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M Fromm

Publications and source records attributed to M Fromm.

At least 91 records · Page 5Linked to original sources
PubMed1994

Effect of acute Yersinia enterocolitica infection on intestinal barrier function in the mouse.

BACKGROUND: Yersinia enterocolitica is an important cause of diarrhea, but little is known about the underlying mechanisms. We therefore studied the impact of acute Y. enterocolitica infection on intestinal barrier function in a mouse model. METHODS: For this purpose CD-1 mice were infected with Y. enterocolitica (serotype 08; 6 x 10(7) viable bacteria), and alternating current impedance analysis was performed on days 1, 2, 3, 5, and 8 after infection. RESULTS: The infection resulted in a decrease in epithelial resistance from 18.0 +/- 0.9 omega.cm2 (controls) to 12.1 +/- 0.5 omega.cm2 (day 1, p < 0.001), from which the animals recovered by day 5. To locate this loss in barrier function, the horizontal distribution of local conductances was measured by voltage scanning, yielding two results. First, conductance was homogeneously distributed across the chamber area, excluding erosions or ulcers among the gross surface area and favoring tight junction opening as the source of barrier dysfunction. Second, the conductance of villus tips was compared with that of the intervillus region (consisting of lateral villus walls plus crypts). On day 1 the former was increased by 74% and the latter by 18%. Then, two other mechanisms of diarrhea were tested, namely malabsorption and secretion. First, the increase in ISC after the addition of 3-O-methylglucose, representing Na(+)-glucose cotransport, was shown not to be impaired. Second, bumetanide-inhibitable ISC, representing electrogenic Cl- secretion, also did not differ between controls and infected animals. CONCLUSIONS: Our data show that epithelial barrier dysfunction plays a role in Y. enterocolitica infection, while Na(+)-glucose cotransport and electrogenic Cl- secretion are unaltered.

Acute Disease↗
PubMed1994

Tight junctions of the human corneal endothelium: morphological and electrophysiological features.

The corneal endothelium controls the hydration and nutrition of the avascular corneal stroma. To analyze the role of the tight junctions (TJ) for these functions, we examined human corneal endothelium by thin-section and freeze-fracture electron microscopy and by impedance analysis. On thin sections, tannic acid was seen to mark the external leaflet of the lateral plasma membrane also beyond the location of the TJ, indicating a significant macromolecular porosity of the TJ. On freeze-fracture images, the TJ surrounded the entire apicolateral plasma membrane but were found to be focally incomplete, suggesting a nonhomogeneous seal of the lateral intercellular space. Impedance analysis revealed a very leaky endothelial layer with a transendothelial resistance of 9.0 +/- 1.4 omega cm2. These findings are consistent with the hypothesized "pump-leak" model of the corneal endothelium: the TJ allow an effective dehydration of the corneal stroma, whereas the interruptions in the TJ network may be the morphological correlate for the passage of nutrients into the corneal stroma. Our data corroborate the assignment of the corneal endothelium to the group of "very leaky epithelia" that exhibit high transport rates of water and solutes against only minimal osmotic gradients.

Cell Membrane↗
PubMed1993

Bovine and porcine large intestine as model epithelia in a student lab course.

A short-circuit current experiment on epithelial ion transport is described that is suitable for student classes in human and animal physiology. Segments of late distal colon from either pig or cow are obtained from the slaughterhouse depending on the animals' daily schedule. Initial tissue preparation already in the slaughterhouse, cold storage, and proper choice of bath solutions are essential prerequisites for success. Students monitor spontaneous transepithelial voltage and short-circuit current (Isc) by use of manually operated voltage clamp units. Two main transport mechanisms are studied, electrogenic Na+ absorption and Cl- secretion. Electrogenic Na+ absorption is studied by measuring the Isc drop after amiloride. Then Cl- secretion is stimulated by theophylline and subsequently inhibited by furosemide. In some experiments K+ secretion can be detected by the blocking effect of mucosal Ba2+. Response of tissues from pig and cow is qualitatively similar but quantitatively different. The equipment is sturdy and inexpensive, can be provided by most departmental workshops, and has been tested for 3 yr in regular lab courses. Observations made during these experiments are closely related to clinical states, such as secretory diarrhea, cystic fibrosis, and hyperaldosteronism, as well as to the mechanisms of clinically used diuretics.

