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Biomedical subjects

G C Hansson

Publications and source records attributed to G C Hansson.

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

The intestinal tract and the pathophysiology of arterial hypertension: an experimental study on Dahl rats.

Salt depleted rabbits and humans excrete an oral sodium load more quickly via the kidneys than an intravenous one. This has been ascribed to the presence of a sodium sensor in the gastrointestinal tract which in some way can influence renal function. The purpose of this study was to investigate this response in the Dahl rats. Renal and faecal sodium excretion was followed in the two strains of rats (normotensive, saltresistant (SR/Jr) and hypertensive, saltsensitive (SS/Jr) rats). After 4 days on a low salt diet they were given NaCl (1.5 mmol k(-1) body wt) either by gavage or intravenously. SR/Jr rats showed an increased renal sodium excretion both after oral and intravenous sodium repletion. The excretion was 2-3 times greater after th oral than after the intravenous administration. The SS/Jr rats augmented their renal sodium excretion only after the oral load, although the sodium excretion was significantly less than in SR/Jr rats. In fact, during the first 8 h after giving sodium orally the renal excretion of sodium was on an average eight times larger in the SR/Jr than in the SS/Jr rats. Renal excretion of sodium was similar in the two strains after intravenous administration. We conclude that the hypertensive SS/Jr rats have great difficulties in excreting an oral sodium load, a phenomenon that may be of importance in the pathophysiology of arterial hypertension in this strain of rats.

Animals↗
PubMed1995

The small intestine, salt intake and arterial hypertension.

The possibility that the small intestine is of importance for the development of arterial hypertension is discussed in relation to the presence of an intestinal sodium sensor and an intestinal natriuretic factor. We propose that an intestinal sodium sensor upon activation releases a factor that evokes renal excretion of sodium. Based on observations made in the Dahl strain of hypertensive rats it is suggested that an attenuation of this mechanism may contribute to the development of arterial hypertension.

Animals↗
PubMed1995

Partial purification of a factor from the feline small intestine causing natriuresis in vivo and inhibiting rubidium uptake into renal cells in vitro.

Small intestine of cat was homogenized under denaturing condition (6.0 M guanidinium chloride) in the presence of protease inhibitors and molecules in the 500 to 10,000 Da mass range were obtained by sequential ultrafiltrations. This material was separated by gel chromatography (Sephadex G-25), where fractions eluted in the mass range of 500-1000 Da inhibited 86Rb uptake into kidney slices in vitro. Furthermore, the same fractions exhibited a natriuretic effect when given intravenously to anesthetized rats. The gel chromatography fractions were further purified by cation exchange chromatography (CM-Sephadex C-25). Fractions eluted with a NaCl solution of around 250 mM inhibited 86Rb uptake into renal cortical tissue in vitro and showed natriuretic activity when tested in vivo in rats. It is proposed that intestine contains a water soluble natriuretic factor with an apparent molecular mass of 500-1000 Da. This material evokes a natriuresis in vivo and inhibits 86Rb uptake in vitro.

Animals↗
PubMed1994

A secreted mucin carrying sialyl-Lewis a from colon carcinoma cells binds to E-selectin and inhibits HL-60 cell adhesion.

Sialyl-Lewis x and a are known as ligands for E-selectin (ELAM-I) involved in leukocyte-endothelial adhesion. L-CanAg (light cancer antigen), secreted by a colon carcinoma cell line COLO 205, is a soluble mucin-type glycoprotein expressing sialyl-Lewis a antigens. L-CanAg was purified from spent culture medium by trichloracetic acid precipitation and Superose 6 gel filtration. With a monoclonal antibody against E-selectin (BBAI) as a positive control, the purified L-CanAg was shown to bind to E-selectin-Fc coated into plastic microtiter wells and to the surface of transiently E-selectin-transfected COS-I cells in a Ca(2+)-dependent way. Immunofluorescent double labelling showed that both BBAI and L-CanAg stained the same cells and morphological co-localization on E-selectin-transfected COS-I cells. Like BBAI, L-CanAg can inhibit leukocyte HL-60 cell adhesion to E-selectin-transfected COS-I cells, and this inhibition can be blocked by a F(ab')2 fragment directed against the sialyl-Lewis a epitope.

