ARCHITECTURE OF ERYTHROCYTE SKELETON--BILAYER INTERFACE
(2002)
Abstract :
DESCRIPTION: (Adapted from Applicant's Abstract) Dr. Shohet hypothesizes that the distance between the spectrin-based membrane skeleton and the bilayer increases in certain types of hereditary spherocytosis due to defects or deficiencies in membrane proteins. This physical uncoupling is thought to increase the motion of spectrin, weaken distance-dependent interactions between the skeleton and the bilayer, and destabilize membrane lipids. To test this hypothesis, Dr. Shohet will pursue the following specific aims: 1.Measure the distances from the two major anchorage sites of the membrane skeleton to the lipid bilayer in intact red cell membranes; i.e., the distance from the spectrin:ankyrin binding region to the bilayer and the distance from the spectrin: band 4.1 binding region to the bilayer. These measurements will be determined by fluorescence energy transfer (FET) and single-photon radioluminescence. Novel methodologic approaches include: alkylation of thiophosphorylated phosphorylation sites to label ankyrin and band 3; a fluoresceinated recombinant 52-amino acid peptide of band 4.1 containing the spectrin binding domain to label spectrin at its docking site on the junctional complex; and a non-fluorescent "dark" acceptor to increase FET distance range. 2. Measure the distance from the more mobile spectrin span region of the membrane skeleton to the lipid bilayer and define the dynamic behavior of this region. First, the average distance from spectrin to the bilayer will be measured utilizing uniformly fluorescently labeled spectrin and the novel technique of total internal reflectance (TIR) developed by the UCSF group to detect fluorophores at distances 100-2000 beneath the membrane surface. Second, the distance from the beta-chain carboxy terminus in the middle of the spectrin oligomer to the bilayer will be measured using a fusion of B-spectrin with the green fluorescent protein from jellyfish. A similar distance estimate will be generated using a fluorescent Fab against the 10th repeat of the a-spectrin subunit. Third, the extent of motion of the central region of the spectrin tetramer will be measured using fluorescence recovery after photo-bleaching combined with total internal reflectance. 3. Test the hypothesis that bilayer coupling is defective in certain spherocytic hemolytic anemias by measuring bilayer:skeletal distances and spectrin motion in pathologic cells. The above distance and dynamics measurements will be made on spherocytic erythrocytes with mutations or deficiencies in ankyrin or band 4. Similar measurements will be conducted on Southeast Asian Ovalocytes which are more rigid than normal cells and which may exhibit increased cytoskeletal-bilayer coupling with reduced spectrin motion. In aggregate, the role of bilayer-skeletal interactions in red cell membrane structure and stability will be more clearly defined.
| Project Number : | 5R01DK016095-25 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | erythrocyte, green fluorescent protein, lipid bilayer membrane, membrane protein, membrane structure, spectrin alkylation, ankyrin, band 3 protein, binding protein, chemical stability, hereditary elliptocytosis, hereditary spherocytosis, intermolecular interaction, membrane transport protein, phosphorylation, protein structure /function Cnidaria, human tissue, luminescence, tissue /cell culture |
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ARCHITECTURE OF ERYTHROCYTE SKELETON--BILAYER INTERFACE
(2001)
| Project Number : | 2R01DK016095-21A2 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON--BILAYER INTERFACE
(2001)
Abstract :
DESCRIPTION: (Adapted from Applicant's Abstract) Dr. Shohet hypothesizes that the distance between the spectrin-based membrane skeleton and the bilayer increases in certain types of hereditary spherocytosis due to defects or deficiencies in membrane proteins. This physical uncoupling is thought to increase the motion of spectrin, weaken distance-dependent interactions between the skeleton and the bilayer, and destabilize membrane lipids. To test this hypothesis, Dr. Shohet will pursue the following specific aims: 1.Measure the distances from the two major anchorage sites of the membrane skeleton to the lipid bilayer in intact red cell membranes; i.e., the distance from the spectrin:ankyrin binding region to the bilayer and the distance from the spectrin: band 4.1 binding region to the bilayer. These measurements will be determined by fluorescence energy transfer (FET) and single-photon radioluminescence. Novel methodologic approaches include: alkylation of thiophosphorylated phosphorylation sites to label ankyrin and band 3; a fluoresceinated recombinant 52-amino acid peptide of band 4.1 containing the spectrin binding domain to label spectrin at its docking site on the junctional complex; and a non-fluorescent "dark" acceptor to increase FET distance range. 2. Measure the distance from the more mobile spectrin span region of the membrane skeleton to the lipid bilayer and define the dynamic behavior of this region. First, the average distance from spectrin to the bilayer will be measured utilizing uniformly fluorescently labeled spectrin and the novel technique of total internal reflectance (TIR) developed by the UCSF group to detect fluorophores at distances 100-2000 beneath the membrane surface. Second, the distance from the beta-chain carboxy terminus in the middle of the spectrin oligomer to the bilayer will be measured using a fusion of B-spectrin with the green fluorescent protein from jellyfish. A similar distance estimate will be generated using a fluorescent Fab against the 10th repeat of the a-spectrin subunit. Third, the extent of motion of the central region of the spectrin tetramer will be measured using fluorescence recovery after photo-bleaching combined with total internal reflectance. 3. Test the hypothesis that bilayer coupling is defective in certain spherocytic hemolytic anemias by measuring bilayer:skeletal distances and spectrin motion in pathologic cells. The above distance and dynamics measurements will be made on spherocytic erythrocytes with mutations or deficiencies in ankyrin or band 4. Similar measurements will be conducted on Southeast Asian Ovalocytes which are more rigid than normal cells and which may exhibit increased cytoskeletal-bilayer coupling with reduced spectrin motion. In aggregate, the role of bilayer-skeletal interactions in red cell membrane structure and stability will be more clearly defined.
| Project Number : | 5R01DK016095-22 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | erythrocyte, green fluorescent protein, lipid bilayer membrane, membrane protein, membrane structure, spectrin alkylation, ankyrin, band 3 protein, binding protein, chemical stability, hereditary elliptocytosis, hereditary spherocytosis, intermolecular interaction, membrane transport protein, phosphorylation, protein structure /function Cnidaria, human tissue, luminescence, tissue /cell culture |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON--BILAYER INTERFACE
(2001)
Abstract :
DESCRIPTION: (Adapted from Applicant's Abstract) Dr. Shohet hypothesizes that the distance between the spectrin-based membrane skeleton and the bilayer increases in certain types of hereditary spherocytosis due to defects or deficiencies in membrane proteins. This physical uncoupling is thought to increase the motion of spectrin, weaken distance-dependent interactions between the skeleton and the bilayer, and destabilize membrane lipids. To test this hypothesis, Dr. Shohet will pursue the following specific aims: 1.Measure the distances from the two major anchorage sites of the membrane skeleton to the lipid bilayer in intact red cell membranes; i.e., the distance from the spectrin:ankyrin binding region to the bilayer and the distance from the spectrin: band 4.1 binding region to the bilayer. These measurements will be determined by fluorescence energy transfer (FET) and single-photon radioluminescence. Novel methodologic approaches include: alkylation of thiophosphorylated phosphorylation sites to label ankyrin and band 3; a fluoresceinated recombinant 52-amino acid peptide of band 4.1 containing the spectrin binding domain to label spectrin at its docking site on the junctional complex; and a non-fluorescent "dark" acceptor to increase FET distance range. 2. Measure the distance from the more mobile spectrin span region of the membrane skeleton to the lipid bilayer and define the dynamic behavior of this region. First, the average distance from spectrin to the bilayer will be measured utilizing uniformly fluorescently labeled spectrin and the novel technique of total internal reflectance (TIR) developed by the UCSF group to detect fluorophores at distances 100-2000 beneath the membrane surface. Second, the distance from the beta-chain carboxy terminus in the middle of the spectrin oligomer to the bilayer will be measured using a fusion of B-spectrin with the green fluorescent protein from jellyfish. A similar distance estimate will be generated using a fluorescent Fab against the 10th repeat of the a-spectrin subunit. Third, the extent of motion of the central region of the spectrin tetramer will be measured using fluorescence recovery after photo-bleaching combined with total internal reflectance. 3. Test the hypothesis that bilayer coupling is defective in certain spherocytic hemolytic anemias by measuring bilayer:skeletal distances and spectrin motion in pathologic cells. The above distance and dynamics measurements will be made on spherocytic erythrocytes with mutations or deficiencies in ankyrin or band 4. Similar measurements will be conducted on Southeast Asian Ovalocytes which are more rigid than normal cells and which may exhibit increased cytoskeletal-bilayer coupling with reduced spectrin motion. In aggregate, the role of bilayer-skeletal interactions in red cell membrane structure and stability will be more clearly defined.
