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K Lindström

Publications and source records attributed to K Lindström.

At least 55 records · Page 3Linked to original sources

Glomerular charge selectivity for horseradish peroxidase and albumin at low and normal ionic strengths.

The classical concept of a negative glomerular charge barrier has recently been questioned, mainly based on the somewhat high clearance for anionic horseradish peroxidase (HRP). The validity of using anionic HRP can be tested by changing the properties of the charge barrier. A rather unequivocal approach is to reduce the ionic composition of the perfusate and hence increase the Debye length. We determined the glomerular clearance for horseradish peroxidase and serum albumin, using isolated rat kidneys perfused at 8 degrees C to reduce the tubular modification of the primary urine. The perfusate contained trace amounts of the neutral 125I-nHRP and the anionic 131I-aHRP and were otherwise identical except for different ionic strengths, 152 mM and 34 mM, respectively. During control, the fractional clearance (theta) was 0.11 +/- 0.015 for nHRP and 0.045 +/- 0.010 for aHRP, with an average clearance ratio (n/a) of 2.8 +/- 0.24. Low ionic strength reduced theta for aHRP to 0.027 +/- 0.006, giving an increased clearance ratio for HRP of 4.2 +/- 0.44. The existence of a negative charge barrier is supported by the experiments. The result obtained during normal perfusion is compatible with a charge density (omega) of 34 mEq L-1, using a model of homogeneously charged membrane. Low ionic strength perfusion reversibly reduced the concentration of fixed charges to 12 mEq L-1, suggesting an almost threefold increase of the glomerular membrane volume. Thus, the glomerular charge barrier should be regarded to have a dynamic gel structure rather than being a rigid membrane.

Animals↗

[3H]SCH 58261, a selective adenosine A2A receptor antagonist, is a useful ligand in autoradiographic studies.

We have characterized the new potent and selective nonxanthine adenosine A2A receptor antagonist SCH 58261 as a new radioligand for receptor autoradiography. In autoradiographic studies using agonist radioligands for A2A receptors ([3H]CGS 21680) or A1 receptors (N6-[3H]cyclohexyladenosine), it was found that SCH 58261 is close to 800-fold selective for rat brain A2A versus A1 receptors (Ki values of 1.2 nM versus 0.8 microM). Moreover, receptor autoradiography showed that [3H]SCH 58261, in concentrations below 2 nM, binds only to the dopamine-rich regions of the rat brain, with a K(D) value of 1.4 (0.8-1.8) nM. The maximal number of binding sites was 310 fmol/mg of protein in the striatum. Below concentrations of 3 nM, the nonspecific binding was <15%. Three adenosine analogues displaced all specific binding of [3H] SCH 58261 with the following estimated Ki values (nM): 2-hex-1-ynyl-5'-N-ethylcarboxamidoadenosine, 3.9 (1.8-8.4); CGS 21680, 130 (42-405); N6-cyclohexyladenosine, 9,985 (3,169-31,462). The binding of low concentrations of SCH 58261 was not influenced by either GTP (100 microM) or Mg2+ (10 mM). The present results show that in its tritium-labeled form, SCH 58261 appears to be a good radioligand for autoradiographic studies, because it does not suffer from some of the problems encountered with the currently used agonist radioligand [3H]CGS 21680.

Adenosine↗

Phylogeny of Rhizobium galegae with respect to other rhizobia and agrobacteria.

