Hemoglobin M Saskatoon (alpha 2 beta 2 63(E7) His----Tyr) in an Algerian family.
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Biomedical subjects
Publications and source records attributed to C Lacombe.
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Transient rheological behavior of blood which involves non newtonian viscosity, elasticity and thixotropy can be modelized with a Maxwell rheological state equation which depends on a structure parameter having dimension of a shear rate. Identification of the model parameters leads to use an exponential apparent shear rate step and to use recursive filters for taking into account the impulse response of the viscometer servo-control device. Typical results for a normal blood sample are given.
We assessed the prevalence of overt and latent primary myeloproliferative disorders in hepatic vein thrombosis. Cultures of bone marrow or peripheral blood mononuclear cells were done in 20 patients with Budd-Chiari syndrome. Erythroid colony formation in the absence of erythropoietin, which is a reliable indicator for a primary myeloproliferative disorder, was seen in 16 patients in whom Budd-Chiari syndrome was due to hepatic vein thrombosis, including 13 women aged 18 to 45 years. Among these 16 patients, the conventional criteria for the diagnosis of a primary myeloproliferative disorder were met in only 2. Primary myeloproliferative disorder, often without peripheral blood changes, is a major cause of hepatic vein thrombosis in young women.
A specific immunological assay was used to demonstrate that the erythropoietic factor produced by the recently described FMuLV-induced murine erythroleukemic cell line IW32 is an authentic erythropoietin (epo). Several independent virus-induced erythroleukemic and myeloblastic cell lines were tested for epo production. Among six erythroleukemic cell lines induced by FMuLV, another (NN10) was shown to produce epo by biological and immunological assays. Four Friend-virus-induced erythroleukemias and four FMuLV-induced myeloblastic cells were negative. The amounts of epo produced were similar in IW32 and NN10 supernatants after 48 h in culture. The in vitro bioassay gave the highest levels (up to 1000 mU/ml), the in vivo bioassay the lowest, and the radioimmunoassay gave intermediate results. NN10 and IW32 cell lines have been induced by two different FMuLV and were shown to be independent by cytogenetic studies. The molecular weights of IW32 and NN10 epo were close to the molecular weight of mouse plasma epo but elution profiles suggested that some differences might exist between these epos. Cloned IW32 and NN10 cells were shown to retain both the ability for erythroid differentiation after incubation with chemical inducers and the ability to produce epo. This demonstrates that malignant erythroid cells were the source of epo production in these cell lines.
The association of B cell chronic lymphocytic leukemia and pure red cell aplasia in a 42 year-old patient led to the discovery of a thymic enlargement. After six weeks treatment including steroids, cyclophosphamide and three courses of plasma exchange without improvement, surgical thymectomy was followed by a reticulocytosis and remission of the red cell aplasia. The tumor was a thymolipoma. Characteristics of this are thymic tumor, his connections with pure red cell aplasia and in vitro differentiation of erythroid progenitors are discussed.
A new abnormal hemoglobin was found in a diabetic Maltese woman by citrate agar electrophoresis. This variant was undetectable by isoelectric focusing. No hematological abnormalities were observed. The structural analysis included isolation of the abnormal beta chain, high pressure liquid chromatography of the corresponding tryptic peptides and then microsequencing of the abnormal T1. These procedures revealed a double abnormality: the presence of a methionyl residue extending the NH2 terminus and a histidine to proline substitution in position NA2.
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A new beta-variant has been detected and structurally defined in a French male, with a life-long history of hemolytic anemia. This variant is moderately unstable and has a low oxygen affinity. The abnormal hemoglobin was not detected by standard electrophoretic procedures. It moved slightly slower than Hb A during isoelectric focusing (IEF). Two minor fractions were also seen; the first migrated just cathodal to Hb F, as did partially oxidized Hb A or hemichrome derivatives of some unstable hemoglobins; the second in the position of free alpha-chains. The abnormal beta-chain was readily separated from both beta A- and alpha A-chains by acid-urea-Triton globin chain electrophoresis. Structural study was conducted simultaneously by fingerprinting and high-performance liquid chromatography (HPLC) of tryptic peptides. A new mutation beta 38(C4)Thr----Pro was found, which was named Hb Hazebrouck.
