Marker features for malignancy in ectocervical cells

Marker features for malignancy have recently been observed in ectocervical cells, even in cells that are visually normal in appearance. This study assessed the statistical significance of these marker features using a mixed-model nested-design analysis of variance (ANOVA). Features in blue intermediate cells from patients with normal cytology, moderate dysplasia, and severe dysplasia/carcinomain situ, nonkeratinizing cells from patients with moderate dysplasia, severe dysplasia/carcinomain situ, and invasive cancer, and dysplastic cells from areas of metaplasia from patients with moderate dysplasia, severe dysplasia/carcinomain situ, and invasive cancer were tested. ANOVA clearly demonstrated that the marker features differentiate between cells of the same cell type originating from patients in different diagnostic categories. In every instance, the differences owing to the diagnostic category were statistically significantly greater than those caused by patient-to-patient variability. Although the discriminating marker features in the intermediate cells were almost exclusively spectral features reflecting staining differences, morphometric features were also marker features in the dysplastic cells.     

Pleiotropic phenomena in autolytic enzyme(s) content, flagellation, and simultaneous hyperproduction of extracellular alpha-amylase and protease in a Bacillus subtilis mutant.

A mutant of Bacillus subtilis 6160 that had been isolated by its hyperproduction of alpha-amylase and protease lacked flagella and motility, and its content of autolytic enzyme(s) was reduced to one-third to one-fourth that of the parent. These phenotypic differences were completely co-transferred by the deoxyribonucleic acid (DNA) of the mutant when five DNA recipient strains of B. subtilis were transformed. The revertants, isolated by motility with a frequency of approximately 10(-7), recovered a normal level of autolytic activity and showed reduced productivity of alpha-amylase and protease. This point mutation allowed normal flagellin synthesis, spore formation, and rate of growth. The comparison of cell envelope of the mutant with that of the parent indicated that there was no significant difference except loss of flagella. Therefore the association at the cell surface of a group of extracellular proteins consisting of alpha-amylase, proteases, flagellin, and autolytic enzymes(s) seem to be coordinately regulated by the gene or seem to be affected coordinately by certain undetected alterations of the cell envelope.     

Glutamate Synthase: Properties of the Reduced Nicotinamide Adenine Dinucleotide-Dependent Enzyme from Saccharomyces cerevisiae

A reduced nicotinamide adenine dinucleotide (NADH)-dependent glutamate synthase has been detected and partially purified from crude extracts of Saccharomyces cerevisiae. The enzyme is specific for NADH, glutamine, and alpha-ketoglutarate (K(m) values of 2.6 muM, 1.0 mM, and 140 muM, respectively) and has a pH optimum between 7.1 and 7.7. The stoichiometry of the reaction has been determined as 2 mol of glutamate synthesized per mol of glutamine consumed. Glutamate synthase can be distinguished from either of the glutamate dehydrogenases of yeast on the basis of its substrate requirements and behavior during agarose gel and ion exchange chromatography. Variations in the specific activity of glutamate synthase, which occur in response to changes in the growth medium, are similar in character to those observed with the nicotinamide adenine dinucleotide phosphate-dependent (anabolic) glutamate dehydrogenase.     

Transformation of Bacillus subtilis in α-Amylase Productivity by Deoxyribonucleic Acid from B. subtilis var. amylosacchariticus

Deoxyribonucleic acid (DNA) of Bacillus subtilis var. amylosacchariticus showed almost the same ability as B. subtilis Marburg to induce transfer of several genetic markers in DNA-mediated transformation. DNA-DNA hybridization data also showed an intimate relationship between the two strains. Genetic elements involved in the production of extracellular alpha-amylase (EC 3.2.1.1.) in B. subtilis var. amylosacchariticus were studied by using DNA-mediated transformation. Two Marburg derivatives, NA20(amyR2) and NA20-22(amyR1), produced about 50 and 10 U of alpha-amylase per mg of cells, respectively, whereas B. subtilis var. amylosacchariticus produced as much as 150 U of the enzyme per mg of cells. When B. subtilis var. amylosacchariticus was crossed with strain NA20-22 as recipient, transformants that acquired high alpha-amylase productivity (about 50 U/mg of cells) were obtained. Genetic analysis revealed that a regulator gene (amyR) for alpha-amylase synthesis was found in B. subtilis var. amylosacchariticus, as in the case of B. natto 1212 (amyR2) and B. subtilis Marburg (amyR1). The allele was designated amyR3; it is phenotypically indistinguishable from amyR2, but is readily distinguishable from amyR1. The presence of amyR3 was not sufficient for an organism to render production of an exceptional amount of alpha-amylase. Extra-high alpha-amylase producers could be obtained by crossing B. subtilis var. amylosacchariticus as donor with strain NA20 as recipient. The transformants produced the same or even greater amounts of the enzyme than the donor strain. Results suggest the presence of another gene that is involved in the production of the exceptional amount of alpha-amylase.     

Continuous hydrocarbon fermentation with colloidal emulsion feed. A kinetic model for two-liquid phase culture

Candida tropicalis was cultured in a chemostat-type fermentor with n-hexadecane, dispersed in water as submicron droplets, as the only carbon substrate. The emulsion as well as the aqueous medium were fed continuously into the fermentor. A Monod-type equation can correlate the specific group rate in the continuous fermentor with the concentration of submicron droplets. The same equation can also be fitted to the data for the conventional-type batch culture in the same fermentor in which an oil phase as well as an aqueous phase existed, if the hydrocarbon concentration in the aqueous phase excluding oil drops is employed as the substrate concentration. This demonstrates that Candida tropicalis takes up only submicron droplets of n-hexadecane as the carbon substrate.     

Altered Protein Metabolism in Infection by the Late tsB11 Mutant of Simian Virus 40

The DNA of the temperature-sensitive mutant tsB11 is replicated at the same rate as the DNA of wild-type virus in infection at the restrictive temperature. The progeny mutant DNA cannot be distinguished from wild-type DNA by gel electrophoresis and is assembled into a nucleoprotein complex with the same velocity sedimentation characteristics as the wild-type complex. Analysis of in vivo protein synthesis by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoprecipitation techniques demonstrated that the capsid components VP1, VP2, and VP3 of the mutant and wild-type virus are synthesized at a similar rate, but VP1 fails to accumulate within cells infected by tsB11. Furthermore, VP1 is located predominantly in the cytoplasmic rather than in the nuclear fraction of extracts from cells infected by the mutant. Immunofluorescent studies localized virion antigen within the nucleolus as well as the cytoplasm. The altered intracellular distribution and stability of VP1 suggest that it may be the mutant protein of tsB11. The synthesis of a 72,000 dalton protein is consistently induced in significant quantity in cells infected by tsB11 at the restrictive temperature. A protein of the same apparent molecular weight is present in smaller quantities in uninfected cells and is only slightly increased in quantity in cells infected by wild-type virus.