Herpes simplex virus type 1 glycoprotein C-negative mutants exhibit multiple phenotypes, including secretion of truncated glycoproteins

A virus-neutralizing monoclonal antibody specific for glycoprotein C (gC) of herpes simplex virus type 1 strain KOS was used to select a number of neutralization-resistant mutants. A total of 103 of these mutants also were resistant to neutralization by a pool of gC-specific antibodies and thus were operationally defined as gC-. Analysis of mutant-infected cell mRNA showed that a 2.7-kilobase mRNA, comparable in size to the wild-type gC mRNA, was produced by nearly all mutants. However, six mutants, gC-5, gC-13, gC-21, gC-39, gC-46, and gC-98, did not produce the normal-size gC mRNA but rather synthesized a novel 1.1-kilobase RNA species. These mutants had deletions of 1.6 kilobases in the coding sequence of the gC structural gene, which explains their gC- phenotype. Despite the production of an apparently normal mRNA by the remaining 97 mutants, only 7 mutants produced a detectable gC polypeptide. In contrast to wild-type gC, which is a membrane-bound glycoprotein with an apparent molecular weight of 130,000 (130K), five of these mutants quantitatively secreted proteins of lower molecular weight into the culture medium. These were synLD70 (101K), gC-8 (109K), gC-49 (112K), gC-53 (108K), and gC-85 (106K). The mutant gC-3 secreted a protein that was indistinguishable in molecular weight from wild-type KOS gC. Another mutant, gC-44, produced a gC protein which also was indistinguishable from wild-type gC by molecular weight and which remained cell associated. Pulse-labeling of infected cells in the presence and absence of the glycosylation inhibitor tunicamycin demonstrated that these proteins were glycosylated and provided estimates of the molecular weights of the nonglycosylated primary translation products. The smallest of these proteins was produced by synLD70 and was 48K, about two-thirds the size of the wild-type polypeptide precursor (73K). Physical mapping of the mutations in synLD70 and gC-8 by marker rescue placed these mutations in the middle third of the gC coding sequence. Mapping of the mutations in other gC- mutants, including two in which no protein product was detected, also placed these mutations within or very close to the gC gene. The biochemical and genetic data available on mutants secreting gC gene products suggest that secretion is due to the lack of a functional transmembrane anchor sequence on these mutant glycoproteins.     

Scanning electron microscope studies of the mucosa of rats infected with Hymenolepis diminuta (Cestoda)

The intestinal morphology of rats given one, 10 or 100 cysticercoids of hymenolepis diminuta was examined by scanning electron microscopy. The presence of this tapeworm causes extensive villous atrophy and fusion. The most extreme changes in mucosal architecture were observed adjacent to the mature proglottides of the worm and in these areas the villi were reduced either to flattened plate-like structures or to low irregularly shaped undulations. The presence of one large H. diminuta resulted in more severe pathological damage than caused by several smaller worms. Colonization of the upper region of the ileum by long filamentous bacteria was also observed in rats infected with H. diminuta.     

Cholinesterase in larvae of the ascidian,Ciona intestinalis,developing from fragments cut from centrifuged eggs

Development Genes and Evolution - UnfertilizedCiona eggs were centrifuged, stratifying their mitochondria and some other cytoplasmic components. Each centrifuged egg had a mitochondria-free,...     

Factors Influencing the Onset of Chronic Respiratory Disease

To investigate the effect of different environmental and personal factors on ventilatory function 10,971 children resident and going to school in four areas of Kent were examined. Details of past respiratory illnesses were obtained by a questionary completed by the parents; the examination included measurement of height, weight, and peak expiratory flow.Area of residence, social class, family size, and a past history of pneumonia, bronchitis, or asthma were found to be associated with differing levels of peak expiratory flow. These four factors acted independently, and the effects were additive. It is suggested that environment in the early years of life can produce adverse changes which may exist throughout life and contribute to the development of chronic respiratory disease.     

Genetic Basis of Colicin E Susceptibility in Escherichia coli I. Isolation and Properties of Refractory Mutants and the Preliminary Mapping of Their Mutations

Colicin E-resistant mutants were isolated in Escherichia coli K-12 which, although still apparently possessing the E receptor and adsorbing colicin, were nevertheless insensitive (refractory) to its effect. Eight phenotypic groups were obtained, but some mutants from three of these groups were all shown to map at gal, whereas a second refractory locus, giving resistance to E1 alone, mapped close to thy. It is suggested that the successful fixation of any of the three distinct colicins of group E may involve a dual role for the cell surface "receptor," the first for the binding of the protein and the second for the correct orientation of the bound molecule relative to the cytoplasmic membrane. The majority of the refractory mutants isolated may derive from changes in components concerned with the second of these receptor functions. Two groups of mutants, however, refractory to only E1 or E2, probably reflect changes in the intracellular transmission systems which specifically mediate the effects of these two colicins, the changes not allowing transmission through the cytoplasmic membrane to the respective targets of the colicins. The E1 adsorption site was shown to be distinct from that for E2 and E3, indicating an early separation of the colicin E transmission systems.