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Biomedical subjects

R Karlsson

Publications and source records attributed to R Karlsson.

At least 55 records · Page 3Linked to original sources
PubMed1995

Identification of the trapped calcium in the gelsolin segment 1-actin complex: implications for the role of calcium in the control of gelsolin activity.

The X-ray structure of the complex of actin with gelsolin segment 1 revealed the presence of two calcium ions, one bound at an intramolecular site within segment 1 and the other bridging the segment directly to actin. Although earlier calcium binding studies at pH 8.0 revealed only a single calcium trapped in the complex (and also in the binary gelsolin-actin complex), it is here shown that two calcium ions are bound under the conditions of crystallization at physiological pH. Mutation of acidic residues in either actin or segment 1 involved in ligation of the intermolecular calcium ion resulted in loss of one of the bound calcium ions at pH < 7, but not at pH 8. Thus the calcium ion trapped in the segment 1-actin complex is that located at the intramolecular site. The implications of this for gelsolin function are discussed.

Actins↗
PubMed1995

Human papillomavirus infection is transient in young women: a population-based cohort study.

The prevalence of human papillomavirus (HPV) infection in cervical cell scrapes from a cohort of 276 young women was determined by a general two-step polymerase chain reaction. HPV infection fluctuated among young women during a 2-year interval. The total prevalence of HPV infection decreased from 21% to 8.3%. The most prevalent HPV types at enrollment were HPV-16 (3.3%) and HPV-6 (2.9%). At follow-up, the most common type was HPV-16 (2.9%), while no HPV-6 was detected. In 2 women only, the same HPV type persisted. Regression of HPV infection was found in 80% of the women. A new HPV type-specific infection was detected in 7.2% of the women and was independently associated with a new sex partner or an abnormal smear since enrollment.

Adult↗
PubMed1995

The silent suffering women--a population based study on the association between reported symptoms and past and present infections of the lower genital tract.

OBJECTIVES: To assess the prevalence of lower genital tract symptoms and the association between reported symptoms and past and present signs of sexually transmitted diseases (STD) in young women. DESIGN: All women belonging to the 19-, 21-, 23- and 25-year age cohorts and living in the catchment area of the community health centre, were invited by mail to take part in a population-based study. The participants answered a structured questionnaire and a gynaecologic examination was performed. Samples for wet smear, cervical Pap smear, HPV DNA determination and Chlamydia trachomatis culture were taken at the gynaecologic examination. The presence of genital warts was noted. A blood sample was analysed for antibodies against C trachomatis and HSV-2. SETTING: The community health care centre was located in Umeå, a city in Northern Sweden. RESULTS: Of the 886 women who were eligible, 611 (70%) participated in the investigation. One out of four women reported symptoms from the lower genital tract. The most commonly reported symptoms were itching, followed by discharge, and soreness. The most commonly reported STD was C trachomatis (15%). The most prevalent present STD was HPV infection (20%) whereas C trachomatis infection could be isolated from 2.7% of the women. Antibodies against C trachomatis and HSV-2 were present among 22% and 6% of the women, respectively. There was a significant correlation between the women's complaint of vaginal discharge and previous C trachomatis infection, lack of lactobacilli and presence of leucocytosis in wet smear. CONCLUSIONS: We have in a population-based study of young healthy women found that one out of four women had some kind of lower genital tract complaint. Itching was the most commonly reported symptom and was associated with pseudohyphae and acetowhite patches. Reported vaginal discharge and soreness were associated with the history of a past C trachomatis infection and signs of a disturbed vaginal flora.

Adult↗
PubMed1994

Real-time competitive kinetic analysis of interactions between low-molecular-weight ligands in solution and surface-immobilized receptors.

