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Biomedical subjects

E García

Publications and source records attributed to E García.

At least 91 records · Page 5Linked to original sources
PubMed1999

[Feasibility and safety of intracoronary ultrasound. Experience of a single center].

BACKGROUND AND OBJECTIVES: Intracoronary ultrasound provides a number of advantages in the quantification and characterization of coronary stenoses with regard to contrast angiography. However, previous studies have reported a 3.5 to 11% complication rate, and a 10-30% failure rate in performing this technique. The purpose of the study is to analyze the feasibility of performing intracoronary ultrasound and the incidence of complications associated with the use of contemporary, state of the art equipment. MATERIAL AND METHODS: The feasibility of performing intracoronary ultrasound, analyzed as the percentage of successes and failures in performing the examination was reviewed, as well as the complication rate associated with the technique in all the procedures carried out between July 1, 1994 and February 29, 1996 in which intravascular ultrasound was attempted. Complications were categorized as related, non-related and uncertainly related to the ultrasound study. RESULTS: 239 vessels were studied with intravascular ultrasound in 209 procedures (74% interventional) performed on 139 patients. Ultrasound examination was feasible in all the diagnostic studies and in 96% of the interventional procedures. The major and minor procedural complication rate was 2.4 and 10.5% respectively. No major complication was related to the ultrasound examination. Three patients experienced minor complications (1.4%) related to the ultrasound study. All three complications occurred in baseline studies during interventional procedures. CONCLUSIONS: Intracoronary ultrasound is feasible and safe in the vast majority of the procedures. Improvements in smaller catheter size and design and larger operator expertise have significantly reduced the complication rate, particularly the most frequent coronary spasm so far. Complications are associated with baseline studies during interventional procedures and with less operator expertise.

Aged↗
PubMed1999

[The clinical management guidelines of the Sociedad Española de Cardiología in acute myocardial infarct].

In the recent years, new possibilities have emerged in the diagnosis and management of acute myocardial infarction with ST segment elevation and its complications. Moreover, a deep transformation has taken place in the health care system organization, particularly in aspects related to care of patients presenting non-traumatic chest pain, both in pre-hospital and hospital areas. All these issues warrant a consensus document in Spain dealing with the role that these important changes should play in the whole management of myocardial infarction patients. This document revises and updates all the main clinical issues of acute myocardial infarction patients from the moment they contact with the health care system outside the hospital until they return home, after staying at the coronary care unit and the general hospitalization ward. All those aspects are considered not only in the uncomplicated myocardial infarction but also in the complicated one. This review also includes a set of recommendations on structural and organisational aspects, mainly referred to the prehospital and emergency levels.

Cardiology↗
PubMed1999

Primary angioplasty versus systemic thrombolysis in anterior myocardial infarction.

OBJECTIVES: This study compares the efficacy of primary angioplasty and systemic thrombolysis with t-PA in reducing the in-hospital mortality of patients with anterior AMI. BACKGROUND: Controversy still exists about the relative benefit of primary angioplasty over thrombolysis as treatment for AMI. METHODS: Two-hundred and twenty patients with anterior AMI were randomly assigned in our institution to primary angioplasty (109 patients) or systemic thrombolysis with accelerated t-PA (111 patients) within the first five hours from the onset of symptoms. RESULTS: Baseline characteristics were similar in both groups. Primary angioplasty was independently associated with a lower in-hospital mortality (2.8% vs. 10.8%, p = 0.02, adjusted odds ratio 0.23, 95% confidence interval 0.06 to 0.85). During hospitalization, patients treated by angioplasty had a lower frequency of postinfarction angina or positive stress test (11.9% vs. 25.2%, p = 0.01) and less frequently underwent percutaneous or surgical revascularization after the initial treatment (22.0% vs. 47.7%, p < 0.001) than did patients treated by t-PA. At six month follow-up, patients treated by angioplasty had a lower cumulative rate of death (4.6% vs. 11.7%, p = 0.05) and revascularization (31.2% vs. 55.9%, p < 0.001) than those treated by t-PA. CONCLUSIONS: In centers with an experienced and readily available interventional team, primary angioplasty is superior to t-PA for the treatment of anterior AMI.

