Isolation and characterization of noradrenalin storage granules of bovine adrenal medulla

A method is described for the preparation of (1) the heavy population of bovine adrenal chromaffin granules ($\text{S}{}_{\mathrm{<mtext>H</mtext>}}\text{(}\text{average sedimentation coefficient}\text{) = 12 400}\text{S}$ in 0.25 M sucrose) essentially free from contamination with mitochondria and other organelles, and (2) a subpopulation of this heavy population which is highly enriched in noradrenalin ($\text{?95\%}$ of the total catecholamine is noradrenalin). The method is based on isopycnic gradient centrifugation using a self-generating gradient of polyvinylpyrrolidone-coated colloidal silica particles (Percoll) in 0.5 M sucrose medium.The isolated population of noradrenalin granules appeared highly electron dense in transmission electron microscopy and revealed a rather narrow size distribution. The specific content of amine and adenine nucleotides (with reference to total granule protein) was markedly higher than for the total population of heavy chromaffin granules. The molar ratio of amines to adenine nucleotides was, however, lower in the noradrenalin granules, i.e. 4.8 vs. 11.9.     

Chironomid communities as water quality indicators

Recent mathematical indices summarizing biological communities of indicators are recapitulated. Improvements of these indices based on weighting according to width of trophic ranges of each species are suggested. Their principle deficiencies, however, are pointed out.
Revised lists of characteristic profundal as well as littoral and sublittoral chironomids in Nearctic and Palearctic lakes show that at least 15 characteristic chironomid species communities can be delineated, 6 in each of the oligotrophic and the eutrophic ranges and 3 in the mesotrophic range. It is proposed that these communities be lettered consecutively in the Greek alphabet from α (alpha) to o (omikron). A key to the 15 divisions based on the species associations in the profundal zone of harmonic lakes is put forward. There is very good correlation between the 15 divisions and the ratios of average total phosphorus to mean lake depth and average chlorophyll a to mean lake depth.
The ratio of chironomids to oligochaetes and the distribution patterns of single species have proven useful in pin-pointing localized areas of pollution. The primary mechanism governing the distribution of chironomid communities in oligotrophic and mesotrophic lakes appears to be the availability of food materials rather than the annual hypolimnetic oxygen concentration. In eutrophic lakes the relationships between organic matter accumulation and oxygen levels are so interdependent as to be inseparable.     

Ceruloplasmin Polymorphism and Parentage Test in Hereford Cattle

In recent years parentage control by means of blood grouping tests (blood and protein systems) has been required for bulls to be registered in the Danish Hereford Herd Book. Because the Hereford breed shows less variation in the blood and protein systems, the probability of excluding an incorrectly stated bull (or cow) is estimated to be some 15 % lower in Hereford than in Danish dairy breeds (RDM, SDM and Jersey).     

The morphogenesis of dimorphic pollen and anthers in Tripogandra amplexicaulis. Light microscopy and growth analysis

A characteristic of the genus Tripogandra is the presence of two dissimilar whorls of anthers. In T. amplexicaulis the short filamented anthers (S anthers) develop fertile, spheroidal pollen, the long filamented ones (L anthers) develop elongated spores, which degenerate prior to anthesis. Meiosis is regular in both types but occurs approximately two days later in the L anthers. Developmental traits of the L spores are a continuous, uniaxial extension and a sigmoid course of growth in volume accompanied by progressive vacuolarization and growth of starch grains and usually no division. The number of spores in the L anthers amounts to only 60 % of that in the S anthers, but the values for anther fresh weight and pollen sac volume of the former eventually exceed those of the latter and the relative growth rate in these variables is consistently higher in the L anthers. The relative rate in dry weight, however, is similar for both anthers and the absolute rate is clearly lower in the L anthers. These growth characteristics are reflected in a greater vacuolarization of both spores and the amoeboid tapetum in the L anthers. Cytological features of male sterile plants with hypotrophy of the tapetum are related to the sterility syndrome of the L anthers and a common hypothesis for the deviating features is presented.     

Bovine Brain Myelin Glycerophosphocholine Choline Phosphodiesterase is an Alkaline Lysosphingomyelinase of the eNPP-Family, Regulated by Lysosomal Sorting

Glycerophosphocholine choline phosphodiesterase (GPC-Cpde) is a glycosylphosphatidylinositol (GPI)-anchored alkaline hydrolase that is expressed in the brain and kidney. In brain the hydrolase is synthesized by the oligodendrocytes and expressed on the myelin membrane. There are two forms of brain GPC-Cpde, a membrane-linked (mGPC-Cpde) and a soluble (sGPC-Cpde). Here we report the characterisation sGPC-Cpde from bovine brain. The amino acid sequence was identical to ectonucleotide pyrophosphatase/phosphodiesterase 6 (eNPP6) precursor, lacking the N-terminal signal peptide region and a C-terminal stretch, suggesting that the hydrolase was solubilised by C-terminal proteolysis, releasing the GPI-anchor. sGPC-Cpde existed as two isoforms, a homodimer joined by a disulfide bridge linking C414 from each monomer, and a monomer resulting from proteolysis N-terminally to this disulfide bond. The only internal disulfide bridge, linking C142 and C154, stabilises the choline-binding pocket. sGPC-Cpde was specific for lysosphingomyelin, displaying 1 to 2 orders of magnitude higher catalytic activity than towards GPC and lysophosphatidylcholine, suggesting that GPC-Cpde may function in the sphingomyelin signaling, rather than in the homeostasis of acylglycerophosphocholine metabolites. The truncated high mannose and bisected hybrid type glycans linked to N118 and N341 of sGPC-Cpde is a hallmark of glycans in lysosomal glycoproteins, subjected to GlcNAc-1-phosphorylation en route through Golgi. Thus, sGPC-Cpde may originate from the lysosomes, suggesting that lysosomal sorting contributes to the level of mGPC-Cpde on the myelin membrane.     

