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Biomedical subjects
Publications and source records attributed to S Nyström.
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Measurements of regional cerebral blood flow (rCBF), analysis of cerebrospinal fluid, auditory brain stem responses (ABR) and oculomotor tests were performed in 19 patients with fibromyalgia. The results from the rCBF measurements showed a normal flow level with slight but significant focal flow decreases in dorsolateral frontal cortical areas of both hemispheres. The ABR results showed signs of dysfunction at least at the brain stem level and the oculomotor tests showed high frequency of pathology. The cerebrospinal fluid analysis showed discrete changes in the cell differential count. Possible explanations for the involvement of the central nervous system in fibromyalgia are discussed.
Isoprenylation is an important posttranslational modification that affects the activity, subunit interactions and membrane anchoring of different eukaryotic proteins. The small, cell-wall-less prokaryote Acholeplasma laidlawii has more than 20 membrane acyl-proteins enriched in myristoyl and palmitoyl chains. Radioactive mevalonate, a precursor to isoprenoids, was incorporated into several specific membrane proteins of 20 to 45 kDa and two soluble proteins of 23-25 kDa, respectively. No acyl proteins and none of the polar acyl lipids became labelled but these are all labelled by radioactive fatty acids. Mevalonate was incorporated mainly into a minor neutral, non-saponifiable lipid which migrated just above a C30-isoprenoid (squalene) on TLC-plates. The isoprenoid chains could not be released by mild alkaline hydrolysis from most of the isoprenylated proteins, although this procedure releases acyl chains from lipids and all acylated proteins. Isoprenylated proteins were enriched in the detergent phase upon partition with the non-ionic detergent Triton X-114. This behaviour is similar to the acyl proteins of this organism and indicates that the isoprenoid chains give the proteins a hydrophobic character.
Mycoplasmas are small bacteria without a cell wall, often found as surface parasites on eukaryotic cells. Of the more than 200 membrane proteins from Acholeplasma laidlawii resolved by two-dimensional PAGE, 23 were covalently modified with acyl chains. These acyl proteins had lower pI values than average and were all labelled by different exogenously supplied radioactive fatty acids attached by O-ester bonds. The fatty acids were selectively incorporated in the order myristic acid (14:0) greater than palmitic acid (16:0) greater than stearic acid (18:0) greater than oleic acid (18:1). However, endogenously synthesised saturated fatty acids, most of which were 16:0, were preferred over the supplied ones. A fraction of the exogenous 14:0 was elongated to 16:0. Absence of saturated fatty acids increased the incorporation of 18:1. The maximum extent of modification was one acyl chain for protein T2, on the exterior surface and two acyl chains for protein D12, spanning them membrane. Exogenously supplied fatty acids were incorporated into membrane lipids in proportion to their occurrence. However, the acylated proteins always contained 8-10 times more saturated chains than did the lipids. When exogenously supplied, all A. laidlawii polar membrane lipids could donate acyl chains to the acylated proteins but the neutral fraction (fatty acids and diacylglycerol) was most efficient. An incorporation into the acylated proteins of labelled cysteine, but not glucose or glycerol, was observed. Acylated proteins with different chains interacted similarly with a Triton X-114 detergent phase, and no full-size proteins (or acylated fragments) were released from cells by proteolytic enzymes. The results indicate an anchoring with peptide segments in addition to the acyl chains. Both 14:0 and 16:0 were attached at one end of both T2 and D12, but the N-terminal methionine of T2 was not acylated. The extent of modification and preference for saturated chains in the A. laidlawii membrane acylated proteins is more similar to eukaryotic than to eubacterial proteins.
