Isolation and Identification of the Phenols of Paul's Scarlet Rose Stems and Stem-Derived Suspension Cultures

The phenols of Paul's Scarlet rose stems and stem-derived cell cultures have been analyzed using C₁₈-reversed-phase high performance liquid chromatography.

Rose stems were found to contain gallic acid, (+)catechin, (−)epicatechin, the dimers (−)epicatechin-(+)catechin and (+)catechin-(+)catechin, a polymeric procyanidin, ferulic acid, and several gallotannins. In contrast, a cell suspension of Paul's Scarlet rose which has been maintained in culture for over 25 years contained only low levels of gallic acid and (−)epicatechin-(+)catechin. The phenol content of a second rose cell line which was started from the same initial isolate in 1957, but which was maintained in a laboratory other than our own was quantitatively and qualitatively similar to the cell line kept in our laboratory for the last 20 years. A third cell line which we started 6 months ago contained a wide variety of phenols, most of which were in common with those of rose stems.

Selective subculturing of smaller cell clumps of our oldest cell line failed to enhance either the quantities or the diversity of phenols which accumulated in these cultured cells. Possible reasons for the failure of selective subculturing to enhance phenol levels in this long-established cell line are discussed.

     

Movements and associations of ribosomal subunits in a secretory cell during growth inhibition by starvation

In Chironomus tentans salivary gland cells, the cytoplasm can be dissected into concentric zones situated at increasing distances from the nuclear envelope. After RNA labeling, the newly made ribosomal subunits are found in the cytoplasm mainly in the neighborhood of the nucleus with a gradient of increasing abundance towards the periphery of the cell. The gradient for the small subunit lasts for a few hours and disappears entirely after treatment with puromycin. The large subunit also forms a gradient but one which is only partially abolished by puromycin. The residual gradient which which is resistant to the addition of the drug is probably due to the binding of some large ribosomal units to the membranes of the endoplasmic reticulum (J.-E. Edstrom and u. Lonn. 1976. J. Cell Biol. 70:562-572, and U. Lonn and J.-E. Edstrom. 1976. J. Cell. Biol. 70:573-580). If growth is inhibited by starvation, only the puromycin-sensitive type gradient is observed for the large subunit, suggesting that the attachment of these newly made subunits to the endoplasmic reticulum membranes will not occur. If, on the other hand, the drug-resistant gradient is allowed to form in feeding animals, it is conserved during a subsequent starvation for longer periods than in control feeding animals. This observation provides a further support for an effect of starvation on the normal turnover of the large subunits associated with the endoplasmic reticulum. These results also indicate a considerable structural stability in the cytoplasm of these cells worth little or no gross redistribution of cytoplasmic structures over a period of at least 6 days.     

Acclimation of different strains of the olive fly, Dacus oleae, to low temperatures

Male and female D. oleae have similar powers of acclimation when exposed to low temperatures. Their torpor thresholds depend upon the temperature to which they have been acclimatised. During slow cooling (i.e. less than 1°C per min) they are capable of some rapid acclimation which enables them to lower their torpor threshold by almost 1°C degree, as compared with when they are chilled quickly. After abrupt transfer from 25°C to a different temperature, acclimation takes some time to be accomplished. At 15°C and above it occurs within 10 days but at temperatures below this, progressive acclimation lowers the torpor thresholds to the very low levels typical of flies overwintering under natural conditions. During this long term acclimation torpor thresholds may change by almost 0.5°C per 1°C change of acclimation temperature.No differences were observed in the ability of either flies from northern and southern Greece, or normal and γ-irradiated laboratory reared flies to acclimate to winter conditions in the field. In all cases, torpor thresholds were progressively lowered in advance of the decline in weekly minimum temperatures.     