PCR-RFLP with nine restriction enzymes was applied to the 16S and 23S rRNA genes of 42 rhizobial and agrobacterial strains to determine the phylogenetic position of Rhizobium galegae and increase the understanding of the evolution of ribosomal operons. The strains were selected based on previous phylogenetic studies. PCR primers were designed so that they amplified a 2.3 kb fragment of the 23S rRNA gene (excluding the B8 loop). Universal primers rD1 and fD1 were used to amplify the full-length 16S rRNA. The RFLP analysis resulted in 27 and 32 different restriction patterns for 16S and 23S, respectively. The RFLP patterns were transformed to genetic distances and dendrograms were constructed from the data using the unweighted pair group method with averages. The shapes of the dendrograms derived from the analysis of the 16S and 23S rRNA genes correlated well, with only a few strains having different positions. The 23S tree generally had deeper branching than the 16S tree, allowing better discrimination between species and strains. The combined data from the two analyses described 36 genotypes. The eight R. galegae strains formed a homogeneous cluster in all dendrograms. The RFLP analysis was confirmed by partial sequence analysis of the 16S rRNA gene (the first 800 bp), which correlated well with full-length 16S rRNA sequence analysis. The 16S data placed R. galegae near the genus Agrobacterium with Agrobacterium vitis as its nearest neighbour, whereas in the 23S and the combined dendrograms it showed closer affinity to the genus Rhizobium.

Base Sequence↗

Characterization of tropical tree rhizobia and description of Mesorhizobium plurifarium sp. nov.

A collection of strains isolated from root nodules of Acacia species in Senegal was analysed previously by electrophoresis of total cell protein, auxanographic tests, rRNA-DNA hydridization, 16S rRNA gene sequencing, DNA base composition and DNA-DNA hybridization [de Lajudie, P., Willems, A., Pot, B. & 7 other authors (1994). Int J Syst Bacteriol 44, 715-733]. Strains from Acacia were shown to belong to two groups, Sinorhizobium terangae, and a so-called gel electrophoretic cluster U, which also included some reference strains from Brazil. Further taxonomic characterization of this group using the same techniques plus repetitive extragenic palindromic-PCR and nodulation tests is presented in this paper. Reference strains from Sudan and a number of new rhizobia isolated from nodules of Acacia senegal, Acacia tortilis subsp. raddiana and Prosopis juliflora in Senegal were included. As a result of this polyphasic approach, the creation of a new species, Mesorhizobium plurifarium, is proposed for a genotypically and phenotypically distinct group corresponding to the former cluster U and containing strains isolated from Acacia, Leucaena, Prosopis and Chamaecrista in West Africa (Senegal), East Africa (Sudan) and South America (Brazil). The type strain of Mesorhizobium plurifarium ORS 1032 has been deposited in the LMG collection as LMG 11892.

Acacia↗

Three phylogenetic groups of nodA and nifH genes in Sinorhizobium and Mesorhizobium isolates from leguminous trees growing in Africa and Latin America.

The diversity and phylogeny of nodA and nifH genes were studied by using 52 rhizobial isolates from Acacia senegal, Prosopis chilensis, and related leguminous trees growing in Africa and Latin America. All of the strains had similar host ranges and belonged to the genera Sinorhizobium and Mesorhizobium, as previously determined by 16S rRNA gene sequence analysis. The restriction patterns and a sequence analysis of the nodA and nifH genes divided the strains into the following three distinct groups: sinorhizobia from Africa, sinorhizobia from Latin America, and mesorhizobia from both regions. In a phylogenetic tree also containing previously published sequences, the nodA genes of our rhizobia formed a branch of their own, but within the branch no correlation between symbiotic genes and host trees was apparent. Within the large group of African sinorhizobia, similar symbiotic gene types were found in different chromosomal backgrounds, suggesting that transfer of symbiotic genes has occurred across species boundaries. Most strains had plasmids, and the presence of plasmid-borne nifH was demonstrated by hybridization for some examples. The nodA and nifH genes of Sinorhizobium teranga ORS1009T grouped with the nodA and nifH genes of the other African sinorhizobia, but Sinorhizobium saheli ORS609T had a totally different nodA sequence, although it was closely related based on the 16S rRNA gene and nifH data. This might be because this S. saheli strain was originally isolated from Sesbania sp., which belongs to a different cross-nodulation group than Acacia and Prosopis spp. The factors that appear to have influenced the evolution of rhizobial symbiotic genes vary in importance at different taxonomic levels.

Acyltransferases↗

A rapid PCR-based DNA test for enterotoxic Bacillus cereus.

