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M Karlsson

Publications and source records attributed to M Karlsson.

At least 73 records · Page 4Linked to original sources
PubMed2000

Expression, purification, and characterization of histidine-tagged mouse monoglyceride lipase from baculovirus-infected insect cells.

Monoglyceride lipase (MGL) has been produced with the baculovirus-insect cell system. The mouse MGL cDNA was subcloned into a baculovirus transfer vector in frame with a sequence encoding an N-terminal stretch of six histidine residues. Purification to apparent homogeneity was obtained by nickel-chelating chromatography. The final yield was 3 mg of pure enzymatically active MGL per liter of Sf9 cell suspension culture. Analysis by SDS-PAGE and mass spectrometry showed that the recombinant histidine-tagged enzyme had the expected molecular mass. With monoolein as substrate, the specific activity and the apparent K(m) were close to those of rat MGL of adipose tissue.

Adipose Tissue↗
PubMed2000

The impact of respiratory variables on mortality in non-ARDS and ARDS patients requiring mechanical ventilation.

OBJECTIVES: Primarily, to determine if respiratory variables, assessed on a daily basis on days 1-6 after ICU admission, were associated with mortality in non-ARDS and ARDS patients with respiratory failure requiring mechanical ventilation. Secondarily, to determine non-respiratory factors associated with mortality in ARDS and non-ARDS patients. DESIGN: Prospective multicentre clinical study. SETTING: Seventy-eight intensive care units in Sweden and Iceland. PATIENTS: Five hundred twenty non-ARDS and 95 ARDS patients. MEASUREMENTS AND RESULTS: Potentially prognostic factors present at inclusion were tested against 90-day mortality using a Cox regression model. Respiratory variables (PaO2/FIO2, PEEP, mean airway pressure (MAP) and base excess (BE)) were tested against mortality using the model. Primary aim: in non-ARDS a low PaO2/FIO2 on day 1, RR (risk ratio) = 1.17, CI (95% confidence interval) (1.00; 1.36), day 4, 1.24 (1.02; 1.50), day 5, 1.25 (1.02; 1.53) and a low MAP at baseline, 1.18 (1.00; 1.39), day 2, 1.24 (1.02; 1.52), day 3, 1.33 (1.06; 1.67), day 6, 2.38 (1.11; 5.73) were significantly associated with 90-day death. Secondary aim: in non-ARDS a low age, RR = 0.77 (0.67; 0.89), female gender, 0.85 (0.74; 0.98), and low APS (acute physiologic score), 0.85 (0.73; 0.99), were associated with survival; chronic disease, 1.31 (1.12; 1.52), and non-pulmonary origin to the respiratory failure, 1.27 (1.10; 1.47), with death. In ARDS low age, RR = 0.65 CI (0.46; 0.91), and low APS, 0.65 (0.46; 0.90), were associated with survival. CONCLUSIONS: No independent significant association was seen between 90-day mortality and degree of hypoxaemia, PEEP, MAP or BE for the first full week of ICU care in either ARDS or non-ARDS. In a sub-group of non-ARDS a lower PaO2/FIO2 and MAP tended to influence mortality where a significant association was seen for 3 of 7 study days. Age, gender, APS, presence of a chronic disease and a pulmonary/non-pulmonary reason for the respiratory failure were associated with mortality in non-ARDS, while only age and APS showed a similar association in ARDS.

APACHE↗
PubMed2000

Occurrence of multiple ribonucleotide reductase classes in gamma-proteobacteria species.

Ribonucleotide reductase (RNR) is central to de novo synthesis of deoxyribonucleotides and essential for all living cells. Three classes have been described; class I is oxygen dependent and represented by two subclasses, Ia (NrdAB) and Ib (NrdEF); class II (NrdJ) is indifferent to oxygen; and class III (NrdDG) is oxygen sensitive. More than one class can be found in an organism, reflecting the oxygen status of its environment. We have investigated, by using PCR and Southern blot, the occurrence of the different classes among species of the gamma-Proteobacteria. Class III are present in all species tested, but the presence of the other classes varies. Some species contain one unique additional enzyme, class Ia, Ib, or II, whereas others contain two additional enzymes, class Ia and Ib, or class Ia and II.

