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Biomedical subjects

O Bertrand

Publications and source records attributed to O Bertrand.

At least 109 records · Page 6Linked to original sources

Automated purification of human protein band 3, the major integral protein of the erythrocyte membrane.

Human erythrocyte protein band 3 was purified from a Triton X-100 extract of white ghosts. This purification, including an ion-exchange chromatography and a group-affinity chromatography, was automated. The apparatus was assembled from commercially available elements and allowed for the recovery of 2 to 3 mg pure band 3 in 2 hr. The purification could be repeated several times a day. The advantages of automation are discussed.

Anion Exchange Protein 1, Erythrocyte↗

Continuous measurement of pulmonary gas exchange during general anaesthesia in man.

We report a system for the continuous measurement of oxygen uptake (VO2) and carbon dioxide output (VCO2) during open-circuit anaesthesia. Gas concentrations were measured by a mass-spectrometer, and expired flow by a pneumotachograph. The values measured by the system were compared in vitro to values produced by a nitrogen-dilution technique. Excellent correlations were found. Continuous measurements were performed in 21 patients anaesthetized for abdominal surgery. Compared to pure intravenous anaesthesia (flunitrazepam-fentanyl), anaesthesia including the administration of nitrous oxide or nitrous oxide and halothane led to more pronounced and sustained decreases in VO2 and core temperature, with a better cardiovascular stability. Two hours postoperatively, VO2 was not different from preoperative values. After a transient increase at the onset of anaesthesia, the respiratory exchange ratio (VCO2/VO2) returned to preoperative values, and then remained subsequently unchanged. Based on these observations, the system described provides an accurate approach to noninvasively monitoring the pulmonary gas exchange in the operating room.

Aged↗

Transient drug-induced abolition of BAEPs in coma.

Combined infusion of high doses of lidocaine and thiopental in a comatose patient induced major latency and amplitude BAEP changes, which progressed to complete BAEP abolition. Responses returned to normal after drugs were discontinued. EEGs during the episodes showed long-lasting periods of activity suppression, but were never isoelectric. BAEPs are resistant to hypothermia and barbiturates, but must be interpreted cautiously in patients treated with a combination of anesthetic drugs that includes lidocaine.

Adult↗

Glycosylation of human fibrinogen and fibrin in vitro. Its consequences on the properties of fibrin(ogen).

As observed for many proteins, glucose has been shown to bind non enzymatically to fibrinogen and to fibrin. The in vitro degree of glycosylation depends upon the concentration of glucose added to fibrinogen and also on the incubation period. This glycosylation induces a decrease in fibrin lysis by plasmin. On the contrary, it does not influence either fibrinogen activity as cofactor in ADP-induced platelet aggregation or the binding of thrombin onto a clot. Despite the 4 day half life of fibrinogen, since clearance of fibrinogen is exponential, it is assumed that very small quantities of fibrinogen may remain for a long time in the circulation leading to the presence of a low level of a highly glycosylated form of fibrinogen. Consequently, poorly degradable fibrin might be derived from this highly glycosylated fibrinogen and thus be responsible for capillary occlusion and also for atherosclerotic complications in the diabetic patient.

Adenosine Diphosphate↗

Mapping of scalp potentials by surface spline interpolation.

Evoked potentials and EEGs record punctate electrical activity at electrode sites. To represent the overall potential distribution on the entire scalp it is necessary to interpolate between these sampled values. Surface splines are mathematical tools for interpolating functions of two variables. In comparison to the classical methods of interpolation, based on linear combination of the potentials of the 4 nearest electrodes, spline methods are smoother, give more precisely located extrema and converge faster toward the 'true' potential surface when the number of recording electrodes is increased. These advantages are at the expense of lengthier computation time.

Brain Mapping↗

Protein band 3 phosphotyrosyl phosphatase. Purification and characterization.

A phosphotyrosylprotein phosphatase has been purified from human red cell cytosol by successive DEAE cellulose, phosphocellulose and Red Procion-H3B-Sepharose chromatography. Overall purification was about 9000 with a yield of 30%. The enzyme was more than 95% pure as judged by SDS polyacrylamide gel. Its molecular weight was 17,000 and maximum activity was observed at pH 5.5. It was active towards both the phosphorylated tyrosine on the cytosolic fragment of the red cell protein band 3 and para-nitrophenyl phosphate. However the effects of ligands differ for the two substrates.

Anion Exchange Protein 1, Erythrocyte↗

Brain-stem monitoring. I. A system for high-rate sequential BAEP recording and feature extraction.

