RESUMEN
Macronuclei isolated from Tetrahymena are contracted in form (average diameter: 10.2 micron) at a final Ca/Mg (3:2)concentration of 5 mM. Lowering the ion concentration to 1 mM induces an expansion of the average nuclear diameter to 12.2 micron. Both contracted and expanded nuclei are surrounded by a largely intact nuclear envelope as revealed by thin-sectioning electron microscopy. Nuclear swelling is accompanied by an expansion of the nuclear envelope as indicated by the decrease in the frequency of nuclear pore complexes from 52.6 to 42.1 pores/micron2 determined by freeze-etch electron microscopy. Contracted nuclear membranes reveal particle-devoid areas (average size: 0.21 micron2) on 59% of their fracture faces at the optimal growth temperature of 28 degrees C. About three-fifths of the number of these smooth areas disappear upon nuclear membrane expansion. Electron spin resonance using 5-doxylstearic acid as a spin label indicates a higher lipid fluidity in contracted than in expa,ded nuclear membranes. Moreover, a thermotropic lipid clustering occurs at approximately 17 degrees C only in expanded nuclear membranes. In contrast to the nuclear membrane-bound lipids, free lipids extracted from the nuclei rigidify with increasing Ca/Mg concentrations. Our findings are compatible with the view that the peripheral layer of the fundamental nuclear protein-framework, the so-called nuclear matrix, can modulate, inter alia, the lipid distribution and fluidity, respectively, in nuclear membranes. We suggest that a contraction of the nuclear matrix's peripheral layer induces a contraction of the nuclear membranes which, in turn, leads to an isothermic lateral lipid segregation within nuclear membranes.
Asunto(s)
Fluidez de la Membrana , Lípidos de la Membrana/fisiología , Proteínas de la Membrana/fisiología , Membrana Nuclear/fisiología , Animales , Magnesio/farmacología , Fluidez de la Membrana/efectos de los fármacos , Membrana Nuclear/efectos de los fármacos , Membrana Nuclear/ultraestructura , Tetrahymena pyriformisRESUMEN
RATIONALE AND OBJECTIVES: The effect of ioxaglate and iopamidol, two recently developed low-osmolality contrast media, on the shear elasticity of erythrocyte membranes was studied at an iodine concentration of 300 mg/mL and at a 20% volume concentration and compared with that obtained for control solutions matched in osmolality. METHODS: The authors used a micromanipulation technique, which consists of visualizing deformations of individual erythrocytes when gently aspirated into the tip of a glass micropipette by an accurately controlled pressure. An erythrocyte membrane shear elasticity modulus mu was then deduced. An increase in mu corresponded to an increase in erythrocyte membrane rigidity. RESULTS: In all cases, the erythrocytes remained discocytic. The shear elasticity modulus of the erythrocyte membrane is found to be (1) in the presence of ioxaglate (4.3 +/- 0.9 microN/m) lower (P < .001) than in the presence of hyperosmolar saline (6.2 +/- 1.3 microN/m) and lower (P < .02) than in the presence of iso-osmolar control (4.7 +/- 0.7 microN/m); (2) in the presence of iopamidol (5.4 +/- 0.7 microN/m) lower (P < .001) than in the presence of hyperosmolar sucrose (6.4 +/- 1.2 microN/m) and higher (P < .001) than in the presence of iso-osmolar control (4.7 +/- 0.7 microN/m); and (3) lower (P < .001) in the presence of ioxaglate (4.3 +/- 0.9 microN/m) than in the presence of iopamidol (5.4 +/- 0.7 microN/m). CONCLUSIONS: Under the experimental conditions used, both contrast media (ioxaglate and iopamidol) modify the erythrocyte membrane shear elasticity modulus less than do the matched hyperosmolar controls. Moreover, ioxaglate makes the erythrocyte membrane less rigid, and iopamidol makes the erythrocyte membrane more rigid than does the iso-osmolar control.