The occurrence of DNA sequences encoding the hemolysin HblA complex and Bacillus cereus enterotoxin BceT, which have recently been confirmed as enterotoxins, was studied in Bacillus spp. To amplify these DNA sequences, PCR primer systems for the B component of hblA and for bceT DNA sequences were developed. The results from the amplification of hblA sequences correlated well with results obtained with the B. cereus enterotoxin (diarrheal type) test kit (RPLA kit), but not with the results of the Bacillus diarrheal enterotoxin visual immunoassay (BDE kit). Except for two thermophilic strains, all strains that were positive in PCR amplification assays with the hblA primers were also positive when tested with the RPLA kit. The hblA DNA sequence was found in 33 strains, and these strains were closely related according to 16S rDNA-RFLP analysis, except B. pasteurii. In PCR amplifications with the bceT primers only the model strain gave a positive signal. It is concluded that screening of the hemolysin HblA complex by the PCR method allows faster detection of enterotoxin production than does testing with the RPLA enterotoxin kit.

Bacillus cereus↗

Cardiovascular risk during early adult life. Risk markers among participants in "Live for Life" health promotion programme in Sweden.

STUDY OBJECTIVE: To study differences in cardiovascular lifestyle risk factors and biological risk markers in early adult life, with special attention to age and sex differences. Lifestyle cardiovascular risk factors included dietary habits, physical inactivity, smoking, alcohol habits, psychosocial strain, and mental stress. Biological risk markers included anthropometric variables, arterial blood pressure, and serum cholesterol concentration. DESIGN: A combined individual and community based preventive programme, including health examinations. SETTING: All communities in the County of Skaraborg in south western Sweden. PARTICIPANTS: Altogether 12,982 men and women aged 30 or 35 years who underwent health examinations over seven years. MAIN RESULTS: In both sexes, biological risk markers studied were worse in 35 year old subjects than in 30 year olds. Furthermore, a larger proportion of men aged 35 years were smokers and were physically inactive compared with 30 year old men. However, dietary habits were better in both sexes in the upper age group. At both ages there were also significant differences between men and women. Women, compared with men, had better dietary habits and lower alcohol consumption but smoked more and experienced greater mental stress and psychosocial strain. All biological risk markers were worse in men than in women at both ages studied. During the observation period, some improvement of the health profile of the participants was observed, indicating a beneficial effect of the intervention programme. CONCLUSIONS: The results indicate that the risk of cardiovascular disease, as assessed from studying lifestyle and biological risk markers, increases early in life, suggesting that preventive measures should start early.

Adult↗

A1 and A2A adenosine receptors and A1 mRNA in mouse brain: effect of long-term caffeine treatment.

The effect of oral treatment with caffeine, in doses that are known to produce marked adaptive effects, was investigated on A1 and A2A receptors in the mouse brain. Caffeine (0.1, 0.3 or 1 g/l) was added to the drinking water and the animals were sacrificed after a 14-day treatment period. Ligand binding to A1 receptors was studied, using quantitative autoradiography, with the agonist [3H]cyclohexyladenosine (CHA) and the antagonist [3H]1,3-dipropyl-8-cyclopentyl xanthine (DPCPX). Caffeine did not remain in the sections during the autoradiography experiments. Caffeine treatment (1 g/l, but not 0.1 or 0.3 g/l) tended to increase [3H]CHA binding to the CA3 subfield of the hippocampus, but in no other region studied. There was no change in the number of A1 receptors since [3H]DPCPX binding to the CA3, cerebral and cerebellar cortex was not influenced by caffeine treatment. There was similarly no change in the ability of CHA to displace [3H]DPCPX binding, suggesting that there are no major changes in the proportion of A1 receptors that are coupled to G-proteins. mRNA for the A1 receptor, measured by in situ hybridization, did not differ significantly between caffeine-treated and control mice in the structures examined. Thus, higher doses of caffeine can cause an increase in A1 agonist binding without a corresponding change in A1 mRNA or in A1 antagonist binding, suggesting that the adaptive changes seen upon prolonged caffeine treatment may be in sites different from A1 receptors. Caffeine (1 g/l) increased A2A receptors in the striatum measured as binding of the agonist [3H]CGS 21680 suggesting that up-regulation of A2A receptors may be an adaptive effect of caffeine intake.

