Regulation of carbon flow in Selenomonas ruminantium grown in glucose-limited continuous culture.

We have applied a model that permits the estimation of the sensitivity of flux through branch point enzymes (D. C. LaPorte, K. Walsh, and D. E. Koshland, J. Biol. Chem. 259:14068-14075, 1984) in order to analyze the control of flux through the lactate-acetate branch point of Selenomonas ruminantium grown in glucose-limited continuous culture. At this branch point, pyruvate is the substrate of both the NAD-dependent L-(+)-lactate dehydrogenase (LDH) and the pyruvate:ferredoxin oxidoreductase (PFOR). The LDH was purified, and it exhibited positive cooperativity for the binding of pyruvate. The LDH had an [S].5 for pyruvate of 0.43 mM, a Hill coefficient of 2.4, and a K' equal to 0.13 mM. The PFOR, assayed in cell extracts, exhibited Michaelis-Menten kinetics for pyruvate, with a Km of 0.49 mM. Carbon flux through the LDH and the PFOR increased 80-fold and 3-fold, respectively, as the dilution rate was increased from 0.07 to 0.52 h-1 in glucose-limited continuous culture. There was nearly a twofold increase, from 6.5 to 11.2 mumol min-1 mg of protein-1 in the specific activity (i.e., maximum velocity) of the LDH at dilution rates of 0.11 and 0.52 h-1, respectively. A flux equation was used to calculate the intracellular concentration of pyruvate; a fourfold increase in pyruvate, from 0.023 to 0.093 mM, was thereby predicted as the dilution rate was increased from 0.07 to 0.52 h-1. When these calculated values of intracellular pyruvate concentration were inserted into the flux equation, the predicted values of flux through the LDH and the PFOR were found to match closely the flux actually measured in the chemostat-grown cells. Thus, the 80-fold increase in flux through the LDH was due to a twofold increase in the maximum velocity of the LDH and a fourfold increase in the intracellular pyruvate concentration. In addition, the flux through the LDH exhibited ultrasensitivity to changes in both the maximum velocity of the LDH and the intracellular concentration of pyruvate. The flux through the PFOR exhibited ultrasensitivity to changes in the maximum velocity of the LDH and hyperbolic sensitivity to changes in the intracellular concentration of pyruvate.     

Biofeedback control of migraine headaches: A comparison of two approaches

In order to assess the relative effectiveness of finger warming and temporal blood volume pulse reduction biofeedback in the treatment of migraine, 22 female migraine patients were assigned to one of three experimental conditions: temporal artery constriction feedback, finger temperature feedback, or waiting list. Biofeedback training consisted of 12 sessions over a 6-week period. All patients completed 5 weeks of daily self-monitoring of headache activity (frequency, duration, and intensity) and medication before and after treatment. Treatment credibility was assessed at the end of Sessions 1, 6, and 12. Results showed that temporal constriction and finger temperature biofeedback were equally effective in controlling migraine headaches and produced greater benefits than the waiting list condition. Power analyses indicated that very large sample sizes would have been required to detect any significant differences between the two treatment groups. No significant relationships were found between levels of therapeutic gains and levels of thermal or blood volume pulse self-regulation skills. Likewise, treatment outcome was not found to be related to treatment credibility. Further analyses revealed that changes in headache activity and medication were associated with changes in vasomotor variability. Because blood volume pulse variability was not significantly affected by biofeedback training, questions about its role in the therapeutic mechanism are raised.This research was supported in part by grants from the Quebec Ministry of Education and the Quebec Ministery of Social Affairs to the first author, and an award from the Medical Research Council of Canada to the second author. The authors are indebted to Drs. Frank Andrasik, Howard Barbaree, Edward Blanchard, Martin Ford, and Patrick McGrath, as well as to two anonymous reviewers, for their helpful comments on an earlier draft of this paper.     