Single strain regression analysis was performed on PDM II medium for E. faecalis with 10, 30 and 120 micrograms gentamicin disks using E. faecalis, strain ATCC 29212 as the reference. This method permits the calculation of zone diameters corresponding to different MIC values for different disk contents. The lack of discrimination between normal low-level resistant strains and high-level resistance using the 10 micrograms disk was confirmed. However, both the 30 micrograms and 120 micrograms disks seemed to provide a separation of the normal low-level gentamicin-resistant population from strains with increased resistance. Since the 30 micrograms disk is used routinely in some countries, there should be no need for an extra high content disk in these laboratories. This was confirmed when 96 clinical isolates of E. faecalis were analysed and the results of routine disk diffusion tests were compared with the MIC values. Two of the strains showed high-level gentamicin resistance (greater than 2000 mg/l) and produced no zone of inhibition. The other 94 isolates showed gentamicin MIC values between 4-16 mg/l, and 72 of the MIC results were 8 mg/l. The zone diameters for these strains ranged between 15 and 25 mm with a mean of 18.2 and a median value of 18 mm. In order to include statistical considerations of the zone size populations for setting of breakpoints, a study of gentamicin zone size distributions was performed for several bacterial species. Inhibition zone diameter values around the 30 micrograms gentamicin disk for 2079 clinical isolates of E. faecalis, 2268 S. aureus, 3201 E. coli and 547 strains of P. mirabilis from different years were plotted as histograms. Tests for agreement with a Gaussian distribution showed that the histograms were slightly peaked and skewed towards higher zone values. Parametric and non-parametric statistical tests were compared and the results showed that means and medians were very similar and that parametric fractile estimations at the lower end of the histogram populations were conservative and could be used in view of a slightly lower rate of false resistance. The 1% parametric fractile of 12 mm was selected as a suitable breakpoint for the identification of normal, low-level resistant isolates of E. faecalis using the standardized disk test of the Swedish Reference Group for Antibiotics.
The study was conducted in a representative Swedish rural-urban population sample of 3,064 persons. The IQ factor was determined directly by a single well established procedure. No systematic association between IQ level and the mean number of children was found.
Septicaemia caused by coagulase negative staphylococci is a problem in the neonatal intensive care units (NICU). The very low birthweight (VLBW) infants are at a special high risk because of their immature host defense. In this study the potential risk factors were compared between the VLBW septicaemia patients and the VLBW infants who had not contracted septicaemia in the NICU. The factor most clearly related with septicaemia was the use of umbilical artery catheters. The strategy against neonatal septicaemia includes restriction of the use of intravascular catheters. Also enhancement of the host defense by immunoglobulin therapy is considered. The coagulase negative staphylococci were multiresistant to antibiotics. The combination of netilmicin and benzylpenicillin covered the bacteria found in septicaemia cases in the NICU, and is now the standard treatment in suspected cases. Coagulase negative staphylococci are treated with vancomycin or netilmicin.
Of the 51 polypeptides detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the plasma membrane of the helical mollicute Spiroplasma melliferum, 21 are acylated, predominantly with myristic (14:0) and palmitic (16:0) chains. This is notably the case for spiralin, the major membrane protein of this bacterium, which contains an average of 0.7 acyl chains per polypeptide, attached very probably by ester bonds to alcohol amino acids. The amphiphilicity of spiralin was demonstrated by the behavior of the protein in charge-shift electrophoresis, its incorporation into liposomes, and its ability to form in the absence of lipids and detergents, globular protein micelles (diameter, approximately 15 nm). The presence of epitopes on the two faces of the cell membrane, as probed by antibody adsorption and crossed immunoelectrophoresis, and the strong interaction between spiralin and the intracytoplasmic fibrils show that spiralin is a transmembrane protein. The mean hydropathy of the amino acid composition of spiralin (-0.30) is on the hydrophilic side of the scale. Surprisingly, the water-insoluble core of spiralin micelles, which is the putative membrane anchor, has a still more hydrophilic amino acid composition (mean hydropathy, -0.70) and is enriched in glycine and serine residues. Taking into account all these properties, we propose a topological model for spiralin featuring a transbilayer localization with hydrophilic domains protruding on the two faces of the membrane and connected by a small domain embedded within the apolar region of the lipid bilayer. In this model, the membrane anchoring of the protein is strengthened by a covalently bound acyl chain.
Completely computer-focused ultrasound imaging has been achieved using the UHB (ultrasound holographic B) method. This is a synthetic aperture method that combines ultrasonic holography and the conventional B-scan method. A wide-angle ultrasound burst (4 MHz) is transmitted into the object and the reflected echoes are measured as a function of the propagation time, but both the amplitude and phase angle are recorded, giving multiple one-dimensional holograms in a cross-sectional plane. This UHB data is rearranged to obtain one-dimensional wave fields, which represent the wave fronts reflected from each depth. The image is then reconstructed by propagating these wave fields backward to their original positions on the image using Fourier transform, a spectrum shift theorem, and inverse Fourier transform. The whole UHB image is obtained by calculating the intensity lines from these wave fields and by scanning these lines one by on the cathode ray tube (CRT) monitor. Following extensive laboratory testing with single and multielement transducers, special 32- and 64-element linear array transducers were fabricated, as was computer software capable of producing a UHB image in 40 seconds. Tissue imaging tests followed by clinical imaging of 30 patients showed that images obtained with this prototype compared favorably with those obtained by state-of-the-art conventional ultrasound scanners. The potential advantages of the new method are enhanced lateral resolution and the incorporation of phase information in tissue characterization. This would be important in diagnostic and intraoperative, especially neurosurgical, imaging.
