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PubMed1976

Duration of hospitalization in "uncomplicated completed acute myocardial infarction". An Ad Hoc Committee review.

The clinical and laboratory findings diagnostic of acute myocardial infarction include at least two of the following: (1) a history of pain consistent with myocardial ischemia, (2) electorcardiographic findings consistent with infarction, and (3) a rise in the serum level of specific cardiac enzymes. By the 4th or 5th day of illness, specific criteria can be applied to assign certain patients to a subset with "uncomplicated completed acute myocardial infarction." These criteria include the absence of evidence of (1) continuing cardiac ischemia, (2) left ventricular failure, (3) shock, (4) important cardiac arrhythmias, (5) conduction disturbances, and (6) other serious illnesses in patients with an established acute myocardial infarction. In terms of prognosis and management, patients in this subset should be regarded as substantively different from patients in other subsets. They should respond favorably to short periods of immobilization and hospitalization than those generally used. They may remain at bed rest (modified in regard to sitting and the use of a commode) for 4 days. Subsequently, mobilization with a program of progressive activity over the ensuing 5 to 10 days should reduce the duration of hospitalization to less than the current average of 17.5 to 20.8 days for patients with acute myocardial infarction. Nine to 14 days should suffice in most instances. Current and future trials may indicate that still earlier mobilization and shorter hospitalization periods can be applied to certain patient groups, but the evidence on this point is incomplete. For the individual patient, many factors will determine the optimal duration of bed rest and hospital stay. The patient's physician must consider the therapeutic benefits that may attend earlier mobilization and shorter hospitalization while weighing potential disadvantages. When the responsible physician does not regularly care for the patient, consultation with an experienced cardiologist is desirable. Patients whose condition is classified as "uncomplicated" may manifest deterioration during their illness and require assignment to a subset with a different prognosis and requiring different forms of treatment. For patients with uncomplicated acute myocardial infarction, as well as those in other subsets, absolute rules for therapy are unwise and application of broader principles by the alert physician is more likely to be beneficial.

Arrhythmias, Cardiac↗
PubMed1993

Lack of DNA enzymatic photoreactivation in HeLa cell-free extracts.

Using a DNA-protein binding assay, we have previously identified and characterized a UV-damaged DNA recognition protein (UVDRP) from HeLa cells [(1991) Nucleic Acids Res. 19, 6413-6418]. In this report, the photoreactivating activity of UVDRP from the yeast, Saccharomyces cerevisiae, and HeLa cells was investigated. Although yeast and human cells are evolutionarily different from each other, both UVDRPs were conserved in the sense of their biochemical characteristics except that the yeast UVDRP also exhibited an enzymatic photoreactivating activity. A mammalian expression vector plasmid DNA carrying the bacterial chloramphenicol acetyltransferase (CAT) gene was UV irradiated in vitro followed immediately by exposure to photoreactivating light, and its transient expression in repair-deficient xeroderma pigmentosum (XP) cells was investigated. More than 80% of the CAT activity was inhibited by UV irradiation, which was partially restored (> 60%) by a partially purified yeast photolyase. In contrast, HeLa cell extracts did not express a photoreactivatable recovery from UV-induced inhibition of the CAT activity tested in the same system. This study has demonstrated the potential use of the DNA-mobility shift assay to investigate enzymatic photoreactivation, and has indicated the absence of the repair mechanism in human cells.

Cell Line↗
PubMed1987

Simultaneous measurement of tocopherols and tocopheryl quinones in tissue fractions using high-performance liquid chromatography with redox-cycling electrochemical detection.

Tocopherols and tocopheryl quinones in lipid extracts of biological samples have primarily been measured using relatively insensitive ultraviolet detection methods. Oxidative electrochemical detection increases both the sensitivity and selectivity when measuring the tocopherols. We have developed an electrochemical detection system which sequentially reduces and oxidizes tocopheryl metabolites eluted from a reversed-phase high-performance liquid chromatography column, achieving sensitivities of about 0.05 pmol for both the tocopherols and their quinones. Using a rapid and mild extraction procedure, endogenous levels of alpha- and gamma-tocopherol as well as their respective quinones were measured in homogenates of chicken liver and muscle, and in dilute preparations of rat liver microsomes. The principle of the detection system could be applied to the determination of tocopheryl dihydroquinones, ubiquinols and ubiquinones with slight modifications to the mobile phase buffer and the electrode potentials of the detector.