| Project Number : | 5R01DK016095-23 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | erythrocyte, green fluorescent protein, lipid bilayer membrane, membrane protein, membrane structure, spectrin alkylation, ankyrin, band 3 protein, binding protein, chemical stability, hereditary elliptocytosis, hereditary spherocytosis, intermolecular interaction, membrane transport protein, phosphorylation, protein structure /function Cnidaria, human tissue, luminescence, tissue /cell culture |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON--BILAYER INTERFACE
(2001)
Abstract :
DESCRIPTION: (Adapted from Applicant's Abstract) Dr. Shohet hypothesizes that the distance between the spectrin-based membrane skeleton and the bilayer increases in certain types of hereditary spherocytosis due to defects or deficiencies in membrane proteins. This physical uncoupling is thought to increase the motion of spectrin, weaken distance-dependent interactions between the skeleton and the bilayer, and destabilize membrane lipids. To test this hypothesis, Dr. Shohet will pursue the following specific aims: 1.Measure the distances from the two major anchorage sites of the membrane skeleton to the lipid bilayer in intact red cell membranes; i.e., the distance from the spectrin:ankyrin binding region to the bilayer and the distance from the spectrin: band 4.1 binding region to the bilayer. These measurements will be determined by fluorescence energy transfer (FET) and single-photon radioluminescence. Novel methodologic approaches include: alkylation of thiophosphorylated phosphorylation sites to label ankyrin and band 3; a fluoresceinated recombinant 52-amino acid peptide of band 4.1 containing the spectrin binding domain to label spectrin at its docking site on the junctional complex; and a non-fluorescent "dark" acceptor to increase FET distance range. 2. Measure the distance from the more mobile spectrin span region of the membrane skeleton to the lipid bilayer and define the dynamic behavior of this region. First, the average distance from spectrin to the bilayer will be measured utilizing uniformly fluorescently labeled spectrin and the novel technique of total internal reflectance (TIR) developed by the UCSF group to detect fluorophores at distances 100-2000 beneath the membrane surface. Second, the distance from the beta-chain carboxy terminus in the middle of the spectrin oligomer to the bilayer will be measured using a fusion of B-spectrin with the green fluorescent protein from jellyfish. A similar distance estimate will be generated using a fluorescent Fab against the 10th repeat of the a-spectrin subunit. Third, the extent of motion of the central region of the spectrin tetramer will be measured using fluorescence recovery after photo-bleaching combined with total internal reflectance. 3. Test the hypothesis that bilayer coupling is defective in certain spherocytic hemolytic anemias by measuring bilayer:skeletal distances and spectrin motion in pathologic cells. The above distance and dynamics measurements will be made on spherocytic erythrocytes with mutations or deficiencies in ankyrin or band 4. Similar measurements will be conducted on Southeast Asian Ovalocytes which are more rigid than normal cells and which may exhibit increased cytoskeletal-bilayer coupling with reduced spectrin motion. In aggregate, the role of bilayer-skeletal interactions in red cell membrane structure and stability will be more clearly defined.
| Project Number : | 5R01DK016095-24 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | erythrocyte, green fluorescent protein, lipid bilayer membrane, membrane protein, membrane structure, spectrin alkylation, ankyrin, band 3 protein, binding protein, chemical stability, hereditary elliptocytosis, hereditary spherocytosis, intermolecular interaction, membrane transport protein, phosphorylation, protein structure /function Cnidaria, human tissue, luminescence, tissue /cell culture |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON: BILAYER INTERFACE
(1995)
Abstract :
Evidence from pathologic red blood cells suggests that the coupling of skeletal proteins with the overlaying membrane is essential for normal cell integrity and dynamic behavior. Although extensive biochemical and electron microscopic studies have provided an initial concept of the skeletal structure, the geometry of the skeleton's relationship to the overlying bilayer is not well under-stood. Our goal is to use biophysical approaches to obtain a detailed perspective of the skeleton:bilayer interface in the intact cell that can eventually be studied under dynamic conditions. Such studies should provide insight into the instability of the membrane in certain hemolytic disorders. To define the molecular geometry of this interface, we plan to measure various skeleton:bilayer distances using fluorescence energy transfer and condensed phase radioluminescence techniques. For this purpose, the following steps will be carried out: (1)Labelling and incorporation of membrane elements: Site-directed fluorescent labelling of spectrin will be achieved using a novel cleavable, fluorescent, photoaffinity crosslinking reagent. In situ specific labelling of actin, band 3, and 4.1 will also be conducted. Fluorescent and tritiated lipophilic compounds will be obtained commercially. The labelled protein and lipid species will then be incorporated in to inside- out vesicles and ghosts. Proper biochemical incorporation of proteins will be assessed by:determination of their binding affinity and reversibility of binding to the membrane, competitive displacement with antibodies or appropriate functional protein fragments, and electron microscopic localization with anti-fluorescein immuno-gold conjugates. Proper functional incorporation of proteins will be assessed by monitoring membrane deformability and fragility by ektacytometry. (2)Measurement of skeleton:bilayer distance in normal cells: Short distances (within 20-80 A) between the fluorescently-labelled skeletal proteins and the plane of the inner leaflet membrane lipids will be measured by fluorescence energy transfer. Longer distances (between 80 to 500 A) will be measured by condensed phase radioluminescence using fluorescently-labelled skeletal proteins and tritiated membrane lipids. (3)Assessment of the effects of physiologic perturbations on the skeleton:bilayer distances: The changing interactions between the skeleton and the overlying bilayer will be examined during various membrane stresses (e.g., shear stress, hypotonic expansion, echinocytogenesis of different types, and mild oxidation with Heinz bodily induction). The localized topography of skeleton:bilayer distances (e.g., in the spikes of echinocytes or adjacent to Heinz bodies) will be measured in individual cells using a laser scanning confocal fluorescence dual channel microscope coupled to an image processing system.