Asunto(s)
Membrana Eritrocítica/efectos de los fármacos , Yopamidol/farmacología , Ácido Yoxáglico/farmacología , Elasticidad , Deformación Eritrocítica/efectos de los fármacos , Membrana Eritrocítica/fisiología , Humanos , Concentración Osmolar , ViscosidadRESUMEN
A viscometric study of blood from insulin-treated diabetic patients is carried out. Patients are divided into three major groups--group I: without or with minimal retinopathy and recent diabetes (n = 37), group II: without or with minimal retinopathy and at least 20 years of diabetes duration (n = 35), group III: with severe retinopathy (n = 27). Each group is also subdivided according to the glycosylated hemoglobin (HbA1c) level, used to assess long-term glycemic control. Finally, the rheological parameters of six groups are compared: three of which have a HbA1c level less than 7.5% [I1 (n = 15), II1 (n = 9), III1 (n = 9)] and three others have a HbA1c level more than 7.5% [I2 (n = 22), II2 (n = 26), III2 (n = 18)]. The most important result concerns the thixotropy index xi t, which reflects the dynamic property of red blood cell (RBC) disaggregability under shear. Strong correlations between this parameter and HbA1c level are found for groups I (r = -0.53, p < 0.001) and III (r = -0.68, p < 0.001), providing evidence of a RBC disaggregability disorder for a poor glycemic control of diabetes. In contrast, such a correlation is not pointed out for the group II. As the value for xi t is not statistically different for groups II1 and II2 and is close to the normal value in both groups, the existence of a rheological protection against the retinopathy could be involved.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Viscosidad Sanguínea , Diabetes Mellitus Tipo 1/sangre , Agregación Eritrocitaria , Hemoglobina Glucada/análisis , Adulto , Retinopatía Diabética/sangre , Elasticidad , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Rheological disorders, such as blood hyperviscosity, red blood cell (RBC) hyperaggregation or hypodeformability, may be responsible for vascular complications in diabetes. As pointed out by many reports, flavonoids are known to act on RBC rheology in relation with venous thrombosis. These disorders were evaluated by viscometry in order to follow the hemorheological impact of a flavonoid (Daflon 500 mg). Hemorheological effects of Daflon 500 mg, 6 tablets per day for 28 days, were tested in 18 diabetic patients by viscometric measurements carried out on 2 ml blood sample withdrawn on EDTA. Measurements, before and after treatment, were compared with usual statistical tests. The following results were obtained: (1) a decrease of blood viscosity, more pronounced at low shear rate; (2) a better RBC disaggregability process under shear. Therefore, the main conclusion was that the hemorheological improvement, observed after Daflon 500 mg treatment, due to a decrease of RBC aggregation, can induce a decrease of blood flow resistance and a reduction in stasis and resulting ischemia.
Asunto(s)
Viscosidad Sanguínea/efectos de los fármacos , Angiopatías Diabéticas/tratamiento farmacológico , Diosmina/uso terapéutico , Agregación Eritrocitaria/efectos de los fármacos , Deformación Eritrocítica/efectos de los fármacos , Flavonoides/uso terapéutico , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
A rheological study was performed on blood samples from 10 insulin-treated diabetics with retinopathy. As part of their treatment, they all received 4 tablets of Daflon 500 mg per day for 30 days. Three complementary rheological criteria were used to characterize blood samples: 1) viscometry, using a Couette viscometer, which produces data on viscosity, shear-thinning, viscoelasticity and tixotropy of blood, 2) aggregametry, using an apparatus based on light reflectometry, using a filtrometer based on the deformations red cells undergo as they pass through narrow pores which produces information on red cell deformability. The main results were: a better red blood cell disaggregability, a decrease in red blood cell aggregation and no change in red blood cell deformability.
