An immunodiffusion activity stain for yeast histidinol dehydrogenase.

A procedure is described for detecting enzymatically active yeast histidinol dehydrogenase-antibody complexes in immunodiffusion agar. The method employs a coupled dye system, consisting of a tetrazolium salt and a phenazine methosulfate intermediate, that produces an insoluble formazan and stains active precipitin lines red. The specificity of the reaction is indicated by its dependence on substrate and by its dependence on an intact HIS4C region, based on observations with mutant forms of the yeast HIS4 multifunctional protein.     

γ-Glutamylamine cyclotransferase

Summary -Glutamylamine cyclotransferase, an enzyme found in a number of animal tissues and cells, catalyzes the conversion of -(L--glutamyl)-L-lysine to free lysine and 5-oxo-L-proline as well as the release of free amines and the formation of 5-oxo-L-proline from a variety of other L--glutamylamines. Among its substrates are both the mono- and di--glutamyl derivatives of putrescine, spermidine and spermine, and a derivative of -(L--glutamyl)-L-lysine in which both the -amino group and the carboxyl group of the lysine moiety are blocked. The enzyme does not act on most -glutamyl--amino acids, nor is it active toward the -lysyl derivatives of L-aspartic acid or D-glutamic acid. Derivatives of -(L--glutamyl)-L-lysine in which the -amino or the -carboxyl function of the glutamyl moiety is blocked also do not serve as substrates. The specificity of -glutamylamine cyclotransferase is in accordance with the proposal that it functions biologically in the latter stages of the catabolism of products of the action of transglutaminases. Some suggestions as to the manner in which -glutamylamine cyclotransferase serves this function are made based on present knowledge of protein degradation.     

Use of in situ hybridization to investigate the regulation of hippocampal corticosteroid receptors by monoamines.

The hippocampus receives major noradrenergic and serotoninergic (5-HT) innervations which interact with corticosteroid-sensitive cells. However, the subregional localization of these actions and the corticosteroid receptor types involved have not been defined and current ligand binding techniques for estimating corticosteroid receptors are hampered by several methodological limitations. We have developed in situ hybridization histochemical techniques to allow specific and sensitive estimation of glucocorticoid (GR) and mineralocorticoid receptor (MR) mRNA expression in rat hippocampus. Investigation of the effects of 5,7-dihydroxytryptamine lesions of 5-HT neurons showed significantly reduced GR and MR mRNA expression in some hippocampal subregions. Both abnormal 5-HT neurotransmission and excessive corticosteroid secretion are associated with major affective disorders, particularly depression. The crucial interaction between these two systems may occur, at least in part, at the level of regulation of hippocampal corticosteroid receptor expression.     

Photosynthesis and transpiration ofPinus radiata D. Don under natural conditions in a forest stand

Summary Gas-exchange ofPinus radiata foliage was measured with climatised cuvettes under natural light in the sun-crown of 8 m tall trees in a forest stand. Measurement began during a period of drought (_WS –8.2 bar, _We –10.5 bar) and continued after elimination of soil moisture-deficit by watering (_WS –0.5 bar, _We –5.5 bar). Soil and air moisture-deficits severely restricted gas-exchange. Watering resulted in an immediate decline in stomatal resistance (r _s ) and an increase in net photosynthesis (P _N ) of 13%. A slower progressive gas-exchange recovery occurred additionally during the 10 days after watering leading to a further decline inr _s to 3 s cm^-1 and an ultimate increase inP _N of 38% when measured under comparable conditions at 8 mb v.p.d. Radiata pine had a high photosynthetic capacity with a measured maximumP _N of 10.2 mg CO₂ dm^-2 h^-1 total needle surface (11.4 mg CO₂ g^-1 DM h^-1).Optimum temperature forP _N in March (late summer) occurred at ca. 18°C. Rate ofP _N was 95% saturated at irradiance of 900 E m^-2 s^-1 and 50% saturated at only 270 E m^-2 s^-1. Radiata pine needles responded directly to changes in atmospheric humidity by adjusting their stomatal diffusive resistance. As a result, between 8 and 18 mb v.p.d.P _N declined by 2.3% mb^-1 increase.     

Analysis of transmembrane proteins from eukaryotic cells

The topography and properties of plasma membrane proteins from mouse L-929 cells are studied by comparing their availability for enzymatic labeling on the external and internal surfaces of the membrane. In order to study the internal surface, phagolysosomes are prepared from cells after they ingest latex particles. The plasma membrane surrounding these seems to have an “inside-out” orientation. The sugars of the membrane glycoproteins in intact phagolysosomes are not available for interaction with lectins or available for periodate-borotritide labeling. A comparison of the lectin-binding proteins lableled by lactoperoxidase-catalyzed iodination on the external cell surface with those labeled on the internal cell surface suggests that a variety of plasma membrane glycoproteins span the lipid bilayer. Using two-dimensional gel electrophoresis it has been shown that selected proteins are labeled at both the internal and external faces of the plasma membrane. Analysis of the 2-D gel electrophoregrams reveals that there are two distinct prominent proteins at 60,000 and 100,000 daltons which are enzymatically iodinated from both sides of the membrane. The partial hydrolysis of the 100,000 dalton protein reveals that different peptides are iodinated when the iodination is performed on intact cells or on the phagolysosomes. These proteins are extensively phosphorylated in cells incubated with inorganic ³²P. We conclude that the phagolysosome is probably oriented in an “inside-out” configuration and that this membrane preparation can be used to study the topographic organization of membrane proteins. The use of oriented membranes, selective labeling of proteins, and affinity separation of proteins in combination with gel electrophoresis to define the position and properties of proteins is discussed.     

