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M Laurent

Publications and source records attributed to M Laurent.

At least 235 records · Page 13Linked to original sources

Modulation of phosphofructokinase behavior by chemical modifications during the immobilization process.

Phosphofructokinase was immobilized within a protein membrane or on soluble protein polymers using glutaraldehyde as cross-linking reagent. The native enzyme was also modified chemically, using the cross-linking reagent alone. A comparative kinetic investigation of these preparations was carried out. The catalytic activity of the chemically modified enzyme and its affinity towards fructose 6-phosphate decreased significantly; the modified enzyme lost its cooperative properties and the allosteric regulation by AMP was affected. When the chemical treatment was performed in the presence of effectors (AMP or ATP) the allosteric transition induced by AMP was restored, suggesting that the cross-linking reagent modified the AMP regulatory sites, albeit no higher-substrate-affinity enzyme conformation was frozen. Molecular data showed that glutaraldehyde produced intramolecular then intermolecular bonds as its concentration increased. When the enzyme was immobilized into protein membranes or on soluble polymers, the enzyme behavior was quite similar: decrease of affinity towards fructose 6-phosphate but no changes in cooperative properties and modifications of allosteric transition induced by AMP. When AMP was present during the immobilisation process, the enzyme immobilized in this way was no longer sensitive to effectors, either AMP or ATP. It showed Michaelian behavior and higher substrate affinity quite similar to that of the native enzyme. The data suggested that a higher-substrate-affinity enzymatic form was most probably stabilized by immobilization.

Adenosine Monophosphate↗

Sulfhydryl groups of yeast phosphofructokinase-specific localization on beta subunits of fructose 6-phosphate binding sites as demonstrated by a differential chemical labeling study.

Yeast phosphofructokinase contains 83 +/- 2 cysteinyl residues/enzyme oligomer. On the basis of their reactivity toward 5,5-dithiobis(2-nitrobenzoic acid), the accessible cysteinyl residues of the native enzyme may be classified into three groups. For titrations performed with N-ethylmaleimide, subdivisional classes of reactivity are evidenced. In each case, the 6 to 8 most reactive cysteines are not protected by fructose 6-phosphate from chemical labeling and do not seem involved in subsequent enzyme inactivation. Differential labeling studies as well as direct protection experiments in the presence of fructose 6-phosphate, indicate that 12 -SH groups/enzyme oligomer (i.e. three -SH groups per binding site) are protected by the allosteric substrate from the chemical modification. Specific labeling by the differential method of the cysteinyl residues protected by fructose 6-phosphate and further separation of the two types of subunits constituting yeast phosphofructokinase, show that the substrate binding sites are localized exclusively on subunits of beta type. Thus, alpha subunits are not implicated directly in the catalytic mechanism of yeast phosphofructokinase reaction.

Binding Sites↗

A spin label study of the erythrocyte membranes in Duchenne muscular dystrophy.

Red blood cells and freshly prepared erythrocyte membranes of 15 patients with Duchenne muscular dystrophy (DMD) as well as age-matched controls were studied by the spin label method. No significant modifications appeared for spin-labelled proteins of ghost membranes. With the two fatty acid spin labels, 5-nitroxide stearate and 16-nitroxide stearate, we have confirmed previous results of Sato et al. concerning the thermal behaviour of the erythrocyte membranes, i.e. no change near the polar part probed by 5-nitroxide stearate and a linearization of the fluidity versus temperature variation around 12 degrees C, as explored by 16-nitroxide stearate. Furthermore we studied in the whole erythrocyte the amplitude of the 5-nitroxide stearate electron spin resonance signal as a function of the microwave power. This saturation effect was observed in 12 out of 15 controls and only in 1 out of 13 DMD cases studied. In erythrocyte membranes labelled with 16-nitroxide stearate the penetration of the label inside membranes was statistically different between DMD and controls. These new findings furnish further arguments in favour of a structural alteration of the phospholipid organization of erythrocyte membranes in DMD. Associated together, these different sets of tests obtained by spin labelling permit good statistical discrimination between DMD and normal subjects.

Adolescent↗

Normal and leukaemic human haemopoietic cells in diffusion chamber. A morphological and functional CFU-C study.

Leukaemic human bone marrow and peripheral blood cells were cultured for 25 d in diffusion chambers implanted into cyclophosphamide treated mice. Normal bone marrow cells were cultured simultaneously. These cells were studied both morphologically and functionally (CFU-C). The leukaemic cells behaved heterogeneously, 2 groups being distinguishable in accordance with their initial in vitro growth pattern (1: no growth or microcluster growth. 2: macrocluster growth). Group I showed progressive cellular death with a diminution of granulocytic progenitors and the appearance of a predominantly macrophagic population. This behaviour resembled that of the control group. The initial microcluster growth pattern remained identical throughout the entire culture period. Group 2, after considerable cellular death up to d 5, showed an explosive proliferation of the granulocytic progenitors and incomplete differentiation (up to myelocyte). The initial macrocluster growth pattern remained identical.

Adolescent↗

[Improvement and normalisation of the QRS complex by stimulation of the bundle of His in complete left branch block].

The asynchronism of ventricular activation resulting from a major degree of left bundle branch block (QRS greater than or equal to 0.12 s) may be suppressed by stimulation of the distal portion of the His Bundle, whether the block be intermittent (3 cases) or permanent (17 cases). The selective stimulation of the His Bundle normalises ventricular depolarisation whilst non-selective stimulation narrows the QRS complex by the fusion of the activation wave fronts from the His Bundle and the interventricular septum. The reestablishment of synchronous ventricular conduction by His Bundle stimulation is generally interpreted as a sign of longitudinal dissociation in the proximal portion of the His Bundle. This results does not, however, exclude the possibility of a very localised lesion at the origin of the left bundle, responsible for a conduction delay, and suppressed by stimulation carried out close to the zone of block (summation effect, electrotonic influence).

Adult↗

[Surgery for carcinoma of the cervix: frozen section examination of the external iliac nodes (author's transl)].

Since 1970, the authros have carried out colpohysterectomy with external iliac node dissection for carcinoma of the cervix at stages 1 and 2. The nodes were subjected to frozen section histological examination. In the presence of signs of invasion, primary iliac block dissection is carried out. Two hundrend and seventy cases have been treated in this way, with only 7 cases of error in the frozen section diagnosis (2,5%). There was 1 false positive, 4 false negatives and 2 doubtful cases with a suspicion of invasion. It is therefore suggested that frozen section histological examination of the external iliac nodes is possible and reliable.

Adult↗

Cellular histamine release, specific and total serum IgE levels in hay fever patients and controls.

The amount of histamine released from blood leucocytes by allergen, the amount of allergen required to release 50% histamine and the total IgE and IgE specific for Dactylis glomerata are compared in nine hay fever patients and five control individuals. The variation with time of total IgE, of specific IgEe and of the percentage of histamine released at a fixed allergen concentration, corresponding to the cellular sensitivity, is shown for one control individual with positive in vitro and skin tests, but without clinical symptoms. For four patients with a low total IgE level the evolution of these parameters during a 1 year treatment period is presented.

Adult↗