Plasma cholesterol and cholesterol absorption were measured in rabbits given either a standard or cholesterol-rich diet which were fed either ad libitum or reduced to 50% of the control ration. The results confirmed the aggravating effect of dietary restriction on the plasma cholesterol response to cholesterol feeding. Hypercholesterolemia was doubled when cholesterol feeding was associated with reduced dietary intake. The percentage of cholesterol absorbed increased significantly in cholesterol-fed rabbits on normal caloric ration, while dietary restriction had no effect on this parameter either with the standard or the cholesterol-rich diet. These data indicate that the mechanisms by which plasma cholesterol increases in response to cholesterol feeding involve increased cholesterol absorption. Nevertheless the aggravating effect of dietary restriction cannot be attributed to increased cholesterol absorption.
The binding of [3H]clonidine on mouse liver plasma membrane was a rapid, saturable and reversible process. It was characterized by two types of population: high affinity receptors with KD of 6.76 +/- 1.02 nM and Bmax of 106.15 +/- 24.05 fmol/mg protein, and low affinity receptors with KD of 63.66 +/- 12.85 nM and Bmax of 818.06 +/- 128.49 fmol/mg protein. Displacement of [3H]clonidine from its binding sites by various ligands indicated that alpha 1--as well as alpha 2--adrenoceptors were involved in the high affinity system. The respective participation of these two types of receptors was discussed.
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IW32 is a recently described murine erythroleukemia cell line that produces an erythropoietic factor similar to erythropoietin by in vivo and in vitro bioassays and without species specificity. Biochemical characteristics of IW32 erythropoietic factor and sheep or mouse plasma erythropoietins were compared. Murine colonies derived from erythroid colony-forming units (CFU-E) in plasma clot culture were used as the bioassay system. Both IW32 erythropoietic factor and sheep plasma erythropoietin were stable in the pH range of 3 to 10, after exposure to denaturing agents (8 mol/L urea, 4 mol/L guanidine hydrochloride, 1% sodium dodecyl sulfate), to a reducing agent (0.1 mol/L 2-mercaptoethanol) and to an oxidizing agent (5 mmol/L sodium metaperiodate). Only the combination of 0.1 mol/L 2-mercaptoethanol and 1% sodium dodecyl sulfate resulted in a significant loss of activity. IW32 erythropoietic factor and murine plasma erythropoietin were similarly precipitated by ethanol and ammonium sulfate. IW32 erythropoietic factor eluted as a single major peak after gel exclusion chromatography, with an estimated molecular weight of 45,000 daltons. Results were identical using supernatants from cultures in the presence of and absence of fetal calf serum. The supernatant of IW32 cells cultured without serum induced erythroid colonies after seven days on normal human bone marrow nonadherent mononuclear cells cultured in serum-free conditions. All these results made it very likely that IW32 cells produce an authentic erythropoietin. This cell line would be very useful for the study of murine erythropoietin.
Twenty-two cases of idiopathic chronic pure red cell aplasia (PRCA) in adults have been studied to evaluate their erythroid progenitors in vitro using the plasma clot technique. Three types of culture growth patterns were observed and classified as follows. Type I: showing a normal number of autologous CFU-E; type II: CFU-E and BFU-E were detectable but constantly decreased; type III: CFU-E and BFU-E were undetectable. The results were reproducible when patients were studied on two or more occasions. A strong correlation was found between the in vitro growth of autologous erythroid colonies and the results of immunomodulating therapy in 18 evaluable patients. A constant response to immunomodulating treatment was observed in type I patients. A constant failure of treatment was observed in type III patients, whereas results of therapy were unpredictable in type II patients. Two patients with chronic PRCA associated with thymoma and three with chronic myeloproliferative disorders were also studied. Patients with PRCA and thymoma behaved in vitro like type I patients. Patients with chronic myeloproliferative disorders exhibited very low numbers or no CFU-E or BFU-E (type II or III). These data support the hypothesis that at least two mechanisms are responsible for PRCA--one immunologically mediated and the other resulting from a stem cell defect. Moreover, they suggest that the study of erythroid progenitors in vitro might be useful in predicting the immunosuppressive therapy effect in adult chronic PRCA.