Asunto(s)
Retinopatía Diabética/sangre , Diosmina/farmacología , Agregación Eritrocitaria/efectos de los fármacos , Deformación Eritrocítica/efectos de los fármacos , Flavonoides/farmacología , Adulto , Anciano , Viscosidad Sanguínea/efectos de los fármacos , Fibrinógeno/metabolismo , Humanos , Persona de Mediana Edad , ReologíaRESUMEN
The Hemorheometer has been adapted to allow the recording of the flow rate during the filtration process. For newtonian fluids, the flow rate variation versus time through the pores is well approximated by Poiseuille's law. For dilute red blood cell suspensions, the same analysis can be applied by introducing the concept of "apparent filtration viscosity" which is higher than the usual viscosity measured by Couette viscometry. The apparent filtration viscosity parameter is related to the deformations undergone by red blood cells as they pass through the narrow pores. Apparent filtration viscosity can be used to obtain a precise determination of the erythrocyte deformability. Measurements performed, for a given blood sample, with pores of different diameters (5 microns, 8 microns and 12 microns) show that the error on the value of apparent filtration viscosity is less than 3%. As a result, the sensitivity of the filtration method allows to discriminate among normal blood samples. High concentrations of erythrocytes or leucocytes are found to modify the apparent filtration viscosity. These factors are apparent in the recorded filtration curves. Their effects on filtration measurements can be easily estimated.
Asunto(s)
Eritrocitos/fisiología , Modelos Biológicos , Reología , Humanos , Factores de TiempoRESUMEN
Plasmatic proteins, namely fibrinogen and globulins, play a major role in red blood cell (RBC) aggregation which is accountable for the three-dimensional structure of blood. Consequently, blood rheological properties linked to this structure must be modified when the protein plasma content changes. This paper gives results and related comments on thixotropic properties of RBC suspensions (0.45 hematocrit) in isotonic solutions containing various amount of fibrinogen to which albumin is added. Thixotropic behavior of these RBC suspensions is studied with a low inertia coaxial cylinders viscometer at a shear rate step of Y = 1 s-1. Rheograms are interpreted in term of thixotropy coefficient. The main conclusion is that albumin improves RBC disaggregability of whole blood, resulting probably from a competitive effect between fibrinogen and albumin in the RBC aggregation process.
Asunto(s)
Agregación Eritrocitaria , Fibrinógeno/fisiología , Albúmina Sérica/fisiología , Viscosidad Sanguínea , Hematócrito , Humanos , ReologíaRESUMEN
Filtration experiments on red blood cell suspensions are usually conducted in a saline buffer solution. As a result, the flow of a particle in a pore is largely dominated by viscous effects, and it is not possible to distinguish between normal and membrane altered cells. A new approach to red cell filtration is proposed here, whereby the cells are suspended in a Dextran solution that has roughly the same viscosity as the internal hemoglobin solution. It is thus aimed to detect alterations of the membrane properties. In order to prove this point, filtration measurements are conducted with a hemorheometer on dilute (hematocrit 8%) suspensions of normal and membrane hardened (diamide treatment) cells, suspended either in a 8 mPa.s Dextran solution or in a 1 mPa.s buffer solution. As expected when the cells are suspended in buffer, there is no detectable difference in the filtration index for normal and treated cells. However, when they are suspended in the 8 mPa.s solution, the filtration index is significantly larger for treated than for normal cells. This shows that filtration in a viscous liquid can be used to measure changes in cell deformability due to membrane modifications.
Asunto(s)
Deformación Eritrocítica/fisiología , Hemofiltración/métodos , Viscosidad Sanguínea/fisiología , Dextranos , Membrana Eritrocítica/fisiología , HumanosRESUMEN
Ultrasound may be used to dissolve arterial and venous thrombi. Its effects depends on the mode of ultrasonic vibration and on the length of the guide wire. The authors studied the in vitro effects of an ultrasonic angioplasty device coupled with a 130 cm long titanium flexible guide wire. The system comprises an automatic scanning function to determine the optimal frequency of resonance and works in the continuous mode of emission. Sixteen thrombi were studied of which 8 were acellular and 8 whole blood. In each group, 4 were treated in association with streptokinase and 4 by ultrasound alone. The ages of the thrombi in each subgroup were 1, 3, 7 and 15 days. All the thrombi were dissolved in 6 minutes or less (3'15" +/- 1'35") at an average optimal frequency of resonance of 19,444 Hz. Ninety six per cent of the debris had a diameter less than 10 microns. Less than 1% of the debris had a diameter larger than 100 microns. These large particles were observed in cellular thrombi and were almost completely absent in dissolved acellular thrombi. They were very fragile. The dissolution of the thrombi was not accelerated by the association of streptokinase. The ultrasonic energy did not induce D-dimer production and its action was probably due to cavitation. Ultrasonic energy could provide an alternative treatment for thrombotic vascular occlusion provided that more flexible guide wires could be designed.
