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J F Stoltz

Publications and source records attributed to J F Stoltz.

At least 37 records · Page 2Linked to original sources

Influence of temperature variation from 5 degrees C to 37 degrees C on aggregation and deformability of erythrocytes.

Erythrocyte aggregation and elongation index (EI) (deformability) are measured at temperatures ranging from 5 degrees C to 37 degrees C by aggregometer MA1, and diffractometer Rheodyne SSD and the LORCA, respectively. The test samples are prepared from blood obtained from healthy subjects in test tubes containing EDTA as an anticoagulant and prior to measurement are placed in a water bath for 30 min, maintained at respective temperatures. The aggregation process is given in terms of primary aggregation time and indices of aggregation. These show significant to highly significant changes with the increase of temperature compared to that at 5 degrees C. The EI increases with the increase of shear stress but shows significant decrease with the decrease of temperature.

Erythrocyte Aggregation↗

In vitro study of intracellular IL-1beta production and beta1 integrins expression in stimulated chondrocytes--effect of rhein.

The purpose of the present study was to investigate the intracellular IL-1beta production and beta1 integrins (alpha4/beta1 and alpha5/beta1) expression on chondrocytes. Chondroytes monolayer (human chondrosarcoma cell line HEM-C55) were incubated for 12, 24 and 48 hours in the presence of Tumor Necrosis Factor-alpha (TNF-alpha, Sigma, France) or recombinant human IL-1alpha (rh-IL1alpha, Becton Dickinson, France). After direct immunolabelling, cells were either analyzed on FACScan flow cytometer (Becton Dickinson, France), or observed under an epi-fluorescence inverted microscope equipped with the CellScan EPR optical scanning acquisition system (IPLab-Scanalytics, USA). We found that the IL-1beta mean fluorescence intensity in flow cytometry and in 3D microscopy was increased in the presence of TNF-alpha or rh-IL-1alpha, and alpha4/beta1 or alpha5/beta1 expression was higher on stimulated cells than on control cells. On the other hand, we have evaluated the in vitro effects of rhein (10(-5) M, Negma, France), an active metabolite of diacerein, on the intracellular IL-1beta and beta1 integrins expressed by stimulated or no-stimulated chondrocytes. The results indicated that rhein leads to a reduction of IL-1beta synthesis whereas a weak decrease of beta1 integrins receptors expression is observed. From this study, it seems that rhein partially reduce cytokine-induced intracellular IL-1beta production, and it has a weak action on alpha4/beta1 or alpha5/beta1 receptors.

Anthraquinones↗

Sodium alginate sponges with or without sodium hyaluronate: in vitro engineering of cartilage.

Studies are underway to design biosystems containing embedded chondrocytes to fill osteochondral defects and to produce a tissue close to native cartilage. In the present report, a new alginate three-dimensional support for chondrocyte culture is described. A sodium alginate solution, with or without hyaluronic acid (HA), was freeze-dried to obtain large-porosity sponges. This formulation was compared with a hydrogel of the same composition. In the sponge formulation, macroscopic and microscopic studies demonstrated the formation of a macroporous network (average pore size, 174 microm) associated with a microporous one (average pore size, 250 nm). Histological and biochemical studies showed that, when loaded with HA, the sponge provides an adapted environment for proteoglycan and collagen synthesis by chondrocytes. Cytoskeleton organization was studied by three-dimensional fluorescence microscopy (CellScan EPR). Chondrocytes exhibit a marked spherical shape with a nonoriented and sparse actin microfilament network. Type II collagen was detected in both types of sponges (with or without HA) using immunohistochemistry. In conclusion, the sponge formulation affords new perspectives with respect to the in vitro production of "artificial" cartilage. Furthermore, the presence of hyaluronate within the alginate sponge mimics a functional environment, suitable for the production by embedded chondrocytes of an extracellular matrix.

Alginates↗

[Quantitative expression of the adhesion receptors VLA-4, VLA-5, L-selectin, MAC-1, and ICAM-1 on the surface of CD34+ cells].

The aim of this work was to quantify by flow cytometry the main adhesion receptors on CD34+ cells. These cells were isolated from bone marrow (BM) or mobilized peripheral blood (PB). The proportions of CD34+/CD49d+ and CD34+/CD49e+ are weaker on PB cells, without quantitative expression variation. This phenotypic variation may induce CD34+ cells exist from BM into circulation, promoting the mobilization. The homing to the BM implicate the CD62L receptor, which expression was found more frequently and stronger on PB cells than on BM. The CD11b, CD18 and CD54 receptors are implicated in CD34+ cells adhesion to BM micro-environment. No significant variation in CD34+/CD11b+ and CD34+/CD18+ cells frequency was noted. Moreover, CD54 receptor was more frequently expressed on PB cells. Quantitative analysis revealed that CD18 was more strongly expressed on BM than on PB cells. This quantitative variation could promote progenitor adhesion by interacting with stromal cells. Finally, quantitative expression of the main receptors on CD34+ cells provides an original option for studying CD34+ cells during the mobilization, the homing or the adhesion to BM micro-environment.

