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O Bertrand

Publications and source records attributed to O Bertrand.

At least 19 recordsLinked to original sources

Oscillatory synchrony between human extrastriate areas during visual short-term memory maintenance.

How do we keep an object in mind? Based on evidence from animal electrophysiology and human brain-imaging techniques, it is commonly held that short-term memory relies on sustained activity in a network distributed over sensory and prefrontal cortices. How does neural firing persist in such a distributed network in the absence of visual input? Hebb's influential but so far unproved proposal, developed more than 50 years ago, is that sustained activation in short-term memory networks is maintained by reverberating activity in neuronal loops. We hypothesized that synchronized oscillatory activity, proposed to provide a dynamic link between distributed areas, could not only coordinate activity in the network but also establish reentrant loops in the system to enable both sustained firing and temporal coincidence of inputs. We show in human intracranial recordings that limited regions of extrastriate visual areas, separated by several centimeters, become synchronized in an oscillatory mode during the rehearsal of an object in visual short-term memory. Synchrony occurs specifically in the beta range (15-25 Hz) and disappears in a control condition. These findings thus confirm experimentally the hypothesis of a functional role of synchronized oscillatory activity in the coordination of distributed neural activity in humans, and support Hebb's popular but unproved concept of short-term memory maintenance by reentrant activity within the activated network.

Action Potentials↗

Effect of gluconic acid as a secondary carbon source on non-growing L-lysine producers cells of Corynebacterium glutamicum. Purification and properties of 6-phosphogluconate dehydrogenase.

We studied the production of L-lysine in Corynebacterium glutamicum ATCC 21543 non growing cells obtained by nutrient limitation. Statistical analysis revealed significant differences in the L-lysine titers of glucose, gluconic acid or glucose-gluconic acid cultures. Higher L-lysine titer obtained in batch cultures with mixed carbon sources or gluconic acid alone were found to be associated with a high 6-phosphogluconate dehydrogenase activity (6PGDH, E.C.1.1.1.44). This enzyme is a pivotal enzyme within the hexose monophosphate pathway, and thus of importance for L-lysine production. 6PGDH was purified and characterized. The purified enzyme migrates as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular mass of 52.5 kDa. The molecular mass of the native enzyme was estimated to be 120 kDa by molecular exclusion chromatography, thus suggesting a homodimeric structure. The amino terminal sequence shows a strong similarity (a match of 86% of the first 20 amino acid) to the 6PGDH from other microorganisms such as, E. coli and B. subtilis. The pI of the dimeric native enzyme and the optimum pH were 6.2 and 8.0, respectively. For the oxidative decarboxylation of 6-phosphogluconate, K(m) of 71 &mgr;M and 43 &mgr;M were obtained for 6-phosphogluconate and NADP(+), respectively.

Journal Article↗

Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells.

The Kell blood-group antigen was originally reported to be a protein expressed in erythroid tissue only. Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. DNA sequencing of the transcript revealed a sequence identical to that obtained from erythroid tissue. In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. Moreover, most tissues analysed exhibited low levels of Kell transcripts. The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. Altogether, the results indicated that KEL expression is not restricted to erythroid tissue.

Antigens, Nuclear↗

New procedures for glycophorin A purification with high yield and high purity.

Glycophorin A (GPA) is the major glycoprotein of the human erythrocyte membrane. It is known to form, in SDS gels as well as in a membrane environment, homodimers, and also heterodimers with the homologous molecule Glycophorin B (GPB). It is shown in this report that the propensity of GPA to dimerize with GPB precludes satisfactory preparation with high yield of pure GPA using classical techniques including SEC and RPLC. It was demonstrated using multiple angle light scattering that GPA is eluted from RPLC columns as dimers. A convenient procedure was devised which allowed us to get pure GPA with high yield. This procedure consists of selectively blocking GPA-GPB heterodimer formation by selective modification of Cysteine 50 of GPB before RPLC.

Amino Acid Sequence↗

Multiple supratemporal sources of magnetic and electric auditory evoked middle latency components in humans.

