Esophageal stimulation by hydrochloric acid causes neurogenic inflammation in the airways in guinea pigs

Hamamoto, Junji, Hirotsugu Kohrogi, Osamu Kawano,Hajime Iwagoe, Kazuhiko Fujii, Nahomi Hirata, and Masayuki Ando.Esophageal stimulation by hydrochloric acid causes neurogenicinflammation in the airways in guinea pigs. J. Appl.Physiol. 82(3): 738-745, 1997.Toinvestigate whether tachykinins are released in the airways in responseto stimulation of the esophagus, we studied the airway plasmaextravasation induced by intraesophageal HCl in the presence or absenceof neutral endopeptidase inhibitor phosphoramidon and NK₁-receptor antagonist FK-888 inanesthetized guinea pigs. The airway plasma leakage wasevaluated by measuring extravasated Evans blue dye in the animalspretreated with propranolol and atropine. Infusion of 1 N HCl into theesophagus significantly increased plasma extravasation in the trachea.Phosphoramidon significantly potentiated plasma extravasation in thetrachea and main bronchi, whereas FK-888 significantly inhibited that extravasation in a dose-related manner. In the capsaicin-treated animals, airway plasma extravasation was completely inhibited even inthe presence of phosphoramidon. Tracheal plasma extravasation potentiated by phosphoramidon was significantly inhibited in the bilateral vagotomized animals. These results suggest that1) tachykinin-like substances arereleased to cause plasma extravasation in the airways as a result ofintraesophageal HCl stimulation and2) there are neural pathwayscommunicating between the esophagus and airways, including the vagusnerve.

     

Yeast Crv4/Ttp1, a predicted type II membrane protein, is involved in an event important for growth, functionally overlapping with the event regulated by calcineurin- and Mpk1-mediated pathways

The Saccharomyces cerevisiae crv mutants (crv1, 2, 3 and 4) exhibit phenotypes, such as calcium resistance and vanadate sensitivity, which are apparently similar to those of calcineurin-deficient mutants. We have cloned and characterized the CRV4 gene that complements all the phenotypes of the crv4 mutant. DNA sequencing revealed that CRV4 is identical to the previously cloned gene TTP1, which encodes a type II membrane protein of unknown function. Deletion of CRV4/TTP1 causes no obvious phenotype except for Ca²⁺ resistance and vanadate sensitivity, but is synthetically lethal in combination with a deletion of MPK1, in a manner which is suppressible by the addition of an osmotic stabilizer. In medium containing sorbitol as an osmotic stabilizer, the cnb1 mpk1 ttp1 triple mutant exhibits a more severe growth defect than does any of the double mutants cnb1 ttp1, cnb1 mpk1 or mpk1 ttp1. A high Ca²⁺ concentration (50 mM) or a constitutively active form of calcineurin partially suppresses the growth defect of the mpk1 ttp1 double mutant. These results indicate that Ttp1 participates in a cellular event essential for growth and morphogenesis, in parallel with the pathways involving Mpk1 MAP kinase and calcineurin. Received: 4 June 1997 / Accepted: 14 July 1997     

Relationship of alcohol-induced changes in Mg2+-ATPase activity of rabbit intestinal brush border membrane with changes in fluidity of its lipid bilayer

We examined the effects of seven n-alkyl alcohols (from n-butyl to n-undecyl alcohol), isoamyl alcohol and benzyl alcohol on the activity of membrane enzyme Mg²⁺-ATPase of the rabbit small intestinal brush border membrane. Their relationships with the changes in the fluidity of the membrane lipid bilayer were examined through studies on the fluorescence anisotropies of diphenylhexatriene (DPH) and its ionic derivatives. Good linear correlations were found both between the partition coefficients of the alcohols and their concentrations causing similar decreases in the activity of Mg²⁺-ATPase and between their partition coefficients and the alcohol-induced changes in fluorescence anisotropies. Within the concentration range of the alcohols tested, including isoamyl alcohol and benzyl alcohol, the decreases in activity of the membrane enzyme Mg²⁺-ATPase clearly corresponded with the decreases in fluorescence anisotropy of DPH, which is thought to be located within the hydrophobic core of the membrane. From these findings, one possible explanation is that inhibition of this enzyme by the alcohols is due to perturbation of the lipid bilayer of the brush border membrane.The authors thank M. Takano, PhD and Y. Tomita, PhD, Department of Pharmacy, University Hospital of Kyoto University, for instruction in preparation of the brush border membrane vesicles. This work was supported in part by grants from the Japanese Ministry of Education, Science and Culture (05671795 and 06304044) and Takeda Science Foundation.     