Amino Acid Sequence↗
PubMed2000

Optimized MLC-beam arrangements for tangential breast irradiation.

BACKGROUND AND PURPOSE: Very large numbers of women are treated with tangential breast irradiation after breast-conserving surgery due to mammary carcinoma. The aim of this study was to improve a conventional treatment plan by modifying the dose intensity in the beams to reduce the absorbed dose outside the planning target volume (PTV) and to reduce the absorbed dose variation inside the PTV diotherapy of mammary carcinoma. MATERIALS AND METHODS: Treatment planning was performed both with conventional technique and with a simple intensity modulation technique for 12 consecutive patients. RESULTS: In all cases a higher degree of dose conformity was obtained with the dose intensity modulation technique. The relative gain was found to be similar for all patients irrespective of the size of the target volume or the irradiated lung volume. CONCLUSION: Simple manual intensity modulation can be used to improve the dose distribution in tangential breast irradiation. With modern accelerators the increased time for this technique is less than 2 min per fraction.

Breast Neoplasms↗
PubMed2000

Analysis of the Look-Locker T(1) mapping sequence in dynamic contrast uptake studies: simulation and in vivo validation.

An alternative to the pulse sequences at present used in dynamic contrast uptake MRI is the dynamic LL-EPI T(1) mapping method. This method generates T(1) estimates in a few seconds, thereby allowing dynamic studies. A particular advantage of the LL-EPI technique is that it provides the opportunity to generate spatial and temporal information about the paramagnetic contrast agent concentration independently of the inflow rate. This paper illustrates, by computer simulations, the accuracy of the estimated 1/T(1) value when using the LL-EPI technique in situations that are not supported by the model. The simulated situations not supported by the model are those in which the longitudinal and transversal relaxation rates change during the T(1) mapping. The most critical moment occurs during a bolus passage of contrast agent when the concentration gradient is large. The computer simulations of the LL-EPI T(1) mapping method in non-supported situations show that in normal perfused capillary tissue the error in the estimated 1/T(1) value is within the absolute error of 0.1 s(-1) in most simulated situations, although in a typical vessel the simulations do indicate that the stated absolute error tolerance of 0.5 s(-1) is exceeded relatively easily. However, this transgression can be rectified by a non-bolus injection of the contrast agent media.

Brain↗
PubMed2000

An abundant TIP expressed in mature highly vacuolated cells.

Aquaporins are water channel proteins found in vacuolar membranes and plasma membranes, and belong to the major intrinsic protein (MIP) family of proteins. In the present study, we purified a 75 kDa MIP protein from a crude fraction of spinach leaf intracellular membranes. Upon urea/SDS-PAGE, the 75 kDa protein appeared as a 21 kDa polypeptide, and the 75 kDa species therefore probably represents a tetramer. The corresponding cDNA was obtained by PCR cloning and had an open reading frame encoding a 25.1 kDa protein. The protein, So-deltaTIP, was most homologous to the tonoplast intrinsic protein (TIP) subfamily of plant MIPs. Using affinity-purified So-deltaTIP-specific peptide antibodies, we investigated the subcellular and tissue distribution of So-deltaTIP. So-deltaTIP was specifically located in the vacuolar membrane. It was abundant in most vacuolated cells in all vegetative organs, but was excluded from the leaf epidermis as well as from the root phloem parenchyma and meristem. In spite of the high sequence homology between delta-TIPs of spinach, Arabidopsis, sunflower and radish, their expression patterns were totally different. However, a comparison of the expression pattern of So-deltaTIP with that of more distantly related TIPs showed similarities with Arabidopsis gamma-TIP, which is expressed in zones of cell elongation/differentiation but excluded from meristematic tissues. Meristematic cells are characterized by many small vacuoles as opposed to elongating and mature cells, which generally harbour a single, large vacuole. Our results indicate that the expression of So-deltaTIP may be induced when the large vacuole is formed.