A series of 40 patients with acoustic neuromas were operated on using the suboccipital or the translabyrinthine approach. Some patients were operated on using both routes. The series extends back to 1977 and contain predominantly large tumours. There was no mortality in the translabyrinthine series of 14 patients. In the suboccipitally treated series of 29 patient, 3 were previously treated using translabyrinthine approach. In this series there was one death directly due to sequelae of neuroma extirpation and in another, additional causes were present. Preoperative facial pareses were frequent. There were several additional pareses in the translabyrinthine series and also several additional in the suboccipital series. Facial pareses were operated on by a plastic surgeon and they did not markedly detract from working capacity. Only 2 patients in the total series had small tumours and useful preoperative hearing. Cochlear function was preserved in them.
Neurosurgical resection of an epileptic focus was performed in eleven patients suffering from drug resistant focal epilepsy. The clinical result was favourable in nine cases and corresponds to the earlier results. The routinely processed biopsy specimens obtained from the brain resections were stained with haematoxylin-eosin and with specific antisera to GFAP, S-100, NSE, laminin, and fibronectin using the peroxidase-antiperoxidase technique. The main pathological finding was gliosis in eight cases, neuronal degeneration in two cases, and a vascular malformation in one case. The anti-GFAP as a specific marker of astrocytes made the astroglial proliferation clearly visible, demonstrating an astroglial scar in four cases and a moderately to strongly increased amount of astroglial cells in another four cases. Anti-S-100 and anti-fibronectin are not as specific markers. They stained both neurones and glial cells with comparable results to that of anti-GFAP but with a lower specificity and sensitivity. Anti-NSE showed decreased amounts of neurones in most of the heavily gliotic lesions and also stained glial cells in some cases. Anti-laminin stained the pial and vascular basement membranes and revealed an increased vasculature in two cases. From these results, it appears that GFAP immunostaining is a highly demonstrative means for the visualization of astrogliosis in epileptic lesions and may be of help in identifying slight focal changes. An exact demonstration of neuronal loss or other neuronal changes still waits for a more specific marker than NSE. A favourable clinical outcome after neurosurgery seems to be associated with the patients showing a clearly gliotic brain lesion in one temporal lobe.
Many integral membrane proteins in Acholeplasma laidlawii are enriched in hydrophilic amino acid residues and covalently modified with fatty acids. In order to understand how these proteins are inserted and anchored in the bilayer, we have cloned several of the major A. laidlawii proteins in Escherichia coli: 900 recombinant clones containing 4-kbp DNA fragments, inserted into the BamHI site of the plasmid pAT 153, were screened with antibodies. With antimembrane antibodies, 26 positive clones were detected, and with a mixture of five different monospecific antibodies, another 7 clones were obtained. Immunological analysis of the colonies in situ verified that antigens for A. laidlawii membrane proteins D12, T2, T3, T4a, and unidentified proteins were produced in separate clones. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting showed several fragments [25 to 94 kilodalton (kDa)] for each of these proteins, some of which were even visible on Coomassie Blue-stained gels. It is concluded that A. laidlawii membrane proteins can be efficiently expressed in E. coli.
Spiralin, a 28-kDa (kilodalton) polypeptide, is the major antigen of Spiroplasma citri and S. melliferum in which it usually represents more than 20% of total membrane protein. The amino acid compositions of the spiralins purified from both spiroplasma species unambiguously show that these proteins are homologous. In addition, several lines of evidence indicate that such a protein is present in the menbrane of S. apis. A 25-kDa polypeptide antigenically related to S. citri spiralin has also been purified from the membrane of the nonhelical variant ASP-1. The spiralin of S. melliferum B88 has been used as a model for extensive characterization. This antigen binds detergent under nondenaturing conditions, can be incorporated into liposomes, and forms protein micelles upon gentle removal of detergent. Digestion of the micelles with trypsin leads to the precipitation of an insoluble material containing a major polypeptide of 3.9 kDa. The amino acid composition of this fragment is different from that of intact spiralin. It is highly enriched in glycine and serine and, as an insoluble peptide, exhibits an unexpectedly high polarity index (PI = 51.4%). Screening for acyl proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodetection in the membrane of S. melliferum indicates that spiralin is actually acylated. This set of properties is evidence that spiralin is an intrinsic membrane protein and strongly suggests that acylation triggers or facilitates integration of the molecule into the lipid bilayer of the spiroplasma membrane.