A computerized system is proposed for the continuous automatic monitoring of BAEPs in the intensive care unit or in the operating room. A new adaptive optimal digital filtering technique has been developed to allow BAEP recording with 200-400 stimulations, i.e., within 10-20 sec only, every 2 min. This method permits a selective cancellation of the major artifacts as soon as they occur: electrical interference and myogenic activities which are commonly encountered in such environments. An algorithm for automatic peak detection (waves I-V) has been implemented, and the whole system has been evaluated in terms of reduction of the intra-individual variability of latencies and amplitudes. Some clinical examples of comatose patient monitoring are briefly presented to illustrate the performance and the reliability of the system.

Brain Stem↗

Brain-stem monitoring. II. Preterminal BAEP changes observed until brain death in deeply comatose patients.

Preterminal BAEP changes were studied until brain death in 8 head-injured patients out of a series of 38 comas monitored by means of a system allowing high-rate sequential recording. Two different modalities of BAEP degradation were disclosed: (1) simultaneous latency increase of all components associated with a decrease of cerebral perfusion pressure (CPP), consistent with ongoing ischaemia of the posterior fossa; (2) deterioration of brain-stem components (waves III-V) with preserved or even enhanced wave I. The latter pattern was not consistently associated with any haemodynamic change and might be related to mechanical factors causing rostro-caudal deterioration of brain-stem function. The time course of BAEP degradation ranged from a few minutes to more than 10 h. In the case of slow preterminal evolution definitely pathological trends were identified even when individual BAEPs were still within normal limits. Such trends would have remained unnoticed in single BAEP records. Hypothermia and anaesthetic drugs were found to induce falsely alarming BAEP changes very similar to those seen during preterminal evolution. Our results suggest that continuous brain-stem monitoring could be helpful for management of comatose head-injured patients.

Blood Pressure↗

ADP-induced platelet aggregation depends on the conformation or availability of the terminal gamma chain sequence of fibrinogen. Study of the reactivity of fibrinogen Paris 1.

Fibrinogen Paris I contains a mutant gamma chain that is longer than the normal chain, resulting in altered fibrin polymerization and cross-linking. Because these functions involve the carboxy-terminal region of the gamma chain, we decided to determine whether fibrinogen Paris I or the isolated Paris I gamma chain supports normal ADP-induced platelet aggregation, a function that requires the ultimate 12 residues of the normal gamma chain (400 through 411). Aggregation of ADP-stimulated normal platelets was defective with fibrinogen Paris I and markedly depressed with the gamma Paris I chain. These findings prompted us to characterize the carboxy-terminal structure of the region of the gamma Paris I chain responsible for this activity. The carboxy-terminal cyanogen bromide (CNBr) peptide of the normal gamma chain (385 through 411) or that from gamma Paris I was isolated by differential adsorption to triethylene-tetramine resin or by reverse-phase high-performance liquid chromatography (HPLC). The CNBr peptide from the Paris I gamma chain was identical to that of the normal gamma chain in its retention time on HPLC, its amino acid composition, and its sequence. Thus, the primary structure of the gamma Paris I chain from residue 384 through 411 is normal, indicating that a peptide insertion has occurred upstream from residue 384, resulting in an impairment of those physiologic functions attributable to the carboxy-terminal end of the gamma chain from position 384 (ie, cross-linking, ADP-induced platelet aggregation, and at least a portion of the gamma chain polymerization site). These observations demonstrate that the gamma chain platelet recognition site in the fibrinogen molecule is necessary but not alone sufficient to support normal ADP-induced platelet aggregation. There appears to be an additional requirement for normal conformation of the gamma chain or availability of its terminal sequence during the interaction of fibrinogen with platelets.

Adenosine Diphosphate↗

Properties of a membrane-bound tyrosine kinase phosphorylating the cytosolic fragment of the red cell membrane band 3 protein.

