Presence of Conjugated 5-Hydroxytryptamine in In Vivo Superfusates of Rat Spinal Cord

The presence of an acid-hydrolyzable conjugate of 5-hydroxytryptamine, presumably 5-hydroxytryptamine-O-sulfate, was demonstrated in in vivo superfusates of rat spinal cord by HPLC with electrochemical detection. In untreated rats, the concentration of the 5-hydroxytryptamine conjugate measured during the basal efflux of 5-hydroxytryptamine did not differ from that measured during the release of 5-hydroxytryptamine evoked by DL-p-chloroamphetamine. Pretreatment of the rats with clorgyline, an inhibitor of the enzyme monoamine oxidase (EC 1.4.3.4), or with probenecid did not alter the concentrations of conjugated 5-hydroxytryptamine measured during the basal efflux of 5-hydroxytryptamine, but did elevate the concentrations of conjugate measured during the evoked release of 5-hydroxytryptamine.     

Concentrations of 3,4-Dihydroxyphenylalanine and Catecholamines and Metabolites in Brain in an Anhepatic Model of Hepatic Encephalopathy

Abstract: Alterations in the catecholaminergic neurotransmitter systems have been shown to occur in hepatic failure and may contribute to development of hepatic encephalopathy. In the present study we used the rat after complete hepatectomy as a model for study of changes that occur in brain in acute liver failure. We attempted to identify processes in the synthesis, storage, and metabolism of catecholamine neurotransmitters that might be changed during liver failure by measuring levels of, together with those of norepinephrine and dopamine, the precursor (3,4-dihydroxyphenylalanine) and the neuronal metabolites of dopamine and norepinephrine (3,4-dihydroxyphenylacetic acid and 3,4-dihydroxyphenylglycol, respectively) in different regions of brains of control rats and of rats after hepatectomy. We found that in most brain regions of hepatectomized rats there were increases in the concentration of 3,4-dihydroxyphenylalanine or of dopamine but decreases in the concentrations of norepinephrine or of 3,4-dihydroxyphenylglycol. The particulate/supernatant ratios of catecholamines are indices of retention of neurotransmitters in storage sites. These ratios were not different in brain regions between control rats and hepatectomized rats, suggesting that vesicular retention of catecholamines in brain was not impaired after hepatectomy. The data suggest that inhibition of dopamine-β-hydroxylase might be a characteristic of hepatic failure.     

Catecholamine Release and Excretion in Rats with Immunologically Induced Preganglionic Sympathectomy

Abstract: Plasma and urinary catecholamines were quantified to assess global sympathoadrenal function in rats with preganglionic lesions caused by antibodies to acetyl-cholinesterase (AChE). Rats were given intravenous injections of normal mouse IgG or murine monoclonal anti-acetylcholinesterase IgG (1.5 mg). Five or 16 days afterward, basal blood samples were taken through indwelling arterial cannulae. A few hours later, the rats were immobilized for 10 min in padded restrainers, and another blood sample was drawn. HPLC determinations showed low basal levels of norepinephrine and epinephrine (<0.2 ng/ml in all rat plasma samples). In control rats, immobilization stress increased levels of plasma catecholamines up to 35-fold. In rats tested 5 days after injection of antibody, the norepinephrine response was much smaller (15% of control), and (he epinephrine response was nearly abolished (5% of control). There was some recovery at 16 days after antibody treatment, but stress-induced catecholamine release was still markedly impaired. Reduced stress-induced release: was not accompanied by major changes in tissue epinephrine or norepinephrine (heart, spleen, adrenal glands, and brain), although adrenal dopamine content dropped by 60%. Urinary excretion was studied in parallel experiments to gain insight into the effects of AChE anti-bodies on basal sympathoadrenal activity. Epinephrine, norepinephrine, dopamine, and selected metabolites were quantified in 24-h urine samples collected at frequent intervals for 30 days after antibody injection. No statistically gnificant changes were detected in the urinary output of dopamine, 3-methoxytyramine, normetanephrine, or 3-methoixy-4-hydroxyphenylglycol. On the other hand, epinephrine and norepinephrine output increased sharply at the time of antibody injection and then fell significantly below control levels. Norepinephrine output returned to normal after 2 weeks, but epinephrine output remained depressed. These results are consistent with previous evidence of widespread and persistent antibody-mediated βmade to the preganglionic sympathetic system.     