| Project Number : | 2R01DK016095-16A2 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, CELL COMPONENTS, CELL MEMBRANE, CELL COMPONENTS, CYTOSKELETON, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, MEMBRANE (BIOLOGICAL) STRUCTURE BIOLOGICAL TRANSPORT, MEMBRANE MODELS, CELL COMPONENTS, VACUOLES-VESICLES, MEMBRANE SURFACE ACTIVITY, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE, PROTEINS, CONTRACTILE AND CYTOSKELETAL PROTEINS, ACTIN, PROTEINS, CONTRACTILE AND CYTOSKELETAL PROTEINS, ANKYRIN, PROTEINS, CONTRACTILE AND CYTOSKELETAL PROTEINS, SPECTRIN, PROTEINS, GLYCOPROTEINS, GLYCOPHORINS, PROTEINS, MEMBRANE PROTEINS, PROTEINS, MEMBRANE PROTEINS, BAND-3 PROTEINS ANIMALS, INVERTEBRATES, ECHINODERMS, IMMUNOLOGY, ANTIBODIES, OPTICS, LIGHT EMISSION, FLUORESCENCE, OPTICS, MICROSCOPY, IMMUNE ELECTRON, RADIOFLUORESCENT PROBES |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON: BILAYER INTERFACE
(1995)
Abstract :
Evidence from pathologic red blood cells suggests that the coupling of skeletal proteins with the overlaying membrane is essential for normal cell integrity and dynamic behavior. Although extensive biochemical and electron microscopic studies have provided an initial concept of the skeletal structure, the geometry of the skeleton's relationship to the overlying bilayer is not well under-stood. Our goal is to use biophysical approaches to obtain a detailed perspective of the skeleton:bilayer interface in the intact cell that can eventually be studied under dynamic conditions. Such studies should provide insight into the instability of the membrane in certain hemolytic disorders. To define the molecular geometry of this interface, we plan to measure various skeleton:bilayer distances using fluorescence energy transfer and condensed phase radioluminescence techniques. For this purpose, the following steps will be carried out: (1)Labelling and incorporation of membrane elements: Site-directed fluorescent labelling of spectrin will be achieved using a novel cleavable, fluorescent, photoaffinity crosslinking reagent. In situ specific labelling of actin, band 3, and 4.1 will also be conducted. Fluorescent and tritiated lipophilic compounds will be obtained commercially. The labelled protein and lipid species will then be incorporated in to inside- out vesicles and ghosts. Proper biochemical incorporation of proteins will be assessed by:determination of their binding affinity and reversibility of binding to the membrane, competitive displacement with antibodies or appropriate functional protein fragments, and electron microscopic localization with anti-fluorescein immuno-gold conjugates. Proper functional incorporation of proteins will be assessed by monitoring membrane deformability and fragility by ektacytometry. (2)Measurement of skeleton:bilayer distance in normal cells: Short distances (within 20-80 A) between the fluorescently-labelled skeletal proteins and the plane of the inner leaflet membrane lipids will be measured by fluorescence energy transfer. Longer distances (between 80 to 500 A) will be measured by condensed phase radioluminescence using fluorescently-labelled skeletal proteins and tritiated membrane lipids. (3)Assessment of the effects of physiologic perturbations on the skeleton:bilayer distances: The changing interactions between the skeleton and the overlying bilayer will be examined during various membrane stresses (e.g., shear stress, hypotonic expansion, echinocytogenesis of different types, and mild oxidation with Heinz bodily induction). The localized topography of skeleton:bilayer distances (e.g., in the spikes of echinocytes or adjacent to Heinz bodies) will be measured in individual cells using a laser scanning confocal fluorescence dual channel microscope coupled to an image processing system.
| Project Number : | 5R01DK016095-19 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | cell membrane, cytoskeleton, erythrocyte, membrane lipid, membrane structure actin, alternatives to animals in research, ankyrin, band 3 protein, erythrocyte membrane, glycophorin, membrane activity, membrane model, membrane protein, spectrin, vesicle /vacuole Echinodermata, antibody, confocal scanning microscopy, fluorescein, fluorescence, immunoelectron microscopy, radiofluorescent probe |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON--BILAYER INTERFACE
(1995)
Abstract :
Evidence from pathologic red blood cells suggests that the coupling of skeletal proteins with the overlaying membrane is essential for normal cell integrity and dynamic behavior. Although extensive biochemical and electron microscopic studies have provided an initial concept of the skeletal structure, the geometry of the skeleton's relationship to the overlying bilayer is not well under-stood. Our goal is to use biophysical approaches to obtain a detailed perspective of the skeleton:bilayer interface in the intact cell that can eventually be studied under dynamic conditions. Such studies should provide insight into the instability of the membrane in certain hemolytic disorders. To define the molecular geometry of this interface, we plan to measure various skeleton:bilayer distances using fluorescence energy transfer and condensed phase radioluminescence techniques. For this purpose, the following steps will be carried out: (1)Labelling and incorporation of membrane elements: Site-directed fluorescent labelling of spectrin will be achieved using a novel cleavable, fluorescent, photoaffinity crosslinking reagent. In situ specific labelling of actin, band 3, and 4.1 will also be conducted. Fluorescent and tritiated lipophilic compounds will be obtained commercially. The labelled protein and lipid species will then be incorporated in to inside- out vesicles and ghosts. Proper biochemical incorporation of proteins will be assessed by:determination of their binding affinity and reversibility of binding to the membrane, competitive displacement with antibodies or appropriate functional protein fragments, and electron microscopic localization with anti-fluorescein immuno-gold conjugates. Proper functional incorporation of proteins will be assessed by monitoring membrane deformability and fragility by ektacytometry. (2)Measurement of skeleton:bilayer distance in normal cells: Short distances (within 20-80 A) between the fluorescently-labelled skeletal proteins and the plane of the inner leaflet membrane lipids will be measured by fluorescence energy transfer. Longer distances (between 80 to 500 A) will be measured by condensed phase radioluminescence using fluorescently-labelled skeletal proteins and tritiated membrane lipids. (3)Assessment of the effects of physiologic perturbations on the skeleton:bilayer distances: The changing interactions between the skeleton and the overlying bilayer will be examined during various membrane stresses (e.g., shear stress, hypotonic expansion, echinocytogenesis of different types, and mild oxidation with Heinz bodily induction). The localized topography of skeleton:bilayer distances (e.g., in the spikes of echinocytes or adjacent to Heinz bodies) will be measured in individual cells using a laser scanning confocal fluorescence dual channel microscope coupled to an image processing system.
| Project Number : | 5R01DK016095-18 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | cell membrane, cytoskeleton, erythrocyte, membrane lipid, membrane structure actin, ankyrin, band 3 protein, erythrocyte membrane, glycophorin, membrane activity, membrane model, membrane protein, spectrin, vesicle /vacuole Echinodermata, antibody, confocal scanning microscopy, fluorescein, fluorescence, immunoelectron microscopy, radiofluorescent probe |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON: BILAYER INTERFACE
(1995)
Abstract :
Evidence from pathologic red blood cells suggests that the coupling of skeletal proteins with the overlaying membrane is essential for normal cell integrity and dynamic behavior. Although extensive biochemical and electron microscopic studies have provided an initial concept of the skeletal structure, the geometry of the skeleton's relationship to the overlying bilayer is not well under-stood. Our goal is to use biophysical approaches to obtain a detailed perspective of the skeleton:bilayer interface in the intact cell that can eventually be studied under dynamic conditions. Such studies should provide insight into the instability of the membrane in certain hemolytic disorders. To define the molecular geometry of this interface, we plan to measure various skeleton:bilayer distances using fluorescence energy transfer and condensed phase radioluminescence techniques. For this purpose, the following steps will be carried out: (1)Labelling and incorporation of membrane elements: Site-directed fluorescent labelling of spectrin will be achieved using a novel cleavable, fluorescent, photoaffinity crosslinking reagent. In situ specific labelling of actin, band 3, and 4.1 will also be conducted. Fluorescent and tritiated lipophilic compounds will be obtained commercially. The labelled protein and lipid species will then be incorporated in to inside- out vesicles and ghosts. Proper biochemical incorporation of proteins will be assessed by:determination of their binding affinity and reversibility of binding to the membrane, competitive displacement with antibodies or appropriate functional protein fragments, and electron microscopic localization with anti-fluorescein immuno-gold conjugates. Proper functional incorporation of proteins will be assessed by monitoring membrane deformability and fragility by ektacytometry. (2)Measurement of skeleton:bilayer distance in normal cells: Short distances (within 20-80 A) between the fluorescently-labelled skeletal proteins and the plane of the inner leaflet membrane lipids will be measured by fluorescence energy transfer. Longer distances (between 80 to 500 A) will be measured by condensed phase radioluminescence using fluorescently-labelled skeletal proteins and tritiated membrane lipids. (3)Assessment of the effects of physiologic perturbations on the skeleton:bilayer distances: The changing interactions between the skeleton and the overlying bilayer will be examined during various membrane stresses (e.g., shear stress, hypotonic expansion, echinocytogenesis of different types, and mild oxidation with Heinz bodily induction). The localized topography of skeleton:bilayer distances (e.g., in the spikes of echinocytes or adjacent to Heinz bodies) will be measured in individual cells using a laser scanning confocal fluorescence dual channel microscope coupled to an image processing system.