Animals↗
PubMed2005

Identification and molecular characterization of the rainbow trout calpains (Capn1 and Capn2): their expression in muscle wasting during starvation.

Calpains are calcium regulated proteases involved in cellular functions that include muscle proteolysis both ante- and postmortem. Here, we describe the molecular characterization of the rainbow trout catalytic subunits of the mu- and m-calpains, respectively. The cDNA sequence for Capn1 encodes a protein of 704 amino acids with a calculated molecular mass of 79.9 kDa. The amino acid sequence shows 66% and 86% identity with the mouse and zebrafish Capn1, respectively. The Capn2 cDNA codes for a protein consisting of 701 amino acid residues with a calculated molecular mass of 78.2 kDa. The protein shows 65% amino acid sequence identity with the mouse and chicken Capn2. The two isozymes of rainbow trout have the characteristic domains: I (propeptide), II (cysteine catalytic site), III (electrostatic switch), and IV (contains five EF-hands). Because starvation induces muscle wasting, the hypothesis of this study was that starvation could affect regulation of the calpain system in muscle. Starvation of rainbow trout fingerlings (15-20 g) for 35 days stimulated the expression of Capn1 (2.2-fold increase, P < 0.01), Capn2 (6.0-fold increase, P < 0.01), and calpastatins (1.6-fold increase, P < 0.05) as measured by quantitative real-time RT-PCR. The mRNA changes led to a 1.23-fold increase in the calpain catalytic activity. The results suggest a potential role of calpains in protein mobilization as a source of energy under fasting condition.

Amino Acid Sequence↗
PubMed2005

Simple method for the separation and detection of native amino acids and the identification of electroactive and non-electroactive analytes.

Detection of native amino acids was accomplished using a capillary electrophoresis (CE)-amperometric detection system, in which a single carbon fiber cylinder (CFC) working electrode instead of a carbon fiber disc (CFD) electrode was mounted in the end part of a poly(dimethylsiloxane) (PDMS)/glass hybrid microchannel. Similar to that using CFD working electrode, here, the electrochemical reduction reaction at the working electrode is also coupled from the separation high voltage system, the coupling degree is related to the in-channel length of the CFC. This property simplifies the fabrication procedure of the working electrode and also provides a convenient and sensitive means for the determination of non-electroactive ions by amperometry. The present detection mode is successfully used to electrochemically detect non-electroactive arginine (Arg), threonine (Thr), glutamic acid (Glu) and electroactive cysteine (Cys). Furthermore, by simply changing the detection potential, we can easily distinguish peak mobilities of electroactive amino acids from that of non-electroactive amino acids.

Amino Acids↗
PubMed2004

Kinetics of interprotein electron transfer between cytochrome c6 and the soluble CuA domain of cyanobacterial cytochrome c oxidase.

Cytochrome c6 is a soluble metalloprotein located in the periplasmic space and the thylakoid lumen of many cyanobacteria and is known to carry electrons from cytochrome b6f to photosystem I. The CuA domain of cytochrome c oxidase, the terminal enzyme which catalyzes the four-electron reduction of molecular oxygen in the respiratory chains of mitochondria and many bacteria, also has a periplasmic location. In order to test whether cytochrome c6 could also function as a donor for cytochrome c oxidase, we investigated the kinetics of the electron transfer between recombinant cytochrome c6 (produced in high yield in Escherichia coli by coexpressing the maturation proteins encoded by the ccmA-H gene cluster) and the recombinant soluble CuA domain (i.e., the donor binding and electron entry site) of subunit II of cytochrome c oxidase from Synechocystis PCC 6803. The forward and the reverse electron transfer reactions were studied by the stopped-flow technique and yielded apparent bimolecular rate constants of (3.3 +/- 0.3) x 10(5) M(-1) s(-1) and (3.9 +/- 0.1) x 10(6) M(-1) s(-1), respectively, in 5 mM potassium phosphate buffer, pH 7, containing 20 mM potassium chloride and 25 degrees C. This corresponds to an equilibrium constant Keq of 0.085 in the physiological direction (DeltarG'0 = 6.1 kJ/mol). The reduction of the CuA fragment by cytochrome c6 is almost independent on ionic strength, which is in contrast to the reaction of the CuA domain with horse heart cytochrome c, which decreases with increasing ionic strength. The findings are discussed with respect to the potential role of cytochrome c6 as mobile electron carrier in both cyanobacterial electron transport pathways.