Animals↗
PubMed1993

Electrogenic Na+ absorption of rat distal colon is confined to surface epithelium: a voltage-scanning study.

There is no quantitative assignment of large intestinal electrogenic Na+ absorption to surface epithelium and crypts so far. We determined the spatial distribution of electrogenic Na+ absorption to crypts and surface epithelium of rat late distal colon using a modified voltage-scanning technique. Voltage deflections resulting from external 30-Hz current were sensed by an extracellular microelectrode stepping at 0.7 Hz above crypt openings or surface epithelium. Local conductances were calculated applying a planar model of electrical field distribution to surface epithelium and a electrostatic disk source model to the crypts. These models were confirmed by methodological experiments where the electrode position was varied in vertical and horizontal direction. Electrogenic Na+ absorption was detected by blocking apical Na+ channels by mucosal 0.1 mM amiloride. Under control conditions surface epithelium contributed 44% (2.0 +/- 0.2 mS/cm2) and crypts 56% (2.6 +/- 0.2 mS/cm2) to the total conductance of 4.6 +/- 0.4 mS/cm2. Electrogenic Na+ absorption was induced by 6 h in vitro incubation in a medium containing 3 nM aldosterone. This caused a short-circuit current (ISC) of 12.1 +/- 0.8 mumol.h-1.cm-2, which was paralleled by a 2.5-fold increase in surface epithelial conductance to 5.1 +/- 0.4 mS/cm2, whereas crypt conductance was not significantly altered (3.0 +/- 0.2 mS/cm2). Amiloride reversed ISC to -0.8 +/- 0.1 mumol.-1.cm-2 and decreased surface epithelium conductance to 2.3 +/- 0.3 mS/cm2 but again had no significant effect on crypt conductance (2.5 +/- 0.3 mS/cm2). Sham incubation (no hormones added) for 6 h neither induced electrogenic transport nor altered local epithelial conductances.(ABSTRACT TRUNCATED AT 250 WORDS)

Aldosterone↗
PubMed1993

Origin of cAMP-dependent Cl- secretion from both crypts and surface epithelia of rat intestine.

Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent Cl- secretion provides the ionic basis for secretory diarrhea. We quantified the spatial distribution of this process by measuring local ion conductance in crypts and surface epithelium or villi of rat late distal colon and ileum. By use of an improved voltage-scanning technique, the tissue was clamped to a 30-Hz sine-wave current and the electrical field above the respective structures was sensed by a stepping glass microelectrode. Under control conditions, crypts and surface epithelium contributed 61 and 39%, respectively, to the total ion conductance of distal colon. Theophylline (10 mM) increased crypt conductance (Gc) by 64% from 2.5 +/- 0.2 to 4.1 +/- 0.3 mS/cm2 and surface epithelium conductance (Gs) by 69% from 1.6 +/- 0.1 to 2.7 +/- 0.1 mS/cm2. These changes in local conductances were completely Cl- dependent, since theophylline had no effect when Cl- was replaced by gluconate. Similar results were obtained when Cl- secretion was elicited by prostaglandin E1 (1 microM) or by dibutyryl-cAMP (DBcAMP, 1 mM). After stimulation, the Cl- channel blocker 5-nitro-2-(3-phenyl-propylamino)benzoic acid (1 mM) decreased both Gc and Gs. In rat ileum, theophylline plus DBcAMP caused an increase in total conductance of 19% only because of its large paracellular conductance. The ratio of scanning signals above villi and intervillous spaces was unaffected, indicating that Cl- conductance is induced in both crypts and villi. We conclude that in distal large intestine cAMP-dependent Cl- secretion is not confined to crypts but is evenly performed also by surface cells. A similar distribution exists in small intestine.

Animals↗
PubMed1993

Control of electrogenic Na+ absorption in rat late distal colon by nanomolar aldosterone added in vitro.