Amino Acid Sequence↗
PubMed2000

Assessment of regional longitudinal myocardial strain rate derived from doppler myocardial imaging indexes in normal and infarcted myocardium.

Myocardial deformation properties may be characterized by regional strain rates (SRs) calculated from Doppler myocardial velocity data. In 10 control subjects and 12 patients with established transmural infarcts, longitudinal median segmental SR, strain, and myocardial velocity were analyzed and compared with the corresponding wall motion score. All segments in control subjects and normal segments in infarct patients showed no significant difference in either systolic or diastolic SR (systolic: -1.27+/-0.39 s(-1) versus -1.23+/-0.24 s(-1), not significant [NS]; and isovolumic relaxation [IVR]: 1.23+/-0.38 s(-1) versus 1.95+/-0.62 s(-1), NS; respectively) and strain (-0.21+/-0.06 versus -0.19+/-0.06, NS). In infarcted segments, peak systolic SR, systolic strain, and early diastolic SR showed the most pronounced reduction (hypokinetic and akinetic) or even inversion (dyskinetic segments: 0.10+/-0.26 s(-1), 0.00+/-0.03, and -1.78+/-0.67 s(-1), respectively; P<.001). In this study, new myocardial deformation indexes were shown to quantitatively describe the function of normal and chronically infarcted regions.

Adult↗
PubMed2000

The cytoplasmic tyrosines of integrin subunit beta1 are involved in focal adhesion kinase activation.

We have previously shown that mutation of the two tyrosines in the cytoplasmic domain of integrin subunit beta1 (Y783 and Y795) to phenylalanines markedly reduces the capability of beta1A integrins to mediate directed cell migration. In this study, beta1-dependent cell spreading was found to be delayed in GD25 cells expressing beta1A(Y783/795F) compared to that in wild-type GD25-beta1A. Focal adhesion kinase (FAK) tyrosine phosphorylation and activation were severely impaired in response to beta1-dependent adhesion in GD25-beta1A(Y783/795F) cells compared to that in wild-type GD25-beta1A or mutants in which only a single tyrosine was altered (beta1A(Y783F) or beta1A(Y795F)). Phosphorylation site-specific antibodies selective for FAK phosphotyrosine 397 indicated that the defect in FAK phosphorylation via beta1A(Y783/795F) lies at the level of the initial autophosphorylation step. Indeed, beta1A-dependent tyrosine phosphorylation of tensin and paxillin was lost in the beta1A(Y783/795F) cells, consistent with the impairment in FAK activation. In contrast, p130(CAS) overall tyrosine phosphorylation was unaffected by the beta1 mutations. Despite the defect in beta1-mediated FAK activation, FAK was still localized to focal adhesions. Taken together, the phenotype of the GD25-beta1A(Y783/795F) cells resembles, but is distinct from, the phenotype observed in FAK-null cells. These observations argue that tyrosines 783 and 795 within the cytoplasmic tail of integrin subunit beta1A are critical mediators of FAK activation and cell spreading in GD25 cells.

Amino Acid Substitution↗
PubMed2000

The catechol-O-methyltransferase (COMT) inhibitor entacapone enhances the pharmacokinetic and clinical response to Sinemet CR in Parkinson's disease.