Asunto(s)
Trombosis/fisiopatología , Ultrasonido , Humanos , Técnicas In Vitro , Terapia Trombolítica/métodos , Trombosis/terapia , Terapia por UltrasonidoRESUMEN
A clinical hemoviscosiSImeter is presented. It is a Couette type viscometer which is automatic and driven by a microcomputer. It was designed and realized in the laboratory. The measurement of blood viscosity requires 1 ml of anticoagulated blood adjusted at 45% of hematocrit. Experiments are performed at a constant temperature (37 degrees C) according to a well-established protocol: gentle mixing of the blood sample followed by a 3 minute rest just before each of the following measurement modes begins. Under steady state flow conditions, 12 viscosity measurements are realized in a range of shear stress rate step of 1 s-1 is applied. The elastothixotropic response of the blood sample is then obtained. Such a hemorheological test, called "hemorheogram", requires 15 minutes. Given to the clinician, the hemorheogram results include the absolute blood viscosities at the shear stress rates of 0.1 s-1, 1 s-1, 10 s-1, 70 s-1, the thixotropy index and the plasma viscosity. Finally, results to be kept in the laboratory files consist of the whole data issued from the patient hemorheogram including the relative blood viscosities (blood viscosity/plasma viscosity) at the same shear strers rate values.
Asunto(s)
Viscosidad Sanguínea , Reología/instrumentación , Humanos , Microcomputadores , Estrés MecánicoRESUMEN
Viscosity values of marrow cells from patients suffering from acute promyelocytic leukemia (acute myeloid leukemia of type 3) have been determined before and during a treatment with all-trans retinoïc acid (active metabolite of the vitamin A, inducing cell differentiation and maturation). Evaluated from aspiration tests into a glass micropipette, the marrow cell viscosity has been compared to that of normal neutrophils. Results seem to point out that the induced hyperleucocytosis is favoured by both a low standard deviation of pathological cell viscosity and a cell viscosity value close to that of normal neutrophils. The systematic study of cell viscosity could therefore help the clinicians to individualize the treatment.
Asunto(s)
Granulocitos/fisiología , Hemorreología , Leucemia Promielocítica Aguda/fisiopatología , Humanos , Leucemia Promielocítica Aguda/patología , Micromanipulación , Estereoisomerismo , Tretinoina/uso terapéuticoRESUMEN
Chemotaxis, the directional locomotion of a cell toward a source of a chemical gradient, is an important phenomenon occurring for mobilizing immune cells at sites of infection and injury. This phenomenon has been simulated in analyzing the movement in vitro of a chemoattracted cell inside a glass micropipette. A microneedle filled with fMLP, N-formyl-methionyl-leucyl-phenylalanine, at a concentration of 9.10(-7) M in 1% gelatin, is inserted in a glass micropipette containing Hanks buffer solution. After diffusion of fMLP in the glass micropipette until a constant gradient is established, the tip of the glass micropipette is moved near a polymorphonuclear neutrophil. Its spontaneous movement inside the micropipette towards the chemotactic source can be observed and quantified. Without any counter-pressure (positive pressure), the cell spontaneously advances with an average velocity of 0.14 +/- 0.04 micron/s. Corresponding to the maximal strength developed by the cell during its motion, the required strength to stop the chemotactic migration has been estimated to be 39 +/- 4 nN. Giving both qualitative and quantitative information on the dynamics of cell motility, this experiment will be helpful in the understanding of some aspects of cell motility in the tissues.