Microlocating lithium in the mouse embryo by use of a (n, α) nuclear reaction

Summary The quantitative imaging of lithium distribution, in histological sections of 15-days old mouse embryos (whose mother had been submitted to Li-treatment), was performed using⁶Li isotope as tracer,⁶Li(n,)³H nuclear reaction for detection, and dielectric track detectors. Despite the particular difficulties of cryosectioning the embryos without disturbing the lithium distribution, the Li regionalization appeared to be very clear-cut. The ectomesodermic tissues were significantly more loaded with lithium than the endodermic ones. This is probably related to the ectomesodermic tissues being also those most sensitive to the teratogenic effect of lithium. The Li-distribution in the embryo brain was almost homogeneous, instead of being heterogeneous as in adult brain. The mean Li-concentration in the embryo brain was not much below the Li concentration in the grey matter of the mother brain, but it was significantly larger than that in the white matter of the mother brain. Our results are discussed in the context of teratogenic effects observed in situ during mammalian development.     

Cellobiose metabolism in Erwinia: genetic study

Summary The study of mutants of Erwinia specifically unable to ferment cellobiose indicates that the mutations are clustered between arg and ile on the chromosome of this organism. In vivo cloning of the genes responsible for cellobiose utilization lead to a plasmid, pBEC2, which complements all Erwinia Clb^- specific mutants. When introduced into wild-type E. coli it allows this organism to use cellobiose, arbutin and salicin; it also complements bglB and bglC mutants of Escherichia coli indicating that arbutin and salicin utilization is due to the products of the pBEC2 cloned genes. From the characterization of mutants pleiotropically affected in the utilization of various carbon sources, including cellobiose, arbutin and salicin, it is proposed that the three--glucosides are substrates of the phosphoenolpyruvate-dependent phosphotransferase system (PTS).     

Characterization of a new class of circular DNA molecules in yeast

This paper describes the isolation of a pure population of covalently closed circular twisted DNA molecules from yeast. These molecules are homogeneous in size, that is consist of monomers of 2.2μ and of multiple length oligomers of n x 2.2μ. While no data rule out the mitochondrial origin of this DNA, its actual intracellular localization remains unknown; it displays the same buoyant density as the main nuclear DNA and therefore is not the heavy nuclear satellite DNA (γ-DNA described by Moustacchi and Williamson (1966)); although circular molecules represent only 1 to 5 % of the total DNA, they can be prepared in sizable and reproducible amounts by a method based on the use of mechanical disruption of yeast cells rather than lysis by snail gut juice.     

Cellular Studies of X-Ray Induced Inhibition of Lens Regeneration

Whole-body X-irradiation of adult newts 0 to 3 days after lentectomy inhibits transformation of the dorsal iris epithelium into a lens in all cases. The first question raised was whether irradiation affects infiltration of the iris area by macrophages, and the phagocytic activities of these cell types in the iris epithelium (prominent phenomena in this system). The number of macrophages infiltrating into the iris epithelium, and their phagocytic activities (indicated by uptake of melanosomes) were not affected by irradiation under those conditions. The second group of experiments concerns the possible effects of irradiation on DNA replication of iris epithelial cells, which become transformed into lens cells in the non-irradiated system. Autoradiographic studies of iris epithelial cells in vivo revealed a significant suppressive effect of irradiation on the frequencies of cells incorporating ³H-thymidine 7 and 14 days after lentectomy. When autoradiography was applied to the primary pure culture of iris epithelial cells at different time intervals after the start of culture and irradiation in vitro , significant and persistent reduction of cell labelling due to irradiation, was demonstrated. Multiplication of spread cells in the iris epithelial culture was strongly and persistently inhibited throughout a period of 2 months. Inhibition of cell labelling and of cell multiplication was always accompanied by reduction in the extent of de-pigmentation of iris epithelial cells. De-pigmentation is one of the requirements for the cells become transformed into lens cells. The possible mechanism of radiation-induced inhibition of lens regeneration is discussed.