The effect of sterols on the metabolism of Pythium species

Summary The growth of several Pythium species is increased between 65 and 100% if cholesterol is added to the growth medium. The optimum concentration is 15 mcg per ml. Mycelium of Pythium ultimum, in which cholesterol is present, incorporates glucose-U-¹⁴C and releases ¹⁴CO₂ at a faster rate than the corresponding sterol free mycelium. In sterol containing cells, more ¹⁴CO₂ is produced from a given amount of absorbed glucose-U-¹⁴C than in sterol free cells, there is thus in sterol containing hyphae a higher level of energy production. This condition can account for the increase in growth due to cholesterol. Only if sterols are present in the cellular membranes of Pythium species is the optimum synthetic capacity reached.     

Phytochrome in etiolated annual rye

Summary During a seven-fold increase in length the content of the coleoptile in photoreversible phytochrome increased four-fold and that of the primary leaf nine-fold. The phytochrome content, during growth, expressed on a fresh- or dry-weight basis did not vary greatly for either organ. Phytochrome per mg dry weight (OD⁷³⁰/mg=0.5) was nearly the same in the leaf as in the coleoptile. Coleoptiles studied had a constant DNA content of 4.1 g per organ. DNA content of the leaf increased with age. Phytochrome per DNA was much higher in the coleoptile than in the primary leaf and increased with growth in each of these organs. Thus, there was not a constant amount of phytochrome per cell in either tissue with increasing age and there was not the same amount of phytochrome per cell in the coleoptile as in the primary leaf at any age.This work was supported in part by U.S. Atomic Energy Commission Contract No. AT (30-I)2373.     

Deoxyribonucleic Acid Synthesis During Microcyst Germination in Myxococcus xanthus

Deoxyribonucleic acid (DNA) synthesis was measured during microcyst germination in Myxococcus xanthus by radioactive thymidine incorporation, autoradiography, and chemical analysis. Microcysts contained an average of 6.6 conserved units of DNA, corresponding to 3 to 4 chromosomes per cell. Correlation of the DNA content and chromosome number of microcysts indicated that the molecular weight of the nonreplicating M. xanthus chromosome is 4.9 x 10(9) daltons. DNA synthesis was initiated 3.5 to 4 hr after induction of germination. From 4 to 6 hr, the rate of synthesis was constant and the accumulation was linear. After a lag period (6 to 6.5 hr), the rate of DNA synthesis increased, reaching a second plateau at 9 hr. From 9 to 11 hr, the rate was again constant and the accumulation was linear. Cellular division during germination showed an unusual kind of synchrony. A model is presented that accounts for chromosomal replication and cell division during microcyst germination.     

Energy Sources for the Absorption of Rubidium by Chlorella

Die aktive (stoffwechselabängige) Aufnahme von Radiorubidium aus 8.10^?5 molarer Lösung durch Chlorella pyrenoidosa wurde in Langzeitversuchen (1–2 Tage) untersucht. Die Annahinc ist, daβ die dafür notwendige Energie in Form von ATP bereitgestellt werden muβ. Es wurde gefunden, daβ Gegenwart von Na- Oder Cl-Ionen für die aktive Rb-Aufnahme nicht notwendig ist; die Rb-Pumpe ist also nicht an eine Na- Oder Cl-Pumpe gekoppelt. Vorbeladene Chlorella gibt Rb über längere Zeiten langsam an das Medium ab. Durch Experimente im Licht bzw. Dunkel und in Gegenwart bzw. Abwesenheit von Luft wurde gezeigt, daβ bei 9000 lux die Beleuchtung mehr Energie für die aktive Aufnahme erbringt als die Atmung. Der Ersatz von Luft durch Sauerstoff hat keine Auswirkung. Die Versorgung mit ATP durch Atmung—nicht aber durch (zyklische) Photophosphorylierung—wird durch 5.10^?4 M DNP weitgehend unterdrückt. Die aktive Aufnahme ist klein, wenn sie sich nur auf Glykolyse stützt. Gleichzeitige Einwirkung von Licht, Luft (Sauerstoff) und DNP auf Chlorella führt zu irreversibler Schädigung und Ausbleichung der Algen. Sie hören auf, Rb aufzunehmen und verlieren vorher absorbiertes Rb wieder. Gründe für diesen Effekt werden diskutiert. Die aktive Aufnahme von Rb wird durch 5.10^?2 M Glukose gefördert, u.z.w. im Licht und im Dunkel, in Luft- und in Stickstoffatmosphäre. In dieser Beziehung unterscheidet sich die aktive Rb- von der aktiven Bromid-Aufnahme. Die Förderung durth Glukose wird durch DNP gehemmt, ausgenommen im Dunkel in Stickstoffatmosphäre. In Gegenwart von DNP wird die Rb-Aufnahme durch 1% CO₂ sowohl im Licht als auch im Dunkel stark herabgedrückt.     