Dihydrofurocoumarin glucosides from Angelica archangelica and Angelica silvestris

A water-soluble root extract of Angelica archangelica subsp. litoralis afforded, in addition to adenosine, coniferin and the two known dihydrofurocoumarin glycosides, apterin and 1′-O-β-d-glycopyranosyl-(S)-marmesin (marmesinin), two new dihydrofuranocoumarin glycosides, 1′-O-β-d-glucopyranosyl-(2S, 3R)-3-hydroxymarmesin, and 2′-β-d-glucopyranosyloxymarmesin. For the latter a 2S-configuration was demonstrated, the stereochemistry at position 1′ remaining undefined. Roots of A. silvestris similarly afforded 1′-O-β-d-glucopyranosyl-(2S, 3R)-3-hydroxymarmesin. By correlation with the aglycone 2S,3R)-3-hydroxymarmesin obtained in this work, the absolute configurations (2S,3R) were established for the known dihydrofurocoumarin diesters smirniorin and smirnioridin.     

Activity of ectoenzymes of the heart breaking up ATP in the period of myocardial reperfusion after ischemia

Nowadays the activity of heart ectoenzymes, breaking up ATP, are determined in the undamaged tissue to eliminate the effect to the ATP of intracellular enzymes. Our aim is to study the activity of ecto-adenosine triphosphatase in the heart damaged by ischemia. It was established that the activity of ectoenzymes, breaking up ATP, may be measured in the tissue damaged by ischemia only in case when the cellular membrane becomes impermeable for the intracellular enzymes. The activity of adenosine triphosphatase is significantly reduced during the myocardial reperfusion after ischemia and appeared as one of the criteria by which we can assume the degree of the heart reperfusion damage.     

Crossed hydrophobic interaction immunoelectrophoresis: An analytical method for detection of amphiphilic proteins in crude mixtures and for prediction of the result of hydrophobic interaction chromatography

Crossed hydrophobic interaction immunoelectrophoresis is an analytical technique in which the principles of quantitative immunoelectrophoresis and hydrophobic interaction are directly combined. Using phenyl-Sepharose as hydrophobic (amphiphilic) matrix we have shown how the method permits detection of amphiphilic proteins in three model systems: native- and trypsinated intestinal brush border aminopeptidase, serum proteins, and detergent-solubilized erythrocyte proteins. In the case of first system the relative amounts of the two forms of the enzyme have been determined using immunochemical quantification. Comparison of the present method with column hydrophobic interaction chromatography reveals concordant results for both serum and erythrocyte proteins. Tests of some alkyl-substituted agaroses show that they work in a manner similar to phenyl-Sepharose.     

A comparative ecological investigation of two related mayfly nymphs

Zusammenfassung Heptagenia sulphurea wird auf Steinen, H. fuscogrisea in der Vegetation von Fließgewässern gefunden (Fig. 2).In Experimenten wählte H. sulphurea Steine und groben Kies. H. fuscogrisea wählte Batrachium als Substrat, nur ein geringer Anteil der Larven fand sich auf Callitriche (Fig. 3).Die Zeit, die zwischen Loslassen und erneutem Anhaften auf dem Substrat im strömenden Milieu verstreicht, war unterschiedlich bei den zwei Spezies. Die auf Steinen lebende Art reagierte schneller als die aus der Vegetation. H. sulphurea verhielt sich, wenn sie von der Strömung ergriffen worden war, ziemlich passiv, doch trug die turbulente Strömung das Tier dem Substrat zu, so hatte es gewöhnlich sicheren Halt.Im Strömungsaquarium bevorzugt H. sulphurea die strömungsexponierten Bereiche, aber viele waren auch im ruhigen Wasser (Fig. 5).Wurde H. fuscogrisea von der Strömung ergriffen, so führte sie Schwimmbewegungen aus. Erreicht sie eine Haltmöglichkeit an strömungsexponiertem Platz, so wird sie oft weitergetragen. Die Tiere lassen sie meist in ruhigem Wasser nieder.Im Strömungsaquarium bevorzugt H. fuscogrisea überwiegend die Regionen ruhigen Wassers (Fig. 5).Die Zeit zwischen Loslassen und Festsetzen war deutlich durch die Temperatur beeinflußt (Fig. 6).Der dichte Haarbesatz an der hinteren Kante der Femur von H. sulphurea wird als Anpassung an das Leben in der Strömung angesehen, weil er die Wirbelbildung herabsetzen kann. Der Haarbesatz entlang der Tibia bei H. fuscogrisea wird als Anpassung zum Schwimmen angesehen, zu dem these Spezies befähigt ist, wenn sie von der Strömung ergriffen wird (Fig. 8).Die Spezies von Steinen ist etwas mehr abgeflacht als die aus der Vegetation (Fig. 7).