2-KETO-SUGAR ACIDS IN GREEN FLAGELLATES: A CHEMICAL MARKER FOR PRASINOPHYCEAN SCALES1,2

The chemical composition of cell walls (thecae) of three taxa of scaly green flagellates (Prasinophyceae) was investigated. The theca of Tetraselmis striata, Tetraselmis tetrathele, and Scherffelia dubia consists mainly of carbohydrate (80% of dry weight), with proteins (5%), calcium (4%), and sulfate (6%) as minor components. The principal sugars (60% of dry weight) are the 2-keto-sugar acids 3-deoxy-manno-2-octulosonic acid (KDO), 3-deoxy-manno-5-O-methyl-2-octulosonic acid (5OMeKDO), and 3-deoxy-lyxo-2-heptulosaric acid (DHA). Arabinose, gulose, galactose, galacturonic acid, and in S. dubia, xylose and rhamnose were also found. Examination of scale preparations from Mantoniella squamata, Mesostigma viride, Pyramimonas amylifera, and Nephroselmis olivacea revealed that the 2-keto-sugar acids were always associated with the presence of typical prasinophycean scales on the cell surface. In contrast, 2-keto-sugar acids were not detected in the cell wall of Chlamydomonas reinhardtii nor in polymer preparations from the culture medium of Chlamydomonas reinhardtii, Dunaliella bioculata, Dunaliella primolecta, Asteromonas gracilis, Hafniomonas reticulate, Pedinomonas tuberculata, Monomastix sp., and Micromonas pusilla. We conclude that 2-keto-sugar acids are chemical markers for prasinophycean scales.     

Tubular inclusions within polyploid nuclei ofGerris najas do not react with a monoclonal anti-β-tubulin antibody

Summary Variable aggregates, composed of tubules with a mean diameter of 19 nm, were found exclusively within polyploid nuclei of the midgut, Malpighian tubes, cyst cells, testis epithelium, and trophocytes ofGerris najas. The nuclear inclusions are always in direct contact with the nucleoplasm, and no other structures are associated with them. They appear most abundant within degenerating nuclei of the midgut surface epithelium, where they form paracrystalline bodies or spindle-shaped inclusions with tapered ends. Smaller fusiform inclusions occur in younger epithelial nuclei but not in the diploid nuclei of regenerative cells. In other tissues, mainly spindle-shaped inclusions can be observed, the longest (4.5 m) in cyst cell nuclei. The mean diameter of the tubules determined from transverse sections, resembles that of cytoplasmic microtubules and was verified statistically. The inclusions within trophocyte nuclei failed to react with monoclonal anti--tubulin antibody, although the antibodies could penetrate the nuclei after extensive lysis of the cells.     

Cloning and electrophysiological analysis of KST1, an inward rectifying K+ channel expressed in potato guard cells.

Potassium uptake by guard cells represents part of the osmotic motor which drives stomatal opening. Patch-clamp measurements have identified inward rectifying K+ channels capable of mediating K+ uptake in guard cells and various other plant cell types. Here we report the molecular cloning and characterization of a voltage-dependent K+ channel (KST1) from potato (Solanum tuberosum L.) guard cells. In situ hybridization shows expression of kst1 in guard cells. Two-electrode voltage-clamp and patch-clamp studies of the gene product after cRNA injection into Xenopus oocytes identified KST1 as a slowly activating, voltage-dependent, inward rectifying K+ channel. The single channel current voltage curve was linear in the range -160 to +20 mV, with a deduced single channel conductance of 7 pS in symmetrical 100 mM K+. This channel type, modulated by pH changes within the physiological range, required ATP for activation. In line with the properties of a K(+)-selective channel, KST1 was permeable to K+, Rb+ and NH4+ and excluded Na+ and Li+. Cs+ at submillimolar concentrations blocked the channel in a voltage-dependent manner. Related studies on potato guard cell protoplasts confirmed the biophysical characteristics of the kst1 gene product (KST1) in the heterologous expression system. Therefore, KST1 represents a major K+ uptake channel in potato guard cells.     

Mortality associated with moderate intakes of wine,beer, or spirits.

OBJECTIVE--To examine the association between intake of different types of alcoholic drinks and mortality. DESIGN--Prospective population study with baseline assessment of alcohol intake, smoking habit, income, education, and body mass index, and 10-12 years'' follow up of mortality. SETTING--Copenhagen city heart study, Denmark. SUBJECTS--6051 men and 7234 women aged 30-70 years. MAIN OUTCOME MEASURE--Number and time of cause-specific deaths from 1976 to 1988. RESULTS--The risk of dying steadily decreased with an increasing intake of wine--from a relative risk of 1.00 for the subjects who never drank wine to 0.51 (95% confidence interval 0.32 to 0.81) for those who drank three to five glasses a day. Intake of neither beer nor spirits, however, was associated with reduced risk. For spirits intake the relative risk of dying increased from 1.00 for those who never drank to 1.34 (1.05 to 1.71) for those with an intake of three to five drinks a day. The effects of the three types of alcoholic drinks seemed to be independent of each other, and no significant interactions existed with sex, age, education, income, smoking, or body mass index. Wine drinking showed the same relation to risk of death from cardiovascular and cerebrovascular disease as to risk of death from all causes. CONCLUSION--Low to moderate intake of wine is associated with lower mortality from cardiovascular and cerebrovascular disease and other causes. Similar intake of spirits implied an increased risk, while beer drinking did not affect mortality.     