Antigens, Neoplasm↗
PubMed1994

Purification and characterization of sialyl-Le(a)-carrying mucins of human bile; evidence for the presence of MUC1 and MUC3 apoproteins.

Purification of sialyl-Le(a)-carrying mucins from primary human bile by trichloroacetic acid precipitation, delipidation, and gel filtration in guanidinium chloride gave three separable fractions, one of which was further purified by affinity chromatography. These fractions, named SBG1 (for soluble bile glycoprotein), SBG2, and SBG3 had molecular masses of > 1100, 800-950, and 100-250 kDa, respectively, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Their mucin characteristics were indicated by a high carbohydrate content, ranging from 74 to 95%. The carbohydrate compositions indicated the presence of very long fucosylated polylactosamine chains. Amino acid analyses showed high abundance of serine and threonine in all three fractions (19-36%), confirming their mucin-like nature. Immunochemical analyses of deglycosylated samples detected the MUC1 mucin apoprotein in SBG2 and the MUC3 protein in SBG1. To our knowledge, this is the first report of a MUC3 mucin being purified. This mucin showed no significant reduction in size upon trypsin treatment or disulfide bond reduction and alkylation. Gel filtration of three samples of secondary bile showed that the size distribution of sialyl-Le(a)-carrying glycoproteins was similar to that found in primary bile, and immunochemical analysis showed that the MUC1 protein was present in all three samples. In one sample an additional fraction was isolated, which was insoluble in 6 M guanidinium chloride, but was solubilized upon reduction and alkylation. mRNAs from gallbladder epithelia were analyzed in Northern blot hybridizations showing that the MUC1 and MUC3 but not the MUC2 mucin apoprotein genes were expressed.

Amino Acid Sequence↗
PubMed1994

Molecular cloning of a cDNA coding for a region of an apoprotein from the 'insoluble' mucin complex of rat small intestine.

The major part of rat small intestinal mucins occurs as an 'insoluble' glycoprotein complex unextractable in 6 M guanidinium chloride unless disulfide bonds are cleaved. One of the trypsin-resistant high-glycosylated domains of this complex (glycopeptide A) was recently isolated. We have now deglycosylated it with HF, injected it into rabbits and the obtained antiserum was used for expression cloning providing a cDNA clone (VR-1A). This clone contained an open reading frame of 235 amino acids composed of two regions. The deduced N-terminal 53 amino acids included seven Cys residues and only one Ser, followed by a region of 182 residues with 64% Ser and Thr but devoid of Cys residues. Analysis of mRNA revealed a transcript of about 12 kb, identical in size to a band labelled with a probe based on the rat mucin-like protein (MLP/Muc2) cDNA. Pulsed-field gel electrophoresis of genomic rat DNA showed identical bands (380 and 500 kb) when blots were sequentially probed with the MLP/Muc2 probe and VR-1A. A panel of mouse x rat hybrids was used to localize the gene corresponding to both VR-1A and Muc2 to rat chromosome 1. The results strongly suggest that the 'insoluble' mucin complex of the rat small intestine is encoded by the Muc2 gene.

Amino Acid Sequence↗
PubMed1994

Inhibitory effects of theophylline, terbutaline, and hydrocortisone on leukotriene B4 and C4 generation by human leukocytes in vitro.