With surface plasmon resonance detection it is possible to measure the binding kinetics between a macromolecule in solution and a receptor immobilized on a sensor surface. The detector response is proportional to the mass of the analyte that binds to the surface, and therefore, a direct observation of a low-molecular-weight (lmw) analyte ( < 5000 Da) interacting with its immobilized binding partner is normally not possible. I describe here a competitive approach in which a lmw analyte and a high-molecular-weight analyte react at the same time with the immobilized receptor. Using this approach it is possible to extend kinetic analysis to lmw analyte-receptor interactions. A qualitative analysis allows rapid affinity ranking of different lmw analytes interacting with the same receptor, and a quantitative analysis of binding data allows the calculation of rate constants for the lmw analyte-receptor interaction. The competitive kinetics approach may therefore be used as an alternative to other affinity techniques for the characterization of lmw ligands, for identification of inhibitors, and for drug screening.

Amino Acid Sequence↗
PubMed1994

Intermolecular contacts in various crystal forms related to the open and closed conformational states of the catalytic subunit of cAMP-dependent protein kinase.

All hitherto solved crystal structures of the catalytic (C) subunit of cAMP-dependent protein kinase can be classified into two groups, those with a closed and those with an open conformation of the ATP-binding lobe. The molecules with the closed conformation are all related by a crystallographic 2(1) axis that connects them into an infinite-chain motif. The motif has only one large contact region that involves many residues, several of them in the ATP-binding lobe, embedded in an extensive network of water molecules. The dominant feature of this region is the hydrophobic interaction between Trp196 and Arg133, Arg134. This motif has been found so far in three different crystal forms, two correspond to ternary enzyme-inhibitor-ATP complexes with mammalian and recombinant C, and one to a binary enzyme-inhibitor complex with recombinant C. The open conformation has been found in two closely related crystal structures, both of cubic symmetry, of the apoenzyme and a binary complex of the mammalian catalytic subunit. In this cubic structure of the binary complex, the hydrogen-bonded intramolecular contacts between Arg18 of the inhibitor and the ATP-binding lobe of the binary and ternary complexes of the recombinant enzyme are missing due to a strong hydrophobic intermolecular contact involving the diiodinated Tyr7. In solution, no crystal contacts prevent these hydrogen bonds involving Arg18 from forming so that it is likely that the binary complex with Tyr7 of the peptide inhibitor iodinated or not, can assume the closed conformation in solution. While the closed structure very likely represents a stable conformation in solution, there is no evidence to suggest that the open conformation represents a unique stable conformational state of the enzyme in solution.

Journal Article↗
PubMed1994

Working on a moving surface--a biomechanical analysis of musculo-skeletal load due to ship motions in combination with work.

Ship motions are recognized by fishermen as a cause of high musculo-skeletal load. In the present study, the motions of a Swedish trawler at sea were registered over three degrees of freedom. Simultaneously, the working postures of a fisherman on board were registered in the sagittal plane during five different working situations: (1) standing erect in still conditions; (2) holding a load of 21 kg in still conditions; (3) standing erect during motion of the ship; (4) standing during motion of the ship, holding a load of 21 kg; and (5) repeatedly lifting and lowering a 21 kg load during motion of the ship. All registrations were sampled using a computer where data were processed in a two-dimensional, dynamic biomechanics model, developed for this particular purpose. Vertical and horizontal forces as well as moments were calculated for seven major joint systems of the body, as was compression at the L4/L5 vertebral level. In situation (3), ship motions were mainly counteracted by motions in the lower extremity and lumbar back, thus inducing increased strain in these parts of the body; other parts of the body were little affected. Holding a load considerably increased the load on most joints. Lifting the load further increased the musculo-skeletal strain and also increased the range of moments in each joint as well as the range of lumbar compression. Moment at the C7 vertebral level was relatively unaffected by ship motions and by handling external loads.

Biomechanical Phenomena↗
PubMed1994

The effect of a new oral hygiene training program on approximal caries in 12-15-year-old Brazilian children: results after three years.

