Identification of residues in the cysteine-rich domain of Raf-1 that control Ras binding and Raf-1 activity

We have identified mutations in Raf-1 that increase binding to Ras. The mutations were identified making use of three mutant forms of Ras that have reduced Raf-1 binding (Winkler, D. G., Johnson, J. C., Cooper, J. A., and Vojtek, A. B. (1997) J. Biol. Chem. 272, 24402-24409). One mutation in Raf-1, N64L, suppresses the Ras mutant R41Q but not other Ras mutants, suggesting that this mutation structurally complements the Ras R41Q mutation. Missense substitutions of residues 143 and 144 in the Raf-1 cysteine-rich domain were isolated multiple times. These Raf-1 mutants, R143Q, R143W, and K144E, were general suppressors of three different Ras mutants and had increased interaction with non-mutant Ras. Each was slightly activated relative to wild-type Raf-1 in a transformation assay. In addition, two mutants, R143W and K144E, were active when tested for induction of germinal vesicle breakdown in Xenopus oocytes. Interestingly, all three cysteine-rich domain mutations reduced the ability of the Raf-1 N-terminal regulatory region to inhibit Xenopus oocyte germinal vesicle breakdown induced by the C-terminal catalytic region of Raf-1. We propose that a direct or indirect regulatory interaction between the N- and C-terminal regions of Raf-1 is reduced by the R143W, R143Q, and K144E mutations, thereby increasing access to the Ras-binding regions of Raf-1 and increasing Raf-1 activity.     

Aedes (Ochlerotatus) surcoufi (Theobald, 1912). Restoration of the binomen; morphological analysis; position in the holarctic "excrucians" complex (author's transl)

The study of the morphology of eggs, larvae, pupae and adults of Aedes excrucians from Germany, Italy and France has led the authors to rise again the binomen Aedes surcoufi (Theobald, 1912) to particularize the european populations. At the same time, the characteristics of the different species of the Aedes excrucians complex are discussed.     

Relationship between internal calcium and outward current in mammalian ventricular muscle; a mechanism for the control of the action potential duration?

In sheep and calf ventricular bundles, increasing the internal calcium by increasing the frequency of voltage-clamping to plateau range potentials increased the time-independent outward current. This effect was more marked with higher [Ca]o, and was reduced if the Ca current blockers Verapamil or D 600 were used. 2. If the internal Ca was increased by the addition of cyanide and reduction of external sodium the outward current was also increased. The frequency-dependent increase in outward current also occurred in this Na-poor (12 mM) solution. 3. Tension measurement on the ventricular bundles showed that a Na-free solution with cyanide did not cause a contracture. On changing from Tyrode to a Na-free solution containing cyanide, and on changing back to Tyrode there was a potentiation of the twitch. 4. In Na-poor solution with cyanide, although no contracture was found, ECa was less positive, suggesting that under these circumstances Ca accumulates at the inner side of the membrane, but not around the myofibrils. 5. The prolongation of the action potential in Cl-free solution is frequency-dependent. A greater prolongation is seen at lower frequencies suggesting that Cl current is relatively more important for repolarization at lower frequencies of stimulation. 6. It is suggested that calcium at the inner side of the membrane sets the level of the background outward current. A feed-back mechanism on this basis is proposed for the control of the action potential duration. Various factors that could influence this basic mechanism are discussed.     

The role of cholecystokinin in radiographic opacification of the gallbladder

Studies of the effect of cholecystokinin (CCK) on hepatic elimination of intravenously administered iopanoate and on gallbladder opacification were performed using nonoperated dogs with intact enterohepatic circulation and normal endogenous bile salt pool. Intravenous administration of CCK (3 units/kg) resulted in a 32% increase in apparent transport maximum (maximum rate of elimination) of iopanoate and earlier and enhanced gallbladder opacification. This increase in apparent transport maximum was abolished by cholecystectomy, indicating that the increase was a result of the release of bile salts from the gallbladder rather than a direct effect of the hormone on the hepatic elimination of iopanoate. The early gallbladder opacification and increased density of contrast material in the gallbladder were related to CCK-induced emptying of the gallbladder and subsequent filling with opacified bile.     

Interfacing digitiezed Disa anemometer output to an Apple II computer

For measuring the velocities in a fluctuating fluid flow, the new Disa 56C00 multichannel constant-temperature anemometer system can be used with the 56N00 signal analysis equipment to provide a digital output of mean, root mean square or correlation values. A simple interface arrangement and appropriate software control for reading data directly into an Apple II computer are described. With these the subsequent calculation of derived parameters, such as displacement and momentum thickness of boundary layers and turbulence intensity, can be quickly and easily completed.