Effect of Lytic Enzymes of Acanthamoeba castellanii on Bacterial Cell Walls

Extracts of Acanthamoeba castellanii (Neff) contain alpha- and beta-glucosidase, beta-galactosidase, beta-N-acetylglucosaminidase, amylase, and peptidase. All of these activities are optimal between pH 3 and 4. These extracts also were found to clarify suspensions of cell walls from nine different gram-positive bacteria, including Micrococcus lysodeikticus. The pH optimum for the lytic activity was between 3 and 4. The extent of lysis of the various cell walls did not correlate with the release of free amino groups and of free N-acetylated sugars from the walls during digestion with these extracts. Suspensions of cell walls of Escherichia coli (a gram-negative bacterium), Cordiceps militaris (a fungus), and Acanthamoeba cysts, as well as of colloidal chitin, were not clarified by incubation with these extracts, although reducing sugars were released from each of these materials. Exhaustive digestion of M. lysodeikticus walls by lysozyme released no free N-acetylglucosamine. The products of exhaustive digestion of this cell wall with Acanthamoeba extracts were free N-acetylglucosamine, free N-acetylmuramic acid, glycine, alanine, glutamic acid, lysine, and N-acetylmuramic acid peptide fragments. These results suggest that the amoeba extracts contain endo- and exo-hexosaminidases, in addition to beta-hexosaminidase and peptide hydrolases.     

Selective purification of the thiol peptides of myosin

1. A method for selective purification of thiol peptides is described. Thiol groups in a protein are treated with radioactive cystine by disulphide-thiol interchange. The labelled cystine peptides in a digest can then be fractionated for peptide ;maps'. Performic acid oxidation of paper strips containing the radioactive peptides followed by further ionophoresis yields the purified cysteic acid peptides. 2. The thiol peptides in a peptic digest of cystine-exchanged myosin were purified in this way, and their amino acid sequences were determined. 3. The conclusion that myosin contains at least 16, and probably between 20 and 22, unique thiol sequences indicates that the molecule consists of two chemically equivalent components.     

The Contractile and Control Sites of Natural Actomyosin

The various contractile and control sites of natural actomyosin gel were studied by comparing the kinetics of ATP hydrolysis with those of gel contraction, measured as an increase in turbidity. Contraction of actomyosin gel seems to require the cooperative reaction of ATP (with Mg) at two different sites. One of these sites catalyzes the hydrolysis of ATP and most probably contributes the driving force for contraction; the binding of ATP to the other site appears to break certain links that retard movement of the gel components. At limiting concentrations of ATP, the rate of contraction seems to depend on the rate of breaking these links as well as on the rate of ATP hydrolysis. But when both sites are saturated, the rate of contraction appears to be limited only by the rate of ATP hydrolysis. In addition to these two contractile sites, there are also two different control sites. At one, the relaxing site, the binding of ATP with Mg inhibits ATP hydrolysis and gel contraction. At the other, the binding of calcium activates contraction by overcoming the inhibitory action of Mg and ATP at the relaxing site. This control system—inhibition by substrate and disinhibition by calcium—can be selectively inactivated by heat and reactivated by dithiothreitol, a disulfide-reducing agent. These observations on the isolated contractile system are discussed in relation to the contraction and relaxation of muscle.     

Protein synthesis by microsomal particles from regenerating rat liver

1. Washed microsome particles from regenerating liver were shown to incorporate [(14)C]leucine into protein more actively than similar preparations from normal liver. 2. The total incorporation in the preparations from regenerating liver increased linearly with the amount of protein incubated, whereas this was not so with preparations from normal liver. 3. The greater activity of regenerating-liver microsomes appeared to be associated with the bound polysomes. 4. The size distribution of polysomes obtained after removal of membrane with deoxycholate was the same in normal and regenerating liver. 5. In general the activity of polysome preparations from normal and regenerating liver was similar. 6. It is concluded that the greater activity of the particles in the microsome fraction from regenerating liver is to be attributed to the ribosomes bound to membrane and that their activity is controlled by factors present in the membrane.     

Effect of exogenous serum gonadotrophin (PMS) on aspects of reproductive development in female domesticated canaries

PMS was injected thrice weekly for four weeks into first winter and second winter canaries during the following periods: September-October, November-December, January-February and February-March. Measures were made of ovary and oviduct weight, brood-patch development (defeathering, vascularity and sensitivity changes), nest-building and egg-laying. PMS caused overy growth in all months: this was greatest in the second year birds in February. Both control and treated ovaries increased in size towards the breeding season. Oviduct growth depends mainly on the size of the treated ovary. Defeathering was produced by PMS in all months: its rate and extent increased towards the breeding season. Vascularity was also produced by PMS. It was least in November-December but after that the rate at which it appeared increased towards the breeding season. By contrast, the effect of PMS on brood-patch skin sensitivity was greater in the autumn group than subsequently. Nest-building occurred in September-October, but was then very slight until February-March, which is the beginning of the breeding season. Eggs were laid in most months; fewer injections were needed to produce egg-laying as the breeding season approached.     

OBSERVATIONS ON LEAF AND BUD FORMATION IN HYDROCHARIS MORSUS-RANAE

Regular sequences of leaf and bud formation occur in several members of the Hydrocharitaceae, including Hydrocharis, in which buds are normally formed in the axil of every second leaf of the phyllotactic spiral. Leaf inception begins by periclinal divisions of the inner cells of the 2-layered tunica. Bud formation, which occurs in the apical meristem itself, immediately following the inception of the subtending leaf primordium, begins by divisions in various planes in the corpus, the 2 tunica layers remaining continuous throughout. The young bud meristem soon gives rise to a lateral bud, before leaf formation begins upon it. Because of these and other features, this species is one of considerable morphogenetic interest. Morphogenesis of the whole plant, and in particular the factors controlling the regular sequence of leaf and bud formation, have been and are being investigated experimentally.