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Biomedical subjects

O Mattsson

Publications and source records attributed to O Mattsson.

At least 19 recordsLinked to original sources
PubMed2001

Two differentially regulated Arabidopsis genes define a new branch of the DFR superfamily.

Two tandem genes were identified on Arabidopsis chromosome II (AtCRL1 and AtCRL2) encoding proteins with homology to members of the dihydroflavonol-4-reductase (DFR) superfamily. The encoded CRL1 and CRL2 proteins share 87% mutual amino acid sequence identity whereas their promoter regions are highly divergent, suggesting differential regulation of the CRL genes. Phylogenetic analysis placed CRL1 and CRL2 in a separate branch of the DFR superfamily. Northern blotting showed strong AtCRL1 induction by abscisic acid (ABA), drought, and heat shock, and high expression level in seeds, thus resembling the expression pattern of late embryogenic abundant ABA-responsive genes. Differential expression of the two genes during plant development was confirmed in plants expressing transcriptional fusions between the two promoters and the Escherichia coli beta-glucuronidase reporter gene. This showed that, whereas high expression of AtCRL1 in mature seeds declines during subsequent vegetative growth, transcriptional activity from the AtCRL2 promoter increases during vegetative growth. Expression of both genes is restricted to vascular tissue. Based upon their homology to proteins involved in lignin synthesis, we propose that AtCRL2 is involved in generating conducting tissue late in development, while AtCRL1 is involved in vascular tissue differentiation and/or synthesis in the germinating embryos.

Journal Article↗
PubMed2001

Gibberellin response mutants identified by luciferase imaging.

The gibberellin (GA) 20-oxidase encoded by Arabidopsis GA5 catalyzes the synthesis of active GAs. GA5 is a regulatory step in GA biosynthesis as GA5 mRNA levels are negatively regulated by its bioactive GA products. A fusion between the GA5 promoter and the firefly luciferase reporter (GA5-LUC) was shown to be similarly regulated, indicating GA feedback of GA5 occurs at the transcriptional level. The fidelity of the GA5-LUC reporter permitted a fusion genetic screen to identify mutants altered in transgene expression. This bioimaging screen identified two types of recessive mutants with increased LUC activity and apparent GA-related growth phenotypes, a dwarf (lue1) and two late flowering mutants (fpa1-3 and fpa1-4). Mutant progeny exhibited altered levels of LUC and of endogenous GA5 and other GA-regulated mRNAs. SSLP-based mapping localized lue1 to chromosome I near the ga2 locus, although complementation analyzes showed that lue1 is not allelic to ga2. Mapping and complementation analyzes showed that the late flowering mutants are allelic to fpa1. This provides genetic evidence for crosstalk between the autonomous and gibberellin-dependent flowering pathways.

Arabidopsis↗
PubMed2000

Arabidopsis map kinase 4 negatively regulates systemic acquired resistance.

Transposon inactivation of Arabidopsis MAP kinase 4 produced the mpk4 mutant exhibiting constitutive systemic acquired resistance (SAR) including elevated salicylic acid (SA) levels, increased resistance to virulent pathogens, and constitutive pathogenesis-related gene expression shown by Northern and microarray hybridizations. MPK4 kinase activity is required to repress SAR, as an inactive MPK4 form failed to complement mpk4. Analysis of mpk4 expressing the SA hydroxylase NahG and of mpk4/npr1 double mutants indicated that SAR expression in mpk4 is dependent upon elevated SA levels but is independent of NPR1. PDF1.2 and THI2.1 gene induction by jasmonate was blocked in mpk4 expressing NahG, suggesting that MPK4 is required for jasmonic acid-responsive gene expression.

Alleles↗
PubMed2000

Arabidopsis ATP A2 peroxidase. Expression and high-resolution structure of a plant peroxidase with implications for lignification.

Lignins are phenolic biopolymers synthesized by terrestrial, vascular plants for mechanical support and in response to pathogen attack. Peroxidases have been proposed to catalyse the dehydrogenative polymerization of monolignols into lignins, although no specific isoenzyme has been shown to be involved in lignin biosynthesis. Recently we isolated an extracellular anionic peroxidase, ATP A2, from rapidly lignifying Arabidopsis cell suspension culture and cloned its cDNA. Here we show that the Atp A2 promoter directs GUS reporter gene expression in lignified tissues of transgenic plants. Moreover, an Arabidopsis mutant with increased lignin levels compared to wild type shows increased levels of ATP A2 mRNA and of a mRNA encoding an enzyme upstream in the lignin biosynthetic pathway. The substrate specificity of ATP A2 was analysed by X-ray crystallography and docking of lignin precursors. The structure of ATP A2 was solved to 1.45 A resolution at 100 K. Docking of p-coumaryl, coniferyl and sinapyl alcohol in the substrate binding site of ATP A2 were analysed on the basis of the crystal structure of a horseradish peroxidase C-CN-ferulic acid complex. The analysis indicates that the precursors p-coumaryl and coniferyl alcohols are preferred by ATP A2, while the oxidation of sinapyl alcohol will be sterically hindered in ATP A2 as well as in all other plant peroxidases due to an overlap with the conserved Pro-139. We suggest ATP A2 is involved in a complex regulation of the covalent cross-linking in the plant cell wall.