Leukotriene B4 (LTB4) and leukotriene C4 (LTC4) are considered to be important mediators in the pathophysiology of asthma. Theophylline, terbutaline, and hydrocortisone are drugs commonly used in the treatment of asthma. In the present study we have investigated the in vitro inhibitory effects of theophylline, terbutaline, and hydrocortisone on LTB4 and LTC4 generation from human leukocytes. After preincubation in the presence of these drugs, the cells were stimulated with the calcium ionophore A 23187 and the supernatants were analyzed for their LTB4 and LTC4 content using reverse-phase high-performance liquid chromatography (HPLC). Total leukotriene (LT) production (the combined amounts of LTB4 and LTC4) was dose-dependently inhibited by pretreatment with theophylline, terbutaline or hydrocortisone. Therapeutic levels of hydrocortisone (5 x 10(-6) M) plus theophylline (5 x 10(-5) M) inhibited LTB4 and LTC4 production in an additive way, as did the combination of hydrocortisone plus terbutaline (5 x 10(-8) M). A statistically significant effect of diminished LTB4 generation was obtained after preincubation with therapeutic levels of theophylline plus terbutaline, but no such effect was seen for LTC4 levels. The in vitro inhibitory effects on LTB4 and LTC4 generation from human leukocytes by theophylline, terbutaline, and hydrocortisone, as well as the additive effect of hydrocortisone plus theophylline or terbutaline, add to our understanding of the therapeutic effects of these drugs in the treatment of bronchopulmonary obstruction.

Calcimycin↗
PubMed1994

Defective inhibition by dexamethasone of leukotriene B4 and C4 production by leukocytes in patients with cystic fibrosis.

To investigate the effects of glucocorticoids on leukotriene (LT) generation in patients with cystic fibrosis (CF), we evaluated calcium ionophore A23187-induced LTB4 and LTC4 production by leukocytes with and without pretreatment with dexamethasone. The CF patients were in good condition and did not have acute infection. There were no significant differences in LTB4 and LTC4 production without dexamethasone pretreatment between the CF patients and controls. However, the ratios of LTB4 and LTC4 production by leukocytes preincubated with dexamethasone to those of leukocytes without dexamethasone pretreatment were significantly higher in the CF patients than in the controls (both p < 0.05). Our data suggest that the response of LTB4 and LTC4 production to dexamethasone is disturbed in patients with CF. The generation of LTs may be enhanced due to a disturbance in glucocorticoid suppression.

Adolescent↗
PubMed1994

Effects of pancreatic and snake venom phospholipase A2 on the generation of leukotriene B4 and C4 by human leukocytes in vitro.

To investigate the effects of phospholipase A2 (PLA2) on leukotriene (LT) generation, we evaluated LTB4 and LTC4 production by human leukocytes after incubation with porcine pancreatic and snake venom PLA2. The pancreatic and venom PLA2 did not induce LTB4 and LTC4 generation by human leukocytes alone. However, both types of PLA2 promoted opsonized zymosan-induced LTB4 production in a dose-dependent manner (from 10(-10) to 10(-8) M), whereas they had no effect on opsonized zymosan-induced LTC4 generation. This promoting effect of pancreatic and venom PLA2 required Ca2+ in the incubation medium and was inhibited by mepacrine (10(-5) M). These findings suggest that increased production of PLA2 in combination with inflammatory stimuli such as endotoxin enhance LTB4 biosynthesis in acute pancreatitis.

Humans↗
PubMed1993

Characterization of two different glycosylated domains from the insoluble mucin complex of rat small intestine.