It has been possible to obtain in a mammalian epithelium of dietetically and surgically untreated animals a dose response of in vitro-added aldosterone (Aldo, 10(-10) to 10(-5) M) on electrogenic Na+ absorption (JeNa). JeNa was measured in the Ussing chamber on stripped rat late distal colon 8 h after in vitro addition of Aldo. Submaximal effects were obtained at 3 nM Aldo; after a lag time of 2 h, short-circuit current (Isc) increased to a maximum of 234 +/- 15 microA/cm2 and dropped after 0.1 mM amiloride to -18 +/- 3 microA/cm2, resulting in JeNa of 9.4 +/- 0.6 mumol.h-1 x cm-1. Net Na+ tracer fluxes and Isc exhibited parallel time courses, so that electroneutral Na+ transport was not induced in late distal colon by acute Aldo. A plot of JeNa vs. Na conductance revealed an electromotive force (ENa) of 126 +/- 1 mV for all Aldo concentrations tested. Kinetic data were as follows: Michaelis constant 1.2 nM, maximal velocity (Vmax) 10.5 mumol.h-1 x cm-2, and Hill coefficient 2.1. In contrast to the large effect in late distal colon, 3 nM Aldo caused JeNa of < 1 mumol.h-1 x cm-2 in early distal colon, proximal colon, and cecum. Antimineralocorticoid sensitivity and ENa did not vary with Aldo concentration or time of the experiment, consistent with a unique mechanism during the early and late response up to 8 h, as well as at mineralocorticoid and glucocorticoid Aldo concentrations. Acute Aldo in a range of 0.1-10 nM fully controls JeNa between zero and Vmax in late distal colon.(ABSTRACT TRUNCATED AT 250 WORDS)

Aldosterone↗
PubMed1993

11-beta-Hydroxysteroid dehydrogenase of rat lung: enzyme kinetic, oxidase-reductase ratio, electrolyte and trace element dependence.

The modulation of the intracellular glucocorticoidal effect on surfactant synthesis of the fetal lung by the metabolic capacity of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) could be an important factor in lung maturation. The kinetic properties of microsomal 11 beta-HSD of the rat lung are characterized with respect to product inhibition, substrate specificity, effect of electrolytes or trace elements, and the dependence of the oxidase reductase (OR) ratio on incubation conditions. With NADP+ product inhibition of the reductase was demonstrated. The most common trace elements and electrolytes exhibited no effect on the activity of 11 beta-HSD. It is shown that the OR ratio was strongly dependent on assay conditions. With optimal assay conditions oxidase activity exceeds reductase activity in adult and fetal rat lung microsomes (OR ratio > 1). Thus, glucocorticoids are mainly metabolized to their inactive forms. The enzyme activity in the adult is about 10 times higher than in the fetal lung. The low enzyme activity in fetal lungs could be the reason why the glucocorticoidal effects on surfactant synthesis are not suppressed despite the predominance of oxidase activity.

11-beta-Hydroxysteroid Dehydrogenases↗
PubMed1993

Epithelial ion transport in the ileal J-pouch after proctocolectomy in the rat.

In ulcerative colitis and polyposis coli, creation of an ileal pouch with ileoanal anastomosis after proctocolectomy has become a common surgical method. The aim of our study was to characterize the adaptation of the epithelial ion transport function in the pouch by using electrophysiologic techniques. Proctocolectomy and ileoanal anastomosis was performed in rats either with (pouch) or without (control) creation of an ileal J-pouch. To characterize the epithelial barrier function, impedance analysis was performed 6 months after surgery. Epithelial resistance was 29 +/- 2 omega.cm2 in controls and was unchanged in the pouch (28 +/- 4 omega.cm2; NS). In contrast, subepithelial resistance increased from 33 +/- 3 omega.cm2 to 54 +/- 5 omega.cm2 (P < 0.01) owing to work hypertrophy of the muscle layers in the pouch. To detect changes in net ion transport, ion tracer fluxes were measured in the Ussing chamber. In control ileum, absorptive net Na and Cl fluxes of similar magnitude were present, indicating that electroneutral NaCl absorption was the predominant ion transport system. Neither Isc nor net Na and Cl fluxes were significantly altered in the pouch. Glucose-coupled Na absorption was measured as the 3-o-methyl-glucose-induced increase in Isc. Km remained unaltered, while Vmax decreased from 7.5 +/- 2.1 mu eq.h-1 cm-2 in controls to 1.7 +/- 0.8 mu eq.h-1 cm-2 (P < 0.05) in the pouch. Then, maximal transport capacity for electrogenic Cl secretion was measured as the Cl-induced increase in Isc blockable by serosal bumetanide (in the presence of theophylline and prostaglandin E1).(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗
PubMed1993

Parameters influencing piezoelectric shock wave lithotripsy of biliary calculi.