OBJECTIVES: Entacapone is a specific, potent, peripherally acting catechol-O-methyltransferase (COMT) inhibitor. It has been shown to improve the bioavailability of plasma levodopa and extend its clinical effect when used as an adjunct to standard levodopa preparations, but there is little experience of the effect of entacapone on controlled release levodopa preparations. METHODS: A double blind, placebo controlled, single dose, randomised, cross over trial was performed in 14 patients with Parkinson's disease with motor fluctuations to investigate the clinical effect of a single dose of entacapone (200 mg) when administered with either standard levodopa-carbidopa (Sinemet) or controlled release levodopa-carbidopa preparations (Sinemet CR). RESULTS: When entacapone was administered with standard Sinemet the duration of the clinical response to standard Sinemet was longer in comparison with the response after placebo (p=0.02). Moreover, in the same patients, entacapone significantly increased the duration of the clinical response to Sinemet CR (p=0.05) without prolonging the latency of response or enhancing dyskinesias. CONCLUSIONS: These data confirm the clinical efficacy of entacapone-standard Sinemet combination. They also indicate that adding entacapone to controlled release levodopa preparations might provide a useful treatment option in patients with Parkinson's disease with motor fluctuations. A double blind clinical trial with a chronically administered entacapone-Sinemet CR combination is, however, required to verify this viewpoint.

Adult↗
PubMed1999

Genetic basis of macrolide and lincosamide resistance in Brachyspira (Serpulina) hyodysenteriae.

Macrolide antibiotic resistance is widespread among Brachyspira hyodysenteriae (formerly Serpulina hyodysenteriae) isolates. The genetic basis of macrolide and lincosamide resistance in B. hyodysenteriae was elucidated. Resistance to tylosin, erythromycin and clindamycin in B. hyodysenteriae was associated with an A-->T transversion mutation in the nucleotide position homologous with position 2058 of the Escherichia coli 23S rRNA gene. The nucleotide sequences of the peptidyl transferase region of the 23S rDNA from seven macrolide and lincosamide resistant and seven susceptible strains of Brachyspira spp. were determined. None of the susceptible strains were mutated whereas all the resistant strains had a mutation in position 2058. Susceptible strains became resistant in vitro after subculturing on agar containing 4 micrograms ml-1 of tylosin. Sequencing of these strains revealed an A-->G transition mutation in position 2058.

Anti-Bacterial Agents↗
PubMed1999

Effect of latanoprost on regional blood flow and capillary permeability in the monkey eye.

OBJECTIVE: To evaluate the effects of latanoprost on regional blood flow and capillary permeability in the monkey eye. METHODS: Anesthetized cynomolgus monkeys were unilaterally treated with a single dose containing 6 pg of latanoprost; or 10 microg of PhXA34 (13,14-dihydro-15R, S-17-phenyl-18,19,20-trinor-prostaglandin F2alpha [PGF2alpha]-isopropyl ester), which contains about 50% latanoprost. Regional blood flow in the eye was measured with radioactively labeled microspheres; capillary permeability was measured by determining the extravascular plasma-equivalent albumin space using 125I-albumin, 131I-albumin, and 51Cr-labeled erythrocytes. RESULTS: Latanoprost or PhXA34 had no or only a slight effect on the regional blood flow when measured 1, 2 1/2, 3, 4 1/2, and 6 hours after dose administration, with the exception of the anterior sclera, in which a moderate increase in blood flow was detected. No effect on capillary permeability to albumin was detected when studied 30 minutes to 2 1/2 hours and 5 to 6 hours after dose administration. CONCLUSION: Latanoprost, a selective prostaglandin F receptor agonist, exerted no or only slight vascular effects for up to 6 hours after dose administration in the monkey eye, with the exception of the anterior sclera, in which a moderate increase in blood flow was detected. CLINICAL RELEVANCE: Naturally occurring prostaglandins may cause marked microcirculatory changes in the eye that could be of clinical concern. Latanoprost, a selective prostaglandin F receptor agonist, seems to be devoid of such effects.

Administration, Topical↗
PubMed1999

Particle trace visualization of intracardiac flow using time-resolved 3D phase contrast MRI.