Animals↗
PubMed2004

ZEC, a zinc finger protein with novel binding specificity and transcription regulatory activity.

A novel 114-kDa zinc finger protein, ZEC, has been found by cDNA cloning and characterized. ZEC was strongly expressed in the testis, liver and kidney, and also in embryonic stem cells. Epitope-tagged experiments indicated nuclear localization of ZEC. ZEC contained 18 C2H2 zinc fingers which were organized in two clusters. A ZEC binding DNA sequence, C/GA/TA/TGGTTGGTTGC, which we have designated the GT box, was identified by random oligonucleotide binding selection assay. The GT box did not contain binding sites for other previously characterized transcription factors and thus represented a potentially novel DNA target sequence. Electrophoretic mobility shift assay (EMSA) showed that both clusters of zinc fingers bound to the same DNA sequence. Site-directed mutagenesis revealed that the core sequence TTGGTT within the GT box was essential to ZEC binding, while DNA sequences outside of the core sequence enhanced this interaction. Furthermore, co-transfection assays demonstrated that ZEC could activate a reporter luciferase gene driven by this DNA sequence.

Amino Acid Sequence↗
PubMed2006

The effect of granulocyte colony stimulating factor on regional and global myocardial function in the porcine infarct model.

BACKGROUND: Stem cell therapy has been shown to attenuate the reduction of left ventricular function following myocardial infarction. Most studies have utilized either a direct injection or intra-coronary infusion of cells, but cytokine mobilization of stem cells in the murine model of acute myocardial infarction has been reported to induce similar improvement in cardiac function. METHODS: An antero-apical infarction was induced in swine by balloon occlusion, followed by the daily administration of granulocyte colony stimulating factor (G-CSF) or placebo for 5 days. We used left ventricular angiograms and 2D echocardiograms to assess global function, and 3D echocardiograms to assess regional function prior to infarction, immediately following infarction, and at 8 weeks. Histologic evaluation was performed after sacrifice at 8 weeks. RESULTS: There was no significant difference in early or late post-infarction left ventricular ejection fraction or in myocardial histology between the two groups. Following G-CSF therapy, however, 3D echocardiography demonstrated that the regional ejection fractions of the infarcted segments showed a 50.3% improvement in the G-CSF pigs compared to a 7.4% deterioration in the untreated pigs (p=0.005). CONCLUSIONS: Global left ventricular ejection fraction remained unchanged, and there is no histologic evidence for infarct attenuation following G-CSF infusion in the porcine infarct-reperfusion model. There was recovery of regional function in the infarcted segment in the G-CSF pigs. These data suggest that bone marrow mobilization in larger species has limited potential as a therapy designed to replace infarcted myocardium or to improve overall cardiac function, although further studies are needed to examine regional effect in the infarct area.

Animals↗
PubMed2006

Kinetics and adsorption behavior of carboxymethyl starch on alpha-alumina in aqueous medium.

The adsorption of carboxymethyl starch (CMS) at the alpha-alumina/aqueous solution interface has been investigated through adsorption studies, electrokinetics mobility measurements, and FTIR spectroscopy. Zeta potential measurements show that the addition of CMS results in a more dramatic increase in the absolute zeta potential in the alkaline region, as well as a shift of the isoelectric point to lower values, indicating the adsorption of CMS from the aqueous solution onto the alumina surface. The positive hydrophilic surface sites of alumina are responsible for the adsorption of CMS molecules. The adsorption of CMS is possible after charge reversal by the addition of excess CMS. Nearly 30 min of contact time are found to be sufficient for the adsorption of CMS to reach equilibrium. CMS adsorption follows a Langmuir isotherm with adsorption capacities of 91.74 mg CMS per gram of alpha-alumina. For the adsorption of CMS, pseudo-second-order chemical reaction kinetics provides the best correlation with the experimental data. FTIR analysis indicated that CMS forms outer complexes with alumina surfaces depending on the shifting of the asymmetric and symmetric bands.