A total of 225 human gallbladder stones were divided into sets of two, three, or four identical calculi (with maximum diameters of 6-26 mm) and submitted to piezoelectric shock wave lithotripsy in vitro in order to investigate the influence of the following parameters on stone disintegration: shock wave intensity, pulse frequency, and various physicochemical parameters such as the volume and viscosity of the fluid surrounding the stone, the hardness of the calculus, and its chemical composition. The fragmentation efficacy increased with enhanced shock wave energy (P less than 0.01). If the volume of the fluid (30 mL) surrounding the calculus was small, the disintegration end-point (defined by maximum fragment diameters less than or equal to 4 mm) was achieved after a lower number of pulses (median: 250 pulses) than in the case of a large fluid volume (80 mL) (500 pulses; P less than 0.01). On the other hand, however, factors such as the shock wave pulse rate (0.9 Hz vs 1.6 Hz), variations in viscosity of a water-jelly mixture surrounding the stone, the chemical composition of the stones as determined by X-ray diffractometry, and stone hardness were found to have no significant influence on the fragmentation efficacy.

Cholelithiasis↗
PubMed1992

Ion transport in the experimental short bowel syndrome of the rat.

The adaptational changes of epithelial ion transport in the short bowel syndrome were studied. Ileal remnants of rats were investigated 8 weeks after 70% proximal small intestinal resection. Pure epithelial resistance measured by impedance analysis decreased from 27 +/- 1 to 21 +/- 1 omega.cm2, and polyethylene glycol 4000 fluxes increased from 2.5 +/- 0.3 to 3.6 +/- 0.3 nmol.h-1.cm-2, indicating increased permeability of the short bowel. Unidirectional flux measurements in control ileum showed absorptive net fluxes of Na+ and Cl- that were assigned to electroneutral NaCl absorption and a short-circuit current that was accounted for by the residual flux (HCO3- secretion). Neither NaCl absorption nor HCO3- secretion were altered in the short bowel. Also, electrogenic Cl- secretion, defined after maximal stimulation by theophylline and prostaglandin E1 was not changed in the short bowel. In contrast, electrogenic Na+/glucose cotransport increased in Vmax from 2.0 +/- 0.3 in controls to 5.0 +/- 1.0 mumol.h-1.cm-2 in the short bowel. Tight junction structure was studied by freeze-fracture electron microscopy. The number of horizontal strands was unchanged, whereas tight junction depth was slightly increased in the short bowel. Microvillus area of short bowels was increased by 20% in villus regions. Under the light microscope, villus height was increased by 30%. In conclusion, the short bowel mucosa undergoes adaptive responses to reduced overall absorptive area by increasing glucose-dependent electrogenic Na+ absorption to 250%, which is partly caused by increased villus and microvillus surface area. Electrogenic Cl- and HCO3- secretion and electroneutral NaCl absorption remained unchanged. The decreased epithelial resistance is caused by mucosal surface amplification.

Adaptation, Physiological↗
PubMed1991

Uncoupled basal sodium absorption and chloride secretion in prairie dog (Cynomys ludovicianus) gallbladder.

1. Prairie dog gallbladders mounted in a Ussing-type chamber and bathed with symmetrical Ringer's solutions exhibited a transepithelial resistance (Rt) of 51 +/- 5 omega cm2, a lumen negative potential difference (Vms) of 11.5 +/- 0.7 mV and a short-circuit current (Isc) of 6.9 +/- 0.3 microEq/hr/cm2. 2. Radioisotopic ion flux experiments revealed that the basal Isc of 6.9 +/- 0.3 microEq/hr/cm2 was mostly accounted for by net Na+ absorption of 3.2 +/- 0.5 microEq/hr/cm2 and net Cl- secretion of 2.9 +/- 0.3 microEq/hr/cm2. 3. In HCO3- free Ringer's, net Na+ flux was virtually abolished, net Cl- flux decreased by 50% and Isc was reduced by 77%. 4. 10(-3) M mucosal amiloride and DIDS reduced Isc by 28 and 24%, respectively. 5. Mucosal NaCl diffusion potentials indicated that the paracellular pathway was cation selective. 6. Thin section electron micrographs showed a single cell population in this epithelium suggesting that net Na+ absorption and Cl- secretion may emerge from the same cells. 7. We conclude that prairie dog gallbladder epithelium is an electrogenic tissue and, in contrast to gallbladders of most other species, simultaneously but independently absorbs Na+ and secretes Cl-.