Asunto(s)
Quimiotaxis de Leucocito , Reología/instrumentación , Vidrio , Humanos , Micromanipulación , N-Formilmetionina Leucil-Fenilalanina , Valores de ReferenciaRESUMEN
Raynaud's phenomenon is mainly linked with cold provoked vasomotor perturbations, but also with rheological alterations since blood viscosity is enhanced by lowering temperature. Several methods are available for studying distal vascularization: peri-ungual capillaroscopy, digital plethysmography and laser-Doppler. Digital arteriography must be reserved to serious ischemia regarding the general anesthesia needed to avoid spasm. All these methods explore especially the vessel wall. Conservely, blood viscosity which has been developed for 25 years investigates the content of the vessel. Since 1965, numerous hemorheological studies pointed out the rheological disorders, especially those concerning plasma and blood viscosity. The most usual viscometry abnormalities revealed erythrocyte hyperaggregation, red cell hypodeformability, blood and plasmatic hyperviscosity. In a comparative study, 46 patients with Raynaud's phenomenon were studied: we performed peri-ungual capillaroscopy, plethysmography and viscosity measurements. The results demonstrated a link between capillaroscopy and thixotropy. Both investigations are never normal at the same time in connectivites and never abnormal at the same time in Raynaud's disease (primary Raynaud's phenomenon). In conclusion hemorheological studies showed nearly normal rheological parameters in Raynaud's disease, but abnormal rheological parameters in secondary Raynaud's phenomenon.
Asunto(s)
Viscosidad Sanguínea , Enfermedad de Raynaud/sangre , Adulto , Femenino , Humanos , MasculinoRESUMEN
In collaboration with the Laboratory of Physiology, the Unity of Biorheology realized hemorheological studies on healthy volunteers who performed four kinds of well-controlled exercises on a cycle ergometer. Blood was always collected just before exercise and just after exercise. Rheological measurements were performed (blood and plasma viscosities, blood thixotropy and blood viscoelasticity) as well as biological (plasma proteins) and biophysical (osmolarity, pH) measurements. In all the cases, results show the importance of a hemoconcentration and its hemorheology consequences.
Asunto(s)
Viscosidad Sanguínea/fisiología , Prueba de Esfuerzo , Hematócrito , Humanos , Postura , ReologíaRESUMEN
Thrombolytic agents and new antiplatelet drugs used in acute myocardial infarction (AMI) could change whole blood viscosity. The aim of this pilot trial is to compare blood viscosity at four shear rate levels among three groups of patients: AMI receiving thrombolysis with alteplase (n: 10), AMI eligible for primary angioplasty with abciximab (n: 10), healthy volunteers (n: 10). Viscosity measurement was obtained in 30 minutes with a Couette hemoviscosimeter. At baseline, blood viscosity level was higher in patients with acute coronary syndromes than in healthy volunteers (72 +/- 32 mPa.s versus 51 +/- 13 mPa.s, p<0.05). After thrombolysis, viscosity was higher at 90 minutes than at third day, paradoxically with fibrinogen elevation (72 +/- 32 mPa.s versus 58 +/- 27 mPa.s, p=0.01). In primary angioplasty with abciximab, viscosity decreased significantly (56 +/- 28 mPa.s versus 43 +/- 13 mPa.s, p=0.01). The effects of ionic contrast agent and abciximab are discussed. In healthy volunteers group, 100 mg aspirin once a day during 7 days reduces blood viscosity at high shear stress. The small size of the study population restricts correlation analysis with major clinical adverse events. A larger trial is necessary to evaluate the predictive value of whole blood viscosity in reocclusive and/or hemorrhagic events in those reperfusion strategies but also in case of thrombolytic agent and abciximab combination.
