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Biomedical subjects

G Nilsson

Publications and source records attributed to G Nilsson.

At least 109 records · Page 6Linked to original sources

On the pH-lowering potential of lactobacilli and mutans streptococci from dental plaque related to the prevalence of caries.

The common method used today to identify persons at risk of dental caries is to estimate the numbers of cariogenic bacteria such as lactobacilli and mutans streptococci in saliva or plaque samples taken from the patient. However, the value of these bacterial counts for explaining and predicting individuals at risk of caries has not been powerful enough. Evaluating one virulence factor such as the acidogenicity of these bacteria might increase their explanatory values for caries. Sixty children aged 14-15 yrs participated in this study. Smooth surface caries and restorations were registered and total plaque samples collected. Counts of lactobacilli and mutans streptococci were estimated, and the pH-lowering potential of both bacteria was measured in an adapted glucose broth. The results showed a weak association between dental caries and lactobacilli, but in the subgroup with this bacterium the explanatory value increased to 14% and in the subgroup with a strong pH-lowering potential it was as high as 27%. For mutans streptococci the associations were weak in all groups.

Adolescent↗

Twenty-four-hour basal and repetitive pentagastrin-stimulated gastric acid secretion in normal and sham-operated rats and in rats after gonadectomy or treatment with estradiol or testosterone.

BACKGROUND: Studies in different species have suggested, but not established, that sex hormones influence gastric acid secretion. We studied how acid output is affected by the sex hormones estradiol or testosterone in vivo and in vitro. METHODS: In gastric fistula rats that were normal, sham-operated, neonatally gonadectomized, or treated with estradiol or testosterone, 24-h basal and pentagastrin-stimulated acid secretion was measured. The in vitro effects of estradiol and testosterone on histamine-induced aminopyrine accumulation in isolated parietal cells were also determined. RESULTS: Basal acid output was similar in the two sexes, but stimulated secretion was significantly higher (34%; P < 0.01) in males. Ovariectomy did not influence acid output, whereas orchidectomy reduced basal (18%; NS) and stimulated 24-h secretion (P < 0.01). Estradiol decreased (23%; NS) the 24-h basal output in females but not in males. Estradiol suppressed stimulated secretion in females (29%, P < 0.01) and males (42%, P < 0.01) during the day. At night the stimulated secretion increased in both females (17%, NS) and males (32%, P < 0.05). A similar pattern was found when rats were treated with testosterone. In vitro, estradiol and testosterone reduced histamine-stimulated aminopyrine accumulation in both female and male isolated parietal cells. CONCLUSIONS: Estradiol and testosterone both appear to influence gastric secretion in rats, and their action differs between day and night, between the sexes, and between basal and stimulated secretion.

Aminopyrine↗

C3a and C5a are chemotaxins for human mast cells and act through distinct receptors via a pertussis toxin-sensitive signal transduction pathway.

Mast cells are known to accumulate at sites of inflammation, however, the chemotaxins involved are undefined. Since most natural leukocyte secretagogues also induce cell migration, and since the anaphylatoxins C3a and C5a are mast cell secretagogues, we hypothesized that both C3a and C5a are also mast cell chemotaxins. Here we report that C3a and C5a are, in fact, potent chemotaxins for the human mast cell line HMC-1. The optimal concentrations, half-maximal effective concentrations (a measure of agonist potency) and the efficacy (response at the optimal concentration) compared with medium control were, for C3a: 10 nM, 0.5 nM, and 256%, respectively; for C5a: 1 nM, 10 pM and 145%. Chemotaxis of HMC-1 cells to both C3a and C5a was blocked by pertussis toxin, suggesting that Gi-coupled receptors are involved in signal transduction. C3a and C5a also induced transient pertussis toxin-inhibitable increases in [Ca2+]i (ED50 = 1 nM for both) that could be homologously but not heterologously desensitized, suggesting that the receptors for C3a and C5a are distinct. These results make C3a the most effective mast cell chemotaxin identified to date. The chemotactic potency described here for C3a is also 100- to 1000-fold greater than for all of its previously described cellular actions. Direct chemoattraction of mast cells by C3a and C5a may help explain the rapid accumulation of mast cells at sites of inflammation.