Die Morphologie der Schleimsekretion im Fruchtknoten vonAptenia cordifolia

Zusammenfassung Der Fruchtknoten vonAptenia cordifolia enthÄlt wÄhrend der Samenentwicklung einen proteinreichen Polysaccharidschleim. Verschieden alte schleimproduzierende Placentarpapillen werden einer elektronenmikroskopischen Analyse unterzogen. Kurz vor dem Einsetzen der Schleimproduktion ist das rauhe ER noch spÄrlich entwickelt. Der Golgi-Apparat ist unauffÄllig und wenig aktiv. Zu Beginn der Schleimbildung sind als hauptsÄchliche Strukturkomponenten hypersekretorische Dictyosomen und ER-umschlossene Vakuolen (storage vesicles) zu beobachten. Es wird angenommen, da\ diese Komplexe aus rauhem ER und vermutlich mitèinander verschmolzenen Golgi-Vesikeln die charakteristischen Synthese-Einheiten für den Polysaccharid-Protein-Schleim darstellen, da sie nachweislich neben Polysacchariden auch Proteine enthalten. Membranfusionen zwischen Vesikeln und dem Plasmalemma deuten auf Exocytose-Prozesse unter Beteiligung des Golgi-Apparates hin. Daneben wird eine holocrine Ausscheidung des in den storage vesicles zunÄchst gespeicherten Polysaccharid-Protein-Schleimes bei Degeneration des Protoplasten vermutet.

---

Morphology of slime secretion in the seed vessels ofAptenia cordifolia

Summary During seed development the gynaeceum ofAptenia cordifolia produces a mucilage rich in carbohydrates and protein. The mucilage-producing placentary papillae are analyzed in different developmental stages by electron microscopy. Just before mucilage production is started, the rough ER occurs but sparsely. At that time the dictyosomes are inconspicuous and of low activity. When mucilage production commences, one can observe hypersecretory dictyosomes and ER-ensheathed vacuoles (storage vesicles) as the main structural components. It is suggested that the complexes of rough ER and probably fused Golgi vesicles are the synthetizing units of the carbohydrate protein mucilage, since in these complexes both components can be identified cytochemically. Fusion sites of plasmalemma and vesicles indicate processes of exocytosis-probably involving the Golgi apparatus. In addition, a holocrine excretion of the mucilage initially enclosed in the storage vesicles via degeneration of the protoplast is assumed.

     

Populations- und syn?kologische Modelle in der Ornithologie

Ohne Zusammenfassung

---

Population models and synecological models in ornithology

     

Muscle water and electrolytes following varied levels of dehydration in man.

In an effort to assess the effects of dehydration on the content of water and electrolytes (Na+, K+, Cl-, and Mg2+) in plasma and muscle tissue, eight men exercised in the heat (39.5 degrees C, 25%). Blood urine, and muscle biopsy samples were obtained before exercise and after the subjects had reduced their body weight by 2.2, 4.1, and 5.8%. On the average, plasma and muscle water (H2Om) contents were found to decline 2.4 and 1.2% for each percent decrease in body weight. Muscle sodium (Na+m) and chloride (Cl-m) content remained unchanged with dehydration, while muscle magnesium (Mg2+m) declined 12% as a result of the 5.8% dehydration. In terms of intracellular concentrations, K+i increased 7.2 and 10.6% at the 2.2 and 4.1% dehydration levels, respectively. Calculations of the resting membrane potential suggest that the water and electrolyte losses observed in these studies do not significantly alter the excitability of the muscle cell membrane.     

Cryoenzymology: the use of sub-zero temperatures and fluid solutions in the study of enzyme mechanisms.

Behind the firm discrimination maintained between active and passive transport lies a definition of energetic coupling as a fusion between an exergonic chemical reaction and an uphill transport. In contrast, energetic coupling between paired chemical reactions tends. to be defined much more loosely, as if the term were merely equivalent to sequential linkage, even though the actual usage may parallel that in transport. This article argues for a sharpening of this definition through integrated consideration of chemi-chemical and chemi-osmotic coupling.Furthermore; it calls attention to the applicability of energetic coupling to both the backward and forward fluxes of the energized transport. When two parallel but distinct active transport systems act on the same solute, one is likely to operate more steeply uphill than the other. The situation then easily arises, and is probably widespread, whereby entry occurs largely by the first process and exodus by the reversal of the second, still energetically linked. In this way cases of chemi-osmoti-chemical coupling probably arise, beyond the one proposed by Mitchell. Presumably the term retention process has in the past unknowingly (and illogically) referred to the second transport process. The “uncoupling” of an active transport does not tend simply to convert it to a facilitated diffusion, and both fluxes are likely to be modified. Accordingly, measure of only one flux will not describe a change in energy transfer.