Journal Article↗
PubMed2006

Environmental impact of toxic metals and metalloids from the Muñón Cimero mercury-mining area (Asturias, Spain).

This paper presents the results of the sampling surveys carried out in order to evaluate the environmental problems associated to La Soterraña, an abandoned Hg mine in Asturias, north of Spain. In particular, this paper overviews the impact of mining and metallurgical activities on terrestrial and aquatic environments. The wastes generated during the mining activity (ore extraction and processing) and later accumulated on the ground, contain great amount of sulphides, becoming potentially acid-generating. Consequently, the mobility of heavy metals and other ecotoxic elements is enhanced. Wastes are generally located close to watercourses and on very steep hillsides, where they are exposed to oxidative weathering, posing a significant risk due to the release of ecotoxic elements to the environment. Background levels were determined at sites, which had not been directly affected by mercury mining. A multielemental geochemical study of mining wastes, soils, stream sediments, water and air samples collected in the area of influence of the old mining and metallurgical works was carried out. Total Hg and As concentrations in soils reach values up to 502 and 19,940 mg kg(-1), respectively, 500 and 2000 times higher than the local background levels. The effects of mining seem to be intense both in waters and stream sediments, as well as in the local atmosphere, whose Hg content is 10 times higher than the background level in the area. Therefore, the target carcinogenic risk was exceeded for As and Hg in La Soterraña site.

Air Pollutants, Occupational↗
PubMed2006

Analysis of nitroxide spin label motion in a protein-protein complex using multiple frequency EPR spectroscopy.

X- and W-band EPR spectra, at room and low temperatures, are reported for nitroxide spin labels attached to cysteine residues selectively introduced into two proteins, the DNase domain of colicin-E9 and its immunity protein, Im9. The dynamics of each site of attachment on the individual proteins and in the tight DNase-Im9 complex have been analysed by computer simulations of the spectra using a model of Brownian dynamics trajectories for the spin label and protein. Ordering potentials have been introduced to describe mobility of labels restricted by the protein domain. Label mobility varies with position from completely immobilised, to motionally restricted and to freely rotating. Bi-modal dynamics of the spin label have been observed for several sites. We show that W-band spectra are particularly useful for detection of anisotropy of spin label motion. On complex formation significant changes are observed in the dynamics of labels at the binding interface region. This work reveals multi-frequency EPR as a sensitive and valuable tool for detecting conformational changes in protein structure and dynamics especially in protein-protein complexes.

Binding Sites↗
PubMedDate not supplied

Optimizing stability in distal humeral fracture fixation.

Restoration of painless and satisfactory elbow function after a fracture of the distal humerus requires anatomic reconstruction of the articular surface, restitution of the overall geometry of the distal humerus, and stable fixation of the fracture fragments to allow early and full rehabilitation. Although these goals are now widely accepted by the orthopaedic community, they may be technically difficult to achieve, especially in the presence of substantial osteoporosis or comminution. Failure, when it occurs, typically occurs at the supracondylar level through loss of fixation in the distal fragments. To prevent such failure and thereby maximize the potential for union and full elbow mobility after a severely fractured distal humerus, 2 principles must be satisfied: (1) fixation in the distal fragment must be maximized and (2) all fixation in distal fragments should contribute to stability between the distal fragments and the shaft. There are 8 technical objectives by which these principles are met: (1) every screw in the distal fragments should pass through a plate; (2) engage a fragment on the opposite side that is also fixed to a plate; (3) as many screws as possible should be placed in the distal fragments; (4) each screw should be as long as possible; (5) each screw should engage as many articular fragments as possible; (6) the screws in the distal fragments should lock together by interdigitation, creating a fixed-angle structure; (7) plates should be applied such that compression is achieved at the supracondylar level for both columns; and (8) the plates must be strong enough and stiff enough to resist breaking or bending before union occurs at the supracondylar level. These can be achieved with parallel plating. Severe metaphyseal comminution and bone loss are managed by supracondylar shortening. From the biomechanical data in the literature, 3 conclusions can be reached concerning fixation of distal humeral fractures. First, the concept that plates need to be placed perpendicular (90 degrees /90 degrees orientation) is unsubstantiated and incorrect. Second, parallel placement of 2 plates in the sagittal plane is as strong or stronger than the 90 degrees /90 degrees orientation. Finally, linking the plates together through the bone, thereby creating the architectural equivalent of an arch, offers the greatest biomechanical stability for comminuted distal humeral fractures. This can be done by interdigitating and locking the screws together as they pass through the distal fragments from the medial and lateral plates placed in the sagittal plane.

