| 1. |
- Lundberg, Peter, et al.
(författare)
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1H NMR determination of urinary betaine in patients with premature vascular disease and mild homocysteinemia
- 1995
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Ingår i: Clinical Chemistry. - 0009-9147 .- 1530-8561. ; 41:2, s. 275-283
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Tidskriftsartikel (refereegranskat)abstract
- Urinary N,N,N-trimethylglycine (betaine) and N,N-dimethylglycine (DMG) have been identified and quantified for clinical purposes by proton nuclear magnetic resonance (1H NMR) measurement in previous studies. We have assessed these procedures by using both one-dimensional (1-D) and 2-D NMR spectroscopy, together with pH titration of urinary extracts to help assign 1H NMR spectral peaks. The betaine calibration curve linearity was excellent (r = 0.997, P = 0.0001) over the concentration range 0.2-1.2 mmol/L, and CVs for replicate betaine analyses ranged from 7% (n = 10) at the lowest concentration to 1% (n = 9) at the highest. The detection limit for betaine was < 15 mumol/L. Urinary DMG concentrations were substantially lower than those of betaine. Urinary betaine and DMG concentrations measured by 1H NMR spectroscopy from 13 patients with premature vascular disease and 17 normal controls provided clinically pertinent data. We conclude that 1H NMR provides unique advantages as a research tool for determination of urinary betaine and DMG concentrations.
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| 2. |
- Lundberg, Peter, et al.
(författare)
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Diffusion of solutes in agarose and alginate gels : 1H and 23Na PFGSE and 23Na TQF NMR studies
- 1997
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Ingår i: Magnetic Resonance in Medicine. - : John Wiley & Sons. - 0740-3194 .- 1522-2594. ; 37:1, s. 44-52
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Tidskriftsartikel (refereegranskat)abstract
- Cells immobilized in gels experience potential metabolic restrictions in the form of reduced diffusion rates of metabolites and ions and their possible selective adsorption on the gel matrix. Diffusion and relaxation characteristics of common solutes in agarose and barium alginate gels were investigated at 37 degrees C by using 1H PFGSE and 23Na TQF NMR spectroscopy. Glucose, glycine, alanine, lactate, sodium ions, and HDO were studied. There were no selective interactions between any of the metabolites and the gel materials but the diffusion coefficients were uniformly reduced. The effects of metabolite diffusion and utilization, in gel beads and threads containing cells, were simulated by using a reaction diffusion model incorporating the measured diffusion coefficients. Metabolism is expected to be very significantly limited by diffusion of solutes to and from the cells that are centrally located within gel threads or spheres of radius approximately 2.0 mm, which is a commonly used size.
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| 3. |
- Lundberg, Peter, et al.
(författare)
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Fructose 3-phosphate and 5-phosphoribosyl-1-pyrophosphate formation in perfused human erythrocytes : 31P NMR studies
- 1994
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Ingår i: Magnetic Resonance in Medicine. - : John Wiley & Sons. - 0740-3194 .- 1522-2594. ; 31:2, s. 110-121
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Tidskriftsartikel (refereegranskat)abstract
- 31P NMR was used to study the formation of fructose 3-phos-phate (F3P) and 5-phosphoribosyl-1-pyrophosphate (PRPP) in perfused human erythrocytes, in the presence of 10 different combinations and concentrations of glucose, inosine, pyru-vate, fructose, and inorganic phosphate (Pi). (1) The cells were immobilized in alginate-coated agarose threads and perfused with a medium containing fructose, and the level of F3P increased continuously over more than 10 h. The net rate of F3P formation was independent of the concentration of 2,3-bis-phosphoglycerate (2,3-DPG) present in the cells. (2) PRPP was formed in high concentrations, relative to normal, in immobilized cells when they were perfused with a medium containing Pi at a low pH (6.6). (3) The 2,3-DPG level decreased simultaneously when the sample was perfused with a medium containing fructose, but without inosine or pyruvate. The measured intracellular pH and free Mg2+ concentration were constant in these experiments. (4) The experiments confirmed the presence of fructose-3-phosphokinase (E.C. 2.7.1.-) and ribose-phosphate pyrophosphokinase (E.C. 2.7.6.1) activity in the human erythrocytes and that the biosynthetic pathways are active in immobilized cells at 37°C. (5) The rates of accumulation of 2,3-DPG and phosphomonoesters (PME) appeared to be strongly correlated.