In membranes of the cell-wall-less prokaryote Acholeplasma laidlawii most proteins are of the integral type. A substantial fraction of these proteins are enriched in hydrophilic amino acid residues. Approximately 20 different major as well as minor proteins were found to be covalently modified with acyl chains. The same set of proteins are acylated when cells are grown in different fatty-acid-supplemented media. In individual proteins the ratio of palmitoyl/oleoyl acyl chains was 12-14 times larger than the acyl chain ratio in polar membrane lipids. The transmembrane protein D12 has close to two acyl chains per molecule. Proteins T2 and T4a, localized in the outer and inner leaflet of the membrane, respectively, occur each as pairs with a difference in relative molecular mass within each pair of approximately 2000. Each of these proteins as well as the other acyl proteins, except the light form of T4a, has close to one acyl chain per molecule. The extent of acylation was increased for certain proteins and decreased for others by treatment with globomycin or phenethylalcohol. The relative amounts of the T2 and T4a pairs were affected by these drugs. It is concluded that the mechanism of acylation is different from that in Escherichia coli lipoprotein and Bacillus penicillinase. The mean hydrophobicity [Kyte & Doolittle (1982) J. Mol. Biol. 157, 105-132] of the A. laidlawii acyl proteins are similar to those of other bacterial acyl proteins but significantly lower than for non-acylated integral membrane proteins, supporting an anchoring function of the acyl chains. The number of membrane acyl proteins in A. laidlawii and two other mycoplasmas are at least twice that in other bacteria.
The efficacy of the antimicrobial removal device in facilitating isolation of bacteria from blood containing antibiotics was evaluated in a multicenter study. Blood specimens from 143 patients who had culture-proven septicemia and had been on antibiotic treatment for at least 24 hours were recultured with and without the aid of the device. Septicemia persisted in 33 of the patients and use of the device improved the isolation rates: 63% of the gram-positive bacteria and 32% of the gram-negative bacteria were isolated only with the aid of the device. Use of the device was an advantage in 10 of 13 cases where the antibiotic concentration in the bottles exceeded the MIC for the infecting microorganism but in only 5 of 19 cases where the concentration was lower than the MIC. It is concluded that the antimicrobial removal device may be useful in patients already on antibiotic treatment, but that its use is limited by the cost and the amount of work involved.
Immunohistochemical and histological investigations were undertaken on 24 surgically-removed pituitary adenomas. By histology (haemalu-eosin staining), 7 chromophobe, 12 acidophil and 5 basophil pituitary adenomas were revealed. For immunohistochemical purposes the peroxidase-antiperoxidase technique was applied. Primary antisera against 10 hormones were used. By immunohistochemistry, 7 prolactin-containing, 2 TSH-containing, 2 GH-containing and 1 beta-endorphin-containing pituitary adenomas were identified. Furthermore, 1 mixed thyrotropic-prolactin human pituitary adenoma was detected. A possible connection between histological and immunocytochemical findings is discussed.
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The present study was designed to chart the distribution of cognitive factors in an adult population. The material consisted of a representative sample of persons aged 18-65 years, living in the county of Stockholm (except for the city of Stockholm) in 1970. The total selection was 3 064 persons. These persons were invited to a hospital for a health examination, covering both physical and mental health. Psychometric tests were included. In the study of cognitive factors the rate of non-response corresponded to 18.7% of the sample (males 20.7%, females 16.6%). The tool of examination was the Dureman-Sälde test battery. It is shown that men score rather higher than women in analytical and spatial but not in verbal ability. Analytical ability declines steadily throughout the age spectrum. Spatial ability shows a declining trend from he age group 36-45 years. Verbal ability is best in the age group 36-45 years and here the reduction in higher ages is very limited. A positive correlation of considerable degree exists between social group and the intelligence factors. This applies particularly to verbal and spatial ability and the overall measures. The connection with logical-inductive ability is quantitatively much less impressive. As regards verbal and spatial ability, there is an age-related tendency to a widened gap between social groups. Standardization for social group in tests is advocated in practical use.