Destruction and reorganization of the receptor membrane in labellar chemosensory cells of the blowfly. Recovery of responses to sugar after destruction

Recovery from destruction by sodium deoxycholate (DOC) was studied with the receptor membrane of the blowfly, Phormia regina. The recovery can be divided into two processes, colchicine dependent and colchicine independent. The colchicine-dependent process was completely depressed by pretreatment with colchicine at 5 mM for 2 min (partially at 0.1 mM for 10 min), but the colchicine-independent one persisted. Vinblastine also caused depression but lumicolchicine did not. Records of responses obtained from the DOC-treated sugar receptor showed long response latencies that gradually became indistinct with recovery. Colchicine also affected this change in response latency after the DOC treatment. These results suggest that the colchicine-dependent recovery process is related to microtubules in the distal process of the receptor cell. The recovery time course and the change in response latency could be quantitatively explained by the simple assumptions that DOC underwent desorption from the receptor membrane (colchicine-independent recovery process) and that regeneration of the disrupted distal process of the receptor cell accompanied recovery in the number of available receptor sites (colchicine-dependent recovery process).     

Interaction of palytoxin and cardiac glycosides on erythrocyte membrane and (Na+ + K+) ATPase

Palytoxin (PTX), at extremely low concentrations (0.01-1 nM), caused K+ release from rabbit erythrocytes. Among the various chemical compounds tested, cardiac glycosides potently inhibited the PTX-induced K+ release. The order of inhibitory potency (IC50) was cymarin (0.42 microM) greater than convallatoxin (0.9 microM) greater than ouabain (2.3 microM) greater than digitoxin (88 microM) greater than digoxin (90 microM). Their corresponding aglycones, even at 10 microM, did not inhibit the K+ release, but competitively antagonized the inhibitory effect of the glycosides. All these cardiotonic steroids inhibited the activity of (Na+ + K+)-ATPase prepared from hog cerebral cortex in narrow concentration ranges (IC50 = 0.15-2.4 microM), suggesting that the inhibition of K+ release is not related to their inhibitory potency on the (Na+ + K+)-ATPase activity, and the sugar moiety of cardiac glycosides is involved in the inhibition. On the other hand PTX, at higher concentrations (greater than 0.1 microM), inhibited the (Na+ + K+)-ATPase activity. However, this inhibitory effect of PTX was not antagonized by ouabain. It is suggested that, compared with ouabain, PTX has additional binding site(s) on the (Na+ + K+)-ATPase.     

The standardization of the candidate national standard anti-D blood-typing serum with the Groupamatic MG50 system

The utility of the Groupamatic MG50 system for quantitative expression of agglutination in terms of continuous response was studied. By use of the characteristics of the design of this system, in which the change in voltage reflects the degree of agglutination, a linear regression relationship between the log ratio of light flux obtained by transformation of the voltage and the log dilution factor of the serum was demonstrated. The availability of the parallel line assay method to the standardization of the blood grouping antisera was also described.     

Mammalian signal peptidase: partial purification and general characterization of the signal peptidase from microsomal membranes of porcine pancreas

Signal peptidase has been enriched extensively from microsomal membranes of porcine pancreas. Microsomal membranes were washed with 1 M KCl and Brij 35, and then solubilized with 1% Nonidet P-40. The solubilized signal peptidase was purified by DEAE-cellulose chromatography and Sepharose CL-6B filtration. Cleavage of pre-human placental lactogen with the partially purified enzyme gave the mature form, whose NH2-terminus was identified as valine. The signal peptidase is heat-labile and approximately 90% of the enzymatic activity was lost at 60 degrees C within 1 min. The pH optimum of the activity was 7 to 8. Chymostatin and o-phenanthroline at concentrations of 2.5 mM inhibited the signal peptidase activity by 62% and 30%, respectively.     

Diazo-reaction positive substance observed in the cortex of Chattonella antiqua

In order to investigate the causative factors responsible for removal of mucous coat from the gill lamellae of young yellowtail, Seriola quinqueradiata by red tide, diazo-reactions were employed for planktons and their media. The concentration of NO2- in the medium containing the raphidophyceae, Chatonella antiqua (ca 2000 cells/ml), was 0.70 +/- 0.05 (mu g/ml +/- SEM). In addition, diazo-reaction positive substances (NOx) which may degenerate the mucous, was highly concentrated in the cortex (perikaryon) of Chattonella antiqua. Morphologically, mucocysts, and chloroplats were likewise present in the cortex. Mucocysts were packed with fine fibrous content. Histochemically, the mucocysts were stained with PAS and had an abundance of nitrogen oxides (NOx). We observed discharge of the fibrous material from the mucocysts. These results suggest that when Chattonella antiqua is passing between the gill lamellae, NOx discharged from the mucocysts may act on the mucous, leading to the degeneration and concomitant removal of the mucous coat from gill lamellae.     

Squid retinochrome. Configurational changes of the retinal chromophore.

The configurations of the retinal chromophore in light and dark reactions of squid retinochrome were investigated by means of high-performance liquid chromatography. Orange light isomerized the chromophore of retinochrome, all-trans-retinal, mainly to the 11-cis configuration in metaretinochrome. Irradiation with shorter-wavelength lights not only accelerates the photoreversal of metaretinochrome to retinochrome but also leads to a slight production of isoretinochrome (13-cis-retinochrome), yielding a photoequilibrium mixture of three kinds of retinochrome. 13-cis- and 9-cis-retinochromes are photosensitive, and are converted into metaretinochrome upon irradiation with orange light. When steadily exposed to orange light in the presence of a trace of retinochrome-protein, all of the all-trans-, 13-cis-, and 9-cis-retinals are catalytically isomerized only to the 11-cis form, although the reaction rate is reduced in the order of the retinals listed above. In the dark, 9-cis-retinochrome, like retinochrome, remains unchanged, but both meta- and 13-cis-retinochromes slowly change to retinochrome. The chromophore of 13-cis-retinochrome changes directly to the all-trans form, whereas the 11-cis chromophore of metaretinochrome goes to all-trans mainly through the 13-cis form. The direct isomerization from 11-cis to all-trans hardly occurs at temperatures as low as 20 degrees C, and shows high values of the activation enthalpy and entropy changes. Based upon these findings, the role of retinochrome in the photoreception of the visual cells is discussed.     

Performance of a perforated plate column as a multistage continuous fermentor

Microorganisms were continuously cultivated in multistage column consisting of ten perforated plate sections to which medium and air were supplied concurrently from the bottom. At steady state the cell concentration in the various stages was gradationally differentiated from the bottom to the top in the direction of medium flow. RNA content per unit cell concentration at each sage was determined. The cells in the lower stages were higher in RNA content than those from the upper stages. Wash out was observed to occur in the column at dilution rates which do not result in wash out in a single stage chemostat system. A study of the flow characteristics revealed that the overall performance of the plate column was equivalent to that of a multistage system, when hole diameter and hole area to column cross sectional area ratio were properly selected. This was true even in highly aerated conditions. These results indicated that the perforated plates in the column hindred intermixing through the plates, and that each stage functioned as an independent stirred vessel. Industrial and research application of this type fermentor was discussed.