Animals↗
PubMed1991

Cl- secretion in epithelial monolayers of mucus-forming human colon cells (HT-29/B6).

HT-29, an undifferentiated human colon cell line, is known to differentiate when cultured without glucose. This study aimed to characterize ion transport in the clone HT-29/B6, which was selected from HT-29 cells differentiated by glucose-free culture. HT-29/B6 cells seeded onto filter membranes grew as polarized monolayers, mainly consisting of mucus-forming cells and exhibiting high transepithelial resistance. Short-circuit current (Isc) of unstimulated HT-29/B6 monolayers in Ussing chambers was 0.1 +/- 0.01 mumol.h-1.cm-2, and conductance was 2.0 +/- 0.2 mS/cm2. Serosal forskolin (FSK; 10(-5) M) induced a sustained Isc of 1.9 +/- 0.1 mumol.h-1.cm-2, associated with a rise of intracellular adenosine 3',5'-cyclic monophosphate (cAMP). Isc was identified as Cl- secretion by tracer studies and by the inhibitory effects of serosal bumetanide and Ba2+. The Cl- channel blockers NPPB and DPC diminished FSK-induced Isc at respective doses of 3 x 10(-4) and 10(-3) M, being effective from either side of the monolayer. Cl- secretion could be triggered by vasoactive intestinal peptide (10(-8) M), prostaglandin E1 (10(-6) M), and dibutyryl cAMP (10(-3) M) as well. In conclusion, HT-29/B6 cells grow as polarized monolayers, forming mucus and secreting Cl- in response to secretagogues. This clone may not only serve as a model for investigation of cellular mechanisms of intestinal Cl- secretion but may also be helpful to elucidate the contribution of mucus cells to this process.

Absorption↗
PubMed1991

In vitro fragmentation of gallstones: comparison of electrohydraulic, electromagnetic and piezoelectric shockwave lithotripters.

To compare the fragmentation efficiency of three different shockwave systems, 63 human gallstone triplets were disintegrated in vitro using an electrohydraulic (MPL 9000, Dornier), an electromagnetic (Lithostar Plus, Siemens) and a piezoelectric (Piezolith 2300, R. Wolf) lithotripter. Since each stone triplet was obtained from the same gallbladder, the concrements of one such set were identical in physicochemical parameters. According to the maximal diameter, the calculi were divided into group A (6 to 15 mm) and group B (16 to 30 mm). Shockwave application was terminated when residual fragments measured 4 mm or less. Forty-five triplets were fragmented at energy settings mainly used in clinical treatment of patients with gallbladder stones (MPL 9000: 20 kV; Lithostar Plus: setting 9 (maximal); Piezolith 2300: setting 3). The fragmentation endpoint was achieved in group A (n = 3 x 36) with the Piezolith 2300 after median 150 (range = 50 to 500) pulses and with the Lithostar Plus after 150 (50 to 750) pulses compared with 500 (50 to 1,500) pulses using the MPL 9000 (p less than 0.01). In group B (n = 3 x 9) the Lithostar Plus (median = 750, range = 250 to 1,250 pluses) required fewer discharges than the Piezolith 2300 (1,250, 250 to 2,500 pulses; p less than 0.05) and the MPL (1,500, 500 to 1,600 [upper limit] pulses; p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Cholelithiasis↗
PubMed1990

pH dependence of protamine action on apical membrane permeability in Necturus gallbladder epithelium.