Adenosine↗

MPN-PCR-quantification method for staphylococcal enterotoxin c1 gene from fresh cheese.

PCR detection methods have been extensively used in diagnostic microbiology. However, a lack of a simple and reliable method for quantification of the PCR products has partly hindered the use of PCR in routine food laboratories. The quantification of PCR products can be done by combining the principles of MPN statistics and PCR technique. We have developed a simple and sensitive MPN-PCR assay for detection and enumeration of enterotoxin C producing Staphylococcus aureus NCTC 10655 from fresh cheese. By amplifying single copy chromosomal enterotoxin C gene fragment, we could detect as little as 20 cfu/g. By Moran's test, most of the DNA dilution series appeared to fulfill the basic mathematical assumptions of ordinary MPN methods. The analysis with MPN-PCR took one day to perform compared with three days analysis time with plate counting. This MPN-PCR method can be readily applied with different primer systems without extensive development work.

Cheese↗

Rat sciatic nerve regeneration through a micromachined silicon chip.

The capacity of regenerating nerve fibres to grow through a perforated silicon chip was tested using the silicone chamber model for nerve regeneration. The chips were fabricated as circular membranes, 4 mm in diameter, thickness 60 microns, with a perforated area, 2 mm in diameter, in the centre. Three types of chips were fabricated utilizing anisotropic etching. The chips were glued with silicone adhesive between two halves of silicone rubber tubing (total length 8 mm, inner diameter 1.8 mm, outer diameter 3.0 mm) which was used to bridge a 4 mm gap between the proximal and distal nerve stumps of a transected rat sciatic nerve. The capacity of regenerating nerve fibres to grow through the holes of the chip was analysed by light and scanning electron microscopy after 4 or 16 weeks of regeneration. Furthermore, the muscle contractility force of the gastrocnemius muscle was measured after 16 weeks of regeneration and compared as a percentage of the contralateral uninjured side. Nerves generated through chips with hole diameters of 10 or 50 microns were morphological and functional failures. The nerve structures distal to chips with hole diameters of 100 microns contained many myelinated nerve fibres in a minifascicular pattern after both 4 and 16 weeks of regeneration. The muscle contractility force was 56% of that of contralateral control muscles.

Animals↗

Ultrasound Doppler vector tomography measurements of directional blood flow.

An experimental system has been developed to verify the possibility of detecting flow activity using a technique called ultrasound Doppler vector tomography. A tomography algorithm is used to reconstruct blood flow vector fields using data from computer-controlled ultrasound continuous-wave Doppler scanning equipment. The result is a picture in which the brightness variations represent the reconstructed values of the curl of the velocity field (inverted delta x v). Continuous ultrasound is transmitted into a region with flow activity and the Doppler-shifted signals are received. To obtain measurement data suited for fan beam tomography, the scanning is performed in a plane from points encircling the region. Reconstructions have been achieved using measurement data from two different flow phantoms. A comparison between the experimental results and simulations shows good conformity.

Blood Circulation↗

The contribution of developmental surveillance to early detection of cerebral palsy.

A retrospective study of the contribution of examinations at Child Health Centres (CHC) for early detection of cerebral palsy (CP) was performed in 23,924 children, born in 1986-90, in the south-western part of Stockholm County. The outcome assessed was age of referral to a habilitation unit. A total of 66 children with CP (2.76/1000) were identified through the register at the responsible habilitation unit. The age of referral was, median 8 months, mean 12.4 months and range 0.5-54 months. A peak of referrals at 8-9 months was produced by CHC checks at 6 months of age. No other examination at the CHC resulted in a significant number of referrals. Nineteen records from CHC concerning moderate and severe cases were reviewed. Out of all examinations with aberrant findings only 11/20 resulted in referrals or further examinations. No child with moderate or severe CP was referred due to developmental screening test findings only.