The supratemporal sources of the earliest auditory cortical responses (20-80 ms) were identified using simultaneously recorded electroencephalographic (EEG) and magnetoencephalographic (MEG) data. Both hemispheres of six subjects were recorded two or three times in different sessions in response to 8000 right-ear 1 kHz pure tones stimuli. Four components were identified: Pa (28 ms), Nb (40 ms), and two subcomponents of the Pb complex, termed Pb1 (52 ms) and Pb2 (74 ms). Based on MEG data, the corresponding sources were localized on the anatomy using individual realistic head models: Pa in the medial portion of Heschl's gyri (H1/H2); Nb/Pb1 in the lateral aspect of the supratemporal gyrus (STG); and Pb2 in the antero-lateral portion of Heschl's gyri. All sources were oriented antero-superiorly. This pattern was clearest in the contralateral hemisphere, where these three activities could be statistically dissociated. Results agree with previous invasive human intracerebral recordings, with animal studies reporting secondary areas involved in the generation of middle latency auditory-evoked components, and with positron emission tomography and functional magnetic resonance imaging studies often reporting these three active areas although without temporal information. The early STG activity may be attributed to parallel thalamo-cortical connections, or to cortico-cortical connections between the primary auditory cortex and the STG, as recently described in humans.

Acoustic Stimulation↗

Oscillatory gamma activity in humans: a possible role for object representation.

The coherent representation of an object has been suggested to be established by the synchronization in the gamma range (20-100 Hz) of a distributed neural network. So-called '40-Hz' activity in humans could reflect such a mechanism. We have presented here experimental evidence supporting this hypothesis, both in the visual and auditory modalities. However, different types of gamma activity should be distinguished, mainly the evoked 40-Hz response and the induced gamma activities. Only induced gamma activities seem to be related to coherent object representations. In addition, their topography depends on sensory modality and task, which is in line with the idea that they reflect the oscillatory synchronization of task-dependent networks. They can also be functionally and topographically distinguished from the classical evoked potentials and from the alpha rhythm. It was also proposed that the functional role of gamma oscillations is not restricted to object representation established through bottom-up mechanisms of feature binding, but also extends to the cases of internally driven representations and to the maintenance of information in memory.

Brain↗

Fibrinogen Alès: a homozygous case of dysfibrinogenemia (gamma-Asp(330)-->Val) characterized by a defective fibrin polymerization site "a".

Congenital homozygous dysfibrinogenemia was diagnosed in a man with a history of 2 thrombotic strokes before age 30. His hemostatic profile was characterized by a dramatically prolonged plasma thrombin clotting time, and no clotting was observed with reptilase. Complete clotting of the abnormal fibrinogen occurred after a prolonged incubation of plasma with thrombin. The release of fibrinopeptides A and B by thrombin and of fibrinopeptide A by reptilase were both normal. Thrombin-induced fibrin polymerization was impaired, and no polymerization occurred with reptilase. The polymerization defect was characterized by a defective site "a," resulting in an absence of interaction between sites A and a, indicated by the lack of fragment D(1) (or fibrinogen) binding to normal fibrin monomers depleted in fibrinopeptide A only (Des-AA fm). By SDS-PAGE, the defect was detected on the gamma-chain and in its fragment D(1). The molecular defect determined by analysis of genomic DNA showed a single base change (A-->T) in exon VIII of the gamma-chain. The resulting change in the amino acid structure is gamma 330 aspartic acid (GAT) --> valine (GTT). It is concluded that the residue gamma-Asp(330) is essential for the normal functioning of the polymerization site a on the fibrinogen gamma-chain.

Adult↗

A murine monoclonal antibody against Kx protein which reacts also with beta-spectrin.

