STUDIES OF NATIVE GLYCOGEN ISOLATED FROM SYNCHRONIZED TETRAHYMENA PYRIFORMIS (HSM)

Native glycogen was isolated from Tetrahymena pyriformis (HSM) by isopycnic centrifugation in cesium chloride density gradients. A density of 1.62 to 1.65 was isopycnic for glycogen. Most of the banded glycogen existed as 35 to 40 mµ particles which had a sedimentation coefficient of 214. These particles were composed of aggregates of 2 to 3 mµ spherical particles. Extraction of glycogen with hot alkali reduced the sedimentation coefficient of native glycogen from 214 to 64.7 and the particle diameter from approximately 40 to 20 mµ and smaller. Cell division was synchronized by a repetitive 12-hour temperature cycle, and glycogen was measured at several times during the cell cycle. The temperature cycle consisted of 9.5 hours at 12°C and 2.5 hours at 27°C. Approximately 90 per cent of the cells divided during the last 1.5 hours of the warm period. The carbohydrate/protein ratio of cells at the end of the cold period was 0.27 and was reduced slightly during the warm period. Glucose was incorporated into glycogen during both periods, although the rate of incorporation was greater during the warm period. No preferential incorporation on the basis of particle size was noted. Incorporation was measured in both native glycogen and KOH-extracted glycogen. Tetrahymena glycogen is compared with rat liver glycogen previously isolated by similar procedures, and the significance of using combined rate-zonal and isopycnic centrifugation for isolating native glycogen is discussed.     

Sterilization of Regenerated Collagen Sutures with β-Propiolactone

Data are presented to show that β-propiolactone when properly applied is a very effective agent for sterilization of regenerated collagen sutures. The chemical sterilization is accomplished with little or none of the loss in strength encountered with heat sterilization. The finished sterile suture is obtained without any harmful residue that might be detrimental to the patient.     

ELECTRON MICROSCOPY OF THE OXYNTIC CELL IN THE GASTRIC GLANDS OF THE BULLFROG (RANA CATESBIANA) : I. The Non-Acid-Secreting Gastric Mucosa

The fine structure of the oxyntic cell from the gastric glands of the bullfrog was studied in lead hydroxide—stained sections of gastric mucosa fixed in buffered osmium tetroxide and embedded in n-butyl methacrylate. The oxyntic cell in non-acid-secreting stomachs (gastric juice pH, 7.4–7.8) is characterized by: (a) numerous closely packed smooth surfaced vesicular and tubular profiles disposed randomly in the cell; some of these elements show interconnections making it possible to identify this component with smooth surfaced endoplasmic reticula of certain other cell types, (b) a small percentage of rough surfaced profiles characteristic of endoplasmic reticula possessing RNP particles on the outer membrane surfaces, (c) a Golgi complex consisting of multiple isolated non-polarized arrays of smooth surfaced parallel elongated profiles and associated vesicular elements, (d) a sparse granular component (140 A) scattered freely in the cytoplasmic matrix, (e) numerous mitochondria with a dense matrix and containing an unusually large number of closely approximated cristae, (f) a number of zymogen granules consisting of either a dense body limited by a membrane or surrounded by a halo of less dense material which is in turn limited by a membrane, and (g) a number of granules (~260 A) containing several smaller granules (~80 A) identified presumably as glycogen. Intracellular canaliculi were not observed. Instead the free surface of the oxyntic cell facing the lumen of the gastric gland shows a complicated plication of the plasma membrane. Intercellular canaliculi are seen frequently between adjacent oxyntic cells. The walls of these canaliculi are made up of folded and ruffled cell membranes. The basal surface of the cell also exhibited this type of configuration. Occasional smooth surfaced profiles are seen communicating with the free surface, the wall of an intercellular canaliculus, or the basal surface of the cell. Although nerve endings were not found in association with oxyntic cells, unmyelinated nerves were observed in the vicinity of the gastric glands.     

ELECTRON MICROSCOPE STUDY OF MITOSIS IN SEA URCHIN BLASTOMERES

The fine structure of cells at different stages of the mitotic cycle was studied in the blastomeres of 6-hour-old embryos of the sea urchin Strongylocentrotus purpuratus. The material was fixed in 1 per cent osmium tetroxide in sea water, buffered with veronal-acetate to pH 7.5, embedded in Araldite, and sectioned with glass knives. The aster, as it forms around the centriole, has the appearance of the endoplastic reticulum, with elements oriented radially from the centrosphere to the periphery of the cell. Anaphase structures described include the kinetochores, with bundles of fine filaments extending toward the centrioles, as well as continuous filaments passing between the chromosomes. Two cylindrical centrioles composed of parallel rods are present in each of the anaphase asters. At late anaphase, elements of the endoplasmic reticulum condense on the surface of the chromosomes to form a double membrane which already at this stage possesses pores or annuli. At telophase bundles of continuous filaments can be seen in the interzonal region. These filaments, as well as those associated with the chromosomes, have a diameter of approximately 15 mµ, and appear physically different from the astral structure.     

Autoradiographies de Coupes d'os compact après marquage aux isotopes in vitro

Résumé Etude qualitative et quantitative du marquage in vitro de coupes d'os compact. Détermination du rôle joué par les processus de diffusion, d'adsorption et d'échange dans ces phénomènes.

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Zusammenfassung Qualitative und quantitative Studie der in vitro-Markierung von Schliffen aus Knochen-Compacta. Bestimmung der Rolle der Diffusions-, Adsorptions- und Austauschprozesse für diese Vorgänge.

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La partie expérimentale de ce travail a pu être réalisée grâce à des subsides fournis par l'European Office of the Air Research and Development Command [contrat n^o AF 61 (514) 1175], par l'Institut Interuniversitaire Belge des Sciences Nucléaires, et par la Commission pour la Science Atomique du Fonds National Suisse de la Recherche Scientifique.