Tissue-specific DNaseI hypersensitive sites in the 5''-flanking sequences of the tryptophan oxygenase and the tyrosine aminotransferase genes.

The genes for tryptophan oxygenase (TO) and tyrosine aminotransferase (TAT) are expressed in a tissue- and development-specific manner and are regulated by glucocorticoids (TO and TAT) and glucagon or its intracellular mediator cAMP (TAT) in rat liver. We have analyzed the chromatin structure of these genes in the vicinity of the 5' ends with regard to DNaseI hypersensitivity and have found DNaseI hypersensitive sites upstream of each of the promoters. Mapping of this region reveals three closely spaced cleavage sites near the TO promoter and a doublet of sites near the TAT promoter. In both genes additional cleavage sites are found further upstream. All hypersensitive sites of both genes are absent in kidney nuclei and therefore appear to be specific for the tissue expressing the genes. A correlation of expression and modified chromatin structure was also observed in a hepatoma cell line expressing TAT but not TO: hypersensitive sites are present in TAT but not in TO chromatin. Upon glucocorticoid induction an additional hypersensitive site is detected approximately 2 kb upstream of the TAT promoter in liver and hepatoma cells.     

Fully automated and fast image analysis of autoradiographs with a TAS-Leitz. Determination of size,Feulgen fluorescence and grain counts of individual nuclei and their evaluation by a simplified cluster analysis

Summary A fully automatic analysis system based on television image analysis was developed to measure simultaneously three parameters in individual nuclei of microscopic autoradiographs prepared from mouse jejunal crypt cell squashes and ascites tumor cell smears: size, Feulgen fluorescence and reflection from silver grains. A dark light camera with an image intensified silicon tube (RCA-ISIT), an automatic scanning stage and an autofocus device were fitted to a Leitz-TAS microscope. The camera permitted localization of Feulgen stained nuclei and measurement of area and light intensity by means of incident of light fluorescence in the red. After automatic changes of the Opak-illuminator silver grains were determined by means of polarized incident light reflected from the grains in the blue. A 25 x oil objective (aperture 0.75) yielded sufficient resolution for measurements. The nadir between the proportions of labeled and unlabeled nuclei was calculated from the data of one specimen on a PDP-computer using a new algorithm based on the minimal variance of the logarithm of reflected light per nucleus. Labeling indices determined by visual grain counting and by automatic analysis of the autoradiographs were well correlated (r=0.87 to 0.92). Visual grain counts/nucleus and reflected light/nucleus correlated well when individual nuclei were compared (r=0.92 to 0.97) or means of labeled nuclei of various specimens prepared during a 5 year period (r=0.90 to 0.93). Quenching of nuclear Feulgen fluorescence was minimal. The optimal labeling range is 30–100 grain counts/nucleus. The time interval between measurements of two specimens was 25 min for a squash of approximately 350 crypt cells within a 3 mm× 3 mm field, and 20 min for a meandering scan with 1,000 ascites tumor cells.     

Changes in phospholipids of Saccharomyces cerevisiae associated with inositol-less death.

Two inositol-requiring strains of Saccharomyces cerevisiae were examined for changes in levels of phospholipids occurring after inositol deprivation. Lack of inositol results in loss of cell viability (inositol-less death) and in very large increases in two phospholipid precursors, phosphatidic acid and CDP-diacylglycerol; the accumulation of other glycerophospholipids continues for a considerable time at normal rates. Phosphatidylinositol accumulation does not occur in the absence of inositol; however, the further metabolism of this lipid continues, with 80 to 90% of this lipid disappearing. This disappearance is matched by increases in the phosphoinositol containing sphingolipids and extracellular glycerophosphoinositol. These changes are not observed when growth is blocked by cycloheximide or by omission of lysine from a lysine auxotroph, most lipids continuing to accumulate long after growth stops. There appears to be no close coordination in the synthesis of the major yeast phospholipids or between protein synthesis and phospholipid synthesis. However, despite very large changes in the composition of yeast phospholipids that can be achieved by altering culture conditions, it appears that the average charge per phospholipid molecule remains fairly constant.     

