Isolation of two endo-beta-N-acetylglucosaminidases from fig latex

Two endo-beta-N-acetylglucosaminidases (mannosyl-glycoprotein 1,4-N-acetamidodeoxy-beta-D-glycohydrolase, EC 3.2.1.96) (type F-I and type F-II) have been isolated from fig latex. At pH 7.0, type F-1 was retained by the DEAE-Sephadex A-50 column, whereas type F-II was not adsorbed by the column. The optimum pH of type F-I was found to be pH 5.9 and type F-II, pH 5.4. Type F-I enzyme hydrolyzes the tri-mannosyl derivatives di-N-acetylglucosaminylasparagine faster than the penta- or hexa-mannosyl compounds. Type F-II hydrolyzes the penta- and hexa-mannosyl derivatives, but not the tri-mannosyl compound.     

A new fiber-forming protein from Tetrahymena pyriformis

A new fiber-forming protein was isolated from the acetone powder of Tetrahymena pyriformis by co-precipitating with skeletal muscle myosin while trials were made to find actin or actin-like protein in Tetrahymena. It has a molecular weight of 38000 D and forms a tetramer (140000 D, 9 S) in physiological conditions. Its isoelectric point (pH 6.7), amino acid composition and antigenic determinant(s) differ significantly from those of non-muscle actin and skeletal muscle actin. It does not undergo G-F conversion while actin does, and does not activate Mg²⁺-ATPase of skeletal muscle myosin. The protein localizes in the oral apparatus and division furrow as revealed by fluorescent antibody method. The protein can be assembled into 14-nm filaments in a reassembly buffer. The in vitro filaments appear to correspond to some filaments included in the oral apparatus and the contractile ring. The fiber-forming protein from Tetrahymena may play important roles in cell motility including cell division.     

Protein synthesis induced by infection with packaged λdv plasmid

Plasmid λdv or imm²¹dv DNA was joined to a λ arm having a cos site. This recombinant plasmid can be packaged in a λ head, and used to infect Escherichia coli K12 cells. The injected DNA molecules become plasmids in cells. By adding these particles to uv-irradiated uvrA cells, the packageable λdv or imm²¹dv plasmids can be induced to synthesize proteins coded by genes on the plasmid genome. The packageable plasmid system is thus suitable for studying on synthesis and regulation of plasmid-coded biopolymers. Analyses of the dv-coded proteins in gel electrophoreses revealed that among several genes carried on the dv plasmid genome, only those genes that are members of the pRoR-tof-cII-O-P operon can be expressed. Evidence has been presented to show that expression of this operon, which is directly correlated with replication of the genome, is only partially allowed in cells perpetuating the dv plasmid. These observations are discussed in connection with the autorepressor model (D. E. Berg, 1974, Virology 62, 224–233; K. Matsubara, 1976, J. Mol. Biol. 102, 427–439) that genetically accounts for the control mechanism of plasmid replication.     

Light-microscopic detection of acidic glycoconjugates with sensitized diamine procedures

Summary To enhance the efficiency and specificity of diamine methods in light microscopy, these methods were sensitized by sliver enhancement in combination with trichloro(ethylene) platinate (KTP). The sensitized diamine methods consisted of a diamine (high or low iron diamine: HID or LID), KTP, borohydride reduction (BH) and a physical development (PD) sequence. The new methods have been successfully applied to routinely prepared tissue sections obtained from rat organs, such as salivary glands, stomach, colon, kidney, lung and trachea. In the tissues subjected to the sensitized diamine methods, weakly diamine-stained histological structures exhibited vivid positive reactions. The combined sensitized diamine methods and selective procedures, such as enzyme digestion and chemical modification, have substantiated that these methods were of sufficient efficiency and specificity.     

Energy dispersive X-ray microanalysis of neurosecretory granules of mouse pituitary on fresh air-dried tissue spreads

Electron microscopy of fresh air dried spreads of unstained posterior lobe tissue from mouse pituitary disclosed neurosecretory granules. Each granule showed a seemingly homogeneous dense core surrounded by a halo and a bounding membrane. The area between granules in the cytoplasm was relatively well preserved. The energy dispersive X-ray microanalysis revealed peaks for sulfur, chlorine and potassium in two granules. The third granule displayed peaks for phosphorus and chlorine. These elements probably contribute to the high electron density of the granules. There was no peak for calcium, in contrast to the dense bodies of human blood platelets.     

