Light-Induced Changes in the Fluorescence Yield of Chlorophyll a In Vivo: II. Chlorella pyrenoidosa

The long-term fluorescence induction in Chlorella pyrenoidosa consists of a fast rise of the fluorescence yield from the level S (of the first wave transient) to a maximum M, followed by slower decay to a terminal stationary level T. The maximum M is attained within 40 seconds from the onset of illumination while the decay to the terminal level T lasts for several minutes. The fluorescence rise (S → M) coincides with an increase in the rate of oxygen evolution, which, however, remains constant during the fluorescence decay (M → T). Poisons of photosynthesis 3, (3,4-dichlorophenyl)-1,1 dimethylurea (DCMU, o-phenathroline) inhibit the fluorescence induction, while uncouplers of photophosphorylation affect the fluorescence time course only when they function at an early stage of the coupling sequence e.g., carbonyl cyanide p-trifluoremethoxy phenylhydrazone, (FCCP, atabrin). Phosphorylation inhibitors affecting only the terminal esterification step (phlorizin) have little effect on the fluorescence kinetics. These results suggest that the fluorescence induction requires the operation of a phosphorylating electron transport and that it is possibly related to the light-induced structural changes which accompany photophosphorylation.     

The Subcellular Origin of Bioluminescence in Noctiluca miliaris

The light emitted by Noctiluca has its origin in 1 to 5 x 10⁴ organelles ("microsources") which are scattered throughout the perivacuolar cytoplasm, and which appear to be the elementary functional units of bioluminescence. Microscopical techniques, image intensification, and microphotometry were employed in their investigation. Microsources are fluorescent, strongly phase-retarding, and range widely in diameter below 1.5 microns. The number of quanta emitted in a flash from a microsource ("microflash") is of the order of 10⁵ photons. However, microflashes show a wide range of intensities, which are correlated with the size of the organelles from which they arise. Each organelle responds repetitively and with reproducible time course to a succession of invading triggering potentials. Reversible changes in the intensity of the flash emitted by the whole cell ("macroflash") occur because of graduations in intensity of microflashes rather than as a result of changes in the number of responsive organelles. The shape of the flash emitted by individual microsources resembles that of the macroflash except for slightly shorter rise and decay times. It is concluded that the macroflash results from somewhat asynchronous, but otherwise parallel summation of microflashes.     

Enzyme-induced Formation of Spheres from Cells and Envelopes of Escherichia coli

Normal and filamentous whole cells and isolated envelopes of Escherichia coli B were exposed to various enzymatic treatments to remove surface layers and to characterize the component(s) conferring rigidity in this organism. Modification of cell rigidity was determined by sphere formation in both whole cells and isolated envelopes. Enzymes capable of converting trypsinized normal or untreated filamentous whole cells and untreated envelopes to spheres included: lysozyme plus ethylenediaminetetraacetic acid, clostridial phospholipase C, and phospholipase D from cabbage. These data suggest that there are at least two components essential for maintenance of cell rigidity in E. coli B. The first is the peptidoglycan (mucopeptide), which is susceptible to lysozyme. The second is a phospholipid which is either covalently linked to the mucopeptide or in close association with it. This phospholipase C-sensitive component is protected more completely in normal than in filamentous whole cells by a protein layer which is easily modified by trypsin treatment to allow enzymatically induced sphere formation to occur.     

Inheritance of quantitative expression of erythrocyte glucose-6-phosphate dehydrogenase activity in the Negro—a twin study

Studies have been conducted on eight sets of monozygous and nine sets of dizygous female Negro twins, both members of whom were heterozygous for G-6-PD deficiency. Twins were studied both by assay of erythrocytic G-6-PD activity and by the methemoglobin elution test (MET). The MET is a procedure which identifies histochemically cells with appreciable G-6-PD activity and permits accurate determination of the percentage of such cells in heterozygotes. Monozygous twins showed significantly less within-pair variation than dizygous twins with both the MET and G-6-PD assay.Concerning the significantly greater agreement in MET results in monozygous twins than dizygous twins, our present working hypothesis is that X-chromosomal inactivation in the Negro female is genetically controlled, rather than random. However, certain alternate hypotheses allowing for random X-inactivation have not been excluded; these include somatic cell selection after random X-inactivation, and cell exchange between identical twins in utero/it. Studies in nontwin related heterozygotes now underway should help differentiate among these various possibilities.In addition to the studies on 17 pairs of female twins heterozygous for G-6-PD deficiency, 26 pairs of nondeficient female Negro twins have been studied by G-6-PD assay. Within-pair variation in monozygous twins was significantly less than within-pair variation in dizygous twins in all cases. The genetic influences detected with the G-6-PD assay in the female twins could theoretically be due to nonrandom X-inactivation, to genetically determined quantitative differences in enzyme activity (e.g., isoalleles), or to both. By appropriate calculations, based on the MET results, we have factored out the effects of X-inactivation on overall enzyme activity in the heterozygous deficient twins. After removal of the effect of X-inactivation, monozygous twins heterozygous for enzyme deficiency continue to show significantly less within-pair variation than dizygous twins. This finding indicates significant genetic influences on quantitative G-6-PD activity other than X-inactivation and other than the deficiency allele. This conclusion has been strengthened by studies on male twins where X-inactivation is not present.Supported by USPHS research grants AM-09381, HE-17544, AM-09919, and HE-03341, by USPHS Career Development Award 1-K3-AM-7959 (Dr. Brewer) and by U.S.A.E.C. Contract (11-1)-1552.     