Protamine reversibly decreases cation permeability and alters the structure of Necturus gallbladder tight junctions. Conflicting results, however, have been published whether or not it also affects apical cell membrane permeability. We investigated this issue more systematically by measuring voltage (psi mc) and fractional resistance (fRa) of the apical membrane at varying concentrations of protamine, K+, and H+ in the bathing solution. At pH 7.6 and [K+] 2.5 mM, (Poler, M.S. and Reuss, L. (1987) Am. J. Physiol. 253, C662) 6 microM protamine caused psi mc to depolarize from -58 to -51 mV and fRa to decrease from 0.74 to 0.67. If we increased pH to 8.1 these effects were even more pronounced. At [K+] 2.5 mM, but not 4.5 mM, psi mc transiently hyperpolarized for about 5 min after adding protamine. Most importantly, if [K+] was 4.5 mM and pH was adjusted to 7.1 (Bentzel et al. (1987) J. Membr. Biol. 95, 9) no significant changes of psi mc and fRa occurred. In any case, at a supramaximal concentration of 200 microM, protamine did not further increase the paracellular response but produced decreasing psi mc and fRa. We conclude that 6 microM protamine decreases K+ conductance of the apical membrane, if it is already tuned high by high pH. At low control K+ conductance as observed at lower pH, protamine action is restricted to the paracellular pathway. Thus, conflicting results were due to different experimental conditions. At a solution pH of 7.1, 6 microM protamine fulfills criteria of a selective tool for reversibly altering structure and function of the tight junction in Necturus gallbladder.

Animals↗
PubMed1990

Aldosterone low-dose, short-term action in adrenalectomized glucocorticoid-substituted rats: Na, K, Cl, HCO3, osmolyte, and water transport in proximal and rectal colon.

The short-term action of aldosterone in physiological concentration on net fluxes of Na, K, Cl, HCO3, osmolytes, and water was examined in the proximal colon and rectal colon of adrenalectomized (ADX) rats in vivo. The measuring time was 12 h, divided in eight periods of 90 min. (a) Aldosterone alone (6 nmol h-1 kg-1) did not stimulate transport in ADX rats. In these experiments plasma [K] increased to fatal values. A basal glucocorticoid substitution of 24 nmol h-1 kg-1 corticosterone caused plasma K to stay constant throughout the experiment, so that epithelial transport was not handicapped by non-specific effects of ADX, but this also did not restore the decreased transport of ADX rats to control values. Under these conditions (absence of aldosterone) in the rectal colon Na and H2O transport was zero, whereas in the proximal colon flux rates were depressed by between 30% and 50%. In contrast, basal glucocorticoid substitution of 18 nmol h-1 kg-1 corticosterone plus infusion of 6 nmol h-1 kg-1 aldosterone caused transport stimulation to values not significantly different from those of non-ADX controls. We conclude that after ADX, aldosterone at physiological concentrations increases transport if, as a prerequisite, a basal glucocorticoid substitution is provided. Transport of Na, K, and H2O is under the total control of aldosterone in the rectal colon but is only moderately altered in the proximal colon.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗
PubMed1990

Tight junction regulation during impaired ion transport in blind loops of rat jejunum.

Epithelial cell tight junction structure in self-filling blind loops of rat jejunum, a model for blind loop syndrome in humans, was analyzed morphometrically along the crypt-villus axis. In control jejunum, the number of strands and junctional depth, including meshwork depth, decreased from crypt to villus tip. In the blind loop, aberrant strands appeared below the meshwork, particularly in crypt cells. Consequently, total junctional depth was greater than in controls. Furthermore, strand number and junctional meshwork depth were increased in blind loops at the villus tip. It is that site along the crypt-villus axis which showed the most shallow junction in control jejunum. This structural change is paralleled by a three-fold increase in epithelial resistance as previously measured by alternating current impedance analysis. Relative Na over Cl permeability (PNa:Cl) was obtained from dilution potential measurements. PNa:Cl was 1.50:1 in control jejunum and 1.35:1 in the blind loop (n.s.). Considering the cation selectivity of the tight junction, the increase in epithelial resistance in blind loops cannot be attributed to a collapse of the lateral intercellular space but is due to changes in tight junctional permeability resulting from structural alteration. The blind loop syndrome represents a further example of diminished epithelial ion transport and concomitant decrease in tight junction permeability, thus supporting the general concept of regulation of the tight junction in response to active transport activity.

Animals↗