The highly glycosylated domains of rat small intestinal mucins were isolated after reduction and trypsin digestion and separated into two populations (A and B) by gel chromatography. The molecular mass values were 650 and 335 kDa, respectively, and the relative yields suggest that the two glycopeptides occur in equimolar proportions. Electron microscopy revealed linear structures with weight average lengths of 230 nm (A) and 110 nm (B) corresponding to a mass/unit length of about 3 kDa/nm. The protein cores (17-19%) contain large amounts of threonine (over 40%), serine (17-24%), and proline (18-19%). Carbohydrate and sulfate account for approximately 80 and 0.5%, respectively, and gas chromatography-mass spectrometry showed that the patterns of neutral and sialic acid-containing glycans are very similar in the two glycopeptides. Both contain a significant amount (7-10 mol %) of single GalNAc residues, the average oligosaccharide is about 4 sugar residues long, and the largest species observed are heptasaccharides. The major neutral and sialic acid-containing oligosaccharides are Fuc1-2Gal1-3GalNAcol and GlcNAc1-6(NeuGc2-Gal1-3)GalNAcol, respectively. Sialic acid is present as both N-acetyl- and N-glycoloyl-neuraminic acid. Repeated extractions of the tissue with guanidinium chloride left approximately 80% of the mucus glycoproteins as an insoluble glycoprotein complex whereas exposure to dithiothreitol or high speed homogenization accomplished complete solubilization. The "subunits" obtained after reduction with dithiothreitol are larger than glycopeptides A and B, and fragments corresponding in size to the latter are obtained after cleavage with trypsin. Most of the mucins from rat small intestine thus occurs as an insoluble glycoprotein complex composed of subunits joined with disulfide bonds. The subunits contain two highly glycosylated regions with different lengths substituted with very similar oligosaccharides.

Amino Acids↗
PubMed1993

Discrimination of MUC1 mucins from other sialyl-Le(a)-carrying glycoproteins produced by colon carcinoma cells using a novel monoclonal antibody.

Earlier studies (Baeckström et al., J. Biol. Chem., 266: 21537-21547, 1991) have revealed that the colon carcinoma cell line COLO 205 produces two different proteins carrying the carcinoma-associated sialyl-Le(a) carbohydrate epitope. One is the MUC1 mucin apoprotein, and the other protein is smaller and has not been characterized in detail. This paper describes a comparison of COLO 205 with three other colon carcinoma cell lines, aided by the use of a novel MUC1-reactive monoclonal antibody, Ma552. Ma552 reacted with H-CanAg, the MUC1 mucin purified from COLO 205, the binding increasing greatly upon partial deglycosylation of H-CanAg with trifluoromethanesulfonic acid. This pattern of reactivity was in contrast with that of the MUC1-reactive monoclonal antibody HMFG-2, which did not recognize H-CanAg without prior deglycosylation. The nature of the epitope of Ma552 was further investigated using a synthetic peptide corresponding to the tandem-repeat sequence of the MUC1 protein. When the peptide was used as an inhibitor of antibody binding to immobilized, partially deglycosylated H-CanAg mucin, Ma552 was inhibited, as was HMFG-2. Using short, immobilized synthetic peptides identical to parts of the MUC1 tandem repeat, the reactivity of Ma552 was mapped to the hexapeptide TRPAPG. Ma552 and C50, a monoclonal antibody reactive with sialyl-Le(a), were used in immunofluorometric assays to analyze gel filtration fractions of extracts and spent media from the colorectal carcinoma cell lines COLO 205, SW1116, LoVo, and LS 174T. Using C50 in a homologous assay, all sialyl-Le(a)-carrying glycoproteins were detected. Among these, mucins based on the MUC1 apoprotein were identified using Ma552 and C50 in a combination assay. The results showed that sialyl-Le(a) was present on more than one glycoprotein not only in COLO 205 but also in SW1116 and LoVo. The Ma552/Eu-C50 assay revealed the presence of sialyl-Le(a)-carrying MUC1 in COLO 205 as expected, as well as in SW1116. The presence of MUC1 in Ma552-reactive fractions was confirmed by deglycosylation followed by assaying with the monoclonal antibody HMFG-2. Furthermore, Northern blots revealed the presence of MUC1 mRNA only in the two Ma552-positive cell lines.

Amino Acid Sequence↗
PubMed1993

An intestinal natriuretic factor.