| Project Number : | 5R01DK016095-17 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | cell membrane, cytoskeleton, erythrocyte, membrane lipid, membrane structure actin, ankyrin, band 3 protein, erythrocyte membrane, glycophorin, membrane activity, membrane model, membrane protein, spectrin, vesicle /vacuole Echinodermata, antibody, confocal scanning microscopy, fluorescein, fluorescence, immunoelectron microscopy, radiofluorescent probe |
|---|
ARCHITECTURE OF ERYTHROCYTE SKELETON-BILAYER INTERFACE
(1995)
Abstract :
Evidence from pathologic red blood cells suggests that the coupling of skeletal proteins with the overlaying membrane is essential for normal cell integrity and dynamic behavior. Although extensive biochemical and electron microscopic studies have provided an initial concept of the skeletal structure, the geometry of the skeleton's relationship to the overlying bilayer is not well under-stood. Our goal is to use biophysical approaches to obtain a detailed perspective of the skeleton:bilayer interface in the intact cell that can eventually be studied under dynamic conditions. Such studies should provide insight into the instability of the membrane in certain hemolytic disorders. To define the molecular geometry of this interface, we plan to measure various skeleton:bilayer distances using fluorescence energy transfer and condensed phase radioluminescence techniques. For this purpose, the following steps will be carried out: (1)Labelling and incorporation of membrane elements: Site-directed fluorescent labelling of spectrin will be achieved using a novel cleavable, fluorescent, photoaffinity crosslinking reagent. In situ specific labelling of actin, band 3, and 4.1 will also be conducted. Fluorescent and tritiated lipophilic compounds will be obtained commercially. The labelled protein and lipid species will then be incorporated in to inside- out vesicles and ghosts. Proper biochemical incorporation of proteins will be assessed by:determination of their binding affinity and reversibility of binding to the membrane, competitive displacement with antibodies or appropriate functional protein fragments, and electron microscopic localization with anti-fluorescein immuno-gold conjugates. Proper functional incorporation of proteins will be assessed by monitoring membrane deformability and fragility by ektacytometry. (2)Measurement of skeleton:bilayer distance in normal cells: Short distances (within 20-80 A) between the fluorescently-labelled skeletal proteins and the plane of the inner leaflet membrane lipids will be measured by fluorescence energy transfer. Longer distances (between 80 to 500 A) will be measured by condensed phase radioluminescence using fluorescently-labelled skeletal proteins and tritiated membrane lipids. (3)Assessment of the effects of physiologic perturbations on the skeleton:bilayer distances: The changing interactions between the skeleton and the overlying bilayer will be examined during various membrane stresses (e.g., shear stress, hypotonic expansion, echinocytogenesis of different types, and mild oxidation with Heinz bodily induction). The localized topography of skeleton:bilayer distances (e.g., in the spikes of echinocytes or adjacent to Heinz bodies) will be measured in individual cells using a laser scanning confocal fluorescence dual channel microscope coupled to an image processing system.
| Project Number : | 5R01DK016095-20 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | cell membrane, cytoskeleton, erythrocyte, membrane lipid, membrane structure actin, alternatives to animals in research, ankyrin, band 3 protein, erythrocyte membrane, glycophorin, membrane activity, membrane model, membrane protein, spectrin, vesicle /vacuole Echinodermata, antibody, confocal scanning microscopy, fluorescein, fluorescence, immunoelectron microscopy, radiofluorescent probe |
|---|
RED CELL MEMBRANE STUDIES
(1992)
| Project Number : | 5P01DK032094-08 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | ADDK |
|---|
| Project Terms : | erythrocyte membrane |
|---|
RED CELL MEMBRANE STUDIES
(1991)
| Project Number : | 5P01DK032094-07 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | ADDK |
|---|
| Project Terms : | MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE |
|---|
RED CELL MEMBRANE
(1991)
| Project Number : | 5P01DK032094-06 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | ADDK |
|---|
| Project Terms : | MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE |
|---|
RED CELL MEMBRANE STUDIES
(1991)
| Project Number : | 5P01DK032094-05 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | ADDK |
|---|
| Project Terms : | MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE |
|---|
RED CELL MEMBRANE STUDIES
(1991)
| Project Number : | 2P01DK032094-04A1 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | ADDK |
|---|
| Project Terms : | MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1990)
| Project Number : | 5T32HL007100-12 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1990)
| Project Number : | 2T32HL007100-11 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1990)
| Project Number : | 5T32HL007100-15 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1990)
| Project Number : | 5T32HL007100-14 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1990)
| Project Number : | 5T32HL007100-13 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
ROLE OF THE MEMBRANE SKELETON IN PATHOLOGIC RED CELLS
(1987)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Spectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 5R01DK016095-15 |
|---|
| ICD : | NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
ROLE OF THE MEMBRANE SKELETON IN PATHOLOGIC RED CELLS
(1987)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Spectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 5R01AM016095-14 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
ROLE OF THE MEMBRANE SKELETON IN PATHOLOGIC RED CELLS
(1987)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Spectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 5R01AM016095-13 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
ROLE OF THE MEMBRANE SKELETON IN PATHOLOGIC RED CELLS
(1987)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Spectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 5R01AM016095-12 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
ROLE OF THE MEMBRANE SKELETON IN PATHOLOGIC RED CELLS
(1987)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Spectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 2R01AM016095-11 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
RED CELL MEMBRANE STUDIES
(1986)
Abstract :
The overall aim of this project is to define the genesis, evolution and eventual senescence of the normal red cell membrane. Long-term objectives which may accrue from these studies are the development of some understanding of the pathophysiologic mechanisms involved in hemolysis and the more general application of model information gathered on red cell membrane development and maturation towards the understanding of those processes in other somatic cells. Four complementary approaches towards our overall aim will be employed: 1) studies to isolate the genes responsible for directing the synthesis of selected red cell membrane proteins; 2) studies to define the nature of the red cell membrane in fetal and neonatal cells, and to compare it to adult cells; 3) studies to examine the nature of free radical or oxidative damage to red cell membranes during their development, circulation and maturation, and 4) studies attempting to define the senescent red cell membrane and the mechanisms of its production. A broad range of methods from the disciplines of molecular biology, biochemistry, immunology, cell biology and biophysics will be utilized in these studies which will be conducted in five laboratories, a supporting "morphologic" core, and two consulting laboratories. It is hoped that this effort to gain some understanding of the development of the red cell membrane, from its birth to its death, will eventually provide insights which will be useful for the management of hemolytic anemias and other clinical disorders where cell membrane development may be disordered.
| Project Number : | 5P01AM032094-03 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | SRC |
|---|
| Project Terms : | MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE, MEMBRANE, MEMBRANE (BIOLOGICAL) STRUCTURE |
|---|
RED CELL MEMBRANE STUDIES
(1986)
Abstract :
The overall aim of this project is to define the genesis, evolution and eventual senescence of the normal red cell membrane. Long-term objectives which may accrue from these studies are the development of some understanding of the pathophysiologic mechanisms involved in hemolysis and the more general application of model information gathered on red cell membrane development and maturation towards the understanding of those processes in other somatic cells. Four complementary approaches towards our overall aim will be employed: 1) studies to isolate the genes responsible for directing the synthesis of selected red cell membrane proteins; 2) studies to define the nature of the red cell membrane in fetal and neonatal cells, and to compare it to adult cells; 3) studies to examine the nature of free radical or oxidative damage to red cell membranes during their development, circulation and maturation, and 4) studies attempting to define the senescent red cell membrane and the mechanisms of its production. A broad range of methods from the disciplines of molecular biology, biochemistry, immunology, cell biology and biophysics will be utilized in these studies which will be conducted in five laboratories, a supporting "morphologic" core, and two consulting laboratories. It is hoped that this effort to gain some understanding of the development of the red cell membrane, from its birth to its death, will eventually provide insights which will be useful for the management of hemolytic anemias and other clinical disorders where cell membrane development may be disordered.
