Anders Raffalt - Academia.edu (original) (raw)

Papers by Anders Raffalt

Research paper thumbnail of Electron transfer patterns of the di-heme protein cytochrome c4 from Pseudomonas stutzeri

Journal of Inorganic Biochemistry, 2009

We report kinetic data for the two-step electron transfer (ET) oxidation and reduction of the two... more We report kinetic data for the two-step electron transfer (ET) oxidation and reduction of the two-domain di-heme redox protein Pseudomonas stutzeri cytochrome (cyt) c(4) by [Co(bipy)(3)](2+/3+) (bipy=2,2'-bipyridine). Following earlier reports, the data accord with both bi- and tri-exponential kinetics. A complete kinetic scheme includes both "cooperative"intermolecular ET between each heme group and the external reaction partner, and intramolecular ET between the two heme groups. A new data analysis scheme shows unequivocally that two-ET oxidation and reduction of P. stutzeri cyt c(4) is entirely dominated by intermolecular ET between the heme groups and the external reaction partner in the ms time range, with virtually no contribution from intramolecular interheme ET in this time range. This is in striking contrast to two-ET electrochemical oxidation or reduction of P. stutzeri cyt c(4) for which fast, ms to sub-ms intramolecular interheme ET is a crucial step. The rate constant dependence on the solvent viscosity has disclosed strong coupling to both a (set of) frictionally damped solvent/protein nuclear modes and intramolecular friction-less "ballistic" modes, indicative of notable protein structural mobility in the overall two-ET process. We suggest that conformational protein mobility blocks intramolecular interheme ET in bulk homogeneous solution but triggers opening of this gated ET channel in the electrochemical environment or in the membrane environment of natural respiratory cyt c(4) function.

Research paper thumbnail of Application of inductively coupled plasma-mass spectrometry (ICP-MS) and quality assurance to study the incorporation of strontium into bone, bone marrow, and teeth of dogs after one month of treatment with strontium malonate

Analytical and bioanalytical chemistry, 2008

The strontium content of serum, bone, marrow, and teeth was determined by inductively-coupled pla... more The strontium content of serum, bone, marrow, and teeth was determined by inductively-coupled plasma mass spectrometry (ICP-MS). Significant correlations were obtained after the data were subjected to quality assurance (QA) performed according to validated procedures. After four weeks of treatment with strontium malonate, strontium levels increased from 76 +/- 9 microg g(-1) in placebo-treated dogs to levels of 7.2 +/- 1.7 mg g(-1), 9.5 +/- 2.7 mg g(-1), and 9.8 +/- 2.7 mg g(-1) in groups treated with 300, 1000, and 3000 mg kg(-1) day(-1), respectively. Strontium induced a highly significant increase in the bone formation marker, bone-specific alkaline phosphatase (BSAP), and an excellent correlation was found with the bone-strontium content. In females, the placebo-treated group showed a decrease in BSAP of 53%, whereas the three strontium malonate-treated groups showed an increase of 60, 276, and 278% for the groups treated with 300, 1000, and 3000 mg kg(-1) day(-1), respectively....

Research paper thumbnail of Strontium Is Incorporated into the Fracture Callus but Does Not Influence the Mechanical Strength of Healing Rat Fractures

Calcified Tissue International, 2011

Strontium ranelate (SrR) is a new agent used in the treatment of osteoporosis and is suggested to... more Strontium ranelate (SrR) is a new agent used in the treatment of osteoporosis and is suggested to reduce bone resorption and increase bone formation. We investigated whether SrR influences the macro-and nanomechnical properties of healing fractures in rats. A closed tibia fracture model was used to study fracture healing in rats after 3 and 8 weeks of healing. Two groups of rats were treated with SrR (900 mg/kg/day) mixed into the food, while two groups served as control animals. The healing fractures were investigated by three-point bending, dual energy X-ray absorptiometry, energy-dispersive X-ray spectroscopy (EDX), and nanoindentation. There was a 100-fold increase (P \ 0.001) in serum Sr after 3 and 8 weeks of SrR treatment. The callus volume was significantly higher in the SrR-treated group than in control animals (P \ 0.01) after 3 weeks of healing. This was accompanied by a significant increase in callus bone mineral content (P \ 0.05). However, after 8 weeks of healing, no difference was found in either callus volume or bone mineral content. SrR did not influence maximum load or stiffness of the fractures after either 3 or 8 weeks of healing. EDX showed that Sr was incorporated into the callus; however, this did not influence the nanomechanical properties. In conclusion, SrR stimulates callus formation but has no effect on callus remodeling. Sr is incorporated into the newly formed callus tissue, but this has no deteriorating effect on the mechanical properties of rat tibial fractures at either the macroscopic or nanoscopic level after 3 or 8 weeks of healing.