Kx is a polytopic membrane protein of human erythrocytes carrying the Kx blood group antigen, which is deficient in rare patients with McLeod syndrome. Kx is disulphide bond linked to the Kell glycoprotein, which is a bitopic type II membrane protein carrying the Kell blood group antigen. Mice immunized with a synthetic peptide predicted to be located on the second external loop of Kx produced a monoclonal antibody called 3E12 which does not recognize red cells with common Kell phenotype by agglutination and flow cytometry. 3E12 recognizes the Kx protein and the spectrin beta-chain on western blots, the affinity for these two proteins being lowered with increasing ionic strength. Linear epitopes recognized by 3E12 are E116EIEKE121 and L484AQELEKE491 on the Kx protein and spectrin beta-chain, respectively. To quantify the relative amount of Kx in Empigen BB extracts of red cell membranes, an ELISA for Kx was set up which showed conclusively that (i) there is less Kx in membranes of K0 individuals (lacking the Kell glycoprotein) than in membranes of common individuals, and (ii) that all common individuals, typed as K+k-, K-k+ and K+k+, have the same amount of Kx on their red cell membranes. When an erythrocyte membrane detergent extract from one K0 individual was chromatographed on an immobilized 3E12 column, a minute amount of authentic Kell glycoprotein was recovered in acid eluted fractions, indicating that at least the K0 individual under study may still produce some Kell protein.

Amino Acid Sequence↗

Immunopurification of the blood group RhD protein from human erythrocyte membranes.

Rh proteins are membrane proteins encoded by genes at the blood group RH locus. They are of paramount importance in transfusion medicine, but their function is still unknown. Biochemical and biophysical studies of these proteins are scarce since only minute amounts of the very hydrophobic Rh proteins, can be purified from human erythrocytes. Recently, a human monoclonal antibody (LOR-15C9) was described as having the unique property to recognize the Rh30 protein carrying the major blood group D specificity (RhD protein), either in a membrane detergent extract or when blotted on a membrane. In this report, we describe one-step purification of the RhD protein from detergent extracts of red cell membranes, based on immunoaffinity chromatography carried out with immobilized LOR-15C9 IgG. The technique yielded RhD protein with high purity which was devoid of other associated proteins (RhAG, CD47, LW and GPB) that comprise the Rh complex in the erythrocyte membrane. By contrast immunoprecipitation performed with the same antibody led to co-isolation of both RhD and RhAG.

Blood Proteins↗

Analysis of speech sounds is left-hemisphere predominant at 100-150ms after sound onset.

Hemispheric specialization of human speech processing has been found in brain imaging studies using fMRI and PET. Due to the restricted time resolution, these methods cannot, however, determine the stage of auditory processing at which this specialization first emerges. We used a dense electrode array covering the whole scalp to record the mismatch negativity (MMN), an event-related brain potential (ERP) automatically elicited by occasional changes in sounds, which ranged from non-phonetic (tones) to phonetic (vowels). MMN can be used to probe auditory central processing on a millisecond scale with no attention-dependent task requirements. Our results indicate that speech processing occurs predominantly in the left hemisphere at the early, pre-attentive level of auditory analysis.

Acoustic Stimulation↗

The repressor which binds the -75 GATA motif of the GPB promoter contains Ku70 as the DNA binding subunit.

Glycophorin B (GPB) is an abundant cell surface glycoprotein which is only expressed in human erythroid cells. Previous functional analysis demonstrated that the repression of the GPB promoter is determined by the binding of a ubiquitous factor which recognizes a GATA motif centered at position -75. In erythroid cells this ubiquitous factor is displaced by the binding of the erythroid-specific factor hGATA1. Here, we have identified the Ku70 protein as a candidate GPB repressor DNA binding subunit through the screening of a human HeLa expression library using the -75 GATA sequence as bait (one-hybrid method). Electrophoretic mobility shift assays demonstrated that the ubiquitous factor that binds the -75 GATA sequence was the Ku70-Ku80 (Ku) heterodimer. Co-transfection experiments demonstrated that overexpression of Ku70 in the K562 erythroleukeamic cell line resulted in transcriptional repression of the chloramphenicol acetyltransferase reporter gene when placed under the control of the wild-type GPB promoter. Conversely, no repression was observed when a mutation that abolished the binding of Ku was introduced in the GPB promoter construct. Altogether, these results indicate that Ku binds in vivo to the -75 WGATAR motif and is involved in negative regulation of the GPB promoter. These findings suggest that, besides its role in many functions, Ku is also involved in transcriptional regulation of erythroid genes.

Antigens, Nuclear↗

The E-box of the human glycophorin B promoter is involved in the erythroid-specific expression of the GPB gene.