Adenosine Triphosphate↗

Alterations in mast cell proteinases and protease inhibitors in the progress of cutaneous herpes zoster infection.

The possible involvement of mast cell proteases in the cutaneous inflammation of herpes zoster was studied histochemically in ten patients. Mast cell tryptase and chymase bioactivities were demonstrated enzyme-histochemically. The localization of protease inhibitors as well as tryptase and chymase proteins in mast cells was established using a sequential double-staining method which first demonstrated bioactive tryptase or chymase, followed by immunohistochemical identification of these antigens. Biopsies were taken from involved vesicular and erythematous skin, as well as from normal healthy-looking skin. Tryptase-bioactive mast cells were significantly lower in number in the upper, but not in the deeper dermis of vesicular skin (68 +/- 37 cells/mm2, mean +/- SD) when compared with either healthy-looking (97 +/- 38) or erythematous skin (105 +/- 36) (t-test, P < 0.005). In contrast, chymase-bioactive mast cells were significantly reduced in number both in erythematous skin (44 +/- 20, P < 0.02) and even more so in vesicular skin (26 +/- 20, P < 0.0005) when compared with healthy-looking skin (64 +/- 27). The percentage of alpha 1-antitrypsin -immunoreactive and alpha 1-antichymotrypsin-immunoreactive mast cells in the upper dermis increased steadily from the values in healthy-looking skin (37.9 +/- 18.8 and 82.5 +/- 21.6 per cent) to those in erythematous (64.4 +/- 16.4 and 93.5 +/- 7.9 per cent) and vesicular skin (75.2 +/- 10.2 and 96.4 +/- 4 per cent). A novel finding was that cells showing tryptase immunoreactivity but no enzyme activity were found in two out of nine erythematous skin specimens and in four out of seven vesicular specimens. In healthy-looking skin, all cells with chymase immunoreactivity also displayed chymase bioactivity, but only 53.2 +/- 24.25 per cent of these mast cells in erythematous lesions and 44.4 +/- 15.9 per cent in vesicular lesions showed chymase bioactivity, suggesting inactivation of chymase by protease inhibitors. These results show prominent alterations in mast cell proteinases and protease inhibitors, indicating that these enzymes participate in the cutaneous inflammation due to herpes zoster.

Adult↗

The fibroblast mitogenic activity released from human basophilic cell line KU812 is separate from tryptase and PDGF expression.

The human leukaemia cell line KU812 has previously been used to study basophil differentiation. In this study the authors analysed the capacity of KU812 to produce the mast cell proteinase tryptase and to synthesize factor(s) mitogenic for fibroblasts. KU812 cells were treated with tetradecanoyl-phorbol-13-acetate (TPA), conditioned medium from the human T-cell line Mo (Mo-CM), or cultured under serum free conditions. After 4 days the cells were analysed for cell growth, differentiation, content of tryptase, and secretion of fibroblast mitogenic activity. Mo-CM and serum starvation increased the expression while TPA treatment down-regulated the expression of Fc epsilon RI-alpha chain. An increase in tryptase content in cell extracts was detected after 4 days of culture in serum-free medium or in the presence of Mo-CM. KU812 conditioned media was found to have a baseline expression of mitogenic activity on normal human foreskin fibroblasts that was increased after serum starvation or after treatment with TPA. Mast cell-derived tryptase has previously been reported to be mitogenic for fibroblasts, but in this study the expression of tryptase did not correlate with the expression of fibroblast mitogenic activity in KU812 cells. Furthermore, affinity-purified lung tryptase did not show any mitogenic activity. Platelet-derived growth factor was also excluded. Although the factor(s) from KU812 cells stimulating fibroblast proliferation have not been identified, our results indicate that basophils may be potential producers of growth factors inducing fibroblast proliferation.