Research paper thumbnail of Strontium Localization in Bone Tissue Studied by X-Ray Absorption Spectroscopy

Calcified Tissue International, 2014

Strontium has recently been introduced as a pharmacological agent for the treatment and preventio... more Strontium has recently been introduced as a pharmacological agent for the treatment and prevention of osteoporosis. We determined the localization of strontium incorporated into bone matrix from dogs treated with Sr malonate by X-ray absorption spectroscopy. A new approach for analyzing the X-ray absorption spectra resulted in a compositional model and allowed the relative distribution of strontium in the different bone components to be estimated. Approximately 35-45% of the strontium present is incorporated into calcium hydroxyapatite (CaHA) by substitution of some of the calcium ions occupying highly ordered sites, and at least 30% is located at less ordered sites where only the first solvation shell is resolved, suggesting that strontium is surrounded by only oxygen atoms similar to Sr(2+) in solution. Strontium was furthermore shown to be absorbed in collagen in which it obtains a higher structural order than when present in serum but less order than when it is incorporated into CaHA. The total amount of strontium in the samples was determined by inductively coupled plasma mass spectrometry, and the amount of Sr was found to increase with increasing dose levels and treatment periods, whereas the relative distribution of strontium among the different components appears to be independent of treatment period and dose level.

Research paper thumbnail of Polymeric strontium ranelate nonahydrate

The title compound, poly[[μ-aqua-tetraaqua{μ-5-[bis(carboxylatomethyl)amino]-3-carboxylatomethyl-... more The title compound, poly[[μ-aqua-tetraaqua{μ-5-[bis(carboxylatomethyl)amino]-3-carboxylatomethyl-4-cyanothiophene-2-carboxylato}distrontium(II)] tetrahydrate], [Sr2(C12H6N2O8S)(H2O)5]·3.79H2O, crystallizes with nine- and eight-coordinated Sr2+ cations. They are bound to seven of the eight ranelate O atoms and five of the water molecules. The SrO8 and SrO9 polyhedra are interconnected by edge-sharing, forming hollow layers parallel to (011). The layers are, in turn, interconnected by ranelate anions, forming a metal–organic framework (MOF) structure with channels along the a axis. The four water molecules not coordinated to strontium are located in these channels and hydrogen bonded to each other and to the ranelates. Part of the water H atoms are disordered. The compound dehydrates very easily and 0.210 (4) water molecules out of nine were lost during crystal mounting causing additional disorder in the water structure.

Research paper thumbnail of Electron transfer patterns of the di-heme protein cytochrome c4 from Pseudomonas stutzeri

Journal of Inorganic Biochemistry, 2009

We report kinetic data for the two-step electron transfer (ET) oxidation and reduction of the two... more We report kinetic data for the two-step electron transfer (ET) oxidation and reduction of the two-domain di-heme redox protein Pseudomonas stutzeri cytochrome (cyt) c(4) by [Co(bipy)(3)](2+/3+) (bipy=2,2'-bipyridine). Following earlier reports, the data accord with both bi- and tri-exponential kinetics. A complete kinetic scheme includes both "cooperative"intermolecular ET between each heme group and the external reaction partner, and intramolecular ET between the two heme groups. A new data analysis scheme shows unequivocally that two-ET oxidation and reduction of P. stutzeri cyt c(4) is entirely dominated by intermolecular ET between the heme groups and the external reaction partner in the ms time range, with virtually no contribution from intramolecular interheme ET in this time range. This is in striking contrast to two-ET electrochemical oxidation or reduction of P. stutzeri cyt c(4) for which fast, ms to sub-ms intramolecular interheme ET is a crucial step. The rate constant dependence on the solvent viscosity has disclosed strong coupling to both a (set of) frictionally damped solvent/protein nuclear modes and intramolecular friction-less "ballistic" modes, indicative of notable protein structural mobility in the overall two-ET process. We suggest that conformational protein mobility blocks intramolecular interheme ET in bulk homogeneous solution but triggers opening of this gated ET channel in the electrochemical environment or in the membrane environment of natural respiratory cyt c(4) function.

Research paper thumbnail of Application of inductively coupled plasma-mass spectrometry (ICP-MS) and quality assurance to study the incorporation of strontium into bone, bone marrow, and teeth of dogs after one month of treatment with strontium malonate

Analytical and bioanalytical chemistry, 2008

The strontium content of serum, bone, marrow, and teeth was determined by inductively-coupled pla... more The strontium content of serum, bone, marrow, and teeth was determined by inductively-coupled plasma mass spectrometry (ICP-MS). Significant correlations were obtained after the data were subjected to quality assurance (QA) performed according to validated procedures. After four weeks of treatment with strontium malonate, strontium levels increased from 76 +/- 9 microg g(-1) in placebo-treated dogs to levels of 7.2 +/- 1.7 mg g(-1), 9.5 +/- 2.7 mg g(-1), and 9.8 +/- 2.7 mg g(-1) in groups treated with 300, 1000, and 3000 mg kg(-1) day(-1), respectively. Strontium induced a highly significant increase in the bone formation marker, bone-specific alkaline phosphatase (BSAP), and an excellent correlation was found with the bone-strontium content. In females, the placebo-treated group showed a decrease in BSAP of 53%, whereas the three strontium malonate-treated groups showed an increase of 60, 276, and 278% for the groups treated with 300, 1000, and 3000 mg kg(-1) day(-1), respectively....

