Seasonal Changes in the Pattern of Assimilatory Enzymes and of Proteolytic Activities in Leaves of Juvenile Ivy

Ivy growing under natural conditions is an interesting plantto study the influence of external (e.g. temperature, light)and internal (e.g. source/sink relations) factors on leaf metabolism.Leaves of this evergreen plant are subject several times toseasonal changes. The contents of selected assimilatory enzymeswere well conserved throughout the winter indicating that ivyleaves are probably able to make use of short periods with highertemperatures and to immediately restart growth in spring. Totalproteins and carbohydrates increased considerably between Februaryand May before the emergence of the new leaf generation. Theincrease in the content of non-structural carbohydrates wasdue to the accumulation of starch, while soluble sugars peakedin winter and decreased in spring. From May onwards, the assimilateswere retranslocated to the emerging young plant parts. Markedseasonal changes in the peptide hydrolase pattern were observed.All exo- and endopeptidases investigated were minimal duringsummer suggesting that the net protein remobilization from olderleaves was not based on an increase in the level of these majorpeptide hydrolases. Source/sink interactions on a whole plantlevel seem to be decisive in the regulation of seasonal changesin the pattern of assimilatory enzymes and of proteolytic activities.Since ivy leaves remain active for several years, the changesmust be reversible and occur repeatedly during the life-spanof a particular leaf.Copyright 1994, 1999 Academic Press Hedera helix L., ivy, peptide hydrolase, assimilatory enzyme, low temperatures, retranslocation     

Effects of the stereochemical orientation of phenethylamines and imidazolines on alpha-adrenergic receptor-mediated DNA synthesis in primary cultured rat hepatocytes.

The hepatic alpha 1-adrenergic receptor mediates a variety of hepatic functions including respiration, glycogenolysis, gluconeogenesis, and growth. We have utilized a rat primary hepatocyte culture system to show that the alpha 1-adrenergic receptor can be activated in a stereoselective manner by a series of phenethylamines and catecholimidazolines resulting in the stimulation of DNA synthesis as determined by [3H]thymidine incorporation. The phenethylamines adhered to the Easson-Stedman hypothesis with a rank order of potency of (-)-(R)-norepinephrine (NE) greater than (+)-(S)-NE greater than the desoxy analog dopamine (DA) for the stimulation of DNA synthesis. However, the 2-substituted catecholimidazolines did not follow this trend and demonstrated an order of potency of the desoxy analog 3,4-dihydroxybenzyl imidazoline (DHT) greater than or equal to (-)-(R)-2-(3,4,alpha-trihydroxybenzyl)imidazoline (TBI) greater than (+)-(S)-TBI. 4-Substituted catecholimidazolines were less potent as inducers of DNA synthesis than the corresponding 2-substituted analogs with an order of potency of (+)-(R)-4-(3,4-dihydroxybenzyl)imidazoline (DBI) greater than (+,-)-(R,S)-DBI greater than (-)-(S)-DBI. When the beta-hydroxyl moiety of NE is replaced with an amino group as in 3,4-dihydroxyphenylethylenediamine, the isomers are less active than the beta-hydroxylated analogs and also demonstrate no stereoselectivity for the stimulation of DNA synthesis. These results demonstrate that the hepatic alpha 1-adrenergic receptor can recognize various isomeric forms of these compounds and that hepatocellular growth can be modulated in a stereoselective manner by phenethylamines and imidazolines.     

The sodium channel from rat brain. Reconstitution of voltage-dependent scorpion toxin binding in vesicles of defined lipid composition

Purified sodium channels incorporated into phosphatidylcholine (PC) vesicles mediate neurotoxin-activated 22Na+ influx but do not bind the alpha-scorpion toxin from Leiurus quinquestriatus (LqTx) with high affinity. Addition of phosphatidylethanolamine (PE) or phosphatidylserine to the reconstitution mixture restores high affinity LqTx binding with KD = 1.9 nM for PC/PE vesicles at -90 mV and 36 degrees C in sucrose-substituted medium. Other lipids tested were markedly less effective. The binding of LqTx in vesicles of PC/PE (65:35) is sensitive to both the membrane potential formed by sodium gradients across the reconstituted vesicle membrane and the cation concentration in the extravesicular medium. Binding of LqTx is reduced 3- to 4-fold upon depolarization to 0 mV from -50 to -60 mV in experiments in which [Na+]out/[Na+]in is varied by changing [Na+]in or [Na+]out at constant extravesicular ionic strength. It is concluded that the purified sodium channel contains the receptor site for LqTx in functional form and that restoration of high affinity, voltage-dependent binding of LqTx by the purified sodium channel requires an appropriate ratio of PC to PE and/or phosphatidylserine in the vesicle membrane.     