Antigens, CD↗

Effects of three Hb-based oxygen-carrying solutions on neutrophil activation in vitro: quantitative measurement of the expression of adherence receptors.

BACKGROUND: Hb-based oxygen carriers (HbOCs) are currently under investigation as RBC substitutes. These solutions have vasoactive properties and may modify the behavior of the RBC in vitro. The effects of these agents on other blood cells have not been extensively investigated. The aim of this study was to evaluate the potential effect(s) of three differently modified HbOCs on the activation of PMNs. STUDY DESIGN AND METHODS: Whole blood from humans was incubated with Hb dextran-benzene-tetracarboxylate, alphaalpha-Hb, or o-raffinose-poly-Hb for 15, 30, and 60 minutes. The expression of adherence receptors CD62L, CD18, and CD11b, which reflects the activation state of the neutrophils, was assessed in vitro by a direct immunofluorescence method using calibration beads measured by flow cytometry. RESULTS: The observed expression of each of the three adherence receptors for each solution at each time point was similar to that noted with the RPMI control. Decreases in CD62L and increases in CD18 expression were noted at 15 minutes of incubation but, when those values were compared to the values obtained with TNFalpha as the positive control of PMN activation, the profile of expression of adherence receptors obtained with HbOC solutions did not conform to the expected activation profile of PMNs. CONCLUSION: Hb solutions modified by a variety of methods, as obtained and employed in this study, did not appear to activate neutrophils in vitro.

Aspirin↗

Quantitative expression of adhesion molecules on granulocyte colony-stimulating factor-mobilized peripheral blood, bone marrow, and cord blood CD34+ cells.

The purpose of this study is to investigate the function of the main adhesion receptors on CD34(+) cells during hematopoietic stem cell transplantation. Expression was quantified by flow cytometry using calibration beads. CD34(+) cells were isolated from either bone marrow (BM), cord blood (CB), or peripheral blood (PB) from study patients and a control group after granulocyte colony-stimulating factor (G-CSF) administration. The study of the CD34(+) cell differentiation showed that CD34(+) cells are mainly CD38(+) and HLA-DR(+), whatever the type of harvest. However, quantitative analysis elicited a weaker expression of CD38 on PB and CB CD34(+) cells in comparison to BM CD34(+) cells. The proportions of CD34(+)/CD49d(+) and CD34(+)/CD49e(+) were smaller on PB cells, without quantitative expression variation. This phenotypic variation promotes CD34(+) cells to exit from BM into circulation, inducing the mobilization. The homing of the CD34(+) cells to the BM involves the CD62L receptor. The expression of this receptor was found to be more frequent and stronger on PB cells than on BM or CB cells. The CD11b, CD18, and CD54 receptors are implicated in CD34(+) cell adhesion to BM microenvironment. No significant variation in CD34(+)/CD11b(+) and CD34(+)/CD18(+) cell frequency was noted. Moreover, the CD54 receptor was more frequently expressed on CB and PB cells. Quantitative analysis revealed that CD18 was more strongly expressed on BM than on PB cells to promote progenitors adhesion by interacting with stromal cells. Finally, the quantitative expression of the main receptors on CD34(+) cells explained cellular functions during the different steps of hematopoietic stem cells transplantation.

ADP-ribosyl Cyclase↗

Investigation of 3-D mechanical properties of blood vessels using a new in vitro tests system: results on sheep common carotid arteries.

In order to investigate the three-dimensional (3-D) mechanical properties of blood vessels, a new experimental device is described allowing in vitro static and dynamic measurements on segments of arteries with high technical performances. Static tests are applied to sheep common carotid arteries. Considering a thick-walled cylindrical model of orthotropic material under large deformations, a classical 3-D approach based on strain energy density is used to calculate the resulting mechanical behavior law in radial and circumferencial directions and stresses distribution throughout the wall thickness. Results are presented with reference to unloaded and zero-stress initial state thanks to simple measurements of inner and outer circumferences. A particular ratio relating the two main stresses (circumferential and longitudinal) is calculated that put into the forth the progressive modifications in the direction of the predominant stress in the wall and the specific radial location where these changes occur. We observe that this point location is a function of the test conditions of the specimen, i.e., stretching length and level of pressure.

Animals↗

[Introduction to vascular engineering].

