Blood plasma investigations by resonance raman spectroscopy: Detection of carotenoid pigments

Summary Using Raman spectroscopy, we demonstrate that low levels of -carotene, lycopene, and xanthophyll give rise to resonance enhanced bands in blood plasma. These results explain the significance of previously unidentified spectral maxima which have been related to the state of health of the blood donor.     

The effect of D-6-methyl-8-[β-isopropylaminoethyl] ergoline-I on the lactation in nursing rats

D-6-Methyl-8-[-isopropylaminoethyl] ergoline-I [VÚFB-10726], beginning from the dose of 0.05 mg/kg p.o., suppresses lactation through the inhibition of prolactin secretion in nursing rats.     

Poly(A) associated RNA from mitochondria and microsomes of rat brain

Summary Rat brain mitochondria contain a significant proportion of poly(A) associated RNA which is higher than that found in microsomes from the same source. When steady state poly(A) RNA of brain mitochondria was analyzed by microelectrophoresis, it displayed a characteristic separation pattern with a large amount of free poly(A).     

Ecdysone 20-hydroxylase from the midgut of the tobacco hornworm (Manduca sexta L.)

Summary An ecdysone 20-hydroxylase enzyme system that converts -ecdysone to 20-hydroxyecdysone was prepared from the midgut of the tobacco hornworm prepupa. This partially purified enzyme is NADPH dependent and is localized in the mitochondrial fraction of the midgut tissue.     

Octopamine,dopamine and noradrenaline content of the brain of the locust,Schistocerca gregaria

Summary The octopamine, dopamine and noradrenaline content of the brain of the locust,Schistocerca gregaria has been determined using sensitive radiochemical-enzymatic assays. Octopamine and dopamine are present in high concentration but the noradrenaline content is only 1/25 that of octopamine. Both reserpine and fusaric acid (a dopamine--hydroxylase inhibitor) produce a significant depletion of the octopamine stores.The author is a M.R.C. of Canada Fellow. Thanks are due to Drs.A. A. Boulton, A. V. Juorio, andP. H. Wu for helpful discussion.     

Biosynthesis ofα- andβ-ecdysone by the CrayfishOrconectes limosus in vivo and by its Y-Organs in vitro

Summary - and -ecdysone were synthesized from labelled cholesterol by premolt crayfish in vivo and by their Y-organs in vitro.     

Effect of hydrogen peroxide on the binding of Merocyanine 540 to human erythrocytes

The fluorescent dye Merocyanine 540 (MC540) is often used as a probe to monitor the molecular packing of phospholipids in the outer leaflet of biomembranes. In a previous study we showed that the increased staining of erythrocytes with a perturbed membrane structure was mainly due to an increase in the fluorescence yield of cell-bound MC540, rather than to an increase of the number of bound molecules. Erythrocytes and ghosts exposed to continuous fluxes of H₂O₂ exhibited pronounced lipid peroxidation. Further, red blood cells subjected to this form of oxidative stress also showed increased staining with MC540. It appeared that this was caused by a strong increase in binding of MC540, together with a slight red shift of the fluorescence emission maximum and a small increase in the fluorescence yield of bound MC540. The changed MC540 binding characteristics were not observed when lipid peroxidation was suppressed by the presence of the antioxidant BHT in the incubation medium. However, open ghosts exposed to H₂O₂ showed no increase of MC540 binding, excluding a direct involvement of lipid peroxidation. Measurement of fluorescence emission spectra and gel filtration studies showed that MC540 can bind to H₂O₂-exposed hemoglobin. Experiments with erythrocytes lysed in hypotonic medium after exposure to H₂O₂ revealed that peroxidation of lipids with H₂O₂ induced a non-specific permeabilization of the plasma membrane to MC540, thereby allowing MC540 to bind to the oxidatively denatured, more hydrophobic hemoglobin. These results indicate that conclusions about packing of phospholipids in the outer leaflet of the membrane based on increased MC540-staining should be drawn with care. Received 27 September 1996; received after revision 5 November 1996; accepted 27 November 1996     

Two classes of metabolites fromTheonella swinhoei are localized in distinct populations of bacterial symbionts

The marine spongeTheonella swinhoei (lithistid Family Theonellidae, Order Astrophorida) has yielded many important, bioactive natural products, most of which share structural features with bacterial natural products. The presence of microbial symbionts inT. swinhoei has been reported, and it was originally suggested that the cytotoxic macrolide swinholide A and many of the bioactive cyclic peptides fromT. swinhoei were all produced by symbiotic cyanobacteria. By transmission electron microscopy, we found four distinct cell populations to be consistently present inT. swinhoei: eukaryotic sponge cells, unicellular heterotrophic bacteria, unicellular cyanobacteria and filamentous heterotrophic bacteria. Purification and chemical analyses of each cell type showed the macrolide swinholide A to be limited to the mixed population of unicellular heterotrophic bacteria, and an anti-fungal cyclic peptide occurred only in the filamentous heterotrophic bacteria. Contrary to prior speculation, no major metabolites were located in the cyanobacteria or sponge cells.     

Effect of various stressors on the levels of lipid peroxide,antioxidants and Na+, K+-ATPase actrivity in rat brain

The level of malondialdehyde (MDA), an index of lipid peroxidation, and the antioxidants superoxide dismutase (SOD) and glutathione (GSH), as well as the activity of Na⁺, K⁺-ATPase, were assessed in whole rat brain after immobilization, anemic hypoxia (NaNO₂) and 72 h starvation. The effect of these stressors on plasma glucose and corticosterone levels was also observed. Hypoxia and starvation stimulated the lipidj peroxide formation in braini as indicated by an increase in the level of MDA, being higher after starvation than hypoxia. Brain SOD activity was also increased in response to hypoxia and starvation while GSH content was only diminished ini hypoxia. However, neither MDA nor antioxidants were affected by immobilization. On the other hand, the activity of brain Na⁺, K⁺-ATPase was significantly increased by immobilization and hypoxia but decreased in starvation. A similar pattern of change was also observed in plasma glucose and corticosterone levels in response to these stressors. These results elucidate differences in the biochemical response of animals towards various types of stress, with increased lipid peroxide formation in hypoxia and starvation.