Electron microscopic cytochemical localization ofα-hydroxyacid oxidase in rat liver

Summary The substrate specificity and the intraperoxisomal localization of -hydroxyacid oxidase in rat liver has been investigated cytochemically by the cerium technique and biochemically with a luminometric assay. Rat liver is fixed by perfusion with a low concentration (0.25%) of glutaraldehyde and vibratome sections are incubated for 60 min at 37°C in a medium containing 3 mM CeCl₃, 100 mM NaN₃ and 5 mM of an -hydroxyacid in 0.1M of one of the following buffers: Pipes, Mops, Na-cacodylate,Tris-maleate, all adjusted to pH 7.8. Ten different -hydroxyacids with a chain length between 2 and 8 carbon atoms were tested. The best results were obtained with glycolic, argininic andl--isocaproic acids. These cytochemical findings were confirmed also biochemically using purified peroxisomal fractions isolated by gradient centrifugation in metrizamide. The pattern of the intraperoxisomal localization of the enzyme was influenced markedly by the type of buffer used for the cytochemical incubation. Whereas in theTris-maleate medium both the cores and the matrix stained with the same intensity, with all other buffers the reaction in cores was more prominent. The staining of cores was abolished by pretreating sections inTris-maleate (pH 7.8) or alkaline pyrophosphate buffers. These observations establish the substrate specificity of -hydroxyacid oxidase in rat liver and demonstrate the delicate association of this enzyme with the crystalline cores and the matrix of peroxisomes in rat liver.Abbreviations -HAOX l-hydroxyacid oxidase - Argininic acid l--hydroxy--guanidinovaleric acid - Pipes piperazine-N,N-bis(2ethane sulfonic acid) - Mops 3(N-morpholino) propane sulfonic acid - Tris tris-(hydroxymethyl)-aminomethane - Luminol 5-amino-2,3 dihydrophthalazine-1,4-dione - GA glutaraldehyde     

A kinetic model of starch hydrolysis by alpha- and beta-amylase during mashing

Kinetics of malt starch hydrolysis by endogeneous alpha- and beta-amylases has been experimentally investigated in laboratory-, pilot- and industrial-scale reactors. The production rates of glucose, maltose, maltotriose and total extract, and the separate alpha- and beta-amylases deactivation rates are measured at varying mashing temperature and different initial starch concentrations and qualities. Based on the experimental results, a model is proposed that takes into account the initial carbohydrates and enzymes dissolution, the starch gelatinization, the separate hydrolytic action of alpha-and beta-amylases on insoluble and soluble starch and dextrins, and the influence of temperature both on enzyme activities and thermal denaturation rate. The model can predict, at the three scales, the final sugars concentrations in the wort for given initial malt concentrations and enzymatic contents, and for a fixed temperature profile during the mashing process.     

Theoretical conformational analysis of phospholipids II. Role of hydration in the gel to liquid crystal transition of phospholipids

To obtain a satisfactory agreement between computed transition temperatures and those determined experimentally, we introduce explicitly water molecules which hydrate the polar headgroup of dipalmitoylphosphatidylethanolamine molecules. The calculated free energy curves as a function of the intermolecular interchain distance and the degree of hydration of the polar groups permit the determination of the transition of the phospholipid system from the gel to the liquid crystalline phase. The detailed structure of the hydration shell is defined using the supermolecular approach.     

The incorporation of selenium and alterations of macro- and trace element levels in individual blood cells following supplementation with sodium selenite and vitamin E

Significant alterations in the selenium content of erythrocytes, thrombocytes, and neutrophil granulocytes were observed following a daily supplementation of 200 μg Se + 100 mg vitamin E during a period of 2 months. The neutrophil granulocytes incorporated more selenium than the thrombocytes. The iron content of the thrombocytes decreased on selenium supplementation, while the opposite was noted for the neutrophil granulocytes. The glutathione peroxidase activity was not significantly changed during the period of observation.     

Improved demonstration of exocytotic profiles in glomus cells of rat carotid body after perfusion with glutaraldehyde fixative containing a high concentration of potassium

Extensive secretion by exocytosis was demonstrated in the glomus (type I) cells of the adult rat after perfusion of carotid bodies with a potassium-rich (high K) glutaraldehyde fixative. Similar secretory profiles were very rare with a glutaraldehyde fixative containing a low concentration of potassium (low K). The increase in the incidence of exocytotic profiles in glomus cells with the high K fixative was highly significant, whereas no statistical difference could be observed in the incidence of coated pits with the different fixatives. Exocytotic profiles were characterized by the following features: (1) they predominated in non-synaptic regions, but were occasionally observed near synapses between two glomus cells; they were not observed near synapses between glomus cells and nerve terminals; (2) extruded electron-dense material associated with coating of the cell membrane was frequent; (3) different stages of dissolution of the extruded granule material was evident. The possible role of exocytosis as a mode of secretion in the glomus cells and the characteristics of the new high K-glutaraldehyde fixative are discussed.     

Les effets de recristallisation sur la paroi des Involutinidae (Foraminifères) triasiques

The different types of recrystallization which can be observed in the Triassic involutinids should be considered in the taxonomic treatment of this foraminiferal family. If the diagenetic modifications of the test are neglected the same form could be placed into different suborders. Our inclusion of the genusTriasina Majzon in the Involutinidae proposed earlier on purely morphologic grounds, seems to be supported by the fact that the same types of recrystallization exist inTrocholina, Involutina undTriasina.     

Book reviews

Ohne Zusammenfassung

---

Book reviews

     

Peptides with NH2-terminal tryptophan in the adenohypophysis

Summary In the mammalian pituitary formaldehyde-ozone treatment induces strong fluorescence in the cells of the pars intermedia and moderate to strong fluorescence in numerous cells of the pars distalis. Maximum excitation is at 370–375 nm and maximum emission at 495–505 nm. The properties of the cellular fluorescence are indistinguishable from those of tryptamine or peptides with NH₂-terminal tryptophan. From chemical analysis such peptides seem to occur abundantly in the mammalian pituitary. The concentration of these peptides agrees very well with the number and fluorescence intensity of the cells in all species studied. Furthermore, the tryptophyl peptides in the various parts of the pig pituitary have a distribution quite parallel to that of the fluorescent cells. As we have failed to detect tryptamine in the pituitary, we conclude that the formaldehyde-ozone-induced fluorescence in the adenohypophysis reflects the presence of tryptophyl peptides.This study was supported by grants from the Swedish Medical Research Council (04X-1007; 04X-3764), the Ford Foundation, Harald and Greta Jeanssons stiftelse and Riksföreningen mot Cancer (660-K73-01X).For brevity occasionally referred to as tryptophyl peptides.