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J Graf

Publications and source records attributed to J Graf.

At least 127 records · Page 7Linked to original sources
PubMed1987

Identification of an amino acid sequence in laminin mediating cell attachment, chemotaxis, and receptor binding.

We have probed for active sites in the B1 chain of laminin using synthetic peptides comprising certain regions of its amino acid sequence as deduced from cDNA clones. An antibody to a 19-mer from domain III inhibited attachment of HT-1080 and CHO cells to laminin, while the peptide itself was inactive. A nearby peptide (CDPGYIGSR) from domain III with homology to epidermal growth factor was synthesized and found to be one of the principle sites in laminin mediating cell attachment, migration, and receptor binding.

Amino Acid Sequence↗
PubMed1987

Cell membrane and transepithelial voltages and resistances in isolated rat hepatocyte couplets.

The basic electrical properties of an isolated rat hepatocyte couplet (IRHC) system have been analyzed using classical techniques of epithelial electrophysiology, including measurement of electric potentials, resistances and intracellular ion activities. Applications of these techniques are discussed with respect to their limitations in small isolated cells. Mean intracellular and intracanalicular membrane potentials ranged from -23.7 to -46.7 and -4.3 to -5.9 mV, respectively. Membrane resistances were determined using an equivalent circuit analysis modified according to the geometry of the IRHC system. Resistances of the sinusoidal (basolateral) and canalicular (luminal) cell membranes and tight junctions averaged 0.15 and 0.78 G omega and 25 m omega, respectively. The cells are electrically coupled via low resistance intercellular communications (approximately 58 M omega). Intracellular ion activities for Na+, K+ and Cl- averaged 12.2, 88.1 and 17.7 mmol/liter, respectively. The basolateral membrane potential reveals a permeability sequence of PK greater than PCl greater than PNa. The luminal potential showed minimal dependence on changes in transjunctional ion gradients, indicating a poor ion selectivity of the paracellular pathway. The electrogenic (Na+-K+)-ATPase contributes little to the luminal and cellular negative electric potential. Therefore, the luminal potential probably results from the secretion of impermeant ions and a Donnan distribution of permeant ions, a mechanism which provides the osmotic driving force for bile formation. By providing the unique opportunity to measure luminal potentials, this isolated hepatocyte system permits study of secretory mechanisms for the first time in a mammalian gland using electrophysiologic techniques.

Animals↗
PubMed1987

Role of basement membranes in cell differentiation.

Extracellular matrices have diverse biological effects, including promoting the growth and differentiation of various cells of epithelial origin. The components of one of these matrices, the basement membrane, are discussed, as well as studies using these components alone or in combination with cells in culture. The particular response observed varies with the cell type examined and appears to be dependent on multiple interactions with components of the matrix. Potential uses for a basement membrane-derived matrix in vitro and in vivo are being developed.

Animals↗
PubMed1987

Na-H exchange regulates intracellular pH in isolated rat hepatocyte couplets.

Intracellular pH (pHi) was measured directly in isolated rat hepatocyte couplets using pH sensitive microelectrodes. The hepatocytes were maintained in a minimal salt buffer without added hormones or serum. Values of pHi (6.99 +/- 0.12, mean +/- SE) were close to their Nernst equilibria. After intracellular acidification with ammonium chloride, pH regulation was inhibited with 1 mM amiloride or by omission of external sodium, consistent with a Na-H exchange mechanism. Mean intracellular buffering power, in the nominal absence of carbon dioxide, was 34.1 +/- 11.4 mM. In the presence of external bicarbonate, amiloride or omission of sodium slowed, but did not completely inhibit recovery from acidification, indicating that additional pHi regulation mechanisms may operate in this preparation. These studies provide a direct measurement of pHi in hepatocyte couplets and indicate that Na-H exchange, together with a bicarbonate dependent system are important mechanisms for pHi regulation in this preparation.

Amiloride↗
PubMed1986

Electrical and molecular coupling between sodium and proton fluxes in basolateral membrane vesicles of rat liver.

