Nature of Escherichia coli B(P1) Yielder Cells at the Time of Infection with Restricted T1

In the infection of Escherichia coli B(P1) with restricted T1, it was shown that yielder cells consist of both special and nonspecial cells. Special or predetermined yielders occurred only among the earliest yielders. In most instances, yielder-cell formation was most easily explained by assuming that the first step was a chance escape of the restricted phage DNA from the degrading enzyme of the restricting cell.     

The inhibition of pepsin-catalysed reactions by products and product analogues. Kinetic evidence for ordered release of products

1. The inhibition of pepsin-catalysed hydrolysis of N-acetyl-l-phenylalanyl-l-phenylalanylglycine by products and product analogues was studied. 2. The non-competitive nature of the inhibition by the product N-acetyl-l-phenylalanine confirms an ordered release of products, and points to a common mechanism (involving an amino-enzyme) for pepsin-catalysed transpeptidation and hydrolysis reactions. 3. N-Acetyl-l-phenylalanine ethyl ester is also a non-competitive inhibitor, but here the inhibition is of the ;dead-end' type. No ethanol is detectable in reaction mixtures, indicating that this ester cannot act as an amino group acceptor in a transpeptidation process. 4. The same is true for N-methanesulphonyl-l-phenylalanine methyl and methyl thiol esters. No methanethiol is liberated when the methyl thiol ester is present as an inhibitor of the hydrolytic reaction, and the hope that such a thiol ester would effectively trap the amino-enzyme was not fulfilled.     

Locus-specific Changes in Cell Wall Composition Characteristic of Osmotic Mutants of Neurospora crassa

The osmotic phenotype of Neurospora crassa is characterized by inhibition of growth at high osmolalities of growth medium. Mutations at six osmotic loci of linkage group I were examined to assess the biochemical and physiological effects of these mutants. Isolated cell walls from 23 osmotic strains were compared with the wild type with regard to quantitative levels of the following components: percentage of total dry weight, total glucose, alkali-soluble glucose, nonglucose carbohydrates, amino acids, glucosamine, galactosamine, and a compound tentatively identified as quinovosamine. The last component has not previously been observed in N. crassa cell walls. Although the cell wall dry weight content of osmotic mutants was not altered, walls isolated from all of the osmotic strains had less alkali-insoluble glucose than those from the wild type. In addition, all of the loci except cut exhibited other consistent differences from the wild type. The os-1, os-3, and os-5 mutants had low levels of alkali-soluble glucose. The os-3 and os-5 mutants had high levels of nonglucose carbohydrates, and flm-2 had a low level of nonglucose carbohydrates. The os-4 mutants had low levels of galactosamine and amino acids and high levels alkali-soluble glucose. An os-1 mutant, B135, produced less of the whole alkali-soluble fraction of the cell wall.     

DEVELOPMENTAL AND ANATOMICAL CHARACTERISTICS OF THIN-HULL MUTANTS OF CARTHAMUS TINCTORIUS [COMPOSITAE]

Histological observations were made on normal and mutant strains of safflower in order to compare the development of the “fibrous” tissues among the various strains. The fibers of the vascular bundles of normal and F₁'s from crosses between normal and thin-hull mutant types had well-developed secondary cell walls, but they appeared reduced in mutant types. The anthers of all types showed a similar pattern of differentiation and maturation up to the final stages of tapetal breakdown when secondary walls and rib formations appeared in the connective regions and endothecial cells of the normal, striped and F₁ types. These formations were absent from the thin-hull mutants. In both types dehiscence took place along a longitudinal fissure at the junction of the pollen sacs of one lobe of the anther. The anther flaps of normal types opened abruptly, thus effectively bringing the pollen into contact with the elongating style. Those of the mutants collapsed in place preventing the release of pollen. Hulls of the mutant strains were thin because cells were not sclerified during differentiation of the pericarp. Striped hulls resulted from the additional localization of secretory canals in the pericarp of the striped mutant.     

The binding of inhibitors to α-chymotrypsin

1. The binding of three competitive inhibitors, N-acetyl-d-tryptophan, N-acetyl-l-tryptophan and N-acetyl-d-tryptophan amide, to alpha-chymotrypsin was studied over the pH range 2.20-9.65 by the technique of equilibrium dialysis. 2. Within the limits of the experimental method, the binding of the uncharged amide inhibitor is independent of pH over the range investigated. 3. The binding of each of the enantiomeric acids is dependent on the ionization of a group on the free enzyme, of apparent pK(a)7.3. 4. It is shown that the ionizing group results in the active site of the enzyme developing a net negative charge above pH7.3. 5. The enzyme groups responsible are tentatively identified, and the significance of the binding constants with respect to the enzymic catalysis is discussed.     