This study evaluated the effects of a new oral hygiene training program on approximal caries in a population of 12-13-year-old Brazilian schoolchildren with a well-established habit of daily toothbrushing with a F dentifrice. Two hundred twenty-two children were randomly allocated into two test groups (I and II) and one control group (III). Group I subjects were trained to establish needs-related oral hygiene habits based on self-diagnosis and a new behavioral principle, the 'linking method', for establishment of habits. The first three visits (20 minutes each) were scheduled at two-day intervals. They were recalled for a monthly check-up during the first 4 months, and then every 3 months for reevaluation of the results based on self-diagnosis. Group II subjects were recalled at the same intervals for detailed oral hygiene instruction on how to clean every tooth surface using dental tape, toothbrush, and fluoride dentifrice. Group I developed significantly fewer (p < 0.001) new approximal manifest (dentin) caries lesions than groups II and III. The mean values (SEM) were 2.3 (0.29), 4.7 (0.59), and 5.3 (0.68), respectively. The conclusions from our study are: In a toothbrushing population using fluoride dentifrices and fluoridated drinking water, the oral hygiene training program with behavioral modification significantly reduced caries incidence on approximal surfaces. Frequent repetition of training in meticulous oral hygiene is almost redundant.

Adolescent↗
PubMed1994

Serum profiles of oxytocin during oral contraceptive treatment.

Serum profiles of oxytocin were studied by means of a continuous blood sampling system in five young healthy women before and during treatment with a combined oral contraceptive. Oxytocin levels were determined by a specific radioimmunoassay in blood samples collected in 10-min fractions from 22.00 to 06.00. The values were further analyzed by the pulse detection program PULSAR. Great individual differences in oxytocin profiles were observed, and in some of the women these differences were also pronounced between the two sampling occasions. All 10 profiles demonstrated irregular peaks which occurred with varying frequency. Although the baseline level of oxytocin increased in all women and the average concentration increased in four of the women during treatment, there was no clear-cut effect on the peak frequency. Based on results from animal experiments, it is suggested that the increase in oxytocin levels may be related to an excitatory effect exerted by estrogen on oxytocin secreting neurons.

Adult↗
PubMed1993

Analysis of active antibody concentration. Separation of affinity and concentration parameters.

Antibody binding to surfaces with differing amounts of immobilised antigen was measured in a biosensor system using surface plasmon resonance detection. Binding rates obtained during the initial binding phase on high density antigen surfaces were proportional to antibody concentration and independent of antigen-antibody affinity. One antibody calibration curve covering the range from 0.5 to 160 nM (0.08-25 micrograms/ml) antibody was valid for IgG antibodies with different antigen specificities. To illustrate the use of this methodology active antibody concentrations were analysed in culture media and in rabbit serum.

Animals↗
PubMed1993

Mutagenesis of human profilin locates its poly(L-proline)-binding site to a hydrophobic patch of aromatic amino acids.

The actin-binding protein, profilin, contains a src-homology (SH) 3-like fold (Schutt, C.E. et al., submitted), and its tight interaction with poly(L-proline) is reminiscent of the binding activity exhibited by SH3-domains. Here we demonstrate that replacements of aromatic amino acids in a hydrophobic patch on the surface of the profilin molecule abolish its poly(L-proline)-binding capacity. However, the location of this hydrophobic patch is found in another region of the molecule than that displaying structural similarities with SH3 domains.

Amino Acids↗
PubMed1993

Mutations in beta-actin: influence on polymer formation and on interactions with myosin and profilin.

Two beta-actin mutants, one with proline 38 replaced with alanine (P38A) and the other with cysteine-374 replaced with serine (C374S), as well as the wild-type beta-actin, were expressed in the yeast, S. cerevisiae, purified to homogeneity, and analyzed in vitro for polymerizability and interaction with DNase I, myosin, and profilin. Both mutations interfered with the polymerization of the actin, and with its interaction with myosin. The C374S mutation had the most pronounced effect; it reduced the polymerizability of the actin, abolished its binding to profilin, and filaments containing this mutation moved at reduced rates in the in vitro 'motility assay'. The ATPase activity measured in solutions containing myosin subfragment 1 was similar for both the mutant and wild-type actins.