Band 3 protein of human erythrocyte membrane is phosphorylated on a tyrosine residue located near the NH2 terminal by an endogenous tyrosine kinase activity (Dekowski, S., Rybicki, A. and Drickamer, K. (1983) J. Biol. Chem. 258, 2750-2753). A tyrosine kinase phosphorylating the band 3 protein in situ has been extracted from ghosts by non-ionic detergent and partially characterized (Phan-Dinh-Tuy, F., Henry, J. and Kahn, A. (1985) Biochem. Biophys. Res. Commun. 126, 304-312). We have studied the properties of the tyrosine kinase activity which remains bound to the ghosts after detergent extraction using the 43 kDa fragment of protein 3 as substrate. This activity, solubilized from the detergent-resistant material at 0.25 M NaCl and concentrated by phosphocellulose and tyrosine-agarose chromatographies, remains linked to high molecular weight complexes. It is specific for tyrosine. Assayed with the purified 43 kDa fragment it requires the presence of Mn2+ which cannot be replaced by Mg2+. Its affinity for 43 kDa fragment is very high with a Km of 3.3 microM. ATP acts as a phosphoryl donor with a Km of 0.55 microM. The tyrosine kinase activity was not modified by insulin, DMSO, phorbol ester and epidermal growth factor, vanadate and xanthine derivatives. Polyamines spermidine and the polylysine are inhibitors in the presence of Mn2+ but not in the presence of Mg2+. Heparin is a competitive inhibitor of ATP. 2,3-Diphosphoglycerate is an inhibitor at physiological concentrations (Ki = 2 mM). Purified red cell actin is not phosphorylated by the tyrosine kinase. These properties distinguish the red cell membrane-bound tyrosine kinase from other tyrosine kinases extracted from normal cells.

2,3-Diphosphoglycerate↗

Added inspiratory work of breathing during CPAP ventilation: comparison of two demand-valve devices with a continuous flow-system.

Measurements of added inspiratory work (AIW) of breathing imposed by three different CPAP systems were performed in 10 patients. One system was a continuous flow system while the two others were demand flow systems separated from respirators (Ohmeda VD 101 and Draeger CPAP 800 devices). AIW was calculated from pressure and flow signals recorded at the mouthpiece level. The AIW calculated with the two demand-flow systems was found to be the same as the AIW calculated with the continuous flow system although the results obtained by the Draeger device were less constant. The results obtained with the Ohmeda device in our patients conflicted with data previously published using a lung model.

Acute Disease↗

Effects of ex-vivo plasma adsorption over protein A sepharose in acute leukemia.

Plasma adsorption over immobilized Staphylococcus aureus Cowan I has resulted in tumor regression in several animal and human trials. Protein A, because of its ability to bind IgGs has been considered as the effective component of Staphylococcus aureus. In 4 patients with acute leukemia, a plasma volume of 1500 cm3 was passed in an ex-vivo system over immobilized SpA-Sepharose and then reinfused. Almost all of the IgGs contained in the plasma volume could thus be removed. Toxic side-effects were mild. No significant clinical improvement could be obtained. Plasma incubation with SpA did not modify blast cell viability or leukemic progenitor cell growth. Along with others, this study shows that Protein A is probably not the mediator of the tumoricidal responses observed in studies using adsorption over Staphylococcus aureus Cowan I. The ex-vivo system prepared for this study could also be used for plasma IgG removal for the treatment of autoimmune or immune-complex related disorders.

Blood Cell Count↗

[Continuous measurement of gas exchange during artificial ventilation].

A system for on-line measurement of respiratory gas exchange in patients undergoing artificial ventilation is described. Fractional concentrations were measured by a mass spectrometer and expired flow by a pneumotachometer; signals processing was carried out by a microcomputer. The accuracy of the measurement of the burning methanol RQ was within 1.6% at FIO2 lower or equal to 0.4. When compared to a reference method (the Douglas bag method), correlation was found to be excellent. The major problem in measuring gas exchange in a ventilated patient is that physiological signals are contaminated by artefacts which may lead to erroneous computations. An algorithm was developed in order to identify and disregard artefacted periods or ventilatory unsteady state in the patients. The technique used for data processing gave reliable continuous measurements of respiratory gas exchange for periods up to 24 h, without interfering with the nursing of the ventilated patients.

Algorithms↗

Pulmonary gas exchange during hemodialysis.

UNLABELLED: Pulmonary gas exchange was continuously measured in 13 mechanically ventilated patients during 24 hemodialyses for acute renal failure. Minute-ventilation was maintained constant by controlled ventilation and gas exchange was continuously measured by a mass-spectrometer system. Three groups were compared: a cuprophan membrane with an acetate dialysate; a polyacrilonitrile membrane (PAN) with an acetate dialysate; and PAN with a bicarbonate dialysate. Arterial PO2 and the O2 alveolar-arterial gradient were the same regardless of the membrane used. [H+] mildly decreased with all dialysates used. Arterial PCO2 decreased only with the acetate dialysate. O2 consumption increased, up to 20 +/- 5% of the initial values during hemodialysis, and remained increased during the two hours following the hemodialysis. Respiratory exchange ratio was lower after than before the hemodialysis. IN CONCLUSION: the maintenance of a constant minute ventilation prevented hemodialysis induced hypoxemia. VO2 increased during hemodialysis.

Acute Kidney Injury↗