Vascular engineering developed through advancing knowledge of the physiological and pathological processes operating in cells and tissues subjected to environmental mechanical stress. We review briefly the more relevant cellular and general mechanism characteristic of vascular tissue examining the performances obtained with prosthetic materials (allografts, synthetic grafts) and current research and development of new vessel substitutes (biohybrids or biovessels).

Biomechanical Phenomena↗

Effects of an angelica extract on human erythrocyte aggregation, deformation and osmotic fragility.

In Chinese traditional medicine, angelica is widely used for its known clinical effects of ameliorating blood microcirculation. But the mechanism of these beneficial effects still remains unclear. In this work the rheological behaviour of human erythrocytes treated by angelica was studied in vitro. Normal RBCs incubated with an angelica extract at different concentrations (5, 10 or 20 mg/ml) for 60 min at 37 degrees C and then their aggregation, deformation and osmotic fragility were measured with different recently developed optical techniques, namely Erythroaggregometer (Regulest, Florange, France), LORCA (Mechatronics, Amsterdam) and Fragilimeter (Regulest, Florange, France). Experimental results show that angelica (20 mg/ml) significantly decreased normal RBCs' aggregation speed (p<0.01) and could inhibit the hyperaggregability caused by dextran 500. However, the strength of normal RBCs aggregates were not influenced by angelica. When a calcium ionophore A23187 (1.9 microM) was used to harden cell membrane, angelica (20 mg/ml) could significantly (p<0.01) protect erythrocytes against the loss of their deformability even it had no effects on normal RBCs deformation. Finally angelica (5 and 10 mg/ml) decreased significantly (p<0.01) normal RBCs osmotic fragility. In conclusion angelica plays a rheologically active role on human erythrocytes, and this study suggests a possible mechanism for angelica's positive effects against certain cardiovascular diseases.

Asteraceae↗

Characterization of heart atrial fibrillation: proposal for a deterministic model.

Atrial Fibrillation (AFib) has been shown (e.g. Framingham Study) to increase stroke risk and to induce low cardiac output in patients. It is generally accepted that the mechanism of Atrial Fibrillation is reentry. In order to ease the study of AFib's, we propose a deterministic model that could be used in the characterization cardiac signals. The model is based on a linear combination of several "Atrial Flutter"-like signals. We therefore obtained AFib-like aspects. To demonstrate its usefulness we performed a power spectrum transform (i.e. scalar Fast Fourier Transform) of this model. As a result we found that this transform could not be used as is, could lead to errors in the interpretation of results. We expect that this model will help us to evaluate the relevance of mathematical transforms. This may lead to new findings in the classification of AFib, as well as to improvements in patients treatment, follow-up and survival.

Atrial Fibrillation↗

Evaluation of the energy of red blood cell agglutination by monoclonal antibodies.

The knowledge of the energy involved in cell-cell interactions has significant implications in biological and medical sciences. Red blood cells (RBC) become mutually adhered when specific (agglutinins) or nonspecific macromolecules bind adjacent cells in an irreversible or reversible form. Flow chamber technique with digital image processing was successfully applied to determine the partial separation, by shear stress, of two RBC agglutinated face to face (doublet) by specific monoclonal antibodies (MAb) and also to evaluate the shear stress required to attain this separation. The work done in separating adhered cells is then easily calculated and serves to evaluate the antibody affinity. In this work, this technique was applied to evaluate the affinity of a monoclonal antibody specific to the blood group A antigen. The specific disaggregation energy gamma (i.e., the work done by the shear stress) required to disrupt a unit of adhered membrane areas between agglutinated cells was calculated. On play back of the image analysis, measurements were taken to determine the force applied to the doublet and the relative separation between both RBC. Values of gamma(d) (from 2.70 to 4.61 x 10(-9) N/cm) were found to be proportional to the density (D = 27 to 55 molecules/microm(2)) of MAb molecules bound on the RBC membrane.

ABO Blood-Group System↗

In vivo effects of Hb solutions on blood viscosity and rheologic behavior of RBCs: comparison with clinically used volume expanders.