Blood Viscosity in Healthy Subjects and Patients with Coronary Heart Disease

Viscosity of whole blood and plasma was measured in 258 apparently healthy subjects of both sexes from 5 to 60 years of age, and in 86 patients with unequivocal evidence of chronic coronary heart disease. Children and young healthy females had the lowest viscosity readings. Healthy young and middle-aged males had significantly higher blood viscosity than females. Patients with coronary heart disease had significantly higher blood viscosity values than healthy groups of the same sex. It is suggested that the higher viscosity of whole blood and of plasma is a contributory factor in development of clinical manifestations of coronary heart disease and possibly of the basic vascular lesion itself.     

Absorption and long distance transport by isolated stele of maize roots in relation to the dual mechanisms of ion absorption

Summary Ion absorption and transport by intact roots, isolated cortex and isolated stele were compared shortly after tissue isolation and after aging. Absorption isotherms in the low and in the high concentration range show that in stripped-stele, which absorbs at a very low rate immediately after isolation, the capacity of system 1 but not system 2 is built up with aging. In agreement with this result analysis of individual fluxes across plasmamembrane and tonoplast reveals that only the influx from the medium into the cytoplasm increases considerably with aging of stele. Changes observed in aging excised roots and in isolated cortex are much less significant. In spite of the increase of absorption with aging by isolated stele, long distance transport, which is essentially passive through freshly stripped stele, decreases with aging. The reported results reflect the marked permeability of the plasmamembrane of fresh isolated stele, and demonstrate the importance of the cortex as a tissue collecting ions for long distance transport. New evidence for the theory of symplasmatic transport of ions into the xylem vessels is thus provided.

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Zusammenfassung Die Ionenaufnahme durch intakte Wurzeln, isolierte Rinde und isolierte Zentralzylinder und der Ferntransport durch intakte und entrind ete Wurzeln wurden verglichen, und zwar kurz nach der Isolierung und nach Altern der Gewebe. Frisch isolierte Zentralzylinder akkumulieren Io nen nur in ganz geringem Maße oder überhaupt nicht. Von den beiden Systemen der metabolischen Ionenaufnahme, die in einem niedrigen (System 1: bis 0,5 meq/l) und in einem hohen Konzentrationsbereich (System 2: 1-50 meq/l) die Geschwindigkeit der Ionenaufnahme durch intakte Wurzeln und isolierte Wurzelrinde bestimmen, entwickelt sich während des Alterungsprozesses in isolierten Zentralzylindern System 1, nicht aber System 2.In Übereinstimmung mit diesem Befund zeigt die Analyse der Einzelfluxe am Plasmalemma und am Tonoplasten, daß nur der Influx aus der Außenlösung in das Cytoplasma beim Altern der Zentralzylinder beträchtlich ansteigt. Veränderungen beim Altern von abgeschnittenen, intakten Wurzeln und isolierter Rinde sind viel weniger ausgeprägt.Obwohl die Ionenaufnahme beim Altern isolierter Zentralzylinder steigt, verringert sich der Ferntransport, der bei frisch isolierten Zentralzylindern rein passiv ist. Die mitgeteilten Ergebnisse zeigen die ausgeprägte Permeabilität frisch isolierter Zentralzylinder und demonstrieren die Bedeutung der Wurzelrinde als ein Gewebe, das Ionen für den Ferntransport sammelt. Auf diese Weise werden neue Anhaltspunkte für die Theorie des symplasmatischen Transportes der Ionen in die Gefäße gewonnen.