Arabidopsis↗
PubMed2000

Fusion genetic analysis of gibberellin signaling mutants.

A fusion genetic strategy was used to identify gibberellin (GA) signaling mutants in transgenic Arabidopsis expressing the beta-glucuronidase (GUS) and firefly luciferase (LUC) reporter genes under control of the GA-responsive GASA1 promoter. Initial analyses determined the spatial and temporal patterns of reporter expression, and showed that reporter induction by GA was antagonized by ABA. gamma-Irradiated M2 progeny with altered reporter activities were identified by LUC bioimaging followed by GUS assays and northern hybridization of the endogenous GASA1 mRNA. Genetic analysis showed that three mutants, which overexpressed both reporters and endogenous GASA1, were caused by recessive (goe1 and goe2, for GASA over-expressed) and semi-dominant (goe3) mutations at different loci. These mutants are altered in their sensitivity to GA and the GA biosynthetic inhibitor paclobutrazol, and in the expression of several GA signaling related genes.

Base Sequence↗
PubMed2000

Calibration of plane-parallel chambers and determination of p(wall) for the NACP and Roos chambers for 60Co gamma-ray beams.

Procedures for the calibration and use of plane-parallel ionization chambers in high-energy electron and photon beams have been given in the international code of practice IAEA TRS-381. In the present work, plane-parallel ionization chambers of the type PTW-34001 Roos and Scanditronix NACP02 have been calibrated using two N(K)-based procedures. For the NACP chamber the difference between the N(D,air) chamber factors determined in an electron beam and in a 60Co gamma-ray beam, respectively, is of the same magnitude as the experimental uncertainty. Results for the PTW Roos chambers, however, do not agree, in accordance with recent findings of other authors. The value determined in a 60Co gamma-ray beam is questioned and the reason for the discrepancy assigned to the correction factor for the perturbation due to the chamber wall, p(wall). New values of p(wall) have been experimentally determined by comparing absorbed dose measurements based on air-kerma and absorbed dose to water calibration procedures. A new p(wall) factor for the Roos chamber in 60Co gamma-ray beams in water (1.009+/-0.6%) was derived as the weighted average of the different determinations. The value is not significantly higher than the p(wall) factor given in TRS-381 (1.003+/-1.5%), but the combined standard uncertainty is reduced. The chamber to chamber variation for six commercial PTW Roos chambers and a Roos prototype was found to be very small.

Air↗
PubMed2000

Influence of sulphuric acid contaminants on Fricke dosimetry.

The sulphuric acid used for the preparation of the Fricke dosimeter solution may contain trace impurities that can affect the yield of ferric ions. Two methods, pre-irradiation or oxidation with hydrogen peroxide, have been proposed to reduce the influence of these impurities. Fricke users sometimes omit this treatment. In the present work Fricke solutions prepared from six different brands and qualities of sulphuric acid were compared in order to study any influences of the acid on the ferric ion yield. It was shown that the use of analytical grade sulphuric acid from one manufacturer resulted in a reduction of the ferric ion yield of about 5% at an absorbed dose of approximately 20 Gy. If this solution were to be used for an absolute dose determination together with epsilon(m) G values from the literature the absorbed dose would be underestimated by the same amount.

Cobalt Radioisotopes↗
PubMed1999

Experimental study on the influence of the central electrode in Farmer-type ionization chambers.

In the IAEA TRS-381 protocol, k(cel) and p(cel) account for the central electrode perturbation during the air kerma chamber calibration and the in-phantom measurements. The values of these correction factors are based mainly on Monte Carlo simulations. In the present work experimental data on k(cel) and p(cel) for the NE-2571 chamber is provided, relative to a graphite electrode. In addition, the relative influence of the 3 mm diameter A-150 central electrode of the NE-2581 chamber is studied. The experimentally determined value of k(cel) for a 1 mm aluminium electrode is 1.008 +/- 0.2%, and of p(cel) in photon and electron beams 0.993 +/- 0.2% and 0.997 +/- 0.2% respectively. The experimental data and the Monte Carlo simulations agree to within 0.2%. No significant influence of the A-150 central electrode diameter on the absorbed dose determination is shown.