Research paper thumbnail of Strontium Is Incorporated into the Fracture Callus but Does Not Influence the Mechanical Strength of Healing Rat Fractures

Calcified Tissue International, 2011

Strontium ranelate (SrR) is a new agent used in the treatment of osteoporosis and is suggested to... more Strontium ranelate (SrR) is a new agent used in the treatment of osteoporosis and is suggested to reduce bone resorption and increase bone formation. We investigated whether SrR influences the macro-and nanomechnical properties of healing fractures in rats. A closed tibia fracture model was used to study fracture healing in rats after 3 and 8 weeks of healing. Two groups of rats were treated with SrR (900 mg/kg/day) mixed into the food, while two groups served as control animals. The healing fractures were investigated by three-point bending, dual energy X-ray absorptiometry, energy-dispersive X-ray spectroscopy (EDX), and nanoindentation. There was a 100-fold increase (P \ 0.001) in serum Sr after 3 and 8 weeks of SrR treatment. The callus volume was significantly higher in the SrR-treated group than in control animals (P \ 0.01) after 3 weeks of healing. This was accompanied by a significant increase in callus bone mineral content (P \ 0.05). However, after 8 weeks of healing, no difference was found in either callus volume or bone mineral content. SrR did not influence maximum load or stiffness of the fractures after either 3 or 8 weeks of healing. EDX showed that Sr was incorporated into the callus; however, this did not influence the nanomechanical properties. In conclusion, SrR stimulates callus formation but has no effect on callus remodeling. Sr is incorporated into the newly formed callus tissue, but this has no deteriorating effect on the mechanical properties of rat tibial fractures at either the macroscopic or nanoscopic level after 3 or 8 weeks of healing.

Research paper thumbnail of Strontium Localization in Bone Tissue Studied by X-Ray Absorption Spectroscopy

Calcified Tissue International, 2014

Strontium has recently been introduced as a pharmacological agent for the treatment and preventio... more Strontium has recently been introduced as a pharmacological agent for the treatment and prevention of osteoporosis. We determined the localization of strontium incorporated into bone matrix from dogs treated with Sr malonate by X-ray absorption spectroscopy. A new approach for analyzing the X-ray absorption spectra resulted in a compositional model and allowed the relative distribution of strontium in the different bone components to be estimated. Approximately 35-45% of the strontium present is incorporated into calcium hydroxyapatite (CaHA) by substitution of some of the calcium ions occupying highly ordered sites, and at least 30% is located at less ordered sites where only the first solvation shell is resolved, suggesting that strontium is surrounded by only oxygen atoms similar to Sr(2+) in solution. Strontium was furthermore shown to be absorbed in collagen in which it obtains a higher structural order than when present in serum but less order than when it is incorporated into CaHA. The total amount of strontium in the samples was determined by inductively coupled plasma mass spectrometry, and the amount of Sr was found to increase with increasing dose levels and treatment periods, whereas the relative distribution of strontium among the different components appears to be independent of treatment period and dose level.

Research paper thumbnail of Polymeric strontium ranelate nonahydrate

The title compound, poly[[μ-aqua-tetraaqua{μ-5-[bis(carboxylatomethyl)amino]-3-carboxylatomethyl-... more The title compound, poly[[μ-aqua-tetraaqua{μ-5-[bis(carboxylatomethyl)amino]-3-carboxylatomethyl-4-cyanothiophene-2-carboxylato}distrontium(II)] tetrahydrate], [Sr2(C12H6N2O8S)(H2O)5]·3.79H2O, crystallizes with nine- and eight-coordinated Sr2+ cations. They are bound to seven of the eight ranelate O atoms and five of the water molecules. The SrO8 and SrO9 polyhedra are interconnected by edge-sharing, forming hollow layers parallel to (011). The layers are, in turn, interconnected by ranelate anions, forming a metal–organic framework (MOF) structure with channels along the a axis. The four water molecules not coordinated to strontium are located in these channels and hydrogen bonded to each other and to the ranelates. Part of the water H atoms are disordered. The compound dehydrates very easily and 0.210 (4) water molecules out of nine were lost during crystal mounting causing additional disorder in the water structure.