Actins↗
PubMed1993

Structure of the mammalian catalytic subunit of cAMP-dependent protein kinase and an inhibitor peptide displays an open conformation.

The crystal structure of a binary complex of the porcine heart catalytic (C) subunit of cAMP-dependent protein kinase (space group P4(1)32; a = 171.5 A) complexed with a di-iodinated peptide inhibitor, PKI(5-24), has been solved and refined to 2.9 A resolution with an overall R of 21.1%. The r.m.s. deviations from ideal bond lengths and angles are 0.022 A and 4.3 degrees. A single isotropic B of 17 A(2) was used for all atoms. The structure solution was carried out initially by molecular replacement of electron density followed by refinement against atomic coordinates from orthorhombic crystals of a binary complex of the mouse recombinant enzyme previously described [Knighton, Zheng, Ten Eyck, Ashford, Xuong, Taylor & Sowadski (1991). Science, 253, 407-414]. The most striking difference between the two crystal structures is a large displacement of the small lobe of the enzyme. In the cubic crystal, the beta-sheet of the small lobe is rotated by 15 degrees and translated by 1.9 A with respect to the orthorhombic crystal. Possible explanations for why this binary complex crystallized in an open conformation in contrast to a similar binary complex of the recombinant enzyme are discussed. This study demonstrates that considerable information about parts of a crystal structure can be obtained without a complete crystal structure analysis. Specifically, the six rigid-group parameters of a poly alanine model of the beta-structure were obtained satisfactorily from a crystal structure by refinement of difference Fourier coefficients based on an approximate partial structure model.

Journal Article↗
PubMed1993

Crystal structure of the catalytic subunit of cAMP-dependent protein kinase complexed with MgATP and peptide inhibitor.

The structure of a ternary complex of the catalytic subunit of cAMP-dependent protein kinase, MgATP, and a 20-residue inhibitor peptide was determined at a resolution of 2.7 A using the difference Fourier technique starting from the model of the binary complex (Knighton et al., 1991a). The model of the ternary complex was refined using both X-PLOR and TNT to an R factor of 0.212 and 0.224, respectively. The orientation of the nucleotide and the interactions of MgATP with numerous conserved residues at the active site of the enzyme are clearly defined. The unique protein kinase nucleotide binding site consists of a five-stranded antiparallel beta-sheet with the base buried in a hydrophobic site along beta-strands 1 and 2 and fixed by hydrogen bonds to the N6 amino and N7 nitrogens. The small lobe secures the nucleotide via a glycine-rich loop and by ion pairing with Lys72 and Glu91. While the small lobe fixes the nontransferable alpha- and beta-phosphates in this inhibitor complex, the gamma-phosphate is secured by two Mg2+ ions and interacts both directly and indirectly with several residues in the large lobe--Asp184, Asn171, Lys168. Asp166 is positioned to serve as a catalytic base. The structure is correlated with previous chemical evidence, and the features that distinguish this nucleotide binding motif from other nucleotide binding proteins are delineated.

Adenosine Triphosphate↗
PubMed1993

Influence of smoking and snuff use on electrolytes, adrenal and calcium regulating hormones.