In 1975, reports were published that suggested that the gastrointestinal tract can "taste" the intake of sodium and in some unknown way influence the kidneys to increase sodium excretion. To test whether the intestine contained a natriuretic factor, intestinal tissue from cats was homogenized and fractionated by ultrafiltration to a molecular range of approximately 500-10,000 Da and separated by gel chromatography (Sephadex G25). The fractions were pooled into four large fractions that were assayed for "natriuretic" activity on anesthetized rats. The fraction containing the material with an apparent molecular mass of 500-1,000 Da augmented renal excretion of sodium and water, whereas the other pooled fractions did not exhibit any consistent natriuretic effect. The "natriuretic" fractions from gel filtration were further purified by ion exchange chromatography using a cation exchanger. The natriuretic activity was eluted from the ion exchange chromatography column at a NaCl concentration of 250 mM. Preliminary experiments on Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR) suggest that the intestinal influence on renal sodium excretion is more pronounced in SHR than in WKY rats.

Animals↗
PubMed1992

Two novel decaglycosylceramides with a blood group A-active tetrasaccharide repeat in the epithelial cells of the small intestine of inbred rats.

A novel type of blood group A-active glycosphingo-lipid was isolated from the epithelial cells of the small intestine of one strain of inbred rats. Electron-impact mass spectrometry of the permethylated and LiAlH4-reduced glycolipid indicated that it is a decaglycosylceramide with a branched oligosaccharide chain. Methylation analysis, gas chromatography-mass spectroscopy of the partially methylated alditol acetates, sequential degradation by exoglycosidases and characterization of the reaction products by TLC immunostaining with appropriate anti-A and anti-H antibodies, and 1H NMR spectrometry resulted in the characterization of a decaglycosylceramide with two variants in a 7/3 ratio. It was termed AA-10. [formula: see text] The major variant has only type 1 chains, whereas the minor one has type 2 chain in the C6-linked branch. This is a novel type of glycolipid with a blood group A-active tetrasaccharide repeat. Genetic analysis demonstrated that AA-10 is inherited as an autosomal dominant trait.

ABO Blood-Group System↗
PubMed1992

N-linked glycopeptides with blood group determinants lacking neuraminic acid from the epithelial cells of rat small and large intestine.

The N-linked type of glycans were prepared as their glycopeptides after pronase digestion of the epithelial cells from the small and large intestine of two inbred strains of rat. These glycopeptides were analysed for sugar composition, for blood-group activity, by 1H-NMR spectroscopy, and after permethylation by electron-impact mass spectrometry. The glycopeptides were of the triantennary and tetraantennary types with intersected GlcNAc. The terminal parts were, in contrast to most N-linked glycans, devoid of neuraminic acid residues. Instead they contained blood-group determinants. Blood-group-H types 1 (Fuc alpha 1-2Gal beta 1-3GlcNAc) and 2(Fuc alpha 1-2Gal beta 1-4GlcNAc) were found in the small and large intestines of both strains, although type-1 predominated. One rat strain (GOT-W) did not express blood-group-A glycopeptides in the small intestine, but the large intestine from the same strain did. The other strain (GOT-BW) expressed blood-group-A determinants in the small intestine. The lack of neuraminic acid residues in the small and large intestine and of blood-group-B activity in the large intestine differed from that found in glycosphingolipids obtained from the same organs.

Animals↗
PubMed1991

Analysis of sialic acid-containing mucin oligosaccharides from porcine small intestine by high-temperature gas chromatography-mass spectrometry of their dimethylamides.

Acetylation of sialic acid-containing oligosaccharides lactonises the sialic acid residue quantitatively for all oligosaccharides studied except for 6'-sialyl-lactose. The modified, unsulphated, sialylated and sulphated oligosaccharides can then be fractionated by anion-exchange chromatography. Ammonolysis of the lactones followed by methylation yielded the dimethylamides, which are amenable to g.l.c.-m.s. and give intense and informative mass spectra. This approach has been used to characterise the sialic acid-containing O-linked oligosaccharides obtained from the mucin glycopeptides of the small intestine of the pig. At least 28 structures were found, having NeuAc or NeuGc 6-linked to the HexNAc attached to the peptide core or to a Hex 3-linked to HexNAc. Four different disialylated oligosaccharides were found having NeuAc or NeuGc on the Hex residue 3-linked to HexNAc and 6-linked to HexNAc.

Animals↗