, temporal lobe epilepsy and dementia were the first domains of application. Now many studies concentrate on cerebral cognitive or pharmacological activation procedure before the tracer injection to capture an aspect of the brain functioning. It is mandatory to clearly state what is expected from the examination: an answer for a given patient and a specific question or a study of an homogeneous group of patients. In order to yield relevant results, such studies must rely on a rigorous methodology.

Cerebrovascular Disorders↗

Genes encoding acidic and basic class III beta-1,3-glucanases are expressed in tomato plants upon viroid infection.

beta-1,3-glucanases are hydrolytic enzymes considered to constitute part of the general array of defense genes induced by pathogen infection in higher plants. We have isolated and characterized two complementary DNA clones, corresponding to new beta-1,3-glucanases from tomato plants (Lycopersicon esculentum) which are expressed upon challenge with citrus exocortis viroid. Amino acid sequence comparison revealed that they are most similar to beta-1,3-glucanases from tobacco, particularly to PR-Q', the unique component of the class III beta-1,3-glucanase. The deduced amino acid sequences of the two tomato beta-1,3-glucanases indicate that, although being highly similar in amino acid sequence, they have different isoelectric points: pI 10.5 for the basic isoform (Tom PR-Q'b) and pI 5.2 for the acidic one (Tom PR-Q'a). The expression of these two beta-1,3-glucanase messenger RNAs (mRNAs) in response to viroid infection and ethephon treatments was examined. mRNAs for these two isoforms are coordinately expressed and induced similarly to mRNAs for other PR proteins, indicating that they are part of a general and coordinate mechanism of response of tomato plants susceptible to viroid infection.

Amino Acid Sequence↗

A gene encoding a novel isoform of the PR-1 protein family from tomato is induced upon viroid infection.

A Lycopersicon esculentum cDNA clone encoding an acidic-type pathogenesis-related protein (PR-1a1) was isolated, sequenced and characterized. It contains an open reading frame of 175 amino acids and the mature protein, after cleavage of the 21 amino acid signals peptide, has a pI of 5.24. The protein shows highest homology (75% identity) with the basic pathogenesis-related prb-1b protein from tobacco. The PR-1a1 gene shows constitutive expression in roots from tomato plants. It is expressed in leaves and stems upon viroid infection, and appears to be induced by ethylene. Comparative studies of this gene and a related basic isoform of PR-1 indicate that the expression of these two members of the PR-1 gene family in tomato may be differentially regulated upon viroid infection.

Amino Acid Sequence↗

Load transmission through the callus site with external fixation systems: theoretical and experimental analysis.

The fracture callus contribution to the total rigidity of external fixator-fractured bone element was analysed. This study was achieved from both theoretical and experimental perspectives. The theoretical study was done using the finite element method with a three-dimensional model. In this model, both the callus and the development of its elastic characteristics were considered. A series of 38 New Zealand-California white rabbits with tibial fractures treated with some external fixation system was used in this study. Such devices could reproduce either rigid or elastic features. The frame dynamization can be obtained at different fracture healing stages. Animals were classified into four series: (1) rigid fixators, (2) dynamized fixators two weeks after surgery, (3) three weeks after surgery and (4) four weeks after surgery. Tension tests were performed to evaluate callus strength. Theoretical results showed significant levels of callus load transmission (85.5%) when the callus elastic modulus is 1/100 of the elastic modulus of intact bone in an external fixator with rigid features. Experimental analysis of the callus obtained with different external fixator systems did not show differences among them. This fact implies that the immature callus theoretically modelled (E = 100 N mm-2) appears early in the rabbit fracture model (before two weeks, when first dynamization was made), for this reason no differences were found between the two types of treatment (rigid and dynamized systems). This fact questions the reliability of the dynamization process in external fixation of fractures and it implies accurate investigations in the clinical field.

Animals↗

A novel extracellular matrix protein from tomato associated with lignified secondary cell walls.

A cDNA clone representing a novel cell wall protein was isolated from a tomato cDNA library. The deduced amino acid sequence shows that the encoded protein is very small (88 amino acids), contains an N-terminal hydrophobic signal peptide, and is enriched in lysine and tyrosine. We have designated this protein TLRP for tyrosine- and lysine-rich protein. RNA gel blot hybridization identified TLRP transcripts constitutively present in roots, stems, and leaves from tomato plants. The encoded protein seems to be highly insolubilized in the cell wall, and we present evidence that this protein is specifically localized in the modified secondary cell walls of the xylem and in cells of the sclerenchyma. In addition, the protein is localized in the protective periderm layer of the growing root. The highly localized deposition in cells destined to give support and protection to the plant indicates that this cell wall protein alone and/or in collaboration with other cell wall structural proteins may have a specialized structural function by mechanically strengthening the walls.

Amino Acid Sequence↗