Windrichtung und Vegetationsdeckung am Nest der Silberm?we(Larus argentatus)

Zusammenfassung An 1047 Silbermöwen-Nestern auf den Watteninseln Mellum und Memmert wurden 1979/80 ermittelt (Abb. 1): Grenzrichtungen der Vegetationsstände 1–3 (1: Vegetationsstand größter Winkelgröße) und Nest-Zugänge 1–3, ihre Winkelgrößen und mittleren Richtungen. Mehr als 80% der Nester waren zu 40–90% von Vegetation umstanden (Abb. 2). Die Winkelgröße des Gesamtvegetationsstandes betrug auf Mellum 212°, auf Memmert 205° (Tab. 1). Die Nestzugänge hatten eine geringere Winkelgröße als die Vegetationsstände (Tab. 1, Abb. 3). Die Silbermöwe bevorzugte demnach Nestplätze, die von Vegetation schützend umgeben sind. Der Vegetationsstand 1 befand sich bevorzugt auf der westlichen (= Hauptwindrichtung), der Zugang 1 auf der östlichen Nestseite (Tab. 2, Abb. 4). Die Verteilung der Gesamtvegetation zeigte entsprechende Vorzugsrichtungen (Abb. 6). Im Vergleich zu 1979 war 1980 eine Verlagerung der Vorzugsrichtung des Vegetationsstandes 1 um 50–60° von West nach Nord festzustellen (Tab. 2, Abb. 4). Ein Vergleich der mittleren Windrichtungen beider Jahre zeigte eine gleichsinnige Richtungsänderung (Abb. 5). Auf den zwei untersuchten Probeflächen Mellums ließ sich eine Auswahl des Nestplatzes im Hinblick auf Sichtschutz gegen Nachbarn nicht nachweisen (Tab. 3). Die mögliche Funktion der nestumgebenden Vegetation als Wind- und Wetterschutz wird diskutiert.

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Vegetation surrounding Herring Gulls'(Larus argentatus) nests in relation to wind direction

Summary The following measurements were taken for a total of 1047 Herring Gulls' nests on the North Sea islands Mellum and Memmert (West Germany) in 1979 and 1980 (cf. Fig. 1 for terminology): limiting directions, angular sizes, and mean directions of the surrounding vegetation-stands 1 to 3 (1 = vegetation-stand with largest angular size), and of the entrances 1 to 3. For more than 80% of the nests, 40–90% of the circumference were surrounded by vegetation (Fig. 2). The mean angular size of the total vegetation-stand was 212° on Mellum and 205° on Memmert (Tab. 1). The nest entrances had a smaller angular size than the vegetation-stands (Tab. 1, Fig. 3). This shows the Herring Gull's preference for nest sites protected by surrounding vegetation.Vegetation-stand 1 was preferably orientated towards the west (= main wind direction), nest entrance 1 towards the east (Tab. 2, Fig. 4). The distribution of the total vegetation showed corresponding preferred directions (Fig. 6). A comparison of the years 1979 and 1980 revealed a shift of 50°–60°, from west to north, for the preferred direction of vegetation-stand 1 (Tab. 2, Fig. 4). A similar shift could be found for the mean wind direction (Fig. 5). For the two study plots on Mellum, a nest site selection favouring visual isolation from neighbours' nests could not be demonstrated (Tab. 3). The possible function of nest-surrounding vegetation as a shelter against wind and weather is discussed.

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Herrn Dr. Friedrich Goethe zur Vollendung des 70. Lebensjahres gewidmet     

The chemical characterization of favin, a lectin isolated from Vicia faba.

We have determined the subunit structure of the glucose- and mannose-binding lectin favin, from Vicia faba. The molecule is composed of two nonidentical polypeptide chains held together by noncovalent interactions. We have determined the complete amino acid sequence of the smaller alpha chain (Mr = 5,571) and shown that it is homologous to the alpha chain of the lectins from lentil and pea and to residues 72 to 120 of concanavalin A (Con A). The larger beta chain (Mr = 20,000) contains carbohydrate and is homologous to the beta chain of lentil, pea, soybean, peanut, and red kidney bean lectins and is homologous to a portion of the Con A molecule beginning at residue 122. Favin also contains a minor component, beta' (Mr = 18,700), that closely resembles the beta chain but lacks carbohydrate and may, on the basis of apparent molecular weight, lack some part of the COOH-terminal region of the polypeptide chain. Although favin is similar to Con A, it, like the lentil and pea lectins, appears to lack residues corresponding to positions 1 to 71 of Con A. Because these residues contribute significantly to the carbohydrate binding site of Con A, the lack of this region in the otherwise homologous lectin favin suggests that the carbohydrate binding site of favin differs from that of Con A or that the region represented by residues 1 to 71 of Con A is located in a different portion (i.e. in the beta chain) of the favin molecule.