A Friend mink cell focus-inducing (Fr-MCF) virus isolated from a Friend tumor cell line was able to induce acute erythroleukemia associated with polycythemia when injected as a Friend murine ecotropic leukemia virus (F-MuLV) pseudotype into adult Swiss and ICFW mice. One virus isolate recovered from leukemic cells and designated as FV-F3 presented the following properties: (i) persistence of the same leukemogenic power when propagated in vivo and in vitro; (ii) in vivo spleen focus-forming (SFFV) capacity; (iii) presence of erythropoietin (EPO)-independent CFU-E in leukemic animals; (iv) expression of a 32 RNA specifically recognized by a SFFV probe, in FV-F3 infected cells; and (v) expression in FV-F3-infected cell of polypeptides in the range of gp52 SFFV. Peptide analysis of these products revealed close similarities with the parental MCF virus. These data suggest that a SFFV genome arose by genetic recombinational events involving MCF virus.
The influence of dietary restriction on cholesterol transport and metabolism was investigated in rabbits given standard or cholesterol-rich diets (0.2 g cholesterol/kg body weight daily) either ad libitum or with 50% caloric ration. Dietary restriction which has only a slight influence in control rabbits markedly aggravated the disturbances induced by exogenous cholesterol. With limited feeding, control rabbits presented a moderate increase in plasma cholesterol, whereas marked aggravation of hypercholesterolemia was observed in cholesterol-fed rabbits. Analysis of the lipoprotein profile showed that the excess of plasma cholesterol with the restricted cholesterol-rich diet corresponded to an increase in the concentration of very low density lipoprotein (VLDL) and low density lipoproteins (LDL) without any additional changes in the composition of these lipoproteins. No significant change appeared in the high density lipoprotein (HDL) concentration. The parameters of cholesterol metabolism were determined, from the curves of [3H]cholesterol radioactivity decrease, using a two-pool model. The increase in cholesterol turnover rate induced by the cholesterol-rich diet was accentuated by dietary restriction, whereas rabbits on standard restricted diet showed a slight decrease. The large increase in the size of both pools A and B in cholesterol-fed rabbits was even more pronounced with limited feeding. Dietary restriction induced additional accumulation of cholesterol in the aortic wall and the grade of the lesions was also aggravated.
A transplantable murine leukemia, primarily induced by a biologically cloned Friend helper virus, was shown to induce polycythemia in recipient ICFW mice. A leukemia cell line (IW.32) was established in vitro from this transplantable leukemia. Sodium butyrate and hemin induced erythroid differentiation in these leukemia cells as has already been shown with other erythroleukemia cells. The supernatant of this cell line was devoid of spleen focus-forming virus activity. However, it induced the incorporation of 59Fe in polycythemic mice and the in vitro differentiation of murine and human cfu-e into erythroid colonies. Therefore, these erythroleukemia cells produced a factor with all the biological properties of erythropoietin. The erythropoietic activity of IW.32 supernatant was higher in vitro [equivalent to 0.5-1 international unit (IU) of erythropoietin per ml] than in vivo (0.15-0.3 IU/ml). This erythropoietin-like activity was stable at 100 degrees C for 3 min, which ruled out the possibility that a virus was responsible for these effects. Preliminary studies demonstrated that the biochemical properties of the IW.32 factor are strongly similar to those of Connaught step 3 erythropoietin, thus supporting the hypothesis that the IW.32 factor is indeed an erythropoietin.
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