Aged↗
PubMed1999

Three approaches to the development of selective bilayer amperometric biosensors for glucose by in situ electropolymerization.

Three different glucose oxidase biosensors for the enzymatic determination of glucose, based on bilayer polymer coatings consisting of polypyrrole (PPy) and poly(o-phenylenediamine), were developed. The electrode substrates are Pt metal, carbon paste and an organic conducting salt (tetrathiafulvalene-tetracyanoquinodimethane), which introduces appreciable variability in the electrode construction process. The three sensors were compared with one another and with bare and PPy monolayer-coated sensors in terms of performance in the determination of glucose in a synthetic serum sample. These configurations provide improved selectivity against the interferences of electroactive species such as ascorbic acid and uric acid, frequently present in biological samples, and the differences between them can be taken advantage of in application to different kinds of samples.

Biosensing Techniques↗
PubMed1999

First molecular characterization of a uridine diphosphate galacturonate 4-epimerase: an enzyme required for capsular biosynthesis in Streptococcus pneumoniae type 1.

Uridine diphosphate galacturonate 4-epimerases (UDPGLEs) are enzymes that convert UDP-glucuronate into UDP-galacturonate. Although the presence of UDPGLEs has been reported in prokaryoic and eukaryotic organisms, the genes coding for these enzymes are completely unknown. The galacturonic acid-containing capsular polysaccharide of Streptococcus pneumoniae type 1 is synthesized through the action of a specific UDPGLE. We have constructed a defined deletion mutant in the cap1J gene (one of the 15 cap1 genes responsible for the synthesis of the type 1 capsule) that exhibited an unencapsulated phenotype. This mutant was unable to synthesize UDPGLE, suggesting that Cap1J was the type 1-specific UDPGLE of S. pneumoniae. Escherichia coli cells harbouring the recombinant plasmid pRMM38 (cap1J) overproduced a 40 kDa protein, characterized as Cap1J on the basis of the N-terminal amino acid sequence analysis, and expressed high levels of enzymatically active Cap1J epimerase. Cap1J was partially purified, although purification to electrophoretic homogeneity inactivated the enzyme irreversibly. The enzyme has the following characteristics: K(m) for UDP-glucuronate, 0.24 mM; pH optimum, 7.5; equilibrium constant (in the direction of UDP-galacturonate formation), 1.3; and an approximate M(r) of 80,000 for the active form. The Cap1J protein exhibited a fluorescence emission spectrum similar to that of NADH. Upon inactivation with p-hydroxymercuribenzoate, the addition of NAD+ and 2-mercaptoethanol were sufficient to reactivate the enzyme. Among several compounds tested, UDP-galactose and UDP-xylose exhibited the highest inhibition of the UDPGLE activity. Inactivation of UDPGLE activity was also observed in the presence of UMP and several reducing sugars. To our knowledge, this is the first example of a thoroughly molecular characterization of a UDPGLE.

Amino Acid Sequence↗
PubMed1999

The molecular characterization of the first autolytic lysozyme of Streptococcus pneumoniae reveals evolutionary mobile domains.