Isolation of plasma membrane glycoprotein from bovine thymocytes

Glycoprotein was isolated from a purified thymocyte membrane preparation by two methods: lithium diiodosalicylate-phenol extraction and hot 75% ethanol extraction. A higher yield of membrane sialic acid was obtained by the latter method. The preparations had similar apparent molecular weights on sodium dodecyl sulfate gel electrophoresis. Both had similar receptor activities against a panel of hemagglutinins, although the 75% ethanol extract was more active on a weight basis. However, there were significant differences in carbohydrate and amino acid compositions of the two thymocyte extracts. The lithium diiodosalicylate-extracted material had much more glucose, ribose, and glycine than the ethanol extract. The glycoprotein preparations from thymocytes were quite distinct from the glycoprotein of bovine erythrocytes in both composition and receptor properties.     

Effects of fusicoccin on Fresh Weight and Chlorophyll Levels in Cucumber Cotyledons

Incubation of cucumber cotyledons with fusicoccin increasedtheir fresh weights and chlorophyll levels and this effect wasenhanced by KCl. Addition of fusicoccin to this combinationincreased fresh weights but decreased chlorophyll levels. Thissuggests that the effects of fusiccocin on these two processesare probably mediated via different mechanisms. (Received January 4, 1982; Accepted March 25, 1982)     

Vertebrate lectins, Comparison of properties of beta-galactoside- binding lectins from tissues of calf and chicken

Beta-galactoside-binding lectins were isolated from various calf tissues and from chicken hearts by affinity chromatography on asialofetuin-Sepharose, and were compared with respect to biochemical characteristics, binding properties, antigenic cross-reactivity, and cellular localization. The lectins are all thiol group-requiring, divalent cation-independent dimers, of apparent monomer mol wt 12,000 (calf lectins) or 13,000 (chicken lectin), and acidic pI. The calf lectins appear essentially identical by dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, amino acid composition, and radioimmunoassay, while the chicken lectin is distinctly different by these criteria. However, all of the lectins competed for the same binding sites on rabbit erythrocytes, and could be inhibited by the same saccharide haptens (notably lactose and thiodigalactoside). Immuno-fluorescence studies on several cultured cell lines revealed that the bovine and chicken lectins had primarily an intracellular cytoplasmic localization. The beta-galactoside-binding lectins of vertebrates appear to be species-specific rather than tissue-specific.     

Prostaglandin E1 effects on resting and cholinergically stimulated lower esophageal sphincter pressure in cats

Intraluminal esophageal manometry with a sleeve catheter was used to compare the magnitude of decrease in lower esophageal spincter (LES) pressure produced by an arterial or venous infusion of prostaglandin E₁ in cats. Arterial PGE₁ produced significantly lower LES pressures than venous PGE₁ (p < 0.05). Maximal decrease of 75% in basal LES pressure occurred with an associated 15% decrease in systolic blood pressure. The site of action of PGE₁ in producing LES hypotension was studied by injection of either edrophonium, or bethanechol during the maximal PGE₁ effects. Bethanechol, which acts directly on sphincteric smooth muscle, produced an increase in LES pressure during both saline and PGE₁ infusion, while the increases in LES pressure seen with edrophonium during saline infusion were blocked during the PGE₁ infusion. From these studies, we conclude that PGE₁ produces LES hypotension in the cat by an inhibitory effect on the cholinergic pathway responsible for maintaining LES tone. These studies pharmacologically reproduce the LES pressure abnormality previously reported in the cat during acid-induced esophagitis and support the hypothesis that PGE₁ may be involved in the pathogenesis of acute acid-induced lower esophageal sphincter abnormalities.     

THE NATURE OF THE TWO PROTEINS OF BRAIN SPECIFIC ANTIGEN 14-3-2

The three isoenzymes of rat brain enolase (2-phospho d -glycerate hydrolase EC 4.2.1.11.) χχ, χγ and γγ were separated by ion-exchange chromatography and were tested for reaction with an antiserum against brain specific antigen 14-3-2. This monospecific antiserum affects the enolase activity of only the χγ and γγ isoenzymes. Immunoelectrophoretic experiments show that the two proteins which react as 14-3-2 both contain γ enolase subunits, and one of these also contains χ enolase subunits. It is concluded that the 14-3-2 antigen and the γ enolase subunit are identical, and that the two proteins which react immunologically as 14-3-2 are the χγ and γγ enolase isoenzymes.