Mechanisms of Na+-H+ exchange in the hepatocyte were studied utilizing isolated basolateral membrane vesicles prepared by two different methods: Evidence was obtained for the existence of molecular coupling of Na+ and H+ fluxes (Na+/H+-antiport) which exhibits saturation kinetics (Km 7 mmol/l Na+) and is inhibited by amiloride (1.0 mmol/l). Although the two membrane preparations showed differences with respect to ionic permeabilities, our data suggest that a relatively high H+ conductance exists in the basolateral plasma membrane. Hence, electrical coupling of conductive H+ and Na+ fluxes in the opposite direction could contribute to net Na+-H+ exchange across the basolateral hepatocyte plasma membrane.

Animals↗
PubMed1986

17 beta-estradiol regulates and v-Ha-ras transfection constitutively enhances MCF7 breast cancer cell interactions with basement membrane.

The interaction of a line of human breast carcinoma cells (MCF7) with basement membrane components, particularly laminin, was altered by exposure of the cells to estrogen as well as by transfection of the cells with the v-Ha-ras oncogene. In both cases, the cells show a greater ability to attach to a laminin substrate, to migrate to laminin, to grow in the presence of a basement membrane matrix, and to cross barriers of reconstituted basement membrane. These responses were associated with an increase in the expression of laminin receptors. It is postulated that the increase in the invasive behavior of the cells treated with estrogen or transfected with v-Ha-ras is related to the increased number of laminin receptors and their interaction with laminin. Estrogen had no discernible effect on the v-Ha-ras transfected cells. It appears that in the MCF7 cells, the malignant phenotype is under hormonal control and that this control is bypassed after v-Ha-ras transfection.

Basement Membrane↗
PubMed1985

Effects of 1 alpha,25-dihydroxycholecalciferol on sodium-ion translocation across chick intestinal brush-border membrane.

By utilizing isolated brush-border vesicles, Na+ transport across the luminal membrane of chick small intestine was found to be a composite of (i) a saturable (Km 10mM-Na+) amiloride-sensitive Na+/H+ antiport and (ii) a potential-sensitive conductive pathway. No evidence was obtained for the existence of a Na+/Cl- symport system. With the exception of the duodenum, luminal Na+ transfer in the entire small intestine was subject to regulation by vitamin D. Repletion of vitamin D-deficient chicks with 1 alpha,25-dihydroxycholecalciferol [1 alpha,25(OH)2D3] significantly decreased net Na+ uptake by isolated membrane vesicles (by approximately 30%). The sterol suppresses the conductive pathway (25-45% inhibition) as well as the Na+/H+ antiport system. Kinetic analysis of the latter revealed that 1 alpha,25(OH)2D3 altered Vmax (from 12.9 to 4.8 nmol of Na+/20s per mg of protein), but did not change Km. Diminution of Na+ transfer, entailing an increase in the electrochemical transmembrane Na+ gradient, provides an explanation of the simultaneously observed stimulatory action of 1 alpha,25(OH)2D3 on Na+-gradient-driven solute transport in chick small intestine. Indirect evidence was obtained that the luminal plasma membrane of chick small intestine displays a definite H+ permeability that is positively affected by 1 alpha,25(OH)2D3.

Amiloride↗
PubMed1985

[The rare radial flexor muscle of the wrist with a special presentation].

The m. flexor carpi radialis brevis is a very rarely observed muscle with various origins and insertions. Our investigation showed a muscle with its insertion into the base of the second and third metacarpal bone. The important role of this muscle in operations on the wrist-joint is discussed.

Aged↗
PubMed1984

Isolated rat hepatocyte couplets: a primary secretory unit for electrophysiologic studies of bile secretory function.