Air↗
PubMed1997

Practical implementation of enhanced dynamic wedge in the CadPlan treatment planning system.

The Varian CadPlan algorithm for computation of relative dose distributions and monitor unit calculations for Enhanced Dynamic Wedge (EDW) fields is based on a combination of open field beam data and Segmented Treatment data Tables. Calculation of dose by the pencil beam convolution model uses scatter kernels and boundary kernels to create the distribution. The principles of the pencil beam convolution model is presented. Comparison of measured and calculated monitor units and relative dose distributions showed good agreement and the deviations are within international accepted tolerans. Test results indicate that the EDW model works satisfactorily for all energies and wedge angles.

Particle Accelerators↗
PubMed1997

Fluence non-uniformity effects in air kerma determination around brachytherapy sources.

In ionization chamber dosimetry close to brachytherapy sources, the non-uniform photon fluence in the vicinity of the air cavity of the ionization chamber must be corrected for. In this paper, experimentally determined relative non-uniformity correction factors are determined for a PTW23331 chamber and a number of Farmer-type chambers. It is concluded that the 'anisotropic' theory of Bielajew agrees better with the experimental results than the 'isotropic' theory of Kondo and Randolph. The experiments show that neither the material choice nor the radius of the central electrode have any significant effect on the non-uniformity correction factors. Experiments on the wall-material dependence in the non-uniformity correction factor, which is predicted by the anisotropic theory, were inconclusive. Parameters for the Farmer-type chambers for calculation of theoretical non-uniformity correction factors are given. Data to derive anisotropic fluence non-uniformity correction factors are given in tabular form. An error in the Kondo and Randolph equations is reported and the corrected equations presented.

Biophysical Phenomena↗
PubMed1996

Structure and organ specificity of an anionic peroxidase from Arabidopsis thaliana cell suspension culture.

The predominant peroxidase (pI 3.5) (E.C. 1.11.1.7) of an Arabidopsis thaliana cell suspension culture was purified and partially sequenced. Oligonucleotides were designed and a specific probe was obtained. A cDNA clone was isolated from an Arabidopsis cell suspension cDNA library and completely sequenced. The cDNA clone comprised 1194 bp and encodes a 30 residue signal peptide and a 305 residue mature protein (Mr 31,966). The sequence of the mature protein is 95% identical to the well-characterized horseradish peroxidase HRP A2 and has therefore been designated ATP A2. Three introns at positions identical to those found in Arabidopsis and horseradish genes encoding cationic peroxidases were identified. RT-PCR analysis revealed root-specific expression.

Amino Acid Sequence↗
PubMed1995

Ionization chamber dosimetry of proton beams using cylindrical and plane parallel chambers. Nw versus Nk ion chamber calibrations.

Determinations of the absorbed dose in a 170 MeV proton beam have been performed using seven ionization chambers of different types: five cylindrical (two FWT IC-18 and three NE-2571, of which one was modified to have the central electrode made of graphite) and two plane parallel (NACP-02 and Roos FK-6). The ionization was converted into absorbed dose in the proton beam according to the generalization of the formalism provided by the IAEA Code of Practice (TRS 277), which enables the use of the same equations for all kinds of beam used in radiotherapy. The absorbed dose obtained with the two IC-18 chambers, a chamber type commonly used as a reference in proton beams, was up to 1.5% lower than that obtained with the Farmer NE-2571 chamber, which was used as the reference in this work when calibration factors in terms of NK were used. To investigate this difference, experimental ND factors for six chambers (the two IC-18 chambers, the NACP-02, the FK-6 and two of the NE-2571 chambers) were determined in a high-energy electron beam. The procedure commonly recommended for plane parallel ion chambers was used for all the chambers, using the same reference chamber, a Farmer NE-2571. In the 170 MeV proton beam all the ND factors yielded consistent absorbed dose determinations within the estimated experimental uncertainties. This finding calls into question the value of the product kattkm for the IC-18 chamber given by the IAEA Code of Practice used in this comparison, and points at possible chamber to chamber variations that theoretical kattkm factors cannot predict. The investigations enabled the determination of the Pwall(60Co) factor of the Roos FK-6 plane parallel chamber, yielding 1.003 +/- 0.5%, and a correction for the effect of the aluminium central electrode of NE-2571 chambers in proton beams, equal to 1.003 +/- 0.4%. Two of the chambers (the plane parallel FK-6 and the modified cylindrical NE-2571) were provided with calibration factors in terms of absorbed dose to water, Nw, at the quality of 60Co by the Primary Standard Dosimetry Laboratory in Germany (PTB). Using the Nw formalism excellent agreement was found with the determination based on the experimental ND, giving support to the implementation of the NW procedure in therapeutic proton beams.