Bone Screws↗
PubMed2005

Effect of polyelectrolyte conditioning on the enhanced dewatering of activated sludge by application of an electric field during the expression phase.

Activated sludge is known to be poorly dewaterable due to its high surface charge density and the extreme solids compressibility, even after polyelectrolyte conditioning. The application of an electric field during pressure dewatering (PDW) of sludge can enhance the dewaterability by the electroosmosis effect. A comparative study was conducted to investigate the additional effect of an electric field, applied during the expression phase, on the dewatering course of polyelectrolyte conditioned sludge, compared to mere PDW. It was found that the application of an electric field markedly improved the dewatering kinetics for all sludge samples, regardless of the conditioning treatment. Although the conditioning polyelectrolyte characteristics and dose had a major effect on the PDW of sludge, the conditioning history did not have a significant effect on the electroosmotic water transport efficiency during the sludge expression phase. By means of on-line streaming potential measurements and fractionated filtrate electrophoretic mobility measurements, it could be demonstrated that even at high polyelectrolyte doses, leading to positively charged sludge flocs, negative surface charges were still present inside the sludge matrix. During the expression of the sludge cake, when liquid is forced to move through the floc inside pores, these negative surface charges hampered PDW, but enhanced electroosmotic dewatering. Electroosmosis is therefore an appropriate technique to remove the water fraction that is associated with these negative surface charges.

Acrylates↗
PubMed1985

Potential role for a guanine nucleotide regulatory protein in chemoattractant receptor mediated polyphosphoinositide metabolism, Ca++ mobilization and cellular responses by leukocytes.

Islet activating protein from Bordetella pertussis toxin which ribosylates certain guanine nucleotide regulatory proteins causes a marked reduction of chemoattractant-elicited responses such as chemotaxis, O2 production and cAMP elevations in human polymorphonuclear leukocytes. The toxin appears to exert its effects by preventing the rapid breakdown of phosphatidylinositol 4,5-bisphosphate induced by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine, thereby inhibiting the increase in intracellular [Ca++] which normally follows chemoattractant stimulation. Responses of leukocytes exposed to Concanavalin A, the Ca++ ionophore A23187, or phorbol myristate acetate were not affected by the toxin. Thus the chemoattractant receptor appears to be coupled to a phosphoinositide specific phospholipase C through a guanine nucleotide regulatory protein. We propose that this complex of receptor-guanine nucleotide regulatory protein-phospholipase C may be applicable to the class of receptors which mobilize intracellular Ca++ by stimulating polyphosphoinositide breakdown.

Bacterial Toxins↗
PubMed1991

Domains and anomalous adsorption isotherms of dipalmitoylphosphatidylcholine membranes and lipophilic ions: pentachlorophenolate, tetraphenylborate, and dipicrylamine.