A biochemical approach to identify proteins with high affinity for choline-containing pneumococcal cell walls has allowed the localization, cloning and sequencing of a gene (lytC ) coding for a protein that degrades the cell walls of Streptococcus pneumoniae. The lytC gene is 1506 bp long and encodes a protein (LytC) of 501 amino acid residues with a predicted M r of 58 682. LytC has a cleavable signal peptide, as demonstrated when the mature protein (about 55 kDa) was purified from S. pneumoniae. Biochemical analyses of the pure, mature protein proved that LytC is a lysozyme. Combined cell fractionation and Western blot analysis showed that the unprocessed, primary product of the lytC gene is located in the pneumococcal cytoplasm whereas the processed, active form of LytC is tightly bound to the cell envelope. In vivo experiments demonstrated that this lysozyme behaves as a pneumococcal autolytic enzyme at 30 degrees C. The DNA region encoding the 253 C-terminal amino acid residues of LytC has been cloned and expressed in Escherichia coli. The truncated protein exhibits a low, but significant, choline-independent lysozyme activity, which suggests that this polypeptide adopts an active conformation. Self-alignment of the N-terminal part of the deduced amino acid sequence of LytC revealed the presence of 11 repeated motifs. These results strongly suggest that the lysozyme reported here has changed the general building plan characteristic of the choline-binding proteins of S. pneumoniae and its bacteriophages, i.e. the choline-binding domain and the catalytic domain are located, respectively, at the N-terminal and the C-terminal moieties of LytC. This work illustrates the natural versatility exhibited by the pneumococcal genes coding for choline-binding proteins to fuse separated catalytic and substrate-binding domains and create new and functional mature proteins.

Amino Acid Sequence↗
PubMed1999

Regional diastolic function in ischaemic heart disease using pulsed wave Doppler tissue imaging.

AIMS: The aim of this study was to determine the utility of pulsed wave Doppler tissue imaging in the evaluation of regional left ventricular diastolic function in patients with ischaemic heart disease. METHODS AND RESULTS: In 30 normal subjects and 43 patients with ischaemic heart disease, Doppler tissue imaging was performed in each of the 16 segments of the myocardium. The following diastolic pulsed wave Doppler tissue imaging parameters were obtained for each segment: (1) regional early diastolic peak velocity (regional e wave cm.s-1); (2) regional late diastolic peak velocity (regional a wave cm.s-1); (3) regional diastolic e/a velocity ratio; and (4) the regional isovolumic relaxation time, defined as the time interval from the second heart sound to the onset of the diastolic E wave. In patients with ischaemic heart disease, each of these parameters was evaluated and compared in ischaemic and normally perfused segments, based on the presence or absence of obstructive lesions of the supplying coronary artery. In patients with coronary artery disease, several differences were observed between diseased and normal wall segments: the mean segmental peak early diastolic velocity (e wave) was reduced (mean +/- SD: 6.4 +/- 2.1 cm.s-1 vs 8.5 +/- 2.8 cm.s-1; P < 0.01); the e/a diastolic velocity ratio was decreased (0.95 +/- 0.3 vs 1.5 +/- 0.6, respectively; P < 0.01) and the regional isovolumic relaxation time was prolonged (104 +/- 36.7 ms vs 69.6 +/- 30 ms; P < 0.01. No differences were observed in any of these parameters between the normally perfused segments of ischaemic patients and normal subjects. Patients with a normal transmitral diastolic Doppler inflow pattern had a mean of 3.7 +/- 2.7 myocardial segments with a local e/a pulsed wave Doppler tissue imaging velocity ratio < 1, fewer than those with an inverted diastolic transmitral Doppler inflow pattern (10.3 +/- 3 segments; P < 0.001). Overall sensitivity and specificity for an inverted local e/a ratio and a local isovolumetric relaxation time > or = 85 ms were of 62% and 72% and 69% and 80%, respectively. CONCLUSION: Regional diastolic wall motion is impaired at baseline in ischaemic myocardial segments, even when systolic contraction is preserved. Pulsed wave Doppler tissue imaging is a useful non-invasive technique which allows the assessment of regional diastolic performance and dynamics of the left ventricular myocardium. Further studies are required to define this role in the evaluation of coronary heart disease.

Blood Flow Velocity↗
PubMed1999

Hepatic expression of growth hormone receptor/binding protein and insulin-like growth factor I genes in uremic rats. Influence of nutritional deficit.

