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Biomedical subjects

O Bertrand

Publications and source records attributed to O Bertrand.

136 records · Page 8Linked to original sources

Studies on the nature of different molecular forms of glucose-6-phosphate dehydrogenase purified from human leukocytes.

Several molecular forms of human glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate:NADP+ 1-oxidoreductase, EC 1.1.1.49) corresponding to different stages of post-synthetic modifications have been purified from human leukocytes. The various enzyme forms were different in their specific activity, their kinetic properties and their isoelectrofusing pattern. The molecular weight of the subunits of the different forms was not modified. The changes in the electrofocusing pattern were not due to modifications of the N-terminal ends, the oxidation of thiol groups or the non-covalent fixation of an acid molecule upon the enzyme. Carboxypeptidase B cleaved a C-terminal lysine from the different enzyme forms and shifted the isoelectric point of the different enzyme active bands towards the acid pH. The different enzyme forms studied here seemed to result from the action upon 'native glucose-6-phosphate dehydrogenase' of 'modifying factors' especially abundant in some leukemic granulocytes. The modifying factors did not seem to be consumed during the 'modification' of glucose-6-phosphate dehydrogenase. Moreover, the storage for one year of unmodified enzyme resulted in changes in its electrofocusing pattern similar to those quickly induced by the 'modifying factors'. Consequently it appears that the modifying factors are catalysts of the modification of special residues of glucose-6-phosphate dehydrogenase. The hypothesis that this modification involves the deamination of asparagine or glutamine residues is put forward.

Amino Acid Sequence↗

Glucose-phosphate isomerase deficiency due to a new variant (GP I Barcelona) and to a silent gene: biochemical, immunological and genetic studies.

A 12-year-old girl of Spanish origin was found to be double heterozygote for a deficient GP I variant (GP I Barcelona) and for a silent GP I gene. The mother was heterozygote for GP I Barcelona and the father was heterozygote for the silent gene. GP I Barcelona was a fast variant (116%) with an increased isoelectric point (9.55), lability to heat and to urea, and shift of the pH curve towards the acidic pH. The other kinetic characteristics were normal. The ratio of enzymatic activity to immunological reactivity was normal in erythrocytes and white blood cells of the father and the mother but decreased to 75% of normal in blood cells of the daughter. The genetic and molecular mechanisms of GP I deficiency of this patient are discussed.

Adult↗

Human leukocyte glucose-phosphate-isomerase purification by affinity elution and immunological study.

The authors have purified glucose phosphate isomerase (GPI) from human leukocytes ; they used as starting material leukemic leukocytes obtained from a patient with hyper-leukocytic acute myeloid leukemia ; it was possible to obtain several milligrams of pure enzyme from a single patient. The purification procedure includes a two step precipitation by ammonium sulfate and one column chromatography on a cation exchanger with specific elution by 6 phosphogluconate, a ligand of GPI ; of the two cation exchangers tested, phosphocellulose was found to lead to a better yield than CM-Sephadex. The end product of purification had a specific activity of 855 UI/mg and gave only one band in sodium dodecyl sulphate polyacrylamide gel electrophoresis. Anti GPI monospecific rabbit serum was obtained with purified enzyme. GPI from the various blood cells of ten normal controls was studied immunologically and the ratio of the enzymatic activity to the immunological reactivity was measured ; this ratio (i.e. the molecular specific activity) was lower in granulocytes than in lymphocytes and still more depressed in platelets and hemolysate. The significance of such differences is discussed.

Blood Platelets↗

A low-cost automated device incorporating a hollow fiber filtration cartridge for large-scale production of ghosts from human erythrocytes.

Readily available elements were used to build an automatic apparatus dedicated to the preparation of erythrocyte ghosts. The apparatus is designed around a low-cost re-usable hollow-fiber filtration cartridge (marketed for therapeutic plasmapheresis). The apparatus is controlled by a simple programmer (based on a diode matrix and low cost timers and liquid level sensors): once the apparatus is loaded with whole red blood cells, washing of cells, as well as hemolysis and washing of ghosts, is performed by the machine in about 4.5 h without any operator intervention. Automatic filter cleaning takes a further 110 min.

Automation↗

Cortical mapping of gamma oscillations in areas V1 and V4 of the macaque monkey.

To characterize the temporal and spatial parameters of gamma activity evoked by visual stimuli in areas V1 and V4 of the monkey cortex, we recorded the electrocorticogram (ECoG) with an implanted array of 28 and 31 subdural electrodes placed over the surface of the operculum in two anesthetized monkeys. This intermediate level of recordings should help to bridge the gap between multiunit and scalp recordings. Both averaged and single-trial responses to small flashed stimuli, for which we varied the retinotopic position, the luminance and the color, were analyzed in the time-frequency domain using a wavelet-based decomposition of the signal. Large gamma oscillations (40-55 Hz), not phase locked to stimulus onset, were observed during the whole stimulus presentation, whereas visual evoked potentials (VEPs) were present mainly at stimulus onset and offset. Cortical mapping showed that both activities were restricted in spatial extent and followed the retinotopic organization of area V1 on the operculum, thus strongly suggesting they were generated in the underlying cortex. Oscillatory burst detection in single trials showed that one to two bursts lasting from 100 ms to 500 ms occurred in the first 500 ms following stimulus onset, and that bursts occurring during the subsequent phases of the response had a smaller amplitude and duration. Finally, we showed that gamma activity was stronger with higher luminances and for red than for green, yellow, or white stimuli of same luminance. In one animal we recorded gamma activity over area V4. This was of lower magnitude than the activity recorded over V1 and was delayed by 40 ms with respect to the beginning of gamma activity in V1, in contrast with the VEPs that were delayed by 20 ms only. Both gamma oscillations and early VEP followed the retinotopic organization of V4 over the prelunate gyrus. The results show that gamma oscillations are dependent upon the same parameters as the VEPs (retinotopic position, luminance, and color). However, the differences in the time course of VEPs and gamma activity (transient vs. sustained) suggests that these two responses may reflect different cell populations, different networks, or different firing modes.

Animals↗

Sustained and transient oscillatory responses in the gamma and beta bands in a visual short-term memory task in humans.

In a visual delayed matching-to-sample task, compared to a control condition, we had previously identified different components of the human EEG that could reflect the rehearsal of an object representation in short-term memory (Tallon-Baudry et al., 1998). These components were induced oscillatory activities in the gamma (24-60 Hz) and beta (15-20 Hz) bands, peaking during the delay at occipital and frontal electrodes, and two negativities in the evoked potentials. Sustained activities (lasting until the end of the delay) are more likely to reflect the continuous rehearsing process in memory than transient (ending before the end of the delay) activities. Nevertheless, since the delay duration we used in our previous experiment was fixed and rather short, it was difficult to discriminate between sustained and transient components. Here we used the same delayed matching-to-sample task, but with variable delay durations. The same oscillatory components in the gamma and beta bands were observed again during the delay. The only components that showed a sustained time course compatible with a memory rehearsing process were the occipital gamma and frontal beta induced activities. These two activities slowly decreased with increasing delay duration, while the performance of the subjects decreased in parallel. No sustained response could be found in the evoked potentials. These results support the hypothesis that objects representations in visual short-term memory consist of oscillating synchronized cell assemblies.

Adult↗