3T3 Cells↗

Characterization of a human basophil-like cell line (LAMA-84).

LAMA-84, a human leucocytic cell line, which upon establishment was described as having megakaryocytic, erythroid and granulocytic characteristics, was analysed for expression of various differentiation markers. In addition to some of the previously described phenotypic characteristics, this cell line was found to express mRNA for several proteins characteristic for basophilic leucocytes and mast cells. The authors show that LAMA-84 cells express mRNA for the mast cell tryptase, the proteoglycan core protein, carboxypeptidase A and the alpha and beta chains of the high affinity IgE receptor (Fc epsilon RI). The authors examined the potential of LAMA-84 to differentiate in serum-free medium or after DMSO or PMA treatment. Depending on the inducing factor, surface expression of the Fc epsilon RI alpha-chain was increased from 20% to 35-50% of the cells and mRNA levels for tryptase were increased in serum-free medium and after DMSO treatment. LAMA-84 was found to express CD13, CDw17, CD29, CD33, CD40, CD45 and CD117. Furthermore, mRNA for the eosinophil/basophil markers Charcot-Leyden crystal (CLC) protein and the major basic protein (MBP), as well as the erythrocyte differentiation marker alpha-globin, was detected. However, the authors observed only trace amounts of mRNA for another erythroid differentiation marker (glycophorin), trace amounts of the megakaryocytic marker GPIIIa, and no detectable level of GPIb alpha. By comparing the expression pattern of a panel of differentiation markers in LAMA-84, and a second human cell line (KU812) expressing a basophil phenotype, it is evident that these cell lines, which presently are the only two cell lines identified with basophilic characteristics, share a large number of phenotypic characteristics.

Antigens, Surface↗

Stem cell factor-dependent human cord blood derived mast cells express alpha- and beta-tryptase, heparin and chondroitin sulphate.

The present study sought to determine the expression of alpha- and beta-tryptase in in vitro differentiated human cord blood derived mast cells. We also analysed the glycosaminoglycan composition and the phenotype of the cells. The major protease in human mast cells is tryptase, and cDNAs for two different human tryptases have been characterized, the so-called alpha- and beta-tryptase. By reverse transcriptase-polymerase chain reaction (RT-PCR) we could show that stem cell factor (SCF)-dependent cord blood derived mast cells express both alpha- and beta-tryptase. Furthermore, the cells were stained with a monoclonal antibody (mAb) against tryptase, and the tryptase was enzymatically active cleaving the substrate Z-Gly-Pro-Arg- methoxy-2- naphthylamide (MNA). The majority of the cord blood derived mast cells could also be stained with mAbs against chymase, cathepsin G and CD68. They also expressed Kit/SCFR (CD117), CD13, CD29 and CD45 on the cell surface. The proteoglycan-derived polysaccharide composition of the cells was estimated to be 25-35% of heparin origin and 65-75% of chondroitin sulphate origin. Hence, the cord blood derived mast cells exhibit a phenotype in common with the so-called MCTC type of human mast cells.

Base Sequence↗

Laser Doppler perfusion imager (LDPI)--for the assessment of skin blood flow changes following sympathetic blocks.

A laser Doppler perfusion imager (LDPI) for evaluation of skin blood flow is presented and its use illustrated in two cases with a presumptive diagnosis of reflex sympathetic dystrophy (RSD) treated with local anesthetic sympathetic blocks. Among the advantages are the good spatial resolution over the area measured (12 x 12 cm) and that measurements can be done without contact with the skin. Generating a complete image takes 4.5 min but if higher temporal resolution is needed continuous measurements at one point are possible by use of another software routine.

Adult↗

Number of mutans streptococci or lactobacilli in a total dental plaque sample does not explain the variation in caries better than the numbers in stimulated whole saliva.