Humans↗
PubMed1992

DNA methylation is involved in maintenance of an unusual expression pattern of an introduced gene.

In one of 30 transgenic tobacco (Nicotiana tabacum) plants, the expression of an introduced beta-glucuronidase (GUS) gene driven by the cauliflower mosaic virus 35S promoter, was found to be repressed as the plant matured, whereas the endogenous GUS activity was unaffected. Plants grown from seeds or regenerated from leaf discs derived from this plant showed a similar temporal pattern of expression. Suspension-cultured cells established from nonexpressing leaves did not express the introduced gene. In these cells, the silent gene could be reactivated by treatment for 5 or 10 days with 5-azacytidine. Overall, demethylation of the genome preceded recovery of the enzyme activity. The increase in the fraction of reactivated cells progressed in two phases. Up to 8 weeks after starting the 5-azacytidine treatment, approximately 2 to 4% of the cells were expressing GUS, followed by a dramatic increase of GUS-expressing cells. Thirteen weeks after starting the 5-azacytidine treatment, the fraction of GUS-expressing cells amounted to 80%. At this time, the original overall level of DNA methylation was reestablished. The degree of DNA demethylation, as well as the magnitude of reactivation, was dependent on the duration of the 5-azacytidine treatment. These results demonstrate that DNA methylation appears to be involved in the regulation of the introduced GUS gene and that this development-dependent pattern of expression can be inherited.

Journal Article↗
PubMed1991

Specific Levels of DNA Methylation in Various Tissues, Cell Lines, and Cell Types of Daucus carota.

The level of DNA methylation in Daucus carota was found to be tissue specific, but no simple correlation between developmental stage or age of tissue and the level of DNA methylation was found. Among three different suspension culture lines from the same variety grown under identical conditions, large differences in the level of DNA methylation were observed. The highest and lowest levels were found in two embryogenic cell lines originating from the same clone. Suspension cells from one of the embryogenic cell lines were fractionated into three morphologically defined cell types using Percoll gradient density centrifugation, and the uniformity of these fractions was evaluated by image analysis. The three cell types showed different levels of DNA methylation. The lowest level was found in the fraction containing the precursor cells of somatic embryos.

Journal Article↗
PubMed1990

Employment of hydrolytic enzymes in the study of the level of DNA methylation.

A new method for the determination of the level of DNA methylation was established. The method involves enzymatic hydrolysis of DNA by nuclease P1 and bacterial alkaline phosphatase, and separation of the resulting deoxyribonucleosides by HPLC. By this method, DNA was hydrolysed completely to the five deoxyribonucleosides and the complete base composition was determined. Pairing bases were shown to occur in similar amounts, and analysis could be performed on as little as 1 microgram of DNA with a high degree of reproducibility. Among other enzymes hitherto used in order to hydrolyze DNA, micrococcal nuclease, phosphodiesterase II and nuclease P1 have been shown to cause deamination of deoxyadenosine, while deoxyribonuclease I, phosphodiesterase I and bacterial alkaline phosphatase have been shown to be sensitive to contamination by RNA, and to release 5-methyldeoxycytidine at a slower rate than the other four deoxyribonucleosides. Neither of these effects was seen with the new method.

Alkaline Phosphatase↗
PubMed1990

Absorbed dose in water. Comparison of several methods using a liquid ionization chamber.

In the present investigation a liquid ionization chamber has been used as a transfer instrument for the quantity absorbed dose in water in a cobalt-60 gamma-ray beam. The characteristics of the liquid ionization chamber are described. The transferred dosimetric information has been compared with absorbed-dose determination using air-ionization-chamber dosimetry, water calorimetry and ferrous-sulphate dosimetry. The agreement between the different measured absorbed-dose values is very good, i.e. within 0.2%. This is an indication that the consistency in the methods used to determine absorbed dose in water is good. The impact of the new standard for air kerma in air, introduced in 1986 by the BIPM, on the air-ionization-chamber dosimetry is investigated. It is shown that any differences in the dosimetry when using the old or the new set of data cancel out for the cobalt-60 beam. The investigation also shows that the value of epsilon mG for the ferrous-sulphate dosimeter recommended in ICRU 35 for electrons can be used also in cobalt-60 beams.

Calibration↗