| Project Number : | 1P01AM032094-01 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | SRC |
|---|
| Project Terms : | MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE, MEMBRANE, MEMBRANE (BIOLOGICAL) STRUCTURE, genetic manipulation |
|---|
RED CELL MEMBRANE STUDIES
(1986)
Abstract :
The overall aim of this project is to define the genesis, evolution and eventual senescence of the normal red cell membrane. Long-term objectives which may accrue from these studies are the development of some understanding of the pathophysiologic mechanisms involved in hemolysis and the more general application of model information gathered on red cell membrane development and maturation towards the understanding of those processes in other somatic cells. Four complementary approaches towards our overall aim will be employed: 1) studies to isolate the genes responsible for directing the synthesis of selected red cell membrane proteins; 2) studies to define the nature of the red cell membrane in fetal and neonatal cells, and to compare it to adult cells; 3) studies to examine the nature of free radical or oxidative damage to red cell membranes during their development, circulation and maturation, and 4) studies attempting to define the senescent red cell membrane and the mechanisms of its production. A broad range of methods from the disciplines of molecular biology, biochemistry, immunology, cell biology and biophysics will be utilized in these studies which will be conducted in five laboratories, a supporting "morphologic" core, and two consulting laboratories. It is hoped that this effort to gain some understanding of the development of the red cell membrane, from its birth to its death, will eventually provide insights which will be useful for the management of hemolytic anemias and other clinical disorders where cell membrane development may be disordered.
| Project Number : | 5P01AM032094-02 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | SRC |
|---|
| Project Terms : | MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE, MEMBRANE, MEMBRANE (BIOLOGICAL) STRUCTURE, genetic manipulation |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1985)
| Project Number : | 5T32HL007100-10 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1985)
| Project Number : | 2T32HL007100-06 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1985)
| Project Number : | 5T32HL007100-07 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1985)
| Project Number : | 5T32HL007100-08 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1985)
| Project Number : | 5T32HL007100-09 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | MR |
|---|
MEMBRANE LIPID AND PROTEIN METABOLISM IN BLOOD CELLS
(1982)
Abstract :
Membrane components of normal and pathologic erythrocytes will be investigated to identify structural and energetic factors which contribute to the maintenance of red cell shape and deformability. Spectrin appears to be membrane polypeptides critical for maintaining normal shape and deformability. Studies on the chemical events occurring during phosphorylation of spectrin by protein kinase will be extended. The sequence of amino acids around the site(s) of phosphorylation will be determined. The mechanism associated with spectrin dephosphorylation during and concommitant determinations on cell morphology and deformability during metabolic depletion of the red cell will be studied in some detail. Membrane polypeptides from pathologic erythrocytes such as hereditary spherocytes, elliptocytes, and pyropoikilocytes will be examined by a variety of electrophoretic and immunologic techniques to try to identify dysfunctional components. The interaction of spectin with erythrocyte actin will be studied in preparations from normal and pathologic cells. Additional studies on the thermal sensitivities of normal and pathologic erythrocytes will be used to probe constituents in defective membranes. The properties of red cells in a murine genetic disorder (sph/sph) involving a spectrin deficiency will be studied further to evaluate the role of spectrin in maintaining a stable membrane and influencing fluidity properties of the membrane.
| Project Number : | 5R01AM016095-07 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | CELL COMPONENTS, CELL MEMBRANE, HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
MEMBRANE LIPID AND PROTEIN METABOLISM IN BLOOD CELLS
(1982)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Sprectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 5R01AM016095-08 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
MEMBRANE LIPID AND PROTEIN METABOLISM IN BLOOD CELLS
(1982)
Abstract :
The work proposed here is designed to define the role of the membrane in the survival and function of red blood cells. The role of the membrane lipid peroxidation in accelerating premature cell lysis will be studied by determination of the susceptibility of normal and pathologic red cells to peroxidation, measurement of physical and chemical changes in red cells occurring after peroxidation, efforts to determine red cell peroxidation in vitro in pathologic conditions, and the measurement of the effects of in vitro peroxidation on the survival of human erythrocytes with an in vivo animal model. In addition, the role of the membrane protein kinase system and its substrates in the regulation of cell shape and membrane function will be studied. This work will include a detailed examination of the properties of spectrin, a series of studies of membrane: macromolecular interactions, and a series of studies on the phosphorylation of membrane protein. These latter experiments will utilize membranes from normal cells, normal maturing reticulocytes, and hereditary spherocytosis cells of both mice and men.
| Project Number : | 2R01AM016095-06 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | CELL COMPONENTS, CELL MEMBRANE, HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY*, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT*, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY)*, PHYSICAL SEPARATION, ELECTROPHORESIS, GEL*, RADIOTRACERS, DOUBLE (MULTIPLE)* |
|---|
MEMBRANE LIPID AND PROTEIN METABOLISM IN BLOOD CELLS
(1982)
| Project Number : | 3R01AM016095-06S1 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
MEMBRANE LIPID AND PROTEIN METABOLISM IN BLOOD CELLS
(1982)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Spectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 5R01AM016095-10 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
MEMBRANE LIPID AND PROTEIN METABOLISM IN BLOOD CELLS
(1982)
Abstract :
The purpose of the studies of red cell and granulocyte membranes, outlined in this proposal, is to define changes in cell membrane protein and lipid metabolism which may influence cell flexibility and permeability. The flexibility and permeability of erythrocytes is crucial to cell survival in vivo and is also particularly relevant to the problem of prolonged erythrocyte storage. Particular attention is focused upon the role of the Red cell membrane protein Spectrin since defects in Spectrin appear to be a common denominator in several clinical Hemolytic disorders.
| Project Number : | 5R01AM016095-09 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM, LIPIDS, MEMBRANE LIPIDS, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOTRACERS, DOUBLE (MULTIPLE) |
|---|
SPECTRIN'S ROLE IN BLOOD CELL SHAPE AND DEFORMABILITY
(1981)
Abstract :
Proteins associated with the erythrocyte membrane appear to play a critical role in the determination of cell shape and deformability, i.e., survival. A detailed study of the membrane protein kinase system and its substrates will be undertaken in order to ascertain its importance in regulating shape and deformability of the cell. Particular attention will be paid to spectrin, its phosphorylation, and subsequent interaction with erythrocyte actin to form the cytoskeletal network of the membrane. The properties of spectrin, the phosphorylation reaction and spectrin-actin complex will be examined from normal and pathological red cells. By selectively modifying and inhibiting the membrane associated enzymes (protein kinases and adenosine triphosphatases), we hope to develop a coherent mechanism for the involvement of these proteins in controlling the erythrocyte shape and its deformability.