Dipalmitoylphosphatidylcholine (DPPC) vesicles acquire negative surface charge on adsorption of negatively charged pentachlorophenolate (PCP-), and lipophilic ions tetraphenylborate (TPhB-), and dipicrylamine (DPA-). We have obtained (a) zeta-potential isotherms from the measurements of electrophoretic mobility of DPPC vesicles as a function of concentration of the adsorbing ions at different temperatures (25-42 degrees C), and (b) studied the effect of PCP- on gel-to-fluid phase transition by measuring the temperature dependence of zeta-potential at different PCP- concentrations. The zeta-potential isotherms of PCP- at 25, 32, and 34 degrees C correspond to adsorption to membrane in its gel phase. At 42 degrees C the zeta-potential isotherm corresponds to membrane in its fluid phase. These isotherms are well described by a Langmuir-Stern-Grahame adsorption model proposed by McLaughlin and Harary (1977. Biochemistry. 15:1941-1948). The zeta-potential isotherm at 37 degrees C does not follow the single-phase adsorption model. We have also observed anomalous adsorption isotherms for lipophilic ions TPhB- and DPA- at temperatures as low as 25 degrees C. These isotherms demonstrate a gel-to-fluid phase transition driven by ion adsorption to DPPC membrane during which the membrane changes from weakly to a strongly adsorbing state. The anomalous isotherm of PCP- and the temperature dependence of zeta-potential can be described by a two-phase model based on the combination of (a) Langmuir-Stern-Grahame model for each phase, (b) the coexistence of gel and fluid domains, and (c) depression of gel-to-fluid phase transition temperature by PCP-. Within the anomalous region the magnitude of zeta-potential rapidly increases concentration of adsorbing species, which was characterized in terms of a Esin-Markov coefficient. This effect can be exploited in membrane-based devices. Comments are also made on the possible effect of PCP, as an uncoupler, in energy transducing membranes.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1997

Molecular dynamics study of water and Na+ ions in models of the pore region of the nicotinic acetylcholine receptor.

The nicotinic acetylcholine receptor (nAChR) is an integral membrane protein that forms ligand-gated and cation-selective channels. The central pore is lined by a bundle of five approximately parallel M2 helices, one from each subunit. Candidate model structures of the solvated pore region of a homopentameric (alpha7)5 nAChR channel in the open state, and in two possible forms of the closed state, have been studied using molecular dynamics simulations with restraining potentials. It is found that the mobility of the water is substantially lower within the pore than in bulk, and the water molecules become aligned with the M2 helix dipoles. Hydrogen-bonding patterns in the pore, especially around pore-lining charged and hydrophilic residues, and around exposed regions of the helix backbone, have been determined. Initial studies of systems containing both water and sodium ions together within the pore region have also been conducted. A sodium ion has been introduced into the solvated models at various points along the pore axis and its energy profile evaluated. It is found that the ion causes only a local perturbation of the water structure. The results of these calculations have been used to examine the effectiveness of the central ring of leucines as a component of a gate in the closed-channel model.

Amino Acid Sequence↗
PubMed2000

Separation of potentially therapeutic peptide hormones by liquid chromatography. Optimisation of the composition and pH of the mobile phase.

The aim of this work is to optimise the proportion of the organic modifier and the pH of the mobile phase, in order to separate a series of peptide hormones with therapeutic interest in the molecular mass range from 500 to 6000. The composition of the mobile phase was optimised by establishing relationships between retention parameters and either the scale of solvent polarity, or the Kamlet-Taft multiparameter solvent scale of the eluent, using linear solvation energy relationships. Likewise, linear correlations between the chromatographic retention and Reichardt's E(T)N parameter were obtained. These relationships allowed an important reduction of the experimental retention data needed for developing a given separation. In addition, a model describing the effect of the correctly measured pH of the mobile phase on retention in LC was established and tested for the series of selected peptides using an octadecylsilica column. The proposed equations permit the prediction of the optimum pH and also permit the determination of the acidity constants of the peptides in the hydro-organic mixtures using a minimum number of measurements.

Amino Acid Sequence↗
PubMed1991

Pararectus retroperitoneal radical nephrectomy.

We describe a new pararectus retroperitoneal approach to the kidney, which allows easy identification and occlusion of the renal vessels before mobilization of the kidney. This approach potentially reduces morbidity, hospital stay and cost. The technique has been used in 4 cases of radical nephrectomy for tumor and in 1 for pyonephrosis.

Carcinoma, Renal Cell↗