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| 4. |
- Lundberg, Peter, et al.
(författare)
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Immobilization Methods for NMR Studies of Cellular Metabolism : A Practical Guide
- 1994
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Ingår i: ImmunoMethods. - : Elsevier. - 1058-6687 .- 1557-766X. ; 4:2, s. 163-178
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Tidskriftsartikel (refereegranskat)abstract
- Nuclear magnetic resonance (NMR) can be used in a nondestructive manner to study cellular metabolism in intact cell samples such as a suspension of cells. However, many different cell types require a well-regulated medium that includes a buffered pH, as well as a continuous supply of oxygen. A series of methods that have been used for the maintenance of the extracellular conditions involves the immobilization of cells, followed by perfusion of the immobilized cell sample. NMR studies can then be performed for extended periods of time, as well as under sterile conditions. We discuss methodology, with perfused erythrocytes and thymocytes as specific examples.
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| 5. |
- Lundberg, Peter, et al.
(författare)
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NMR studies of erythrocytes immobilized in agarose and alginate gels
- 1992
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Ingår i: Magnetic Resonance in Medicine. - : John Wiley & Sons. - 0740-3194 .- 1522-2594. ; 25:2, s. 273-288
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Tidskriftsartikel (refereegranskat)abstract
- 31P and 13C NMR were used to study the energy metabolism in perfused, human erythrocytes. The erythrocytes were immobilized in agarose threads, Ca- or Ba-alginate beads, and Ba-alginate-coated agarose threads. Erythrocytes were easily washed out from the agarose threads, but not from alginate-containing gels. Various small molecules, such as hypophosphite, dimethyl methylphosphonate, and methylphosphonate, were taken up from the perfusion medium in a normal manner. In addition, the 2,3-bisphosphoglycerate (2,3-DPG) chemical shifts were sensitive to the oxygen partial pressure suggesting that O2 molecules were diffusing through the gel and modifying the binding of 2,3-DPG to hemoglobin. A combination of inosine and pyruvate stimulated the synthesis of 2,3-DPG, but only if inorganic phosphate was present in the perfusion medium. Inosine only resulted in a dramatic rise in the intracellular sugarphosphate concentrations. Furthermore, [2-13C]glucose was converted to [2-13C]lactate by immobilized cells at a rate which was comparable to that in a control suspension. In summary, immobilization in Ba-alginate-coated agarose threads was an efficient way of trapping human erythrocytes for whole cell NMR investigations.
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| 6. |
- Shou, Keyun, et al.
(författare)
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Effect of red blood cell shape changes on haemoglobin interactions and dynamics : a neutron scattering study
- 2020
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Ingår i: Royal Society Open Science. - : ROYAL SOC. - 2054-5703. ; 7:10
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Tidskriftsartikel (refereegranskat)abstract
- By using a combination of experimental neutron scattering techniques, it is possible to obtain a statistical perspective on red blood cell (RBC) shape in suspensions, and the inter-relationship with protein interactions and dynamics inside the confinement of the cell membrane. In this study, we examined the ultrastructure of RBC and protein-protein interactions of haemoglobin (Hb) in them using ultra-small-angle neutron scattering and small-angle neutron scattering (SANS). In addition, we used the neutron backscattering method to access Hb motion on the ns time scale and angstrom length scale. Quasi-elastic neutron scattering (QENS) experiments were performed to measure diffusive motion of Hb in RBCs and in an RBC lysate. By using QENS, we probed both internal Hb dynamics and global protein diffusion, on the accessible time scale and length scale by QENS. Shape changes of RBCs and variation of intracellular Hb concentration were induced by addition of the Na+-selective ionophore monensin and the K+-selective one, valinomycin. The experimental SANS and QENS results are discussed within the framework of crowded protein solutions, where free motion of Hb is obstructed by mutual interactions.
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