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F Hug

Publications and source records attributed to F Hug.

11 recordsLinked to original sources
PubMed1998

Fibronectin synthesis by human tubular epithelial cells in culture: effects of PDGF and TGF-beta on synthesis and splicing.

BACKGROUND: Enhanced synthesis of extracellular matrix proteins including fibronectin (FN) is associated with the development of sclerosis. In this context we studied FN synthesis by tubular epithelial cells in response to transforming growth factor-beta (TGF-beta) and platelet-derived growth factor (PDGF). METHODS: FN protein synthesis by human tubular epithelial cells in culture (TEC) was measured by biosynthetic labeling and ELISA. Splicing of FN was assessed by RT-PCR and by Northern blotting. RESULTS: Cultivated TEC synthesized and released FN, the majority of which was deposited as an unsoluble protein and a minor portion (10 to 15%) was released into the supernatant. TGF-beta and, to a lesser degree, PDGF, up-regulated FN synthesis. All three FN splice variants (EDA, EDB, and IIICS) were produced. PDGF did not influence the splicing. TGF-beta preferentially up-regulated the EDA splice variant, but had no effect on the splicing of the other domains. CONCLUSIONS: PDGF and TGF-beta both up-regulate FN synthesis of TEC. TGF-beta, but not PDGF, also changed the quality of the de novo synthesized FN, and thus has a different role in the development of sclerosis.

Cells, Cultured↗
PubMed1996

Modulation of Cl- secretion in rat distal colon by genistein, a protein tyrosine kinase inhibitor.

The protein tyrosine kinase inhibitor, genistein, caused an increase of short-circuit current (Isc) across the rat distal colon in forskolin-pretreated tissues, suggesting a synergistic interaction of the drug with cAMP-dependent secretion. In the absence of forskolin, genistein had a dual effect on Isc, it increased Isc in tissues with a low baseline, but decreased Isc in tissues with a high baseline Isc. The secretory effect of genistein was dependent on the presence of Cl- and was blocked by inhibitors of Cl- secretion like bumetanide, an inhibitor of the Na(+)-K(+)-Cl- cotransporter, or 5-nitro-2-(3- phenylpropylamino)-benzoate (NPPB), a Cl- channel blocker. Unidirectional flux measurements revealed that genistein inhibited Na+ and Cl- absorption and induced net Cl- secretion. The protein tyrosine phosphatase inhibitor vanadate suppressed the secretory effect of genistein. In contrast, genistein caused an inhibition of carbachol-induced, i.e. Ca(2+)-mediated secretion. Whole-cell patch-clamp experiments confirmed the synergistic effect of genistein on cAMP-induced Cl- currents. In the presence of forskolin, genistein caused a depolarization concomitant with an increase in membrane inward current. In addition, genistein caused an inhibition of a basal K+ conductance and inhibited the Ca(2+)-dependent K+ conductance stimulated by carbachol. These results suggest a complex role of the protein tyrosine kinase pathway in the control of colonic Cl- secretion, an antagonistic action on the cAMP pathway and a synergistic action on the Ca2+ pathway as revealed by the opposing effects of genistein. The physiological importance of this regulation remains to be clarified.

Animals↗
PubMed1996

Modulation by fish oil diet of eicosanoid-induced anion secretion in the rat distal colon.

Eicosanoids are involved in the mediation of inflammatory and allergic processes in the gut. In order to evaluate a potential beneficial effect of the diet, the effect of mediators of inflammation and of a sensitization against egg albumin on anion secretion across the colon was tested using rats fed on a diet containing 15% fish oil as compared to 15% olive oil as donor animals. Feeding on a fish oil diet significantly reduced the response to bradykinin or phospholipase C, known agonist of prostaglandin-induced secretion, by about 50%. The increase in short-circuit current (Isc) induced by the phospholipase A2 stimulator, melittin, or by distension of the gut wall were only insignificantly inhibited by 15-30%. Administration of egg albumin to the mucosas from animals sensitized against egg albumin induced an indomethacin- and tetrodotoxin-sensitive increase in Isc. This response was, however, only insignificantly (30%) reduced by the fish oil diet. In conclusion, the effect of fish oil diet depends on the stimulus used for activation of prostaglandin release. This suggested that different pools of arachidonic acid are differentially affected by the diet or that certain stimuli for phospholipases are strong enough to overcome the effect of a reduced substrate availability. Consequently, a diet rich in polyunsaturated n-3 fatty acids may only play an adjuvant role for the therapy of inflammatory or allergic intestinal diseases.

Acid-Base Equilibrium↗
PubMed1996

Cyclic AMP-dependent regulation of K+ transport in the rat distal colon.