The flow patterns in the human heart are complex and difficult to visualize using conventional two-dimensional (2D) modalities, whether they depict a single velocity component (Doppler echocardiography) or all three components in a few slices (2D phase contrast MRI). To avoid these shortcomings, a temporally resolved 3D phase contrast technique was used to derive data describing the intracardiac velocity fields in normal volunteers. The MRI data were corrected for phase shifts caused by eddy currents and concomitant gradient fields, with improvement in the accuracy of subsequent flow visualizations. Pathlines describing the blood pathways through the heart were generated from the temporally resolved velocity data, starting from user-specified locations and time frames. Flow trajectories were displayed as 3D particle traces, with simultaneous demonstration of morphologic 2D slices. This type of visualization is intuitive and interactive and may extend our understanding of dynamic and previously unrecognized patterns of intracardiac flow.

Heart↗
PubMed1999

The correlation between R2' and bone mineral measurements in human vertebrae: an in vitro study.

The aim of this study was to investigate whether MR imaging of trabecular bone structure using magnetic inhomogeneity measurements is related to the amount of bone mineral in human vertebrae. Weight, bone mineral content (BMCDXA), bone mineral per area (BMADXA) and bone mineral density (BMDCT) were determined in 12 defatted human lumbar vertebrae (L2-L4) by weighing, dual X-ray absorptiometry (DXA) and CT. Inhomogeneity caused by susceptibility differences between trabecular bone and surrounding water was studied with MR imaging at 1.5 T using the GESFIDE sequence. The pulse sequence determines the transverse relaxation rate R2(*) and its two components, the non-reversible transverse relaxation rate (R2) and the reversible transverse relaxation rate (R2'; i. e. relaxation rate due to magnetic susceptibility) in a single scan. Voxel size was 0.9 x 1.9 x 5.0 mm. Positive significant correlations between R2' and weight, BMCDXA, BMADXA and BMDCT were observed (r > 0.61 and p < 0.05 for all). Unexpectedly, R2 was also positively correlated with weight, BMCDXA and BMDCT (r > 0.66 and p < 0.05 for all), but not with BMADXA. Thus, R2' measurements are related to the amount of bone mineral, but they also provide information which is not obtainable from bone mineral measurements.

Absorptiometry, Photon↗
PubMed1999

Orientation of trabecular bone in human vertebrae assessed by MRI.

The aim of the study was to assess the relative number of bone trabeculae in different orientations by using magnetic inhomogeneity measurements by MR imaging. Twelve defatted human vertebrae (L2-L4) were studied by MR imaging and CT. In the MR measurements the reversible transaxial decay rate, R2', was determined using the GESFIDE sequence. The relative contribution to R2' of bone trabeculae oriented along the x (R2'x), y (R2'y) and z (R2'y) axes was assessed, by rotation of the specimen in the magnetic field. The results were validated by CT measurements of trabecular structure at a resolution of 0.2 x 0.2 x 1 mm. using custom-made software. R2' ranged from 4.9 to 32 s-1. After separation, theoretical R2'x ranged from 2.3 to 10.7 s-1, R2'y ranged from 2.6 to 14.0 s-1 and R2'z ranged from 4.7 to 17.9 s-1. The number of bone trabeculae per millimeter identified in axial CT images ranged from 0.15 to 0.38 and from 0.039 to 0.22 per millimeter in sagittal images. The difference between axial and sagittal CT images was statistically significant and due to anisotropic voxels. The correlation between R2' and the number of bone trabeculae per millimeter was statistically significant (r = 0.83, p < 0.001 for x orientation: r = 0.63 for y orientation, and r = 0.59 for z orientation, p < 0.05 for both). The relative amount of bone trabeculae in different orientations can be assessed in vitro using R2' measurements.

Aged↗
PubMed1999

Initiation of Escherichia coli ribosomes on matrix coupled mRNAs studied by optical biosensor technique.