Hepatocyte couplets were isolated by collagenase perfusion from rat liver. Between adjacent cells, the bile canaliculus forms a closed space into which secretion occurs. As in intact liver, Mg2+-ATPase is localized at the canalicular lumen, the organic anion fluorescein is excreted, and secretion is modified by osmotic gradients. By passing a microelectrode through one cell into the canalicular vacuole, a transepithelial potential profile was obtained. In 27 cell couplets the steady-state intracellular (-26.3 +/- 5.3 mV) and intracanalicular (-5.9 +/- 3.3 mV) potentials were recorded at 37 degrees C with reference to the external medium. Input resistances were determined within the cell (86 +/- 23 M omega) and in the bile canalicular lumen (32 +/- 17 M omega) by passing current pulses through the microelectrode. These data define electrical driving forces for ion transport across the sinusoidal, canalicular, and paracellular barriers and indicate ion permeation across a leaky paracellular junctional pathway. These findings indicate that the isolated hepatocyte couplet is an effective model for electrophysiologic studies of bile secretory function.

Adenosine Triphosphatases↗
PubMed1983

Canalicular bile salt-independent bile formation: concepts and clues from electrolyte transport in rat liver.

Studies on canalicular electrolyte transport are reviewed with reference to the concept that hepatocellular inorganic ion secretion may provide an osmotic drive for canalicular water flow. Cellular transport of electrolytes and of some nonelectrolytes appears directly or indirectly (cotransport or potential-sensitive transport) related to the activity of Na+-K+-ATPase of the sinusoidal cell membrane, but the role of the enzyme in regulating bile flow remains undetermined. Bile secretion of the isolated rat liver continues in the absence of either Na+, K+, Cl-, or HCO-3 when these ions are replaced in the perfusion medium by other permanent ions. Transepithelial salt concentration gradients, established experimentally, cause transient changes of bile flow and dissipate very quickly. Isotopic ion equilibration between sinusoids and bile proceeds faster than between sinusoids and liver cells. Both observations indicate extensive electrolyte diffusion through a paracellular shunt pathway. This pathway appears preferentially permeable to cations, and it restricts permeation of molecules of the size of sucrose (no apparent diffusion or effects of solvent drag) or bile acids (no backleak). In promoting canalicular osmotic water flow, transepithelial concentration gradients of NaCl are less effective than those of sucrose, revealing a reflection coefficient of NaCl of 0.3. By perfusion with hypertonic medium containing sucrose, bile flow is reduced. Bile production against this opposing osmotic gradient is accomplished by an increase in biliary organic anion concentration. Inorganic ion concentrations essentially conform to a Gibbs-Donnan distribution across the canalicular epithelium, established by the presence of impermeant anions in bile. Hence, the luminal electrical potential is expected to be negative with respect to the sinusoids. It is concluded that biliary secretion of endogenous organic anions is the major osmotic driving force for canalicular bile salt-independent bile flow and that transport of inorganic ions into bile results mainly from diffusion and solvent drag.

Animals↗
PubMed1983

[Scanning electron microscopic study of hyalin joint cartilage].

Guinea pigs (pirbright white) were subjected to fixed mechanical stress in an endless-belt experiment. Shoulder, elbow, hip, and knee joints were taken and the articular cartilage was prepared for examination with the scanning electron microscope. Ultrastructural findings of cartilage surfaces were described, using different techniques for preparation and fixation. Mechanically stressed animals showed defects in their superficial 'chondrosynovial membrane' with visible fibre networks.

Animals↗
PubMed1983

[Muscular changes caused by suction drainage. Scanning electron microscopy, transmission electron microscopy, light microscopy and morphometric studies of dorsal muscles of the rat (Rattus rattus) using drainage].

Changes in the dorsal muscles of 22 rats (Wistar), resulting from surgical drain, have been studied under different morphological aspects. The lumen of drains were obstructed after a short time. In the drains without using low pressure, erythrocytes and fibrin network were seen by scanning microscopical investigation. Patent muscle fibres could be analyzed in the lumen of drain using negative pressure. They prevent the functional drain system. By the effect of surgical drains the fibres of dorsal muscles had become swollen. This fact could be analyzed by morphometrical technique. Some changes in fibres have been demonstrated by histological methods.

Animals↗