Expression of monoclonal antibody-defined cell surface antigens during rat brain development

Using single-cell suspensions of mechanically dissociated, prenatal BDIX-rat brain cells (13th, 15th, and 21st days after fertilization) for immunization, we have established a collection of 37 monoclonal antibodies (Mabs) directed against neural cell surface determinants. The developmental-stage-dependent expression of cell-surface antigens recognized by these Mabs was analyzed both on plasma membranes isolated from whole brains of BDIX rats (prenatal days 13-22 and adults) using an indirect 125I solid-phase radioimmunoassay, and on intact BDIX-rat brain cells (prenatal days 13-22) using a fluorescence-activated cell sorter. Different types of developmental stage-dependent profiles of Mab binding were found, these being indicative of the presence of neural cell surface determinants whose expression increases, decreases, or does not change with brain development. Some of the Mab-binding profiles showed transient changes as a function of developmental stage. These Mabs are currently being used for the characterization, reproducible identification, and isolation of neural cell subpopulations of the developing rat brain, with the aim of investigating the cell type dependence and developmental (differentiation) stage dependence of malignant transformation following pulse exposure to the carcinogen N-ethyl-N-nitrosourea at defined stages of brain development.     

Electrostatic control by lipids upon the membrane-bound (Na+ + K+)-ATPase. II. The influence of surface potential upon the activating ion equilibria

Sarcoplasmic reticulum Ca2+-ATPase has been reconstituted with excess lipid (25-150 g egg phosphatidylcholine per g sarcoplasmic reticulum protein) by a procedure combining the use of a non-ionic detergent with cholate dialysis. The reconstituted vesicles were analyzed by sucrose density fractionation and freeze-fracture electron microscopy. At the lowest lipid to protein ratios some vesicles containing aggregated protein were observed. At a lipid to protein ratio of 150:1 (w/w) only 30-40% of the reconstituted protein sedimented through 7% (w/v) sucrose. The remainder of the latter preparation was characterized by a high Ca2+-uptake capacity and a coupling ratio of 1.6 mol Ca2+ transported per mol ATP hydrolyzed. Intramembranous particles in this preparation occurred isolated in the membrane. In most cases only one particle could be seen on a fracture face. Cross-linking with cupric phenanthroline indicated that protein-protein contacts were drastically reduced by reconstitution. It is concluded that aggregation of intramembranous particles is not required for optimal Ca2+-transport function. The dispersed preparation obtained by a combined reconstitution and sucrose density fractionation procedure is useful for further characterization of the Ca2+ pump.     

Effects of culture density on the kinetics of germ tube formation in Candida albicans

The relationship between culture density or phase of growth at 24.5 degrees C and the ability of Candida albicans to form germ tubes when shifted to 37 degrees C was investigated. Evidence is presented demonstrating germ tube production from liquid synthetic medium cultures at all phases of growth. Previous studies reported that only cells from stationary phase cultures were competent to form germ tubes. Comparisons between exponential and stationary phase cultures indicate more rapid and more synchronous germ tube production from cells growing in the exponential phase.     

An evaluation of four methods for measuring cholesterol absorption by the intestine in man

Critical comparisons have been made in 12 patients of four methods for measuring cholesterol absorption from the intestine. Methods I-III depend on the use of labeled cholesterol (intravenously or continuous labeling orally) in conjunction with sterol balance measurements; Method IV can be carried out with only a single test dose containing labeled cholesterol plus labeled beta-sitosterol. In the latter technique absorption is calculated as the loss of cholesterol relative to beta-sitosterol during intestinal transit. Method III (isotopic steady-state method) proved to be undependable because of uncertainties in determining the existence of an isotopic steady state. However, Method IV gave good agreement with Methods I and II, and it appears to have certain practical as well as theoretical advantages. Although Method IV requires collections of stools for up to 8 days, it is nevertheless the most rapid and the simplest of all the methods for estimating absorption. It can also be used in certain situations, such as in fur-licking animals, when Methods I and II are inadequate. Therefore, this method would seem to be a valuable addition to other isotopic techniques for estimating cholesterol absorption in man.     

Effects of dietary cholesterol on the regulation of total body cholesterol in man

Studies on the interaction of cholesterol absorption, synthesis, and excretion were carried out in eight patients using sterol balance techniques. Absorption of dietary cholesterol was found to increase with intake; up to 1 g of cholesterol was absorbed in patients fed as much as 3 g per day. In most patients, increased absorption of cholesterol evoked two compensatory mechanisms: (a) increased reexcretion of cholesterol (but not of bile acids), and (b) decrease in total body synthesis. However, the amount of suppression in synthesis was extremely variable from one patient to another; one patient had no decrease in synthesis despite a large increment in absorption of dietary cholesterol, and two patients showed a complete suppression of synthesis. In the majority of cases the accumulation of cholesterol in body pools was small because of adequate compensation by reexcretion plus reduced synthesis, but in a few patients large accumulations occurred on high cholesterol diets when absorption exceeded the compensatory mechanisms. These accumulations were not necessarily reflected in plasma cholesterol levels; these increased only slightly or not at all.     