The molecular basis for GH resistance in chronic renal failure is unknown. It may partly reside in a decreased number of hepatic GH receptors and subsequently reduced IGF-I synthesis. To investigate the hepatic expression of GH receptor/binding protein (GHBP) and IGF-I genes in chronic renal failure, mRNA levels and the concentrations of its splicing variants were measured by Northern Blot in male 5/6 nephrectomized rats (NX, n = 9), aged 26 +/- 1 days, and three groups of sham-operated rats: (1) fed ad libitum (SAL, n = 9); (2) pair-fed with NX (SPF, n = 7); and (3) pair-fed with NX in terms of protein ingestion but calorically supplemented up to the intake of SAL (SPF+, n = 8). NX rats had severe renal failure, serum urea nitrogen 106 +/- 11 mg/dl (mean +/- SEM), and were growth retarded. GH receptor/GHBP gene expression was detected as two bands of 4.7 and 1.2 kb, respectively. The amount of mRNA was lower (P < 0.0001) in NX than SAL, either when both bands were considered together or separately. No differences were found between NX, SPF, and SPF+. Serum GHBP concentrations were higher (P < 0.01) in NX rats than the other groups. For the IGF-I gene, two bands of 7.5 and 1.8-0.8 kb were identified. Expression of IGF-I gene was reduced (P < 0.05) in NX in comparison with SAL, this reduction being more marked for the 7.5 kb transcript (amount of mRNA equal to 56.6 +/- 2.6 vs 84.8 +/- 6.2% of values found in SAL rats). There were no differences between NX and SPF. Normalization of caloric intake in SPF+ resulted in partial recovery of the 7.5-kb band and did not modify the 1.8-0.8 kb mRNAs. Circulating IGF-I levels were no different among the four groups. These data confirm that expression of liver GH receptor/GHBP and IGF-I genes is markedly decreased in uremic rats. Nutritional deficiency and not uremia itself seems to be the main causal factor, with protein deficit playing a major role.

Animals↗
PubMed1999

Evaluation of the in vitro and in vivo performance of two sustained-release lithium carbonate matrix tablets. Effect of different diets on the bioavailability.

Two sustained-release (SR) lithium carbonate (Li) matrix tablets, which use a hydrophilic (HP) matrix of hydroxypropylmethylcellulose (Methocel 4K MP) and a lipid (L) matrix of hydrogenated castor oil (Cutina HR) as sustaining agents, have been studied. In vitro performance through dissolution tests in different media was established. The L and HP formulations were affected by the composition of the dissolution media, and liberation was complete in 8 hr using a variable-pH medium that simulates the gastrointestinal (Gl) pH. Liberation was better described by the diffusional model of the square root of time for the L matrix and by zero-order kinetics for the HP matrix. Absolute bioavailability (BA) and food-induced changes on BA of both formulations were studied. The in vivo study design was a 4 x 4 Latin square involving 12 subjects who received two tablets of a 300-mg dose of SR formulations while fasting or with a standardized normal, high-fat, or high-fat/high-protein meal. The results for both formulations showed no differences in the disposition parameters and mean residence time when the tablets were administered with any type of diet. Changes in rate of absorption were found when both types of tablets were administered with any class of diet. The analysis of the ratio Cmax/AUC (area under the curve) evidenced that changes in Cmax were attributable to a higher rate of absorption for the HP matrix and to a higher amount absorbed for the L matrix. In the last, high-fat and high-fat/high-protein diets produced higher AUCs than under fasting condition. The SR Li tablets formulated with hydrogenated castor oil were affected more by high-fat food, probably because of the increase of pancreatic and biliary secretions promoted by the meal, which would affect the matrix itself. The HP matrix was also affected, but to a lesser extent. The magnitude of the change in Cmax observed with this matrix probably is not important from a clinical point of view. Absolute BA was very low for the lipid matrix; in addition, since it is more seriously affected by food, probably it is not a good choice for a drug such as lithium. The in vivo behavior of the HP matrix makes it advisable to invest in efforts to achieve increased BA. Comparing in vitro and in vivo results, the focus should be achieving sustained, but complete, in vitro liberation in not more than 3 hr, with simulation of the transit time through the stomach and small bowel since lithium ion is only absorbed to this point.