Egg Membrane Lytic Activity of Sperm Extract and its Significance in Relation to Sperm Entry in Hydroides hexagonus (Annelida)

Previous electron microscope studies indicated that the individual spermatozoön of Hydroides hexagonus forms a hole in the vitelline membrane by means of lysis. Other observations established that the hole is real, being visible in living material during sperm entry. During the present investigation sea water extracts from frozen-thawed sperm were tested for lytic effect on the membrane. In normal living eggs the membrane appears as a single thick envelope, but in electron micrographs of sections it is seen to consist of a narrow outer border layer, a wide principal or middle layer, and a narrow inner border layer. After immersion in sperm extract the outer border layer elevates but does not dissolve, the middle layer liquefies and disappears, and the inner border layer seems not to change. This is interpreted as lysis of the middle layer. The extract exerted the same effect on fertilized and unfertilized eggs. In electron micrographs the sections treated with extract greatly resemble that part of the membrane which has been penetrated by the individual spermatozoön. It is concluded that the individual spermatozoön, too, exerts a lytic effect. Together, the present and two earlier studies are considered clearly to demonstrate that in Hydroides the individual spermatozoön does indeed make an entry hole in the egg membrane by applying lytic material to that part of the membrane in its own vicinity.     

A FLORA FROM THE CHESTER SERIES (UPPER MISSISSIPPIAN) OF SOUTHERN ILLINOIS

New localities in southern Illinois have yielded a flora consisting of 9 taxa. The plant remains occur in sandstones of the Chester Series and consist of casts, molds, and flattened impressions. The most important elements in the flora are: Archaeocalamites radiatus, Lepidodendron volkmannianum, Stigmaria ficoides, Rhynchogonium fayettevillense, and an unnamed Trigonocarpus. Stems probably referable to Lepidodendron veltheimii and Dictyoxylon also occur, as do numerous casts of the Knorria and Bergeria types. The range of features found in specimens of Lepidodendron volkmannianum allows some revision to be made of several species previously described by White in the Wedington Sandstone flora of northwest Arkansas, with which the present flora is closely comparable. The difficulties in making precise correlations with Upper Mississippian floras in other parts of the world are discussed. The necessity for critical re-examination of existing described collections and of undertaking more extensive field-work in the Mississippian of N. America is emphasized.     

The interaction of α-chymotrypsin with isosteric substrates of different charge type

1. The synthesis of three substrates of alpha-chymotrypsin of closely similar steric requirements but different charge type is reported. 2. The interaction of these compounds [SS-dimethyl-(l-3-carboxymethyl-3-acetamido)propyl sulphonium iodide, l-2-acetamido-5-methylhexanoic acid methyl ester and N-acetyl-l-glutamic acid alpha-methyl ester] with alpha-chymotrypsin has been studied. 3. For the charged substrates, values of k(0) are two orders of magnitude smaller than, and values of K(m) two orders of magnitude larger than, the corresponding values for the uncharged isostere. 4. The results are interpreted in terms of the known specificity of the enzyme, and the relationship between binding and kinetic specificities is discussed.     

Microporosity of the substratum regulates differentiation of MDCK cells in vitro

Summary We have analyzed the ability of the physical substratum to modulate both the ultrastructural and protein synthetic characteristics of the Madin-Darby canine kidney (MDCK) renal cell line. When MDCK cells were seeded on Millipore Millicell CM microporous membrane cell culture inserts they demonstrated a more columnar organization with an increase in cell density sixfold greater than the same cells seeded on conventional plastic substrata. After 1 wk postseeding on the microporous membrane a partial basal lamina was noted, with a contiguous basement membrane being apparent after 2 wk. One-dimensional sodium dodecyl sulfate gel electrophoresis was used to analyze detergent-solubilized proteins from MDCK cells maintained on plastic substrata vs. microporous membranes. When proteins were pulse-labeled with [³⁵S]methionine, a 55 kDa protein was evident in the cytosolic extract of cells grown on collagen, laminin, and nontreated plastic substrata; but this labeled protein was not evident in similar extracts from cells grown on collagen and laminin-coated microporous membranes. To test if the polarized, basement-membrane secreting phenotype of the MDCK cells could be generated on a microporous membrane without pretreatment with any extracellular matrix (ECM) components, cells were seeded on the Millipore Millicell HA (cellulosic) microporous membrane. This type of substrata does not need a coating of ECM components for cell attachment. A partial basement membrane was formed below cells where the basal surface of the cell was planar, but not in areas where the cell formed large cytoplasmic extensions into the filter. This led us to the conclusion that the microporous nature of the substrata can dictate both ultrastructural and protein synthetic activities of MDCK cells. Furthermore, we suggest that both the planar nature of the basal surface and the microporosity of the substrate are corequisites for the deposition of the basement membrane.     

Ultrastuctural and cytochemical study of spermatogenesis in Scrobicularia plana (Mollusca,Bivalvia)

The ultrastructure of the mature spermatozoa and spermatogenesis of the bivalve Scrobicularia plana are described. Support cells extend from the basal lamina to the lumen of the testis and are laterally connected to the germinal epithelium. Germ cells present intercellular bridges and flagella since the spermatogonial stage. While spermatogonia and spermatocytes appear connected to support cells by desmosome-like junctions, elongated spermatids are held at the acrosomal region by support cell finger-like processes. During spermiogenesis, the acrosomal vesicle differentiates from a golgian saccule and then migrates to the nuclear apex. A microtubular manchette arising from centrioles surrounds the acrosomal vesicle, the nucleus, and the mitochondria at the time these three organelles start their elongation, disappearing after that. The mature spermatozoon of S. plana lacks a distinct midpiece because the mitochondria extend from the region of the pericentriolar complex along the nucleus anteriorly for approximately 1.4 μm. The features of this bivalve type of modified spermatozoon are compared with those of other animal groups having similar modifications.