Age Factors↗

Novel group within the kingdom Crenarchaeota from boreal forest soil.

We report here the results of phylogenetic analysis of archaeal 16S rRNA gene sequences amplified by PCR with Archaea-specific primers with mixed-population DNA extracted directly from forest soil used as a template. Nucleotide signature and phylogenetic analyses show that the sequences obtained belong to the domain Archaea and form a new cluster. Its phylogenetic position suggests that sequences are from a previously undescribed terrestrial group within the kingdom Crenarchaeota.

Archaea↗

A model for quality assessment in primary health care using the tracer condition technique with insulin treated diabetes as one of the tracers.

OBJECTIVE: To find a method for quality assessment in primary health care and to apply this method on a defined disease. DESIGN: General practitioners and hospital doctors worked out a programme for evaluating the quality of outpatient care of patients with insulin treated diabetes as one of five tracers. An expert committee comprising experienced general practitioners and hospital doctors compared two health centres with one outpatient medical department. Data were collected during a defined period combined with a one-year retrospective view, official statistics on medical care, and a questionnaire completed by the patients. SETTING: Two primary health care centres and one outpatient medical department in southwestern Sweden. PATIENTS: 243 insulin treated diabetic patients attending the hospital and 87 insulin treated patients attending the health centres. MAIN RESULTS: There were no difficulties for the expert panel to agree on which indicators were to be studied. The expert panel concluded that there was no difference between the quality of the two levels of care studied. CONCLUSIONS: Medical audit was a useful method for evaluating medical quality. Co-operation between hospital doctors and general practitioners was of great value for evaluation of the quality of primary health care.

Adult↗

Characterization of two lipopolysaccharide types isolated from Rhizobium galegae.

Lipopolysaccharides (LPS) of Rhizobium galegae, a symbiotically nitrogen-fixing species of root-nodule bacteria, were isolated by the phenol-water method from strain HAMBI 1461, the LPS of which resembled enterobacterial smooth type LPS, and from strains HAMBI 1174 and HAMBI 1208, the LPSs of which resembled rough type LPS. The results of PAGE analysis of LPSs, Bio-Gel P2 gel filtration of polysaccharide fractions and the presence of deoxysugars and 4-O-methyl-deoxysugar both in the rough and smooth LPSs suggested that rough LPS contained a short O-antigenic polysaccharide for which we propose the name short O-chain LPS. Accordingly, the smooth LPS is called long O-chain LPS. Despite of the differences in the structure of LPS of R. galegae, all strains were equally effective in nodulating their hosts. The short O-chain LPS of R. galegae showed many features similar to those of phylogenetically related agrobacteria.

Carbohydrates↗

The selective adenosine A2A receptor antagonist SCH 58261 discriminates between two different binding sites for [3H]-CGS 21680 in the rat brain.

We have used quantitative autoradiography to further examine two previously described binding sites for [3H]-CGS 21680 in cortical regions and in striatum, respectively. The striatal binding sites largely represent classical adenosine A2A receptors whereas the cortical sites show characteristics that differ from those of recognised adenosine receptors. A recently developed non-xanthine A2A receptor antagonist SCH 58261 displaced the binding of [3H]-CGS 21680 from the A2A receptors in striatum with an estimated Ki value of 2.4 nM, but was more than 1000-fold less potent in displacing its binding from cortex. Conversely, the adenosine analogue 2-chloro-NECA was found to be some 10 times more potent in displacing CGS 21680 from the cortical binding sites than from A2A receptors. The results provide additional evidence that CGS 21680 binds not only to classical A2A receptors, but also to sites that differ from defined adenosine receptors. They also suggest that effects of CGS 21680 observed in the presence of SCH 58261 might reveal the functional significance (if any) of these sites.

Adenosine↗