"New membrane" formation in Amoeba proteus upon injury of individual cells. Electron microscope observations

Changes in the plasma membrane complex following the injury of single cells of Amoeba proteus were examined with the electron microscope. Two types of injury were employed in this study; cells were either pinched ("cut") in half or speared with a glass microneedle, and quickly fixed. Speared cells, when fixed in the presence of the ruthenium violet (a derivative of ruthenium red), revealed the presence of an extra trilaminar structure outside of each cell. This structure, called the "new membrane," was separated from the plasma membrane complex by a distance of less than a micron. The trilaminar structure of the new membrane strikingly resembled the image of the plasma membrane in all cells examined, except for its increased width (30%). This new membrane appeared nearly to surround the injured amebae. Attempts were made to demonstrate the possible origin of the new membrane, its reality, and its sensitivity to calcium. Also, some evidence is shown concerning the role of the small dense droplets (100–1200 A in diameter) normally present in the cytoplasm of amebae. Their frequent contact with the plasma membrane of the cell as the result of injury is interpreted as indicating their involvement in the formation and expansion of the plasma membrane.     

Selection for increased abdominal bristle number in Drosophila melanogaster with concurrent irradiation

Summary Replicate lines, each, with one hundred pairs of parents selected at 50% intensity, were derived from an inbred line. For twenty generations three lines were selected without irradiation and five with 1000 r X-rays per generation given to both females and males. After adjustment for level of crowding in the cultures, the final mean was 1.3 bristles higher in females and 1.0 bristles higher in males in the irradiated lines than in the unirradiated lines. In terms of phenotypic standard deviations in each sex in the base population, these total responses were 0.74 and 0.59 respectively. Radiation can induce mutations useful in increasing responses in selection programmes, but the average response attributable to radiation is small, and heterogeneity between replicate lines is to be expected.This work was carried out while B. H. was on leave from the New South Wales Department of Agriculture, and held a Commonwealth Research Studentship. Our thanks to Robin Hall and Tricia Brown for technical assistance, to Dr. B. L. Sheldon, C.S.I.R.O. Division of Animal Genetics, for providing the inbred line, and to the Radio-Therapy Unit, Royal Prince Alfred Hospital, for the use of their equipment.     

‘Milk’ secretion for embryogenesis in a viviparous cockroach

The brood sac of viviparous Diploptera punctata is a typical insect integumentary gland which secretes a ‘milk’ containing protein and carbohydrate to nourish the developing embryos. During gestation the secretory cells proliferate organelles of protein synthesis and secretion and brood sac wet weight, protein content, synthetic activity and secretory output increase five- to six-fold ; a maximum of 0.4 mg protein was collected in 24 hr from one brood sac in a later stage of gestation. Following parturition, when secretory activity ceases, these parameters fall markedly, and the secretory cells decrease their mass by autophagic regression. Acid phosphatase has been located histochemically in autolysomes and assayed in brood sac homogenates; activity reaches a maximum five days after parturition.     

Substituted Sialic Acid Prosthetic Groups as Determinants of Viral Hemagglutination

Inhibitors of hemagglutination by type A2 influenza virus and a recently isolated strain of type B influenza virus were separated by sucrose density gradient centrifugation and agarose gel filtration from horse serum. Using selected reagents, it was demonstrated that the active substituent on the horse serum inhibitor of A2 influenza virus was 4-O-acetyl-N-acetylneuraminic acid; however, the active substituent on the inhibitor of the influenza B virus was shown to be N-acetylneuraminic acid (NANA). Sodium metaperiodate treatment of a component of horse serum resulted in a 10 to 15-fold enhancement of inhibitory activity against the type B virus, whereas the A2 inhibitor was completely destroyed. Since this enhancement did not occur with influenza B viruses isolated prior to 1965, it was considered that this sensitivity to an oxidized NANA glycoside may have been a reflection of an antigenic change which occurred at that time. The use of different virus strains and selected chemical reagents to define the important sialic acid prosthetic groups active in inhibition was described.     