Previous studies performed on the glycophorin B (GPB) expression demonstrated that this gene is expressed in erythroid cells only and that the ubiquitous factor Ku70 is involved in the process. Here, we investigated the contribution of the -70 E-box sequence toward the GPB promoter expression. We found that the E-box bound two factors, the USF1/USF2 protein and an unidentified ubiquitous protein which was named factor U. Site-directed mutagenesis performed on the -70 E-box showed that the USF factor had an activating effect in CAT assays. Conversely, mutation of the -70 E-box that impaired the binding of factor U led to a positive CAT activity in nonerythroid cells and thus to the loss of the erythroid-specific expression of the GPB gene. This indicates that, in addition to the Ku70 factor, the extinction of the GPB promoter expression in nonerythroid cells depends also on the repressing effect of the factor U.

Antigens, Nuclear↗

Structure and expression of the mouse homologue of the XK gene.

The human Kx blood group antigen is carried by a 37,000 M(r) apparent molecular mass membrane polypeptide which is deficient in rare individuals with the McLeod syndrome. The X-linked human XK gene is transcribed in many tissues including adult skeletal muscle and brain, sieges of disorders observed in McLeod syndrome. We report here the cloning of the orthologous mouse XK mRNA. Comparison of XK from human and mouse revealed 80% sequence similarity at the amino acid level. The mouse XK gene is organized in two exons and is expressed in many tissues, but its expression pattern is slightly different from that of the human gene. The presence in mouse erythrocyte membrane of a 43,000 M(r) Kx-related protein was demonstrated by immunoblotting with a rabbit antiserum directed against the human protein. With non-reduced samples, a 140,000 M(r) species was detected instead of the 43,000 M(r) protein, suggesting that, as demonstrated in the Kx polypeptide might be complexed with another protein in mouse red cells, presumably the homologue of the human Kell protein of 93,000 M(r).

Amino Acid Sequence↗

Oscillatory gamma activity in humans and its role in object representation.

We experience objects as whole, complete entities irrespective of whether they are perceived by our sensory systems or are recalled from memory. However, it is also known that many of the properties of objects are encoded and processed in different areas of the brain. How then, do coherent representations emerge? One theory suggests that rhythmic synchronization of neural discharges in the gamma band (around 40 Hz) may provide the necessary spatial and temporal links that bind together the processing in different brain areas to build a coherent percept. In this article we propose that this mechanism could also be used more generally for the construction of object representations that are driven by sensory input or internal, top-down processes. The review will focus on the literature on gamma oscillatory activities in humans and will describe the different types of gamma responses and how to analyze them. Converging evidence that suggests that one particular type of gamma activity (induced gamma activity) is observed during the construction of an object representation will be discussed.

Journal Article↗

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Journal Article↗

A ring-shaped distribution of dipoles as a source model of induced gamma-band activity.

As opposed to slow waves, spontaneous and stimulus-induced oscillations in the gamma-band show no polarity reversal in cortical depth, which cannot be explained by the classical equivalent current dipole model usually proposed as a model of pyramidal cell synaptic activity. Here we propose a ring-shaped distribution of dipoles as a source model for these fast oscillations. This distribution generates a field potential that does not reverse through cortical depth. Such a geometry could correspond to horizontally oriented dendritic fields. Moreover, this distribution generates a potential field, but no, or weak, magnetic field on the scalp surface, which corresponds to the observation that visually-induced gamma-band oscillations are detectable in EEG data, but not in simultaneously recorded MEG data.

Brain↗

An evaluation of dipole reconstruction accuracy with spherical and realistic head models in MEG.

MEG forward problem has been solved for about 2000 dipoles placed on the brain surface using a very fine 3-layer realistic model of the head and the boundary element method (BEM). For each dipole, spherical models, one-layer realistic BEM models and coarser 3-layer realistic BEM models, were used to reconstruct the dipole. It was found that the localization bias induced by using a spherical model of the head increased from 2.5 mm in the upper part of the head to 12 mm in the lower part, on average. It was also found that, for the same computing time, a 3-layer model of the head gave on average 2 mm better localization errors than a one-layer model of the head. Orientation errors of less than 20 degrees could only be retrieved with a 3-layer realistic model. Localization and orientation errors highly depended on the dipole position in the brain.

Brain↗