For practical reasons the numbers of mutans streptococci (MS) and lactobacilli (LB) in plaque are commonly estimated from saliva samples. The saliva counts are considered to be a reasonable indicator of the entire dentition's total microbial load. However, the value of salivary counts for explaining and predicting caries have been found to be low. There was therefore reason to compare the relationships between caries on the one hand and, on the other, the number of MS or LB in plaque and in saliva, respectively, in order to assess their relative merits for explaining the variation in caries, both in a total material and in subgroups with less favourable oral hygiene. Sixty children aged 14-15 years participated in the study. Caries and plaque were registered and the number of MS and LB was estimated in total plaque and in stimulated saliva samples. The results showed that the number of MS or LB in plaque did not explain the variation in caries to a greater degree than did the salivary counts. For both bacteria the explanatory values increased, as expected, in subgroups with less favourable oral hygiene, but not even at this higher level of explanation was there any difference between plaque and saliva. The LB count was a stronger explanatory variable than the MS count. It was concluded that the number of MS and LB, estimated in total plaque as well as in saliva samples, is not a useful tool in prediction.

Adolescent↗

Multiple high-affinity binding sites for

Binding of [3H]serotonin (5-HT) to membranes prepared from Arctic charr brain homogenates was most consistent with a one-site model for [3H]5-HT binding, with KD and Bmax values of 5.7&plusmn;0.3 nmol l-1 and 60.7&plusmn;7.3 fmol mg-1 protein, respectively. Similarly, 5-HT displacement of [3H]5-HT was best explained by a monophasic model with an apparent Ki of 4.3&plusmn;0.7 nmol l-1. The ability of a number of synthetic 5-HT receptor ligands to displace [3H]5-HT was studied. 8OH-DPAT was found to interact with three [3H]5-HT binding sites, whereas buspirone, TFMPP, spiperone and mianserin all distinguish two sites. In the presence of 300 nmol l-1 buspirone, 8OH-DPAT and mianserin distinguished two [3H]5-HT binding sites, whereas spiperone interacted with only one. Moreover, 8OH-DPAT differentiated three [3H]5-HT binding sites even in the presence of 0.5 mmol l-1 GTP, making it unlikely that these sites represent different affinity states of G-protein-coupled receptors. GTP had no effect on apparent Ki values for 8OH-DPAT, but reduced the Bmax value of the high-affinity site by 60 %. GTP had a similar effect on the saturation binding curve for [3H]5-HT, reducing Bmax by approximately 50 %, whereas KD was unaffected. The results provide evidence for at least three different high-affinity [3H]5-HT binding sites, one of them showing a pharmacological profile strikingly similar to that of the mammalian 5-HT1A receptor.

Journal Article↗

Brain and body oxygen requirements of Gnathonemus petersii, a fish with an exceptionally large brain

Vertebrates have repeatedly been noted for having remarkably constant ratios of brain to body O2 consumption, the brain using 2-8 % of resting body O2 consumption, suggesting that evolution has put strict limits on the energetic cost of brain function. Only man, with a value of 20 %, is an exception to this rule. However, the results presented here suggest that, in the electric fish Gnathonemus petersii, the brain is responsible for approximately 60 % of body O2 consumption, a figure three times higher than that for any other vertebrate studied, including man. The exceptionally high energetic cost of the G. petersii brain appears to be a consequence both of the brain being very large and of the fish being ectothermic. It was also found that G. petersii has a high ability to utilise O2 at low levels. Thus, during falling [O2], this species was found to maintain both its O2 uptake and its electric discharge rate down to an ambient O2 level of 0.8 mg l-1 (at 26 &deg;C), although it was unable to tolerate an [O2] below 0.3 mg l-1. During severe hypoxia (<0.8 mg l-1), G. petersii attempted to gulp air from the water surface. These results establish a new record for the energetic cost of a vertebrate brain and they show that the species possessing such a brain has a high capacity for utilising O2 at very low ambient concentrations.