| Project Number : | 5R01AM023077-03 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, HEMATOLOGY STUDY SECTION, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE, PROTEINS, MEMBRANE PROTEINS, PROTEINS, MEMBRANE PROTEINS, SPECTRIN BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), CHEMICAL STRUCTURE, STEREOCHEMISTRY, CONFORMATIONS, MUSCLE PROTEINS (AND CONTRACTILE PROTEINS), ACTIN, PHOSPHATASES, ADENOSINE TRIPHOSPHATASE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, ULTRAVIOLET, CHEMISTRY, STOICHIOMETRY, HUMAN, CLINICAL, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), OPTICS, DICHROISM CIRCULAR, PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOAUTOGRAPHY, RADIOTRACERS |
|---|
SPECTRIN'S ROLE IN BLOOD CELL SHAPE AND DEFORMABILITY
(1981)
Abstract :
Proteins associated with the erythrocyte membrane appear to play a critical role in the determination of cell shape and deformability, i.e., survival. A detailed study of the membrane protein kinase system and its substrates will be undertaken in order to ascertain its importance in regulating shape and deformability of the cell. Particular attention will be paid to spectrin, its phosphorylation, and subsequent interaction with erythrocyte actin to form the cytoskeletal network of the membrane. The properties of spectrin, the phosphorylation reaction and spectrin-actin complex will be examined from normal and pathological red cells. By selectively modifying and inhibiting the membrane associated enzymes (protein kinases and adenosine triphosphatases), we hope to develop a coherent mechanism for the involvement of these proteins in controlling the erythrocyte shape and its deformability.
| Project Number : | 5R01AM023077-02 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, HEMATOLOGY STUDY SECTION, MEMBRANE, ERYTHROCYTE GHOST AND MEMBRANE, PROTEINS, MEMBRANE PROTEINS, PROTEINS, MEMBRANE PROTEINS, SPECTRIN BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL, SPHEROCYTOSIS HEREDITARY (GENERAL), CHEMICAL STRUCTURE, STEREOCHEMISTRY, CONFORMATIONS, MUSCLE PROTEINS (AND CONTRACTILE PROTEINS), ACTIN, PHOSPHATASES, ADENOSINE TRIPHOSPHATASE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, ULTRAVIOLET, CHEMISTRY, STOICHIOMETRY, HUMAN, CLINICAL, MAMMALS, RODENTS, MYOMORPHA, MICE (LABORATORY), OPTICS, DICHROISM CIRCULAR, PHYSICAL SEPARATION, ELECTROPHORESIS, GEL, RADIOAUTOGRAPHY, RADIOTRACERS |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1980)
| Project Number : | 5T32HL007100-05 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | TEC |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1980)
| Project Number : | 5T32HL007100-03 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | TEC |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1980)
| Project Number : | 5T32HL007100-04 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | TEC |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1980)
| Project Number : | 1T32HL007100-01 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | TEC |
|---|
BLOOD BANKING SCIENCES AND RELATED PROGRAMS
(1980)
| Project Number : | 5T32HL007100-02 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | TEC |
|---|
CLINICAL CANCER EDUCATION PROGRAM
(1978)
Abstract :
The purpose of this project is to increase the effectiveness of clinical cancer education for both the academic and lay communities served by the University of California, San Francisco. This will be done through the Cancer Research Institute (CRI) of the University by two major mechanisms: First, by strengthening the existing teaching programs of the CRI and coordinating other cancer education efforts within the University; and secondly by making the clinical and research resources of the CRI more available to the scientific, academic and lay communities associated with this campus. These goals will be achieved by: (l) improving the quality and scope of education in cancer therapy and followup by providing support to academic teachers in Oncology medicine and nursing, (2) support of educational programs on campus as well as in the community, (3) establishing a central educational reference area (library) to provide videotaped lectures, scientific and lay literature, and reference search services, (4) support undergraduate, graduate and postgraduate travel for educational and information exchanges, (5) support clinical cancer conferences for physicians and nurses in Northern California.
| Project Number : | 5R25CA017995-02 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CEC |
|---|
| Project Terms : | CANCER (CLINICAL) EDUCATION COMMITTEE, EDUCATION, HEALTH EDUCATION, CANCER EDUCATION (PREVENTION AND CONTROL) EDUCATION, AUDIOVISUAL AND OTHER TRAINING AIDS, EDUCATION, COMPUTER ASSISTED INSTRUCTION, EDUCATION, CURRICULUM, EDUCATION, HEALTH OCCUPATIONS, PREMEDICAL, EDUCATION, POSTGRADUATE, HEALTH CARE SERVICES, COMMUNITY HEALTH SERVICES, HEALTH SCIENCES PROFESSIONS, NURSING ONCOLOGY, INFORMATION DISSEMINATION, TELEVISION, INFORMATION SYSTEMS, INFORMATION RETRIEVAL, INFORMATION SYSTEMS, LIBRARIES, NEOPLASMS DIAGNOSIS, PUBLICATIONS |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1977)
Abstract :
This research is devoted to three major areas on the membrane biochemistry and function of blood cells. These are: 1) the study of membrane protein kinases and the phosphorylation of membrane proteins to examine their possible role in the regulation of cell shape and perhaps membrane permeability; 2) the study of membrane lipid peroxidation in the generation of damaged membranes in hemolytic anemias such as thalassemia and in the remodeling of white cell membranes during phagocytosis; and 3) the study of abnormalities of lipid composition and renewal in unusual congenital hemolytic anemias. In addition to these primary areas of interest, the laboratory is also involved in the use of a "hybrid" erythrocyte model consisting of membranes of one cell resealed around the hemoglobin of another in order to study the influence of abnormal hemoglobins upon membrane function.
| Project Number : | 5R01AM016095-05 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, CELL COMPONENTS, CELL MEMBRANE, HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HEMOLYTIC CONGENITAL (GENERAL), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS STRUCTURE, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP, phosphorylation CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY*, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT*, PHYSICAL SEPARATION, ELECTROPHORESIS, GEL*, RADIOTRACERS, DOUBLE (MULTIPLE)* |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1977)
| Project Number : | 3R01AM016095-05S1 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
CLINICAL CANCER EDUCATION PROGRAM
(1977)
Abstract :
The purpose of this project is to increase the effectiveness of clinical cancer education for both the academic and lay communities served by the University of California, San Francisco. This will be done through the Cancer Research Institute (CRI) of the university by two major mechanisms: First, by strengthening the existing teaching programs of the Cancer Research Institute and coordinating other cancer education efforts within the university; and, secondly, by making clinical and research resources of the CRI more available to the scientific, academic and lay communities associated with this campus. Specifically, these goals will be achieved by: 1) improving the quality of education and patient care in the CRI through the support of full- time academic teachers in Cancer Medicine and Nursing; 2) utilizing the facilities of the University of California Medical T.V. Network to record and disseminate research and clinical education activities taking place within the CRI; 3) upgrading services provided by the library of the Cancer Research Institute making its information retrieval services and those of the UCSF Medical Library more widely available; 4) establishing a clinical cancer syllabus and a monthly publication of the CRI; 5) establishing undergraduate summer student support for clinical study and research in cancer; 6) utilizing and expanding an existing conputer-based examination question book for self-assessment examinations; 7) providing for undergraduate, graduate and consultant travel for educational purposes and information- exchange and 8) supporting clinical cancer conferences for physicians and nurses in Northern California.