Analysis of Variance↗
PubMed1999

A marine snail neurotoxin shares with scorpion toxins a convergent mechanism of blockade on the pore of voltage-gated K channels.

kappa-Conotoxin-PVIIA (kappa-PVIIA) belongs to a family of peptides derived from a hunting marine snail that targets to a wide variety of ion channels and receptors. kappa-PVIIA is a small, structurally constrained, 27-residue peptide that inhibits voltage-gated K channels. Three disulfide bonds shape a characteristic four-loop folding. The spatial localization of positively charged residues in kappa-PVIIA exhibits strong structural mimicry to that of charybdotoxin, a scorpion toxin that occludes the pore of K channels. We studied the mechanism by which this peptide inhibits Shaker K channels expressed in Xenopus oocytes with the N-type inactivation removed. Chronically applied to whole oocytes or outside-out patches, kappa-PVIIA inhibition appears as a voltage-dependent relaxation in response to the depolarizing pulse used to activate the channels. At any applied voltage, the relaxation rate depended linearly on the toxin concentration, indicating a bimolecular stoichiometry. Time constants and voltage dependence of the current relaxation produced by chronic applications agreed with that of rapid applications to open channels. Effective valence of the voltage dependence, zdelta, is approximately 0.55 and resides primarily in the rate of dissociation from the channel, while the association rate is voltage independent with a magnitude of 10(7)-10(8) M-1 s-1, consistent with diffusion-limited binding. Compatible with a purely competitive interaction for a site in the external vestibule, tetraethylammonium, a well-known K-pore blocker, reduced kappa-PVIIA's association rate only. Removal of internal K+ reduced, but did not eliminate, the effective valence of the toxin dissociation rate to a value <0.3. This trans-pore effect suggests that: (a) as in the alpha-KTx, a positively charged side chain, possibly a Lys, interacts electrostatically with ions residing inside the Shaker pore, and (b) a part of the toxin occupies an externally accessible K+ binding site, decreasing the degree of pore occupancy by permeant ions. We conclude that, although evolutionarily distant to scorpion toxins, kappa-PVIIA shares with them a remarkably similar mechanism of inhibition of K channels.

Animals↗
PubMed1999

Characterization of the type 8 capsular gene cluster of Streptococcus pneumoniae.

The complete nucleotide sequence of the capsular gene cluster (cap8) responsible for the biosynthesis of the capsular polysaccharide of Streptococcus pneumoniae type 8 has been determined. The cap8 gene cluster, located between the genes dexB and aliA, is composed of 12 open reading frames. A 14.7-kb DNA fragment embracing the cap8 genes was sufficient to transform an unencapsulated type 3 S. pneumoniae strain to a strain with the type 8 capsule. A possible scenario for the evolution of pneumococcal types 2 and 8 is outlined.

Base Sequence↗
PubMed1999

Construction of a new Streptococcus pneumoniae-Escherichia coli shuttle vector based on the replicon of an indigenous pneumococcal cryptic plasmid.

The nucleotide sequence of a cryptic plasmid (pRMG1) isolated from a type 1 Streptococcus pneumoniae has been determined and two recombinant plasmids, pRMGE1 and pRMGE2, bearing the pRMG1 replicon have been constructed. pRMGE2 is a shuttle vector for Escherichia coli and S. pneumoniae. The important characteristics of this cloning vector are: a size of 5.5 kb including a 1.4 kb fragment of pRMG1 (containing a double-stranded replication origin and an open reading frame encoding a putative replication initiation protein), a multicloning site, two antibiotic resistance markers for selection of plasmid containing cells, and blue-white colony screening in E. coli for identification of insert-containing plasmids.

Base Sequence↗
PubMed1999

Functional organization of the gene cluster involved in the synthesis of the pneumococcal capsule.