Notizen über endopolyploide Kerne im Bereich der Samenanlage von Angiospermen

Zusammenfassung Es bestätigt sich die Beobachtung, daß gerade in der Samenanlage nicht nur sehr häufig hohe Polyploidiegrade erreicht werden, sondern daß sich die Struktur in diesen Kernen zumeist sehr wesentlich von der in den diploiden unterscheidet.In den hochendopolyploiden Kernen des mikropylaren und des chalazalen Endospermhaustoriums vonLoasa papaverifolia findet sich eine den pflanzlichen Riesenchromosomen ähnliche Struktur, wobei die Endochromosomen wohl im heterochromatischen Abschnitt, aber nicht — oder nur über einen außerordentlich kurzen Teil — im euchromatischen zusammenhalten.In den entsprechenden Kernen vonPlantago psyllium sind die Endochromosomen stets als mehr oder weniger gewundene Fäden erkennbar, die entweder im Kernraum annähernd gleichmäßig verteilt sind, oder aber selten radiär gebaute Gruppen in der Anzahl 3n bilden; die SAT-Endochromosomen sind stets zu drei radiär gebauten Gruppen vereinigt. Außerdem erfährt die Kernstruktur eine weitere Abwandlung durch die unterschiedliche Dicke der Endochromosomen.Von den 25 Pflanzen, deren Antipodenkerne untersucht wurden, besitzen 17 hochendopolyploide Kerne; bei 11 zeigen sie Ausbildung bemerkenswerter Strukturen, wie Riesenchromosomen, Bündel, lichtmikroskopisch deutliche Spiralen oder lockere Areale, wobei manche nur einen bestimmten Strukturtypus ausbilden, andere dagegen mehrere.In den Kernen des Suspensorhaustoriums vonGagea lutea weisen die als zarte Fäden erkennbaren Endochromosomen in den niedrigen Polyploidiestufen eine leichte Gruppierung auf, während sie sich in den hochendopolyploiden Kernen zu Riesenchromosomen zusammenschließen.     

Über die Histologie der Intumeszenzen und die Cytologie zweierIpomoea-Arten

Zusammenfassung Ipomoea Batatas undIpomoea purpurea besitzen Chromozentrenkerne; bei ersterer entspricht die Chromozentrenzahl mit durchschnittlich 83 ungefähr der Chromosomenzahl (2n=90), bei letzterer ist sie mit durchschnittlich 60 Chromzentren doppelt so groß (2n=30). In den Dauergeweben der Wurzel bzw. der Knolle sowie in der Achse beider Arten finden sich keine Anzeichen von Endopolyploidie; die Gewebe bleiben also diploid. — Die Entwicklung der Intumeszenzen, die ebenfalls diploid bleiben, geht entweder von den Nebenzellen oder ihnen benachbarten Zellen der Epidermis, oder von der subepidermalen Zellschicht aus; im letztgenannten Fall finden sich die ersten Teilungen dann stets unterhalb einer Spaltöffnung. Die Intumeszenzen sind feste, aus der Epidermis und der ersten Mesophyllschicht hervorgegangene Wucherungen, die bei der Batate eine Länge von ungefähr 1,5 mm erreichen, beiIpomoea purpurea var. morning glory sogar 2 mm lang werden. — Der Endodermis ergrünter Achsen vonIpomoea Batatas fehlt einCasparyscher Streifen, bei im Dunkeln getriebenen Sprossen hingegen wird ein solcher ausgebildet.     

Expression of the pro-apoptotic gene gadd153/chop is elevated in liver with aging and sensitizes cells to oxidant injury

Aging is generally accompanied by reduced tolerance to oxidative stress and altered responsiveness to proliferative signals. We have shown that hepatocytes derived from aged rats (24-26 months) exhibit greater sensitivity to H(2)O(2) treatment and reduced proliferation following epidermal growth factor (EGF) treatment than cells of young adult rats (5-6 months). Here we examined the effects of aging and calorie restriction (CR) on expression of the oxidative stress-inducible and pro-apoptotic gene gadd153 (chop) in these hepatocytes, and we investigated its influence on sensitivity to oxidants. We show that aging was associated with elevated expression of gadd153, both basally and in response to H(2)O(2) treatment. CR, which attenuates age-associated declines in stress tolerance, prevented the age-related increase in gadd153 expression. EGF treatment also resulted in gadd153 induction in old cells. This effect was absent in young cells and in old cells of CR rats. gadd153 induction by EGF was reactive oxygen species-dependent and correlated with heightened sensitivity to subsequent H(2)O(2) treatment, suggesting that elevated Gadd153 contributes to the greater sensitivity of EGF-pretreated old cells to oxidative stress. Additional support for this hypothesis was provided by experiments with Rat1 fibroblasts in which conditional expression of Gadd153 conferred increased sensitivity to H(2)O(2). We propose a model whereby the diminished ability of old hepatocytes to overcome an EGF-triggered reactive oxygen species load leads to induction of the proapoptotic gene gadd153, which, in turn, sensitizes the cells to oxidant injury. Our findings point to gadd153 expression levels as an important factor in liver aging.     