| Project Number : | 1R25CA017995-01 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | CEC |
|---|
| Project Terms : | CLINICAL CANCER EDUCATION COMMITTEE, EDUCATION, HEALTH EDUCATION, CANCER EDUCATION (PREVENTION AND CONTROL) EDUCATION, AUDIOVISUAL AND OTHER TRAINING AIDS, EDUCATION, COMPUTER ASSISTED INSTRUCTION, EDUCATION, CURRICULUM, EDUCATION, HEALTH OCCUPATIONS, PREMEDICAL, EDUCATION, POSTGRADUATE, HEALTH CARE SERVICES, COMMUNITY HEALTH SERVICES, HEALTH SCIENCES PROFESSIONS, NURSING ONCOLOGY, INFORMATION DISSEMINATION, TELEVISION, INFORMATION SYSTEMS, INFORMATION RETRIEVAL, INFORMATION SYSTEMS, LIBRARIES, NEOPLASMS DIAGNOSIS, PUBLICATIONS NEOPLASMS RELATED CONTROL TAG |
|---|
MEMBRANE LIPID METABOLISM
(1977)
| Project Number : | 5K04AM037237-04 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
MEMBRANE LIPID METABOLISM
(1977)
| Project Number : | 5K04AM037237-05 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
MEMBRANE LIPID METABOLISM
(1977)
| Project Number : | 7K04AM037237-01A1 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
MEMBRANE LIPID METABOLISM
(1977)
| Project Number : | 5K04AM037237-02 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
MEMBRANE LIPID METABOLISM
(1977)
| Project Number : | 5K04AM037237-03 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1977)
Abstract :
The objectives of the continued studies of cell membranes outlined in this proposal are to define the role of the membrane in the survival and function of blood cells. In particular, studies of changes in membrane lipid and protein metabolism which influence cell flexibility and permeability are proposed. Four major areas of interest are underway as follows: 1) Biochemical and biophysical studies to explore the organization of red cell membrane lipid and protein; 2) Biochemical studies to explore the role of membrane kinase and ATP in the regulation of red cell shape and deformability in both normal and pathologic red cells; 3) Studies with a "hybrid" membrane-hemoglobin model to explore the possible role of hemoglobin interactions with the membrane in influencing its function; 4) Studies on the influence of membrane lipid peroxidation on biochemical and physical characteristics of membranes and the possible role of cellular hemoglobin in modulating these reactions. Conventional methods include lipid analyses (TLC and GLC), protein analysis by polyacrylamide gel electrophoresis and enzyme analyses. Additional methods employed will include the active and passive labeling of membrane constituents followed by exchange studies of inside-out and rightside-out membrane vesicles with plasma; a novel method for resealing membrane ghosts around exogenous hemoglobin; red cell membrane fusion with lipid liposomes to modify lipid environments; measurement of the effects of protein kinase and peroxidation in red cell membranes by physical methods including nuclear magnetic resonance, electron spin resonance, and optical rotatory dispersion; and doubly labeled in vivo cell survival studies utilizing both hemoglobin (Cr51) and membrane (P32-phosphatidylethanolamine) labels.
| Project Number : | 2R01AM016095-04 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, CELL COMPONENTS, CELL MEMBRANE, HEMATOLOGY STUDY SECTION, LIPIDS METABOLISM BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD AND RE DISORDERS CONGENITAL (HEREDITARY), BLOOD AND RE DISORDERS, ANEMIA HYPOCHROMIC MICROCYTIC, BLOOD CELLS, ERYTHROCYTES, HEMOLYSIS, HEMOPROTEINS, HEMOGLOBIN, HEMOPROTEINS, HEMOGLOBINOPATHIES, SICKLE CELL ANEMIA, HEMOPROTEINS, HEMOGLOBINOPATHIES, THALASSEMIA, PEROXIDES, HYDROGEN PEROXIDE, PHOSPHOTRANSFERASES, ATP:PROTEIN PHOSPHOTRANSFERASE, PURINE NUCLEOTIDES, ADENINE NUCLEOTIDES, ATP BIOPHYSICS (GENERAL)*, CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY*, CHEMISTRY, ANALYTICAL METHODS, SPECTROMETRY, NMR*, DYES, FLUORESCENT DYES AND PROBES, HUMAN, TISSUES, FLUIDS ETC. FROM NON-RELATED SOURCES OUTSIDE IMMEDIATE PROJECT*, PHYSICAL SEPARATION, ELECTROPHORESIS, GEL*, RADIOTRACERS, DOUBLE (MULTIPLE)* |
|---|
IN VITRO CHARACTERIZATION OF CELLS IN HODGKIN'S DISEASE
(1977)
Abstract :
The purpose of this project is to define the biologic properties of Hodgkin's disease cells. Our major method of approach will be to extend our observations of Hodgkin's disease cells in long-term tissue culture. We will continue to study cells currently in culture and will employ established methods for the separation of heterogeneous cell populations. From these populations clonally-derived cell lines will be established in long term culture. The malignancy of the Hodgkin's cell will be verified by cytogenetic, transplantation and further cloning studies. The possible lymphocytic or monocytic (histiocytic) origin of the Hodgkin's cell will be assessed by determining its cytochemical, functional and immunological properties. From these isolated Hodgkin's cells, we intend to identify and characterize tumor-associated antigens. We will then attempt to amplify the immunogenicity of these tumor-associated antigens by neuraminidase treatment. All of these studies are designed to gather information which will facilitate earlier diagnosis and immunotherapy of Hodgkin's disease.
| Project Number : | 5R01CA015182-02 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | PTHB |
|---|
| Project Terms : | NEOPLASMS DIAGNOSIS, NEOPLASMS IMMUNOLOGY, TUMOR ANTIGENS, NEOPLASMS OF BLOOD AND RE SYSTEM, HODGKIN'S DISEASE, PATHOLOGY STUDY SECTION BLOOD AND RE SYSTEM, MACROPHAGES, HISTIOCYTES, BLOOD CELLS, B LYMPHOCYTES, BLOOD CELLS, T LYMPHOCYTES, CARBOHYDRASES, NEURAMINIDASE, GLOBULINS, GAMMA GLOBULINS, IMMUNOGLOBULIN(S), IMMUNOLOGY, ANTIGENS, SURFACE ANTIGENS (GENERAL), NEOPLASMS OF BLOOD AND RE SYSTEM, LEUKEMIA LYMPHOCYTIC, NEOPLASMS OF BLOOD AND RE SYSTEM, LYMPHOMA, SUGAR ACIDS, SIALIC ACIDS, cell sorting CELL INGESTION, PHAGOCYTOSIS, HUMAN, CLINICAL, IMMUNITY, IMMUNOSUPPRESSION, ALLOANTISERA, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, IMMUNOFLUORESCENCE*, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, RADIOIMMUNOASSAY*, IMMUNOLOGY, COMPLEMENT, MAMMALS, LAGOMORPHS*, MAMMALS, MICE*, MAMMALS, UNGULATES, GOATS*, NEOPLASMS RELATED CONTROL TAG, NEOPLASMS TRANSPLANTATION, OPTICS, MICROSCOPY, ELECTRON SCANNING*, TISSUE (CELL) CULTURE, CLONE CELLS, WI-38 CELLS, TISSUE COMPATIBILITY-TRANSPLANT, TRANSPLANTATION HETEROLOGOUS |
|---|
IN VITRO CHARACTERIZATION OF CELLS IN HODGKIN'S DISEASE
(1977)
Abstract :
The purpose of this project is to define the biologic properties of Hodgkin's disease cells. Our major method of approach will be to extend our observations of Hodgkin's disease cells in long-term tissue culture. We will continue to study cells currently in culture and will employ established methods for the separation of heterogeneous cell populations. From these populations clonally-derived cell lines will be established in long term culture. The malignancy of the Hodgkin's cell will be verified by cytogenetic, transplantation and further cloning studies. The possible lymphocytic or monocytic (histiocytic) origin of the Hodgkin's cell will be assessed by determining its cytochemical, functional and immunological properties. From these isolated Hodgkin's cells, we intend to identify and characterize tumor-associated antigens. We will then attempt to amplify the immunogenicity of these tumor-associated antigens by neuraminidase treatment. All of these studies are designed to gather information which will facilitate earlier diagnosis and immunotherapy of Hodgkin's disease.