The optical biosensor technique, based on the surface plasmon resonance (SPR) phenomenon, has been used to study the initiation of protein synthesis by E. coli ribosomes on surface coupled mRNA. mRNA was first periodate oxidized and then hydrazide coupled to the surface of a CM5 sensor chip. The formation of initiation complexes on the surface coupled mRNA was monitored in real-time with a BIACORE 2000 instrument. Mature 70S*mRNA*fMet-tRNA(Met) initiation complexes were assembled on mRNA by sequential introduction of the 30S and 50S subunits supplemented with appropriate initiation factors and fMet-tRNA(Met). We show that the formation of 70S*mRNA complexes on the surface coupled mRNA proceeds efficiently only in the presence of tRNA. Moreover, 70S*mRNA*fMet-tRNA(Met) complexes formed with fMet-tRNA(Met) are more stable than similar complexes formed with deacylated tRNAs. The efficient formation and slow dissociation of mature 70S*mRNA*fMet-tRNA(Met) initiation complexes are most easily explained by the stabilization of the interaction of the ribosomal subunits by fMet-tRNA(Met). This work demonstrates the feasibility of the BIACORE technique for studying the initiation of protein synthesis.

Base Sequence↗
PubMed1999

Emended descriptions of indole negative and indole positive isolates of Brachyspira (Serpulina) hyodysenteriae.

Two type/reference strains of Brachyspira (B.) hyodysenteriae, 14 Belgian and German indole negative, and 14 Belgian, German and Swedish indole positive field isolates of strongly beta-haemolytic intestinal spirochaetes were compared by pulsed-field gel electrophoresis (PFGE) patterns, biochemical reaction patterns, 16S rDNA sequences and MIC determinations of six antibacterial substances. Three tests for indole production, including a spot indole test, were compared with congruent results. All field isolates were classified as B. hyodysenteriae due to a high genetic and phenotypic similarity with the type strains. The Belgian and German indole negative isolates had identical and unique PFGE patterns for the tested restriction enzymes MluI and SalI, as well as identical 16S rDNA sequences, and they could not be differentiated by any of the methods used. Seven unique PFGE patterns were achieved from the 14 indole positive field isolates. The patterns were identical and unique for epidemiologically related isolates. Type/reference strains and isolates without known relation to other tested isolates showed unique banding patterns. The MICs of tylosin, tiamulin, erythromycin, clindamycin, carbadox and virginiamycin were determined in broth for all isolates. In contrast to Belgian and German isolates, the majority of the Swedish field isolates were susceptible to tylosin, erythromycin and clindamycin. Probable pathways of infection for some of the Swedish isolates were determined. The PFGE patterns of epidemic clones of B. hyodysenteriae remained stable for a period of up to 8 years. In vivo development of resistance to macrolide and lincosamide antibiotics due to use of tylosin was clearly indicated for two epidemic clones.

Animals↗
PubMed1999

Phantom and in vivo study of the Look-Locher T1 mapping method.

This paper describes and tests the LL-EPI method for obtaining quantitative T1 estimates in a few seconds thereby allowing dynamic T1 studies. It is shown that the method works even when there is an inflow into the imaged volume, e.g., in a vessel. No calibration is needed. The method has been tested in a phantom study with several different scan parameter set-ups, with and without inflow. The method shows robustness and individual scan parameters and inflow rates do not influence the ability to calculate the Gd-DTPA concentration. Linearity prevail between the measured 1/T1 and the Gd-DTPA concentration in the range 150 < T1 < 2500 ms. In a dynamic Gd-DTPA phantom study, it was shown that the dynamic LL-EPI T1 mapping technique was three times more sensitive than the signal from a T2*-weighted EPI sequence. In an in vivo study, dynamic T1 mapping of the Gd-DTPA uptake in a meningioma was performed. Inspection of the uptake curves indicates that the method is feasible in clinical perfusion studies.

Brain↗