Little is known about the effects of snuff use on health. We have investigated electrolyte levels, adrenocortical and calcium regulating hormones in three groups of healthy young men, including 18 non-tobacco users, 21 snuff users and 19 smokers with similar age and body mass index. Smoking and snuff use was positively associated with alcohol and coffee consumption and inversely related to physical activity. Compared to non-tobacco users, smokers had significantly increased levels of serum sodium and magnesium, plasma calcitonin, urinary cortisol and potassium levels and decreased serum sex hormone-binding globulin as well as serum and urinary creatinine values. However, only decreased sexual hormone-binding globulin and urinary creatinine and increased serum phosphate and urinary potassium levels were seen in snuff users. Among tobacco users we noted that smokers differed from snuff users in that they had higher serum sodium (1.4 mmol/l, p < 0.01), plasma calcitonin (3.3 pmol/l, p < 0.05) and urinary cortisol (41 nmol/24 h, p < 0.05) but lower serum creatinine (5.8 mumol/l, p < 0.01). We conclude that chronic snuff use appears to have less influence on hormone and electrolyte balance than does smoking, and that some of the abnormalities seen in smokers do not seem to be mediated by nicotine.

Adrenal Cortex↗
PubMed1993

Introducing a biosensor based technology for real-time biospecific interaction analysis.

This report describes a system for real-time biospecific interaction analysis, using biosensor technology based on the optical phenomenon surface plasmon resonance. The biospecific interface is a sensor chip consisting of a thin gold film deposited on a glass support and covered with a hydrogel matrix. One component of the interaction being studied is attached covalently to the hydrogel, and other interactants are passed over the chip in solution. The interaction is followed in real time in terms of changes in the mass concentration of biomolecules at the sensor surface. Surface concentrations down to 10 pg/mm2 can be measured. The technique does not require molecular labels such as isotopes or spectroscopic markers, and purification of interacting components can often be avoided. Repeated analyses can be performed on the same sensor chip. With this system, the same general procedure can be used for a wide range of different applications, including concentration determination, kinetic measurements and multi-site binding studies. The sensitivity of the technique can be adjusted by choice of reagents and experimental procedure: determination of specific proteins in serum down to 20 ng/ml and macromolecular association constants from 10(7) M-1 up to 4 x 10(11) M-1 are documentated examples. No other single analytical system has the same versatility and general applicability to biospecific interaction analysis. The system is developed and marketed by Pharmacia Biosensor AB, Sweden.

Biosensing Techniques↗
PubMed1993

Pressure pain thresholds in patients with craniomandibular disorders before and after treatment with acupuncture and occlusal splint therapy: a controlled clinical study.

Fifty-five patients (46 women and 9 men) with craniomandibular disorders and a history of pain of at least 6 months' duration participated in this trial. The patients were randomly assigned to three groups: one group to receive acupuncture; one group to receive occlusal splint therapy; and one group to act as controls. Pressure pain threshold, clinical dysfunction score, and visual analog scale measures were used to evaluate patients before, immediately after, and 6 months after treatment. A moderate, but statistically significant, correlation was found between pressure pain threshold and the number of tender spots in the masticatory muscles (tau = -.43; P < .001), degree of tenderness in the masticatory muscles (tau = -.43; P < .001), clinical dysfunction score (tau = .32; P < .001), and the visual analog scale (tau = -.25; P < .01). The short-term results showed a statistically significant improvement in all evaluations for both treatment groups. No significant differences were found in the control group. The improvements resulted in significant differences between the control and each treatment group immediately after treatment. At the 6-month follow-up, no significant differences in pressure pain threshold or clinical dysfunction score were found in the two treatment groups compared with the short-term results.

Acupuncture Analgesia↗
PubMed1992

Characterization of yeast-expressed beta-actins, site-specifically mutated at the tumor-related residue Gly245.

The tumorigenic cell line HUT14 expresses a beta-actin carrying a mutation at position 245. In this study, two mutant beta-actins with amino acid changes at position 245 replacing the wild-type glycine by an aspartic acid and a lysine residue, respectively, were produced in the yeast Saccharomyces cerevisiae, purified to homogeneity and characterized with respect to polymerization behaviour and interaction with myosin. The major functional effect of these mutations appears to be an impaired polymerization, while the interaction with myosin seems less influenced. In addition, the results also suggest the presence of a Ca(2+)-binding site in the region of residue 245 in actin.

Actins↗