| Project Number : | 5R01CA015182-03 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | PTHB |
|---|
| Project Terms : | BLOOD CELLS, B LYMPHOCYTES, NEOPLASMS CHARACTERISTICS, CELLULAR LEVEL STUDIES (GENERAL), NEOPLASMS IMMUNOLOGY, TUMOR ANTIGENS, NEOPLASMS OF BLOOD AND RE SYSTEM, HODGKIN'S DISEASE, PATHOLOGY STUDY SECTION BLOOD AND RE SYSTEM, MACROPHAGES, HISTIOCYTES, BLOOD CELLS, T LYMPHOCYTES, CARBOHYDRASES, NEURAMINIDASE, GLOBULINS, GAMMA GLOBULINS, IMMUNOGLOBULIN G, IMMUNOLOGY, ANTIBODY FORMATION, NUCLEIC ACIDS REPAIR, DNA REPAIR (ENZYME), NUCLEIC ACIDS SYNTHESIS, DNA, SUGAR ACIDS, SIALIC ACIDS HUMAN, CLINICAL, IMMUNITY, IMMUNOSUPPRESSION, ALLOANTISERA, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, IMMUNOFLUORESCENCE*, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, RADIOIMMUNOASSAY*, MAMMALS, LAGOMORPHS*, MAMMALS, RODENTS, MYOMORPHA, ATHYMIC MICE (NUDE)*, NEOPLASMS TRANSPLANTATION, OPTICS, MICROSCOPY, ELECTRON SCANNING*, TISSUE (CELL) CULTURE, CLONE CELLS* |
|---|
IN VITRO CHARACTERIZATION OF CELLS IN HODGKIN'S DISEASE
(1977)
Abstract :
The purpose of this project is to define the biologic properties of Hodgkin's disease cells. Our major method of approach will be to extend our observations of Hodgkin's disease cells in long-term tissue culture. We will continue to study cells currently in culture and will employ established methods for the separation of heterogeneous cell populations. From these populations clonally-derived cell lines will be established in long term culture. The malignancy of the Hodgkin's cell will be verified by cytogenetic, transplantation and further cloning studies. The possible lymphocytic or monocytic (histiocytic) origin of the Hodgkin's cell will be assessed by determining its cytochemical, functional and immunological properties. From these isolated Hodgkin's cells, we intend to identify and characterize tumor-associated antigens. We will then attempt to amplify the immunogenicity of these tumor-associated antigens by neuraminidase treatment. All of these studies are designed to gather information which will facilitate earlier diagnosis and immunotherapy of Hodgkin's disease.
| Project Number : | 1R01CA015182-01 |
|---|
| ICD : | NATIONAL CANCER INSTITUTE |
|---|
| IRG : | PTHB |
|---|
| Project Terms : | BLOOD AND RE NEOPLASMS, HODGKIN'S DISEASE, NEOPLASMS DIAGNOSIS, PATHOLOGY STUDY SECTION |
|---|
HEMATOLOGY
(1976)
| Project Number : | 5T01HL005677-09 |
|---|
| ICD : | NATIONAL HEART, LUNG, AND BLOOD INSTITUTE |
|---|
| IRG : | HTA |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1975)
| Project Number : | 1R01AM016095-01 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, CELL COMPONENTS, CELL MEMBRANE, LIPOGENESIS BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD CELLS, LEUKOCYTES, GRANULOCYTES (GENERAL), CELL INGESTION, PHAGOCYTOSIS, FATTY ACIDS, HEMATOLOGY STUDY SECTION, HYDROLASES, ACYLASES, NUTRITIONAL ABNORMALITIES, HYPOVITAMINOSIS E, PEROXIDES, HYDROGEN PEROXIDE, cell age HUMAN, NONCLINICAL*, HYDROGEN, TRITIUM*, PHYSICAL SEPARATION, CHROMATOGRAPHY, THIN LAYER*, RADIOLABELED CARBON* |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1975)
| Project Number : | 5R01AM016095-02 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, CELL COMPONENTS, CELL MEMBRANE, LIPOGENESIS BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD CELLS, LEUKOCYTES, GRANULOCYTES (GENERAL), CELL INGESTION, PHAGOCYTOSIS, FATTY ACIDS, HEMATOLOGY STUDY SECTION, HYDROLASES, ACYLASES, NUTRITIONAL ABNORMALITIES, HYPOVITAMINOSIS E, PEROXIDES, HYDROGEN PEROXIDE, cell age HUMAN, NONCLINICAL*, HYDROGEN, TRITIUM*, PHYSICAL SEPARATION, CHROMATOGRAPHY, THIN LAYER*, RADIOLABELED CARBON* |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1975)
| Project Number : | 3R01AM016095-02S1 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1975)
Abstract :
The studies included in this project are primarily concerned with the role of lipids in membrane function in red blood cells. In addition to the continuing, previously reported, areas of interest which included the consequences of peroxidation of membrane lipids, the nature of membrane remodeling during hemolysis, and the role of ATP in membrane renewal, new areas of interest have been developed. These include: (1) the role of membrane protein phosphorylation in the maintenance of cell shape; (2) the preparation ofa useful "hybrid" sickle cell model which enables us to study the membrane independent from the hemoglobin; (3) the nature of hemoglobin: water interactions during the sickling process; and, (4) the development of a calorimetric method for directly measuring metabolic events during red cell glycolysis.
| Project Number : | 5R01AM016095-03 |
|---|
| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
|---|
| IRG : | HEM |
|---|
| Project Terms : | BLOOD CELLS, ERYTHROCYTES, CELL COMPONENTS, CELL MEMBRANE, LIPOGENESIS BIOLOGICAL TRANSPORT, MEMBRANE PERMEABILITY AND TRANSPORT, BLOOD CELLS, LEUKOCYTES, GRANULOCYTES (GENERAL), CELL INGESTION, PHAGOCYTOSIS, FATTY ACIDS, HEMATOLOGY STUDY SECTION, HYDROLASES, ACYLASES, NUTRITIONAL ABNORMALITIES, HYPOVITAMINOSIS E, PEROXIDES, HYDROGEN PEROXIDE, cell age HUMAN, NONCLINICAL*, HYDROGEN, TRITIUM*, PHYSICAL SEPARATION, CHROMATOGRAPHY, THIN LAYER*, RADIOLABELED CARBON* |
|---|
MEMBRANE LIPID METABOLISM IN BLOOD CELLS
(1975)
| Project Number : | 3R01AM016095-03S1 |
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| ICD : | NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES |
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| IRG : | HEM |
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ISOLATION AND CHARACTERIZATION
(1974)
| Project Number : | 1R01CA015182-010001 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| Project Terms : | BLOOD AND RE NEOPLASMS, HODGKIN'S DISEASE, cell sorting BLOOD AND RE SYSTEM, MACROPHAGES, HISTIOCYTES, BLOOD CELLS, B LYMPHOCYTES, BLOOD CELLS, T LYMPHOCYTES, NEOPLASMS IMMUNOLOGY HUMAN, NONCLINICAL*, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, IMMUNOFLUORESCENCE*, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, RADIOIMMUNOASSAY*, MAMMALS, RODENTS, HAMSTERS*, TISSUE (CELL) CULTURE, CLONE CELLS*, TISSUE COMPATIBILITY-TRANSPLANT, TRANSPLANTATION HETEROLOGOUS |
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ISOLATION AND CHARACTERIZATION OF TUMOR ASSOCIATED ANTIGENS
(1974)
| Project Number : | 1R01CA015182-010002 |
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| ICD : | NATIONAL CANCER INSTITUTE |
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| Project Terms : | BLOOD AND RE NEOPLASMS, HODGKIN'S DISEASE, NEOPLASMS IMMUNOLOGY, TUMOR ANTIGENS, NEOPLASMS, HOST-NEOPLASM CARBOHYDRASES, NEURAMINIDASE, CELL SURFACE ACTIVITY (GENERAL), IMMUNITY, CELLULAR IMMUNITY (GENERAL), IMMUNITY, HUMORAL IMMUNITY, NEOPLASMS IMMUNIZATION (IMMUNOTHERAPY), SUGAR ACIDS, SIALIC ACIDS HUMAN, NONCLINICAL*, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, IMMUNOELECTROPHORESIS*, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, IMMUNOFLUORESCENCE*, IMMUNOLOGICAL TESTS AND IMMUNOASSAY, RADIOIMMUNOASSAY*, MAMMALS, RODENTS, HAMSTERS* |
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