Heart structure and ventricular ultrastructure of hemoglobin- and myoglobin-free icefish Channichthys rhinoceratus

Summary The structural and ultrastructural characteristics of the heart of Channichthys rhinoceratus, an antarctic teleost devoid of respiratory pigments, are described and compared with those obtained from the red-blooded related species Notothenia rossii.The heart of the icefish is characterized by a spongy myocardium supplied with a highly developed arterial coronary system. This vasculature includes a subepicardial system and an extensive intratrabecular capillary network. Arterial hilar network and Thebesian vessels may also be present. The bulbus arteriosus shows unusually large spheroid structures located in the middle layer of the wall.Both white- and red-blooded species display comparable myocardial cell morphology and organelle distribution. However, the mitochondrial cristae of the former are more densely packed and the sarcolemma possesses numerous caveolae. A large proportion of non-contractile cells is also found in the icefish ventricular wall.     

Morphogenesis of Newcastle Disease Virus in Chorioallantoic Membrane

Chick embryo chorioallantoic membrane, infected with the Blacksburg strain of Newcastle disease virus, was examined with an electron microscope to investigate the sequence of viral-induced host cell alterations. These were evident mostly in the endodermal epithelial cells lining the allantoic sac and were divided arbitrarily into three stages. Stage 1 was characterized by commencement of cell hypertrophy and hyperplasia and presence of fewer cytoplasmic inclusion bodies normally found in the cells; in stage 2, juxtanuclear nucleocapsid-glycogen aggregates appeared, and there were increased numbers of microvilli; stage 3 was characterized by increased cytoplasmic density and evidence of viral assembly and release. The morphological features of viral assembly and the virion are also described.     

Changes in nitrogen contents and in proteolytic activities in different parts of field-grown wheat ears (Triticum aestivum L.) during maturation

Aminopeptidase, carboxypeptidase and neutral endopeptidase activitieswere analyzed in glumes and in kernels of field-grown wheat(Triticum aestivum L.) during ear development. Kernels harvestedon two dates were subdivided into outer pericarp, cross cells,endosperm and embryo. In developing parts with a net nitrogeninflux (young glumes, embryo, endosperm) the aminopeptidaseactivity is high, but in nitrogen-mobilizing tissues (senescingglumes, Outer pericarp) this activity decreases. Carboxypeptidaseis most active in fully expanded tissues. Neutral endopeptidaseshows the highest activity in the nitrogen mobilizing partsand extremely low activity in the embryo and the endosperm. (Received July 15, 1978; )     

Dipeptide backbone conformation and antibody recognition of a viral octapeptide epitope.

The peptide YKGTMDSG (Tyr-Lys-Gly-Thr-Met-Asp-Ser-Gly) represents an important antigenic determinant from the glycoprotein G2 of the pathogenic Rift Valley fever virus. By preparing a series of single-residue substitution peptides, the importance to antigenicity of individual residues within this octapeptide has been determined. Here, we investigated a simple and rapid computational analysis to test for correlations between the observed antigenicity of the substitution analogue peptides and the calculated conformational preferences in local regions of the peptides. Conformational energy analyses were carried out on all dipeptide combinations represented in the wild-type octapeptide and in the single-residue substitution analogue peptides. Conformational similarities and differences between wild-type and substitution dipeptide pairs were determined. The results of these computational analyses were then compared with the data on the relative antigenicity of the wild-type octapeptide and the substitution analogues. This comparison revealed a positive correlation. Substitution peptides showing changes in antigenicity possessed significant changes in the calculated backbone conformation relative to wild type in the dipeptides encompassing the residue substitution. Substitution peptides showing no change in antigenicity similarly showed no significant changes in dipeptide conformation. The potential utility of dipeptide conformational energy analyses and this preliminary structure-activity correlation are discussed.     

The lactate dehydrogenase of the icefish heart: biochemical adaptations to hypoxia tolerance

Cardiac lactate dehydrogenase from the hemoglobin- and myoglobin-free antarctic icefish has been purified by affinity chromatography. Structural and kinetic properties of the enzyme were found close or identical to those of its skeletal muscle counterpart and other M-type lactate dehydrogenases. A model involving a dual oxidative-anaerobic metabolism of the icefish heart is proposed.     

Genetic variability of proteins from plasma membranes and cytosols of mouse organs

The genetic variability of membrane proteins (structure-bound proteins) and cytosol proteins (water-soluble proteins) was investigated in two inbred strains of the mouse, C57BL/6J and DBA/2J. Membrane proteins and cytosol were isolated from the brain and liver of the mouse. The proteins were separated by two-dimensional electrophoresis. A high number of genetic variant proteins (brain, 30; liver, 72) was found in the cytosol. Most of these variants represented changes in the amount of proteins. Electrophoretic mobility changes occurred only in about 1% (brain, 6; liver, 9) of all protein spots of a two-dimensional pattern. In contrast to the cytosol proteins, no genetic variation was detected among the membrane proteins, not even for the quantitative characteristics of the protein spots. The results obtained for the two classes of proteins suggest that the degree of variability in the amount of proteins is related to the degree of variability in the structure of proteins.