Examination of Market Foods for Coliform Organisms

Food specimens (490) in nine categories were examined for total aerobic plate count and numbers and types of coliform organisms, including the enteropathogenic Escherichia coli (EEC). The total counts were compared with various suggested standards, and a limit of 100,000/g appeared to be a realistic goal, except for certain food types with a high level of natural flora. Plate counts in VRB were compared to counts obtained by isolation by enrichment in LST Broth, and the latter method produced a higher percentage of isolations. The presence of E. coli was determined by use of EC Medium incubated at 44.5 +/- 0.1 C. Only 40.4% of the positive EC tubes, however, contained E. coli. It appeared that a limit of 10 coliform organisms per g as a suggested standard could be met with several types of foods. Isolation of EEC was obtained only three times, or in 0.6% of the specimens.     

Correlation of Transmitter Release with Membrane Properties of the Presynaptic Fiber of the Squid Giant Synapse

Depolarization of the presynaptic terminal by current produced a postsynaptic potential (PSP) which increased with increasing presynaptic polarization and then reached a plateau. Iontophoretic injection of tetraethylammonium ions (TEA) into the presynaptic axon near the terminal produced a prolonged presynaptic spike. The resulting PSP is increased in size and its time course closely followed that of the presynaptic spike. The presynaptic fiber no longer exhibited rectification and strong depolarizations revealed that the PSP reached a maximum with about 110 mv depolarization. Further depolarization produced a decrease in PSP amplitude and finally transmission was blocked. However, a PSP then always appeared on withdrawal of the depolarizing current. Under the conditions of these experiments, the PSP could be considered a direct measure of transmitter release. Bathing the TEA-injected synapse with concentrations of tetrodotoxin (TTX) sufficient to block spike activity in both pre- and postsynaptic axons did not greatly modify postsynaptic electrogenesis. However, doubling TTX concentration reversibly blocked PSP. Thus the permeability changes to Na and K accompanying the spike do not appear necessary for transmitter release. Some other processes related to the level of presynaptic polarization must be involved to explain the data. The inhibition of transmitter release by strong depolarizations appears to be related to Ca action. A membrane Ca current may also be necessary for normal transmitter release.     

GIANT CENTRIOLE FORMATION IN SCIARA

Although somatic tissues of Sciara contain 9-membered centrioles, germ line tissues develop giant centrioles with 60–90 singlet tubules disposed in an oval array. Some 9-membered centrioles still may be seen in second instar spermatogonia. Each of these centrioles is associated with a larger "daughter" or secondary centriole at right angles to it. Most centrioles of second instar spermatogonia consist of 20–50 singlet tubules arranged in an oval, sometimes associated with an even larger secondary centriole. The more recently formed centriole of a pair is distinguishable from its partner by a concentric band of electron-opaque material inside its tubules. If a pair of centrioles at right angles to each other is pictured as a "T" formed by two cylinders, the secondary centriole is always the stem of the T; the primary centriole is the top. The two centrioles are oriented at the pole of the mitotic spindle so that the tubules of the primary centriole are parallel to the spindle axis. Each daughter cell receives a pair of centrioles and, during interphase, each of these centrioles gives rise to a new daughter centriole. A Golgi area of characteristic morphology is found in association with centrioles shortly after two new ones have formed. We conclude that in Sciara a centriole may give rise to a daughter morphologically different from itself. Whether the daughter is a 9-membered or giant centriole depends on the tissue type and stage of development.     