Streptococcus pneumoniae is a major human pathogen and its capsular polysaccharide has been shown to be the main virulence factor. The molecular organization of the genes governing the formation of this capsule was not studied until the 1990s. The capsular clusters (cap) of eight of the 90 known pneumococcal types have now been studied. The cap operon, located between the dexB and aliA genes, is arranged as a central region comprising the genes coding for the specific-type polysaccharide, flanked by open reading frames that are mostly common to all of the serotypes. The biochemical functions of 24 genes required for capsular polysaccharide biosynthesis have been elucidated but the precise role of the flanking regions in capsular formation is unknown. The natural genetic transformation characteristic of pneumococci, the arrangement of the cap locus and the abundance of transposable elements at this locus favor the genetic variability of the capsule in this microorganism. These well-documented observations together with the finding that some genes located outside the cap cluster may also participate in capsule formation increase the complexity of pneumococcal infection control.

Bacterial Capsules↗
PubMed1998

Characterization of the galU gene of Streptococcus pneumoniae encoding a uridine diphosphoglucose pyrophosphorylase: a gene essential for capsular polysaccharide biosynthesis.

The galU gene of Streptococcus pneumoniae has been cloned and sequenced. Escherichia coli cells harboring the recombinant plasmid pMMG2 (galU) overproduced a protein that has been shown to correspond to a uridine 5'-triphosphate:glucose-1-phosphate uridylyltransferase (uridine diphosphoglucose [UDP-Glc] pyrophosphorylase) responsible for the synthesis of UDP-Glc, a key compound in the biosynthesis of polysaccharides. A gene very similar to the S. pneumoniae galU has been found in a partial nucleotide sequence of the Streptococcus pyogenes genome. Knockout galU mutants of type 1 pneumococci are unable to synthesize a detectable capsule. An identical result was found in type 3 S. pneumoniae cells in spite of the fact that these bacteria contain a type-specific gene (cap3C) that also encodes a UDP-Glc pyrophosphorylase. Since eukaryotic UDP-Glc pyrophosphorylases appear to be completely unrelated to their prokaryotic counterparts, we postulate that GalU may be an appropriate target for the search of new drugs to control the pathogenicity of bacteria like pneumococcus and S. pyogenes.

Amino Acid Sequence↗
PubMed1998

Immunoglobulin G concentration and neonatal survival of goat kids delivered in a pen or on open range.

To determine whether kids born on open range (subjected to stress due to management practices) had lower IgG concentrations and a higher mortality risk than kids born in a pen, serum IgG concentrations in 63 one-day-old kids were determined. Median Serum IgG concentrations one day after birth tended to be lower (p = 0.09) in surviving kids born in the open than that of surviving kids delivered in pen (115 vs. 1119 mg dl-1). However, survival risk (71 and 82% for kids born on the open range and pen, respectively; n = 79) were not significantly affected by site of kidding. Median IgG concentration of surviving kids born on a pen was not significantly different (p = 0.13) compared to non-surviving kids (1118 vs. 115 mg dl-1). No differences were detected in either median serum IgG levels or death rates between male and female kids. Kids with IgG concentrations < 800 mg dl-1 showed lower survival risks than those with higher concentrations. We concluded that typical management practices of kids delivered on open range at a goat operation under extensive conditions in northern Mexico tended to reduced colostrally acquired serum immunoglobulin levels, but had no effect directly on mortality.

Animal Husbandry↗
PubMed1998

Molecular structure of the gene cluster responsible for the synthesis of the polysaccharide capsule of Streptococcus pneumoniae type 33F.

The organization and nucleotide sequence of the capsular gene cluster involved in the biosynthesis of the type 33F capsular polysaccharide of Streptococcus pneumoniae have been determined. The complete type 33F operon (cap33f) is composed of 14 potential open reading frames where the last ten genes are group-specific. Putative functions have been assigned to several gene products by sequence comparison with the proteins included in the databases. A functional promoter located immediately upstream from the first gene of the cap33f gene cluster has been demonstrated. A 20 kb DNA fragment containing the cap33f genes and the operon promoter was sufficient to transform a S. pneumoniae type 3 unencapsulated mutant to the type 33F capsule.

Bacterial Capsules↗