Secondary and Tertiary Responses of the Induced Bactericidin from the West Indian Spiny Lobster, Panulirus argus

West Indian spiny lobsters, Panulirus argus, synthesized a hemolymph bactericidin after being injected with killed suspensions of gram-negative bacillus EMB-1 isolated from the normal gut of this lobster. To study differences between the primary response and secondary response, animals were given a primary antigen injection of EMB-1 followed by a second injection of the same antigen 22 to 51 days later. As a rule, secondary bactericidal responses were enhanced over the primary in a manner reminiscent of specific anamnesis in mammalian immunoglobulin synthesis. Immunological memory was also suggested when tertiary responses were compared to secondary and by the persistence of residual titers for many days or weeks without additional antigenic stimulation.     

Alternate Requirement for Vitamin B12 or Methionine in Mutants of Pseudomonas denitrificans, a Vitamin B12-producing Bacterium

Experiments are described which indicate that Pseudomonas denitrificans, an organism that overproduces vitamin B(12), uses the B(12) pathway exclusively for methionine synthesis.     

Histochemische Untersuchungen über den Involutionsmechanismus der Milchdrüse

Zusammenfassung Es wurden mit histochemischen Methoden Milchdrüsen von Meerschweinchen in physiologischer und durch Kastration während der Laktationsperiode hervorgerufener Involution untersucht. Nachgewiesen wurden alkalische und saure Phosphatase (APh und SPh), unspezifische Esterase, Sukzinodehydrase, SH-Gruppen, Nukleinsäuren und Lipide. — Die Aktivität der SPh nimmt während der Involution zu. Die Bedeutung dieses Enzyms für den Rückbildungsprozeß des Drüsengewebes wird diskutiert. — Das Vorkommen der APh-Aktivität in Drüsenendstücken im Involutionsstadium kann auf die Beteiligung dieses Enzyms an der Rückresorption der mit der Milch ausgeschiedenen Substanzen hinweisen. — Zwischen der Aktivität der unspezifischen Esterase und dem Gehalt sowie der Lokalisation von Lipiden besteht eine Abhängigkeit. — Es konnten keine Unterschiede in der Sukzinodehydraseaktivität und dem DNS-Gehalt aufgezeigt werden. — Die Verminderung von SH-Gruppen und RNS hängt mit dem Aufhören der Sekretionsproduktion durch die Zellen der Drüsenendstücke der Milchdrüsen zusammen.

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Histochemical studies on the involution mechanism of the mammary gland

Summary Histochemical methods were used to study mammary glands of guinea pigs in the course of physiologic involution and that induced by castration during lactation. Alkaline and acid phosphatases, unspecific esterase, succinic dehydrogenase, SH groups, nucleic acids and lipids were determined. Acid phosphatase activity was found to be increased in mammary glands, subject to involution. The participation of the enzyme in the involutionary process of the gland tissue is discussed. The distribution of alkaline phosphatase in the secretory sections of the gland during involution would suggest the participation of the enzyme in the reabsorption of the substance secreted with milk. A correlation existing between the activity of unspecific esterase, the level and distribution of lipids in the mammary gland could be established. No differences were detected in the activity of succinic dehydrogenase and DNA level. A decrease in SH groups and RNA content is related to cessation of milk secretion.

     

Shear and the Melting of DNA: An Especially Sensitive Portion of the Escherichia coli Genome

The melting point of DNA is shown to be a function of shear stress. The higher the molecular weight of the DNA, the further its melting point is lowered by a given shear rate. During lysis of E. coli, a part of the DNA is especially shear sensitive, so that its melting curve in the presence of shear shows a low-melting region prior to the main transition. Lysis and dilution of the cell contents destroys the extra shear sensitivity, perhaps because the DNA dissociates from the cell membrane or from some other large subcellular structure. Such a structure would impart increased shear sensitivity to the associated region of the genome.