Ultrastructural localization of cell wall teichoic acids in Streptococcus faecalis by means of concanavalin A

Some teichoic acids are known to be partially substituted by α-D-glucopyranosyl residues such as the teichoic acids of Streptococcus faecalis NCIB 8191. They will, therefore, bind specifically the phytohemagglutinin concanavalin A. Concanavalin A labelled with mercury or colloidal gold coated with concanavalin A has been used to mark isolated cell walls in order to localize the teichoic acids at the ultrastructural level. Besides these two direct marking techniques, the indirect concanavalin A-peroxidase technique (localization of peroxidase by the diaminobenzidine method followed by postosmication) has been applied to thin sections of premarked cells. All three methods gave almost identical results, namely, a dense and homogeneous distribution of the cell wall teichoic acids. In control experiments total inhibition was achieved in the presence of methyl-α-D-mannopyranoside. After trichloroacetic acid or alkali extraction of the teichoic acids from isolated walls no marking could be detected.     

Translation of pure feather keratin mRNA in a wheat embryo cell-free system

Highly purified feather keratin mRNA, prepared by dissociation of mRNP particles in Na dodecyl sulphate, was translated in a wheat embryo cell-free system with similar efficiency to rabbit globin mRNA and RNA purified from cucumber mosaic virus. The only detectable products of translation of the keratin mRNA were keratin chains, which were identical to native keratin chains as judged by several different criteria. These results support previous conclusions that the keratin mRNA can be obtained in a pure state.     

Ultrastructural analysis of the human cerebral ventricular system

Summary The ultrastructural organization of the human fetal choroid plexus was assessed with scanning electron microscopy. The membranous modifications of choroidal ependymal cells differ remarkably between 11 and 20 weeks of intrauterine development and suggest a variable functional capacity at different times of ontogenesis. Based upon existing data coupled with the ultra-architectural organization of cilia, clavate and linear microvilli are seen with scanning electron microscopy, a multiple functional role is hypothesized for choroidal ependymal cells.supported by USPH grant NS 08171.career development awardee K04 GM 70001     

Observations on freeze-fractured membranes of a Trypanosome

Pure preparations of Trypanosoma brucei, free from plasma and cellular components were isolated from rat blood, and concentrated into loose pellets by low-speed centrifugation. Pellets were either processed for thin sectioning as a control for general morphology, or glycerol-treated after glutaraldehyde fixation for preparation of freeze-fracture replicas. Concentration of cells of 50,000–100,000/mm² of sectioned or fractured surface facilitated identification of fracture faces of the cell body, invaginated flagellar pocket and flagellum. Particle distribution and A and B faces of these regions of the cell are described. A collar of B face particles occurs around the neck of the flagellar pocket, possibly associated with a junction controlling ingress of ingested materials to coated vesicles formed along the membrane defining the pocket. A and B faces of the flagellum and adjoining surface of the cell body have shown that the only intra-membrane specialization corresponding to the miniature ‘maculae adherentes’ described previously in thin sections is probably an uninterrupted series of small clusters (3–6) of 80 Å particles on the A face of the flagellar membrane. It is proposed that these arrays represent attachment points for strands linking the axoneme and paraxial rod to the flagellar surface, and are not directly concerned with the physical adhesion of the flagellum to the cell body surface—a linkage that appears to be established within the extracellular gap between these apposed surfaces of the cell. The potential use of freeze-etching in further study of the external antigens of the infective cell is discussed.     

A mathematical model of the nutrient dynamics of phytoplankton in a nitrate-limited environment

Insofar as saturation kinetics are applicable to the growth of phytoplankton in laboratory experiments and to growth in nature, the computer modeling of intracellular nutrient partitioning in populations of cells can lead to better understanding of the dynamics of natural populations. A three-compartment mathematical model was developed to represent a phytoplankton population having the capability to store nitrogen in a nitrate-limited environment. Parameters were estimated by fitting the model to data from two chemostat experiments reported by Caperon (1968). The model was used to simulate growth dynamics observed in chemostat and batch experiments. The model demonstrated the changes which may occur in the nitrogenous constituents of a phytoplankton population with time and environmental conditions. The model also demonstrates three phenomena which have been observed in field and laboratory experiments but which are not represented by the customary Monod model: (1) uptake rates may significantly exceed not growth rates, (2) high growth rates may be encountered at very low environmental nitrate concentrations, and (3) the ratio of internal nitrogen to population size may change significantly during a study period. It is suggested that the amount of nitorgen in storage may be used as an indicator of the physiological state of a monospecific population. Parameters for the one-compartment Monod model were estimated by customary methods form data generated by the three-compartment model. It was shown that difficulties encountered in estimating the yield coefficient and the decay coefficient may be attributed to the intracellular storage phenomenon. It was also demonstrated that the one-compartment Monod model was inadequate to accurately represent population growth in chemostat experiments when intracellular storage is a significant factor.     

Deductive analysis of a protein-synthesis mutant of Escherichia coli

A mutant of E. coli, T ^s68b, selected as unable to grow at 43 C, is unable to synthesize proteins at 43 C, though it can carry out this function at 30 C. This defect is shown to be in the protein-synthetic rather than the RNA-synthetic machinery by an analysis of the response of the strain to infection with the RNA bacteriophage f2. An analysis of the capacity for RNA synthesis and the polyribosome content of these cells at 44 C indicates that the defect resides in the elongation step of protein synthesis. No defects could be detected in vitro. The results are discussed in light of similar data on other mutants and in relation to the general approach of analyzing complex mutants selected with ill-defined phenotypes.This work was supported by Grant GM14368 from the National Institute of Health.     

Aldehyde dehydrogenase isozymes in rat hepatoma-mouse fibroblast hybrids

A study of aldehyde dehydrogenase in rat hepatoma cells and rat hepatoma-mouse fibroblast hybrids revealed that the hepatoma cells had activity comparable to that found in whole rat liver and that the enzyme activity was suppressed in early hybrids and reappeared following chromosome loss. Starch gel electrophoresis and heat inactivation studies showed that a new form of enzyme was produced in the hybrids, possibly a heteropolymorphic combination between the HTC enzyme and a previously repressed mouse form. Staining methods for starch gel electrophoresis and histochemical detection of aldehyde dehydrogenase are described.This work was supported by grants from the Damon Runyon Foundation (DRG 1088s) and the Public Health Service (1-R01-Ca 12310-02).