Caveolin-induced activation of the phosphatidylinositol 3-kinase/Akt pathway increases arsenite cytotoxicity

The inhibitory effect of caveolin on the cellular response to growth factor stimulation is well established. Given the significant overlap in signaling pathways involved in regulating cell proliferation and stress responsiveness, we hypothesized that caveolin would also affect a cell's ability to respond to environmental stress. Here we investigated the ability of caveolin-1 to modulate the cellular response to sodium arsenite and thereby alter survival of the human cell lines 293 and HeLa. Cells stably transfected with caveolin-1 were found to be much more sensitive to the toxic effects of sodium arsenite than either untransfected parental cells or parental cells transfected with an empty vector. Unexpectedly, the caveolin-overexpressing cells also exhibited a significant activation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, which additional studies suggested was likely due to decreased neutral sphingomyelinase activity and ceramide synthesis. In contrast to its extensively documented antiapoptotic influence, the elevated activity of Akt appears to be important in sensitizing caveolin-expressing cells to arsenite-induced toxicity, as both pretreatment of cells with the PI3K inhibitor wortmannin and overexpression of a dominant-negative Akt mutant markedly improved the survival of arsenite-treated cells. This death-promoting influence of the PI3K/Akt pathway in caveolin-overexpressing cells appeared not to be unique to sodium arsenite, as wortmannin pretreatment also resulted in increased survival in the presence of H(2)O(2). In summary, our results indicate that caveolin-induced upregulation of the PI3K/Akt signaling pathway, which appears to be a death signal in the presence of arsenite and H(2)O(2), sensitizes cells to environmental stress.     

The Drug Diazaborine Blocks Ribosome Biogenesis by Inhibiting the AAA-ATPase Drg1

The drug diazaborine is the only known inhibitor of ribosome biogenesis and specifically blocks large subunit formation in eukaryotic cells. However, the target of this drug and the mechanism of inhibition were unknown. Here we identify the AAA-ATPase Drg1 as a target of diazaborine. Inhibitor binding into the second AAA domain of Drg1 requires ATP loading and results in inhibition of ATP hydrolysis in this site. As a consequence the physiological activity of Drg1, i.e. the release of Rlp24 from pre-60S particles, is blocked, and further progression of cytoplasmic preribosome maturation is prevented. Our results identify the first target of an inhibitor of ribosome biogenesis and provide the mechanism of inhibition of a key step in large ribosomal subunit formation.     

Analyses of dose-response curves to compare the antimicrobial activity of model cationic alpha-helical peptides highlights the necessity for a minimum of two activity parameters

To assess and compare different model Leu-Lys-containing cationic alpha-helical peptides, their antimicrobial activities were tested against Escherichia coli as target organism over a broad peptide concentration range. The natural cationic alpha-helical peptides magainin 2 and PGLa and the cyclic cationic peptide gramicidin S were also tested between comparison. The dose-response curves differed widely for these peptides, making it difficult to rank them into an activity order over the whole concentration range. We therefore compared five different inhibition parameters from dose-response curves: IC(min) (lowest concentration leading to growth inhibition), IC(50) (concentration that gives 50% growth inhibition), IC(max) (related to minimum inhibition concentration and minimum bactericidal concentration), inhibition concentration factor (IC(F); describing the increase in concentration of the peptide between minimum and maximum inhibition), and activity slope (A(S); related to the Hill coefficient). We found that these parameters were covariant: two of them sufficed to characterize the dose dependence and hence the activity of the peptides. This was corroborated by showing that the dose dependences followed the Hill equation, with a small, constant aberration. We propose that the activity of antimicrobial peptides can readily be characterized by both IC(50) and IC(F) (or A(S)) rather than by a single parameter and discuss how this may relate to investigations into their mechanisms of action.