Journal Article↗

Central monoaminergic responses to salinity and temperature rises in common carp

Juvenile common carp, Cyprinus carpio, were exposed to increased levels of salinity (1 % NaCl) at 25 &deg;C and 30 &deg;C. Levels of the monoamine neurotransmitters dopamine (DA) and serotonin (5-HT) and their metabolites dihydroxyphenylacetic acid and 5-hydroxyindoleacetic acid were determined in different brain parts. Whereas the elevated temperature only resulted in higher levels of the metabolites, increased salinity caused increased levels of DA and 5-HT as well. Increased levels appeared after the first day of exposure and most effects were further enhanced after 1 week in 1 % NaCl. Increases in DA and 5-HT levels were most pronounced in the hypothalamus, which is the major integrative centre controlling the release of hormones. Thus, one possible role of these changes in neurotransmitter metabolism could be to control the release of prolactin and cortisol, two major hormones involved in the regulation of ion homeostasis in teleosts.

Journal Article↗

Stem cell factor, but not flt3 ligand, induces differentiation and activation of human mast cells.

We have examined the effect of human ligand for the flt3/flk2 tyrosine kinase receptor on the differentiation of human mast cells in suspension cultures. We also explored the effect of flt3 ligand (FL) on the human mast cell line HMC-1 and mRNA expression of flt3/flk2 on in vitro developed human mast cells and HMC-1. The growth of cord blood mononuclear cells in suspension cultures was increased when cells were cultured in the presence of FL compared with cells cultured in the presence of stem cell factor (SCF). When SCF and FL were combined, the total cell growth was increased further. Our data show that Fl by itself neither induced differentiation of mast cells nor acted in the SCF-dependent differentiation of human cord blood-derived mast cells (CBMC). Furthermore, no effects of FL were found on the proliferation of HMC-1 cells or the induction of early-immediate response genes in HMC-1 cells. In addition, neither HMC-1 cells nor CBMC expressed mRNA for flt3. As has been shown before, SCF and FL have little biological effect on their own but synergize well with a number of other hematopoietic growth factors. This study shows that a major difference between FL and SCF is that only SCF affects the differentiation and activation of mast cells.

Base Sequence↗

Secondary structure and biophysical activity of synthetic analogues of the pulmonary surfactant polypeptide SP-C.

Native pulmonary-surfactant-associated lipopolypeptide SP-C, its chemically depalmitoylated form and several synthetic analogues lacking the palmitoylcysteine residues were analysed for secondary structure in phospholipid micelles and for biophysical activity in 1,2-dipalmitoyl-sn-glycero-3- phosphocholine/phosphatidylglycerol/palmitic acid (68:22:9, by wt.). Compared with the native molecule, with the entire poly-valyl part in a known alpha-helical conformation, depalmitoylated SP-C was found to be still mainly alpha-helical, but with an approx. 20% decrease in the helical content. A synthetic hybrid polypeptide where the entire poly-valyl alpha-helical part of native SP-C had been replaced with the amino acid sequence of a transmembrane helix of bacteriorhodopsin is also predominantly alpha-helical. In contrast, synthetic SP-C analogues lacking only the palmitoyl groups, by replacement of the palmitoylcysteine residues with cysteine, phenylalanine or serine, or lacking the positively charged amino acids by replacement with alanine, are considerably less alpha-helical than both native and depalmitoylated SP-C. The data indicate that the SP-C palmitoyl groups are important for maintenance of the alpha-helical conformation in parts of the polypeptide, and that the poly-valyl alpha-helical conformation is not fully formed in synthetic SP-C polypeptides. Furthermore, the helical structure of both native and depalmitoylated SP-C in dodecylphosphocholine micelles is very resistant to thermal denaturation, exhibiting ordered structure at 90 degrees C. The alpha-helical content grossly parallels the peptide-induced acceleration of the spreading of phospholipids at an air/water interface and the increase of surface pressure. The data suggest that the alpha-helical conformation itself, rather than just the covalent structure, is of prime importance for the biological function of synthetic pulmonary-surfactant peptides.

Amino Acid Sequence↗