BACKGROUND: Hb-based oxygen carriers (HbOCs) have vasoactive effects that are still poorly understood. Factors known to have vasoactive effects, such as plasma, whole-blood viscosity, and the rheologic behavior of RBCs, are modulated by HbOCs in vitro, but few in vivo studies have been performed. STUDY DESIGN AND METHODS: Rabbits were phlebotomized (30%) and resuscitated with unmodified stroma-free Hb (SFHb), dextran-tetracarboxylate-Hb (Dex-BTC-Hb), O-raffinose-polymerized Hb (OrpHb), HSA, or hydroxyethyl starch 200 (HES). Plasma viscosity was assessed with a capillary viscometer and whole-blood viscosity with a rotational viscosimeter. RBC aggregation kinetics were determined by analysis of back-scattered light in a rotating device. RESULTS: As compared to that in the control RBC suspension, resuscitation with SFHb, OrpHb, or HSA decreased plasma and whole-blood viscosity as well as RBC aggregation; resuscitation with Dex-BTC-Hb increased whole-blood viscosity at low shear rates as well as RBC aggregation, whereas that with HES decreased whole-blood viscosity but increased RBC aggregation. CONCLUSION: HbOCs have different rheologic effects in vitro and in vivo. There are marked differences among the Hb solutions in their in vivo effects on viscosity and RBC rheologic behavior (especially at low shear rates encountered in the venous circulation and the microcirculation), which may be related to the chemical modifications applied to hemoprotein. These results could contribute to an understanding of the vasoactive effects of HbOCs.

Adult↗

A theoretical approach of the measurement of osmotic fragility of erythrocytes by optical transmission.

The osmotic fragility of the erythrocyte membrane to hypotonic solutions is investigated theoretically. The fragility curves exhibit a strong transmittance rise. This variation is assumed to result from changes in the scattering properties of erythrocytes under dialysis resulting from swelling and hemolysis. The refractive indices of erythrocytes are obtained through the Lorentz-Lorenz relation based on hemoglobin and water contents. The scattering cross sections (needed to calculate the collimated transmittance) and the forward scattered intensity (needed to calculate the incoherent transmittance) are expressed according to the simple algebraic relations of the anomalous diffraction approximation. It is shown that swelling (or shrinking) has no influence on the collimated transmittance. Hemolysis alone causes the abrupt sigmoidal increase of the collimated transmittance with time. The possible transmittance increase (decrease) observed during swelling (shrinking) is due to incoherent transmittance and depends on the detecting solid angle value of the experimental setup.

Hemolysis↗

[Leukocyte adhesion on a fibrinogen-coated surface under static conditions: experimentation and creation of a model].

The adhesion of polymorphonuclear leukocytes (PMNs) on the vascular endothelium is a complex process that occurs during different biological and pathological events and involves numerous molecules. The adhesion cascade is induced after PMN stimulation by various molecular or cellular signals. Fibrinogen is one of the substrates for CD11b/CD18 B2-integrins expressed at the PMN surface; fibrinogen-neutrophil binding is induced by inflammatory reactions. In order to understand this process, we have carried out studies on the basis of preliminary experiments on red blood cells and synthetic particles. The modelization of quiescent PMNs adhesion on a fibrinogen substrate was investigated with a sedimentation cell chamber. Two different physiological conditions were tested: the activated state of PMN by a synthetic pro-inflammatory activator (FMLP). The activated state of PMNs was both quantified by flow cytometry and controlled by fluorescence microscopy. The results suggest that quiescent neutrophils deposit in accordance with the ballistic deposition model. This random adsorption model differs from random sequential adsorption (RSA) in that the cells arriving at the surface are able to roll along cells previously adsorbed introducing the notion of gravitational attraction of cells. The preliminary results obtained with stimulated PMN do not allow to choose between one of this two deposition models. Nevertheless, the qualitative and quantitative effects of FMLP on neutrophils were demonstrated by modifications of adhesion molecules expression.

Antigens, CD↗

The role of 3D-microscopy in the study of chondrocyte-matrix interaction (alginate bead or sponge, rat femoral head cap, human osteoarthritic cartilage) and pharmacological application.

The potentialities of a new non-invasive optical scanning microscopy technique were evaluated through 3D analysis of chondrocyte-matrix interactions. Five different 2D or 3D culture systems were used: (1) MonoLayer (ML) of human chondrosarcoma cell line; (2) rat or human chondrocytes encapsulated in Alginate Bead (AB); (3) human chondrocytes encapsulated in Alginate Sponge (AS); (4) Rat Femoral Head Cap (RFHC); (5) slices of knee human Osteoarthritic Cartilage (HOAC). Chondrocytes ML, AB, RFHC were incubated for 24 h in vitro in the presence of recombinant human interleukin1-beta (rhIL1-beta) and the effects on cytoskeleton organisation (F-actin filament), Focal Adhesion Kinase (FAK) expression (tyrosine kinase), collagenase B expression (metalloprotease) were studied. Furthermore, the production of intracellular IL1-beta by LPS- or rhIL1-beta-stimulated chondrocytes was shown to be partly suppressed by rhein (active metabolite of diacerhein) in all culture systems. This high resolution light microscopy gave complementary information that could be important for a better understanding of the interaction of chondrocytes with the extracellular matrix in a variety of culture devices.

Actins↗