Asunto(s)
Viscosidad Sanguínea , Infarto del Miocardio/sangre , Adulto , Estudios de Factibilidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Infarto del Miocardio/tratamiento farmacológico , Proyectos Piloto , Terapia TrombolíticaRESUMEN
Interactions between the endothelium and erythrocytes may contribute to the vascular complications of sickle cell disease (SCD). Endothelium-derived nitric oxide (NO) plays a major role in the regulation of vasomotor tone in response to wall shear stress (WSS) variations and pharmacologic stimuli. However, little is known about endothelial NO production in patients with steady-state SCD. We investigated endothelial NO production in response to flow or vasoactive agonists in 16 homozygous patients with steady-state SCD and 15 controls. Flow-mediated dilation (FMD), arterial diameter changes in response to 100% oxygen inhalation, blood viscosity, and calculated WSS were determined in all patients and controls. At baseline, WSS was higher in SCD patients than in controls, whereas arterial diameter was similar. In patients with SCD, FMD was impaired (1.73% +/- 0.44% vs 3.97% +/- 0.24% in the controls, P <.001) and vasoconstriction in response to 100% oxygen was abolished. Using venous occlusion plethysmography, forearm blood flow (FBF) was evaluated in response to acetylcholine, nitro-monomethyl-L-arginine (L-NMMA), and sodium nitroprusside (SNP) in subgroups of 9 controls and 7 patients with SCD. Acetylcholine induced a significantly greater FBF increase in the patients (9.7 +/- 2.9 mL/min/100 mL of forearm volume vs 2.5 +/- 1.5 mL/min/100 mL in the controls, P <.001), whereas responses to L-NMMA and SNP were similar. These results suggest that endothelial dysfunction may prevent the arterial diameter of patients with SCD from adapting to chronic or acute shear stress elevations. This may contribute to the pathophysiology of vaso-occlusive crisis in patients with SCD.
Asunto(s)
Anemia de Células Falciformes/fisiopatología , Endotelio Vascular/fisiopatología , Vasodilatación/fisiología , Acetilcolina/farmacología , Adolescente , Adulto , Anemia de Células Falciformes/diagnóstico por imagen , Anemia de Células Falciformes/patología , Arteria Braquial/diagnóstico por imagen , Arteria Braquial/patología , Arteria Braquial/fisiopatología , Estudios de Casos y Controles , Endotelio Vascular/diagnóstico por imagen , Endotelio Vascular/patología , Femenino , Humanos , Masculino , Óxido Nítrico/farmacología , Óxido Nítrico/fisiología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico Sintasa/fisiología , Oxígeno/administración & dosificación , Oxígeno/farmacología , Pletismografía , Flujo Sanguíneo Regional , Estrés Mecánico , Ultrasonografía , Vasodilatación/efectos de los fármacos , omega-N-Metilarginina/farmacologíaRESUMEN
Ultrasonic energy may be used for dissolution of venous or arterial thrombi. However, its effects may depend on the mode of ultrasonic vibration and on the length of the probe. We investigated the in vitro effects of an ultrasonic angioplasty device coupled with a 130 cm-long flexible titanium probe, with an incorporated automatic optimal frequency of resonance scanning function and continuous mode of emission. Sixteen clots were treated of which eight were whole blood and eight cell-free. In each of these groups, four were treated in association with streptokinase and four by ultrasound alone. The ages of the clots in these subgroups of four were 1, 3, 7, and 15 days. All thrombi were dissolved in 6 min or less (3'15" +/- 1'35") at a mean optimal frequency of resonance of 19,444 Hz. Ninety-six percent of the debris were less than 10 mu. Fewer than 1% of the particulates were larger than 100 mu. These large particulates were observed in disrupted whole blood clots and were almost non-existent in disrupted cell-free clots. They were very fragile. Clot dissolution was not speeded by adding streptokinase to ultrasound. Ultrasound did not induce D-Dimer production, and its effect was most likely to be due to cavitation. Ultrasound energy could represent an advance for thrombotic vascular occlusion therapy, provided that more flexible probes can be devised.