Genetic transformation of Streptococcus thermophilus by electroporation

A rapid and convenient electroporation procedure was developed for the genetic transformation of intact cells of Streptococcus thermophilus with various species of plasmid DNA. Transformation frequency was influenced by the capacitance and voltage selected for electric pulsing, the pH and composition of the electroporation medium and the molecular mass of the transforming DNA. Electroporation is a simple and effective technique to introduce plasmid DNA into S. thermophilus and useful in the development of recombinant DNA technology for this important industrial microorganism.     

Eccentric and concentric torque-velocity characteristics of the quadriceps femoris in man

The primary purpose of this investigation was to study the eccentric and concentric torque-velocity characteristics of the quadriceps femoris in man using a recently developed combined isometric, concentric and eccentric controlled velocity dynamometer (the SPARK System). A secondary purpose was to compare the method error associated with maximal voluntary concentric and eccentric torque output over a range of testing velocities. 21 males (21-32 years) performed on two separate days maximal voluntary isometric, concentric and eccentric contractions of the quadriceps femoris at 4 isokinetic lever arm velocities of 0 degree.s-1 (isometric), 30 degrees.s-1, 120 degrees.s-1 and 270 degrees.s-1. Eccentric peak torque and angle-specific torques (measured every 10 degrees from 30 degrees to 70 degrees) did not significantly change from 0 degrees.s-1 to 270 degrees.s-1 (p greater than 0.005) with the exception of angle-specific 40 degrees torque, which significantly increased; p less than 0.05). The mean method error was significantly higher for the eccentric tests (10.6% +/- 1.6%) than for the concentric tests (8.1% +/- 1.7%) (p less than 0.05). The mean method error decreased slightly with increasing concentric velocity (p greater than 0.05), and increased slightly with increasing eccentric velocity (p greater than 0.05). A tension restricting neural mechanism, if active during maximal eccentric contractions, could possibly account for the large difference seen between the present eccentric torque-velocity results and the classic results obtained from isolated animal muscle.     

Glycogen synthesis via the indirect gluconeogenic pathway in the periportal and via the direct glucose utilizing pathway in the perivenous zone of perfused rat liver

The isolated liver from 24 h fasted rats was perfused in a non-recirculating manner in the ortho- and retrograde direction with erythrocyte-containing (20% v/v) media to provide adequate oxygenation of the liver. Glucose and/or gluconeogenic precursors were added as substrates. Glycogen formation was determined biochemically and demonstrated histochemically. With glucose as the sole exogenous substrate glycogen was deposited in the perivenous area, with gluconeogenic precursors it was formed in the periportal zone during ortho- and retrograde flow. When glucose and gluconeogenic compounds were offered together, glycogen was deposited in both zones. The results corroborate the model of metabolic zonation predicting that periportal glycogen is synthesized indirectly from gluconeogenic precursors while perivenous glycogen is formed directly from glucose.     

SCE variability in lymphocytes and fibroblasts

Summary To determine whether the sister chromatid exchange (SCE) distributions obtained in lymphocytes and fibroblasts from different individuals are comparable, a controlled study was set up. Peripheral blood and skin biopsies were taken on the same day from five individuals living for years under the same environmental conditions. All samples were treated in the same fashion, and the SCEs were scored in 50 metaphases of peripheral blood lymphocytes and of skin fibroblasts in an early and in a late passage. A repeat blood sample was taken from the same five indivuduals 1 year later. Based on the results obtained in this first part of the study, five randomly chosen healthy blood donors were sampled at different times and studied in the same fashion. Each chromosome was identified, and the SCE scores were tabulated per chromosome over 50 metaphases. The statistical analysis consisted of fitting log linear models to these scores and examining the best fit by determining the exceedance probabilities (observed significance level). For lymphocytes, the results indicated that the SCE distributions depended only on the chromosome examined, and not on BrdU-exposure time, individuals, or time of sampling. Treatment with ethyl methane sulfonate (EMS) increased the number of SCEs proportionally on all chromosomes. Analysis of the SCE scores on lymphocytes and fibroblasts of the five individuals and on their low and high passage fibroblast cultures revealed the necessity of including higher order interactions in order to fit a suitable model to the data. Therefore comparison of the SCE scores of lymphocytes with those of fibroblasts or comparison of scores on fibroblasts from different individuals could not be done. In practice, to compare samples or individuals, it suffices to score the SCE on a limited number of chromosomes (e.G., the A group) of 50 metaphases.     

Individual differences and variability in the timing of motor activity during walking in insects

The uniformity of the neural physiology of an animal population is a fundamental, rarely tested assumption in most neurophysiological work. In this study, the variability of the timing between the movements of pairs of legs during free walking in cockroaches was assessed. Phases (a measure of timing) of motor bursts in muscles of legs in the American cockroach, Periplaneta americana, were calculated for insects walking straight over a flat, level surface. Student's t, Wallraff, Mann Whitney and Watson U² two-sample tests were used to compare the phases of motor bursts of the same pairs of legs in different insects. The comparisons showed that in spite of the homogeneity both of the animal population and of the conditions under which the insects walked, most of the inter-leg phases of the animals that were compared were significantly different statistically. Further testing of greater numbers of insects using analysis of variance to test for population uniformity confirmed that the insects we tested were not members of a single statistical population with respect to the timing of motor bursts of the legs during walking. We infer that this unexpectedly large variability in a population thought to be relatively homogeneous reflects subtle but biologically significant differences between animals. The possible sources of these differences and their consequences for the study of behavior and its physiological basis are discussed.     

Mechanism of action of a yeast activator: direct effect of GAL4 derivatives on mammalian TFIID-promoter interactions

We have analyzed interactions between the mammalian TATA factor (TFIID) and derivatives of the yeast activator GAL4. The interaction of the TATA factor on the adenovirus E4 promoter with GAL4 binding sites adjacent to the TATA site was qualitatively altered in response to GAL4 binding. Alterations in the TFIID interactions were observed with two GAL4 derivatives that stimulated hybrid E4 promoter activity in vitro but not with a third derivative that bound to DNA but showed no activation. These results indicate that TFIID is a direct target for a GAL4 activation domain and suggest a simple general model for the activation mechanism.