A Drosophila Axin homolog, Daxin, inhibits Wnt signaling.

The vertebrate Axin protein, the product of the mouse fused gene, binds to beta-catenin to inhibit Wnt signaling. We have identified a homolog of Axin in Drosophila, Daxin. Using double-stranded RNA interference, we generated loss-of-function phenotypes that are similar to overexpression of the Drosophila Wnt gene wingless (wg). Overexpression of Daxin produces phenotypes similar to loss of wg. In addition, we show that Daxin overexpression can modify phenotypes elicited by wg and another Drosophila Wnt gene, DWnt-2. Using immunoprecipitation of endogenous Daxin protein from embryos we show that Daxin interacts with Armadillo and Zeste-white 3. The loss-of-function and overexpression phenotypes show that Daxin, like its mammalian counterpart, acts as a negative regulator of wg/Wnt signaling.     

Mandibular movement patterns relative to food types in common tree shrews (Tupaia glis)

Although common tree shrews have long been considered a model system for early eutherian mastication, little information on mandibular movement patterns relative to specific food types has been reported. Detailed analysis of mandibular movement patterns when related to resulting attrition facets may permit more accurate extrapolations regarding the dietary habits of primitive mammals. Marker beads were sewn to chins of five animals that were placed in a restraint system and filmed while they fully masticated mealworm larvae and standardized pieces of banana, almond, and commercial cat chow. These sequences were divided into early, middle, and late thirds of food reduction. Mandibular positions from both frontal and lateral perspectives were digitized frame by frame to yield plots of orbits in three dimensions as well as graphic display of displacements, velocities, and accelerations. Plot coordinates were averaged to generate composite orbital shapes. Significant (p < 0.01) findings included: (1) shortest orbital durations and greatest peak closing velocities and accelerations in early third of reduction; (2) smallest maximum gape, smallest maximum lateral excursion from midline, and longest duration of powerstroke relative to orbital duration in late third of reduction; (3) shortest orbital durations and smallest maximum gape during mastication of chow; (4) greatest maximum lateral excursion during mastication of mealworm larvae; and (5) smallest peak closing accelerations during mastication of banana. Significant differences were also found among subjects for all parameters examined. Capacity for complex jaw movement may have been critical for allowing primitive molars to be used for trituration of a variety of food types, and may have preceded evolution of more specialized molar forms.     

A Comparison of Protein Extraction Methods Suitable for Gel-Based Proteomic Studies of Aphid Proteins

Protein extraction methods can vary widely in reproducibility and in representation of the total proteome, yet there are limited data comparing protein isolation methods. The methodical comparison of protein isolation methods is the first critical step for proteomic studies. To address this, we compared three methods for isolation, purification, and solubilization of insect proteins. The aphid Schizaphis graminum, an agricultural pest, was the source of insect tissue. Proteins were extracted using TCA in acetone (TCA-acetone), phenol, or multi-detergents in a chaotrope solution. Extracted proteins were solubilized in a multiple chaotrope solution and examined using 1-D and 2-D electrophoresis and compared directly using 2-D Difference Gel Electrophoresis (2-D DIGE). Mass spectrometry was used to identify proteins from each extraction type. We were unable to ascribe the differences in the proteins extracted to particular physical characteristics, cell location, or biological function. The TCA-acetone extraction yielded the greatest amount of protein from aphid tissues. Each extraction method isolated a unique subset of the aphid proteome. The TCA-acetone method was explored further for its quantitative reliability using 2-D DIGE. Principal component analysis showed that little of the variation in the data was a result of technical issues, thus demonstrating that the TCA-acetone extraction is a reliable method for preparing aphid proteins for a quantitative proteomics experiment. These data suggest that although the TCA-acetone method is a suitable method for quantitative aphid proteomics, a combination of extraction approaches is recommended for increasing proteome coverage when using gel-based separation techniques.     

EFFECT OF AMPHETAMINES ON SPEECH DEFECTS IN THE MENTALLY RETARDED

In a double-blind study, 106 mentally retarded patients with speech defects were given 15 mg. of d-Amphetamine daily, or a placebo, for a three-month period. Speech defect types were: Stuttering, immature production, oral inaccuracy, lisp, psychotic, mongoloid, aphasia, deafness, and cleft palate. Only the stutterers showed obvious improvement in comparison with the placebo group. Three severe, long-term stutterers showed such dramatic improvement that their whole course in life has been changed.