ACTION OF TRITON X-100 ON CHLOROPLAST MEMBRANES : Mechanisms of Structural and Functional Disruption

Addition of Triton X-100 to chloroplast suspensions to a final concentration of 100–200 µM causes an approximate tripling of chloroplast volume and complete inhibition of light-induced conformational changes, light-dependent hydrogen ion transport, and photophosphorylation. Electron microscopic studies show that chloroplasts treated in this manner manifest extensive swelling in the form of vesicles within their inner membrane structure. Triton was adsorbed to chloroplast membranes in a manner suggesting a partition between the membrane phase and the suspending medium, rather than a strong, irreversible binding. This adsorption results in the production of pores through which ions may freely pass, and it is suggested that the inhibition of conformational changes, hydrogen ion transport, and photophosphorylation by Triton is due to an inability of treated chloroplast membranes to maintain a light-dependent pH gradient. The observed swelling is due to water influx in response to a fixed, osmotically active species within the chloroplasts, after ionic equilibrium has occurred. This is supported by the fact that chloroplasts will shrink upon Triton addition if a nonpenetrating, osmotically active material such as dextran or polyvinylpyrrolidone is present externally in sufficient concentration (>0.1 mM) to offset the osmotic activity of the internal species.     

ULTRASTRUCTURE OF THE SPOON TYPE SYNAPTIC ENDINGS IN THE NUCLEUS VESTIBULARIS TANGENTIALIS OF THE CHICK

The fine structure of the "spoon" type synaptic endings of the chick tangential nucleus was studied with the electron microscope. These endings often measure ~18 µ in length by ~3–4 µ in width. The axoplasm of the endings contains very few synaptic vesicles, a large number of neurofilaments oriented parallel to the long axis of the nerve fiber, and microtubules and numerous mitochondria. The synaptic membrane complex shows areas of localized occlusion of the synaptic cleft with the formation of an external compound membrane. It has not been decided whether these areas have a disc shape; their length measures between 0.04 and 0.47 µ. The five-layer pattern characteristic of an external compound membrane is shown in specimens fixed with formalin—OsO₄, glutaraldehyde—acrolein—OsO₄, and acrolein KMnO₄ but it does not appear in the glutaraldehyde-OsO₄-fixed specimens. The over-all thickness of the external compound membrane varies depending upon the fixative used. The synaptic clefts in the regions between the external compound membrane discs are widened and measure ~300 A. A condensation of dense material occurs in pre- and postsynaptic cytoplasms all along the synaptic membrane complex. The morphological relationships described in the spoon endings are suggestive of electrical transmission.     

MEASUREMENT OF INFLUENZA VIRUS-ANTIBODY REACTION BY QUANTITATIVE ELECTRON MICROSCOPY

Measurement of the weight of individual virus particles from untreated and antibody-treated populations was made by quantitative electron microscopy. The weight of antibody bound depended on the concentration of antibody in solution. One population of viruses exposed to an antibody concentration which resulted in 95% inhibition of hemagglutination showed a mass increase of 55%, corresponding to an absolute increase of 9.0 x 10^-17 g in the median value. Another population, whose hemagglutination inhibition assay was 64%, showed a 39% increase in mass corresponding to an absolute median increase of 7.3 x 10^-17 g. The larger viruses in each population bound a greater absolute amount of antibody than did the smaller ones, but the latter bound relatively more antibody in proportion to their mass. No cross-reactivity was found between the antibody to influenza A/PR8 and the influenza strain B/LEE. Influenza A/PR8 controls exposed to nonspecific gamma-globulin displayed a significant weight loss, at least in part owing to loss from the core, as judged from the electron micrographs.