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N Kamo

Publications and source records attributed to N Kamo.

104 records · Page 6Linked to original sources

Physicochemical studies of taste reception. III. Interpretation of the water response in taste reception.

The model membrane composed of a Millipore filter paper and the total lipids from bovine tongue epithelium or phosphatidylcholine from egg yolk simulated well the water response of a living taste cell, The water response observed with the model membrane adapted to various salt solutions was interpreted in terms of changes in electric potential at the membrane-solution interface, i.e. the water response was attributed to the e.m.f. change produced by diffusion of the electrolytes dissolved in (or adsorbed on) the membrane surface into the bulk solution. The water response of the frog tongue was also investigated by measuring the neural response of the glossopharyngeal nerve. The results obtained were consistent with the mechanism proposed in the present paper. The response of the frog to Ca2+ was examined under the condition where the water response was suppressed, and it was concluded that the water response of the frog is different from the response to Ca2+.

Animals↗

Pysicochemical studies of taste reception. V. Suppressive effect of salts on sugar response of the frog.

The tast responses of frog to various kinds of sugars were measured quantitatively by use of the glossopharyngeal nerve activity under an appropriate condition where the water response was completely suppressed. The concentration dependences of response of frog tongue to D-fructose, D-glucose, and sucrose were almost the same, D-galactose, however, elicited a much larger response in comparison with the other sugars in the whole range of concentrations examined. The sugar response was suppressed extensively by the presence of small amount of salts in the stimulating sugar solution. The suppressive effects of NaCl, KCl, MgCl2, MgSO4, and K4Fe(CN)6 were examined with a fixed concentration of sugar. The results obtained with these salts, added in various concentrations, fell on a single curve when the data were plotted against the ionic strength in the stimulating solution. The present results were consistent with the notion that the taste receptor potential for salts or acids is attributable to a change in the phase boundary potential at the membrane-solution interface as proposed in the previous papers of this series.

Animals↗

Electrode sensitive to sulfa drugs.

An electrode sensitive to sulfa drugs was constructed by using the iron(II)-bathophenanthroline chelate embedded in a liquid membrane. Rapid and Nernstian responses were exhibited against solutions of sulfamerazine and sulfisomidine ranging between 10(-3) and 10(-1) M in concentration. High selectivity was observed in the presence of urea, glycine, aminopyrine, or p-amino-benzoic acid. These chemicals are known to interfere in the usual colorimetric analysis of sulfa drugs.

Chemical Phenomena↗

Phasic and tonic components of gustatory response in the frog.

Characteristics of phasic and tonic responses of the frog glossopharyngeal nerve to various salts were examined under a variety of conditions. The results obtained are summarized as follows: 1) The salt concentration of adapting solutions affected greatly the phasic component of the responses to NaCl, KCl, MgCl2, and CaCl2, whereas the tonic component was independent of the adapting condition. 2) Either the phasic or tonic component was preferentially suppressed under appropriate conditions. 3) An abrupt rise of temperature of stimulating solution produced a phasic response. Magnitude of the phasic response induced by a rise of temperature was a function of both species and concentration of salts in the stimulating solution. 4) Binding of Hg2+ to the receptor and elimination of the bound Hg2+ by mercaptoethanol gave similar large phasic response. A slow elimination of the bound Hg2+ led to no response. It was concluded that a phasic response appears only when environments such as salt concentration or temperature are changing and that its magnitude is related to the rate of changes, whereas a tonic response represents information of the receptor membrane in an equilibrium state. Discussion on the obtained results was made under an assumption that the phasic response is attributed to a dynamic conformational change of the receptor membrane.

Adaptation, Physiological↗

Ion selective electrode for 2,4-dichlorophenoxyacetic acid.

An electrode responsive to 2,4-D (2,4-dichlorophenoxyacetic acid) was constructed by dissolving tetrazolium derivatives as an ion exchanger in a liquid membrane. The electrode exhibited rapid and Nernstian response to solutions of 2,4-D over the concentration range 10(-1) to 10(-4)M. The presence of diverse substances such as acetate, benzoate, and 3-indoleacetate showed no appreciable effect on the electromotive force of the electrode.

2,4-Dichlorophenoxyacetic Acid↗

Physico-chemical studies of taste reception. IV. Response of individual phospholipid membrane to a variety of chemical stimuli.