1. The effect of agonists of the cyclic AMP pathway and of 293B, a chromanole-derived K+ channel blocker, on K+ transport in the rat distal colon was studied by measuring unidirectional fluxes, uptake, and efflux of Rb+ in mucosa-submucosa preparations and by patch-clamp of crypt epithelia from isolated crypts. 2. 293B concentration-dependently inhibited basal and forskolin-stimulated short-circuit current. In isolated crypts 293B blocked a basal K+ conductance but had no effect on cyclic AMP-evoked depolarization induced by the opening of apical Cl- channels. When the effect of cyclic AMP on Cl- conductance was prevented by substituting Cl- with gluconate, an inhibition of total cellular K+ conductance by forskolin and a membrane-permeable cyclic AMP analogue was unmasked. 3. Unidirectional ion flux measurements revealed that 293B suppressed the increase in JRbsm induced by forskolin. This, together with the inhibition of cyclic AMP-induced anion secretion indicates that the drug blocks K+ channels, presumably both in the apical and the basolateral membrane. Forskolin caused not only inhibition of K+ absorption, but also stimulation of K+ secretion. The inhibition was diminished, but not blocked, in the presence of inhibitors of the apical H(+)-K(+)-ATPase, vanadate and ouabain. Forskolin stimulated serosal, bumetanide-sensitive Rb+ uptake, whereas mucosal, ouabain/vanadate-sensitive uptake remained unaffected. 4. Efflux experiments revealed that forskolin caused a redistribution of cellular K+ efflux reducing the ratio of basolateral versus apical Rb+ efflux. 5. These results suggest that intracellular cyclic AMP exerts its effects on K+ transport by several mechanisms: an increase in the driving force for K+ efflux due to the depolarization induced by opening of Cl- channels, a stimulation of the basolateral uptake of K+ via the Na(+)-K(+)-Cl(-)-cotransporter, and a decrease of the ratio of basolateral versus apical K+ conductance leading to an enhanced efflux of K+ into the lumen and a reduced K+ efflux to the serosal compartment.

Animals↗
PubMed1995

L-fucose residues on cellulose-based dialysis membranes: quantification of membrane-associated L-fucose and analysis of specific lectin binding.

Contact of mononuclear human leukocytes with cellulose dialysis membranes may result in complement-independent cell activation, i.e. enhanced synthesis of cytokines, prostaglandins and an increase in beta 2-micro-globulin synthesis. Cellular contact activation is specifically inhibited by the monosaccharide L-fucose suggesting that dialysis membrane associated L-fucose residues are involved in leukocyte activation. In this study we have detected and quantitated L-fucose on commercially-available cellulose dialysis membranes using two approaches. A sensitive enzymatic fluorescence assay detected L-fucose after acid hydrolysis of flat sheet membranes. Values ranged from 79.3 +/- 3.6 to 90.2 +/- 5.0 pmol cm-2 for Hemophan or Cuprophan respectively. Enzymatic cleavage of terminal alpha-L-fucopyranoses with alpha-L-fucosidase yielded 7.7 +/- 3.3 pmol L-fucose per cm2 for Cuprophan. Enzymatic hydrolysis of the synthetic polymer membranes AN-69 and PC-PE did not yield detectable amounts of L-fucose. In a second approach, binding of the fucose specific lectins of Lotus tetragonolobus and Ulex europaeus (UEAI) demonstrated the presence of biologically accessible L-fucose on the surface of cellulose membranes. Specific binding was observed with Cuprophan, and up to 2.6 +/- 0.3 pmol L-fucose per cm2 was calculated to be present from Langmuir-type adsorption isotherms. The data presented are in line with the hypothesis that surface-associated L-fucose residues on cellulose dialysis membranes participate in leukocyte contact activation.

Acrylic Resins↗
PubMed1993

Stimulation of mononuclear cells by contact with cuprophan membranes: further increase of beta 2-microglobulin synthesis by activated late complement components.

Contact of mononuclear cells (MNC) with cuprophan membranes in vitro causes an increase in beta 2-microglobulin (beta 2m) synthesis. Since in vivo the dialyzer membrane is rapidly coated with plasma proteins, contact activation of MNC was tested in the presence of normal human serum (NHS). After contact with cuprophan, deposition of C5b-9 on the cells was seen, followed by an increase in beta 2m synthesis and cytokine release, exceeding that seen after contact activation in the absence of serum. Inactivated serum or serum deficient in C8 did not increase beta 2m production, indicating that the additional activation was due to complement C5b-9. The results suggest that there are two cuprophan-related mechanisms of cell activation: one by contact of cells with the membrane, the other by the complement activation products. Both might synergistically contribute to an increased beta 2m synthesis in hemodialysis patients.

Blotting, Northern↗
PubMed1975

Thermal inactivation of cholera phages.

Thermal inactivation of seven cholera phages have been tested over the temperature range between 50 degrees to 70 degrees C. It was found that the phages vary widely in their heat sensitivity, Mukerjee's phages III being the most sensitive of the whole group. With all the phages over the temperature range studies, the inactivation curve seem to follow the pattern of virus thermal inactivation in general, the inactivation proceeding initially at a rapid rate, which in about 15 minutes time, gradually changes to a slower rate, each component tending to follow kinetics of the first order. The difficulty of explaining this phenomenon on the basis of population heterogeneity has been discussed.

Bacteriological Techniques↗