ADP binding to (Na + K)-activated ATPase

[¹⁴C]ADP binding to EDTA-washed ox brain cell membranes was increased by Na⁺, but decreased by K⁺, Mg²⁺ and Ca²⁺. Na⁺ abolished the effect of K⁺ on ADP binding by a competitive mechanism, but could not reverse the inhibitory action of Mg²⁺ and Ca²⁺. It is concluded that the cation-induced changes in ADP binding reflect properties of (Na⁺ + K⁺)-activated ATPase.     

A blood-borne factor stimulating hepatic biosynthetic process in hepatectomized rabbits

By measurement of energy charge (ATP+1/2ADP / ATP+ADP+AMP) and mitochondrial phosphorylative activity in liver tissue, it was possible to estimate the states of ATP generation and ATP utilization of normal and regenerative liver. In the liver of partially hepatectomized rabbits, a characteristic decrease in energy charge and increase in mitochondrial phosphorylative activity as compared with those of normal liver were noted, consistent with the notion that ATP-consuming reactions such as protein and nucleic acid syntheses are enhanced in the regenerative process. When normal or hepatectomized rabbits were singly circulated extracorporeally, the mitochondrial phosphorylative activity increased with a concomitant rise in energy charge. In cross-circulation between normal and hepatectomized rabbits, the energy charge of the normal partner decreased markedly with a concomitant elevation in mitochondrial phosphorylative activity, while those in the remnant liver of the hepatectomized partners were not significantly different from those of singly circulated hepatectomized rabbits. The decreased energy charge levels of the normal partner were prevented by the premedication of insulin without a significant effect to the hepatectomized partner. It is suggested that there is a humoral factor stimulating the hepatic energy-requiring biosynthetic process in the blood of hepatectomized rabbits.     

Gene-culture waves of advance

Major genetic and cultural changes may have been coupled during hominid evolution. Since hominids have had a wide geographical distribution for about one million years, any mutant gene or cultural innovation that became established had to spread from its origin. A pair of nonlinear diffusion equations is derived which models the propagation of a mutant gene and a cultural innovation. Both are assumed to originate in the same locality along a linear habitat. The mutant gene and its allele are semidominant, and the two cultural choices are transmitted according to what I call the logistic attraction-repulsion model. The genes influence cultural choice, and the two interact to determine fitness. Of particular interest is the case in which mutant gene and cultural innovation are mutually dependent, neither being able to spread without the other. Each equation of the pair is similar in form to Fisher's equation, with a linear function of the other dependent variable replacing the constant coefficient in the reaction term. The partial differential equations are solved numerically to obtain the asymptotic speeds. Their form also suggests an heuristic argument which has proved useful, but I have been unable to obtain any analytic results. The waves of the system are shown to be of two types, synchronous and asynchronous. When genes and culture are mutually dependent, synchronous travelling waves can exist. However, their existence is dependent on initial conditions, and the speed of propagation is slow.     

Light- and electron-microscopic study of the endolymphatic sac of the tree frog,Hyla arborea japonica

Summary The endolymphatic sac of the tree frog and its crystals were observed by light- and electron microscopy. Scanning electron microscopy revealed that the crystals have a faceted body and two pointed ends. Light- and transmission electron microscopy revealed that the endolymphatic sac is composed of many small chambers. In their lumina, numerous ghosts of crystals that resulted from decalcification were observed. The ghosts were demarcated by a linear dense material or embedded in a flocculent substance. The epithelium of the endolymphatic sac is simple squamous or cuboidal and peculiar cytoplasmic granules are found in most cells. The granules are surrounded by a limiting membrane and have varying electron density. Some granules contain a core and/or tubular structures. Vacuoles containing large ghosts are also found in the epithelial cells. These ghosts were quite similar to those in the lumen and sometimes coexist with cell debris. The fine structure of the endolymphatic sac and its crystals is discussed.