Variations in the membrane potential across model membranes made of Millipore filter paper and various single phospholipids were measured in response to salt, acid and distilled water. The phospholipids used were phosphatidylcholine (c), spingomyelin (SM), phosphatidylethanolamine (PE), and phosphatidylserine (PS). Results were compared with those obtained with the model membrane made of the total lipids extracted from bovine tongue epithelium, which simulated well the receptor potential observed with intact tast organs. The membrane potential of PE- and PS-membranes increased monotonously with increase of the concentration of 1:1 type salt, while that of PC- and SM-membranes exhibited no appreciable change in 1:1 salt solutions. Application of CaC12 to the membranes brought about a varity of response depending on the species of lipids used. PE- and PS-membranes showed a larger change in the membrane potential than PC- and SM-membranes when pH of the solution was varied. Fe-3+ was strongly absorbed on the surface of PC and SM-membranes, while Fe-3+ bound to PE- and PS-membranes was easily removed by an application of salt solution. A transient increase in the membrane potential was observed when distilled water was applied to the membrane adapted to an appropriate salt solution, which was similar to the water response observed in taste cells. PC- and SM-membranes responded to water when the membrane adapted to either NaC1 or CaC12, but PS-membrane responded only when the membrane was adapted to a solution containing CaC12. PE-membrane did not respond to water in any cases examined. The membrane prepared with a mixture of two species of phospholipids responded neither to salt nor to water, while the membranes prepared with the total lipids or a mixture of three species of lipids in appropriate ratio responded to both. The water response of the total lipids membrane vanished in a high temperature medium, while the water response of PC-membrane retained in all temperature ranges examined, i.e. between 20 degrees and 62 degrees C. The results obtained suggest that a mosaic structure, where each domain has different functions against various chemical stimuli, is formed on the surface of the model membrane made of the total lipids.

Animals↗

Significance of substrate hydrophobicity for recognition by an oligopeptide transporter (PEPT1).

Our previous paper [(1999) Bioconjugate Chem. 10, 24-31] pointed out that hydrophobicity of substrates/inhibitors plays an important role in the recognition by an oligopeptide transporter (PEPT1) expressed in the human intestinal epithelial cell line Caco-2. To determine the significance of that hydrophobicity, we have now synthesized dipeptide analogues conjugating the epsilon-amino group of Lys in Val-Lys with aliphatic carboxylic acids: acetic acid (C2), propanoic acid (C3), pentanoic acid (C5), hexanoic acid (C6), and decanoic acid (C10). The affinities of these conjugates were estimated by their inhibition of the accumulation rate of Gly-Sar, a well-established substrate for PEPT1. With the increase in length of the hydrocarbon chain of the conjugates, i.e., in the hydrophobicity of the conjugates, the inhibition strengthened. Dixon-Webb plot analysis of the inhibition by the C10-conjugated dipeptide showed competitive inhibition. The trans-stimulation effect of Val-Lys conjugated to C10 or C5 on the uptake of Ceftibuten was observed using rat brush border membrane vesicles. This findings showed that these conjugates are transportable substrates. These results confirmed that the hydrophobicity of substrates/inhibitor is one of the factors in the recognition by PEPT1.

Animals↗

Conjugation of dipeptide to fluorescent dyes enhances its affinity for a dipeptide transporter (PEPT1) in human intestinal Caco-2 cells.

Dipeptide transporters in small intestine have a very wide substrate specificity, so that the transporter sometimes serves as a carrier for peptide-like compounds. We have synthesized dipeptide analogues conjugated at an epsilon-amino group of Lys in Val-Lys or Lys-Sar with fluorescent compounds such as fluorescein isothiocyanate and coumarin-3-carboxylic acid. Uptakes of these peptide analogues were examined by measuring intracellular accumulations into monolayers of the human intestinal epithelial cell line Caco-2 expressing the dipeptide transporter PEPT1. Kinetic analysis and effects of addition either of uncoupler (protonophore) or by Gly-Sar, one of the good substrates of PEPT1, revealed that fluorescent dipeptides were taken up by passive diffusion. In contrast, these analogues remarkably inhibited the Gly-Sar uptake by Caco-2 cells. Among the fluorescent analogues synthesized in this paper, Val-Lys(Flu) was the most potent competitive inhibitor against the Gly-Sar uptake with an inhibition constant of 5 microM. This value is the smallest among those ever reported: Val-Lys(Flu) has the highest affinity for PEPT1 among chemicals ever reported. The importance of the hydrophobic part of the substrate was pointed out.

Biological Transport↗

Cimetidine enhances the antiproliferative effect of 5-fluorouracil on colon carcinoma SW620.

We investigated the effects of cimetidine on p-glycoprotein and on the antiproliferative effect of various anticancer drugs which are recognized by p-glycoprotein. We used colon carcinoma SW620 cells and their doxorubicin resistant SW620 Ad300 derived cells, the latter express p-glycoprotein and have multidrug resistance phenotype. To assess the effect of cimetidine on the efflux activity of p-glycoprotein, we examined its effects on the accumulation of rhodamine123 which is recognized by p-glycoprotein. Cimetidine had no effect on rhodamine123 accumulation on either cell line. Cimetidine did not enhance the antiproliferative effect of any of the anticancer drugs investigated. This indicates that cimetidine was not recognized by p-glycoprotein. However, cimetidine remarkably enhanced the antiproliferation effects of 5-fluorouracil on SW620 cells, not those of 5-fluorouracil on SW620 Ad300, whereas cimetidine itself showed little effect on antiproliferation. This finding indicates that combination therapy using 5-fluorouracil and cimetidine might be useful for cancer patients.

ATP Binding Cassette Transporter, Subfamily B, Mem↗