Polyamine als Stimulatoren von Genaktivitäten Eine autoradiographische Studie an Riesenchromosomen von Chironomus thummi

Zusammenfassung Inaktive Riesenchromosomen der Speicheldrüsen ausgewachsener Chironomus thummi-Larven zeigten nach 60 min Inkubation mit 0,05 N Putrescin-, Spermidin-oder Spermin-Lösung erneute Syntheseaktivitäten, die sich im Einbau von radioaktiv markiertem Uridin autoradiographisch zu erkennen geben. Diese Beobachtung wird im Zusammenhang mit der bekannten genaktivierenden Wirkung von Mg⁺⁺ diskutiert. Auf der Basis zahlreicher biochemisch analoger Wirkungen von Polyaminen und Mg⁺⁺ wird ein ursprünglich für die genaktivierende Wirkung von Mg⁺⁺ postuliertes Modell auf Polyamine übertragen.

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Stimulation of gene activities by polyamines an autoradiographic study with giant chromosomes of Chironomus thummi

Summary Inactive salivary gland giant chromosomes of full grown Chironomus thummi larvae showed after 60 min of incubation in 0.05 N putrescine, spermidine or spermine solutions autoradiographically demonstrable incorporation of radioactiv labelled uridine caused by a reactivation of RNA synthesis. This observations is discussed in relation to the known gene activating effect of Mg⁺⁺, and, since polyamines and Mg⁺⁺ have numerous effects in common, it is suggested that polyamines affect gene activity in a way similar to Mg⁺⁺.

     

Die Rolle der Fimbrien bei der eigenartigen Sternbildung von Agrobacterium luteum

Zusammenfassung Die bemerkenswerte Sternbildung von Agrobacterium luteum Stamm A61 (Moll et al., 1967; Ahrens, 1968) ist auf polare Fimbrien zurückzuführen, die um die Sternaggregate ausgebreitet sind. Es wird angenommen, daß sich Schwärmzellen an den Fimbrien verfangen und durch Kontraktion rasch zum Stern gezogen werden. Nach elektronenoptischen Präparaten sind die Fimbrien bis zu 10,5 lang und können die beobachtete Sternbildung gut erklären. Agrobacterium luteum Stamm B14 besitzt ebenfalls lange polare Fimbrien. Beide Stämme tragen peritriche Geißeln, deren Struktur sich deutlich von den dünnen und unregelmäßigen Fimbrien unterscheidet.

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The function of fimbriae in the peculiar star formation of Agrobacterium luteum

Summary The remarkable way of star formation in Agrobacterium luteum strain A61 (Moll et al., 1967; Ahrens, 1968) was found to be due to polar fimbriae which are spread around the star-shaped aggregates. It is assumed that swarmers adhere to the fimbriae and, by contraction, are swiftly pulled towards the star. As seen in electron microscopic preparations, the fimbriae are up to 10,5 long and may well explain the star formation observed. Agrobacterium luteum strain B 14 also possesses long polar fimbriae. Both strains have peritrichous flagella, the structure of which being clearly distinct from the delicate and irregular fimbriae.

     

Dietary beta-sitosterol as an internal standard to correct for cholesterol losses in sterol balance studies

In the course of carrying out sterol balance studies in 19 patients, we gathered the following evidence that, in some but not all patients, considerable amounts of neutral sterols are "lost" during their passage through the intestinal tract. (a) Since plant sterols are largely nonabsorbable in man, they should be totally recovered in the feces; yet in many patients significantly less plant sterol than expected was recovered, the loss amounting to as much as 56% of daily intake. (b) In two patients in whom cholesterol-(14)C and beta-sitosterol-(3)H were instilled into the terminal ileum, from which neither sterol is absorbed, the feces contained 25% less of each isotope than was instilled. (c) In four patients fed radioactive cholesterol daily until the isotopic steady state was closely approximated, 28-50% of the isotope could not be accounted for. On the other hand, in five patients fed radioactive bile acids until the isotopic steady state was approximated, input equalled output as predicted. Since the amount of -sitosterol absorbed in man is limited (5% or less), this sterol can be used as an internal standard for upward correction of the figure obtained for the amount of neutral steroids excreted. The use of beta-sitosterol for this purpose is based on three considerations: (a) it passes through the intestine in the same physicochemical state as cholesterol; (b) it accompanies cholesterol at every step of its isolation and chromatographic measurement; and (c) it is lost to the same extent as cholesterol. Excretion data for fecal neutral steroids can therefore be corrected for irregular fecal flow as well as for the "unexpected loss" referred to. This loss seems to be due not to errors in stool collection or to technical errors, but to intestinal bacterial degradation of neutral 3beta-OH,Delta(5)-sterols to products not recognized as steroids in the analytical methods used.