The Ontogeny of Social Behavior and Body Coloration in the African Cichlid Fish Haplochromis burtoni

The ontogeny of behavioral patterns and body coloration are described for the African cichlid fish, Haplochromis burtoni. The development of particular color patterns correlates directly with appearance of behavioral patterns which “use” those color patterns. There is a flexible timetable associated with these events which depends on both social and environmental factors.     

Sesquiterpene coumarins from the roots of Ferula assa-foetida.

From the chloroform extract of the roots of Ferula assa-foetida, two sesquiterpene coumarins designated assafoetidnol A and assafoetidnol B were isolated, in addition to six known compounds, gummosin, polyanthin, badrakemin, neveskone, samarcandin and galbanic acid. The structures of the sesquiterpenes new compounds were established by spectroscopic methods.     

Ortho-aminobenzoic acid as a fluorescent probe for the interaction between peptides and micelles

Ortho-aminobenzoic acid (o-Abz) has been used as a fluorescent probe in internally quenched fluorescent peptides for continuous protease assays. We investigated the fluorescent properties of the probe in order to verify if it can be used to monitor the interaction of peptides with micelles. Abz-aminoacyl-monomethyl amides (Abz-Xaa-NHCH(3), where Xaa=Arg, Phe, Leu and Glu) were synthesized. Quantum yield, spectral position, anisotropy and lifetime decay were analyzed in the presence and absence of sodium dodecyl sulfate (SDS) micelles. Significant changes in the fluorescence parameters were observed for Abz-Arg-NHCH(3) in comparison to Abz-Glu-NHCH(3), indicating a strong electrostatic component in the compound's interaction with the negative charged micelles. The change in fluorescence parameters, observed when the probe is bound to hydrophobic amino acids Abz-Phe-NHCH(3) and Abz-Leu-NHCH(3), is probably due to insertion of those compounds into micelles. Abz-NHCH(3) fluorescence is less affected by the presence of micelles, indicating that the occurrence of interaction is dependent on the properties of the amino acid to which the fluorophore is attached. The quenching data with acrylamide confirmed these results. Titration curves allowed the estimation of association constants between Abz compounds and SDS, according to a single partition model. Although the results cannot be strictly applied to the titration with charged compounds, it was verified that the association constant for the isolated Abz-NHCH(3) is significantly lower than those for Abz-Phe-NHCH(3) and Abz-Leu-NHCH(3). It is concluded that the Abz group is a sensitive and convenient fluorescent probe to monitor peptide binding to amphiphilic aggregates. That conclusion is supported by measurements with the peptide Abz-Leu-Arg-Phe-NH(2).     

Genetic constitution of germ cells in intervarietal and interspecific chimeras of Brassica induced by in-vitro grafting

The characteristics of intervarietal and interspecific chimeras synthesized by the graft-culture method were determined by morphology, anthocyanin pigmentation pattern, and crossing. In an intervarietal chimera between YR-ranpou (green cabbage) and Ruby ball (red cabbage) in Brassica oleracea, a segregation phenomenon was noted in which seeds giving rise to purple and green plants were both produced in a single capsule in F₁ progeny from crosses of chimeras with YR ranpou, the anthocyanin-free graft partner type. The degrees of segregation varied, reflecting the structure of the chimeras. YR ranpou-dominant chimeras produced capsules in which seeds gave rise to green plants at a high frequency, while Ruby ball-dominant chimeras produced capsules in which seeds in one capsule gave rise to purple plants at a high frequency. Mixed chimeras produced capsules with green plants or purple plants more regularly than did other chimeral types. Furthermore, a chimeral type in which seeds gave rise to green and purple plants was found in 3.2% of the total crosses. Segregation patterns in the progenies corresponded with the chimeral types. Chlorophyll-deficient variation (resulting in variegation or the production of albino plants) was found at a frequency of 2.6%. These results show that chimeric tissues are actually in a mixed state and that either the ovary develops from more than two cells or else that variation occurs in the germ-cell layer. In interspecific chimeras between Ruby ball and Komatsuna (B. campestris) various types of chimeras generally showed low pollen fertility, few capsules, and low seed-setting. Progenies from selves (geitonogamy), open crosses and crosses with the two parental species produce a predominantly homogeneous genotype showing either the Ruby ball or the Komatsuna type. Only two crosses produced four interspecific hybrids which expressed variations in their morphological and isozymic characters.