Metamorphosis of cranial design in tiger salamanders (Ambystoma tigrinum): A morphometric analysis of ontogenetic change

While ontogenetic analyses of skull development have contributed to our understanding of phylogenetic patterns in vertebrates, there are few studies of taxa that undergo a relatively discrete and rapid change in morphology during development (metamorphosis). Morphological changes occurring in the head at metamorphosis in tiger salamanders (Ambystoma tigrinum) were quantified by a morphometric analysis of cranial osteology and myology to document patterns of change during metamorphosis. We employed a cross-sectional analysis using a sample of larvae just prior to metamorphosis and a sample of transformed individuals just after metamorphosis, as well as larvae undergoing metamorphosis. There were no differences in external size of the head among the larval and transformed samples. The hyobranchial apparatus showed many dramatic changes at metamorphosis, including shortening of ceratobranchial 1 and the basibranchial. The subarcualis rectus muscle increased greatly in length at metamorphosis, as did hypobranchial length and internasal distance. A truss analysis of dorsal skull shape showed that at metamorphosis the snout becomes wider, the maxillary and squamosal triangles rotate posteromedially, and the neurocranium shortens (while maintaining its width), resulting in an overall decrease in skull length at metamorphosis. These morphometric differences are interpreted in light of recent data on the functional morphology of feeding in salamanders. Morphological reorganization of the hyobranchial apparatus and shape changes in the skull are related to the acquisition of a novel terrestrial feeding mode (tongue projection) at metamorphosis. Metamorphic changes (both internal and external) that can be used to judge metamorphic condition are discussed.     

Evidence against the existence of acetylated Sudan Black B

Summary The nature of acetylated Sudan Black B (aSBB) has been investigated, and it has been found, by thin layer chromatography, that each fraction of aSBB has an R _f which is the same as that of a similar fraction of Sudan Black B (SBB). However, aSBB has been found to have fewer fractions, 9–12 than SBB, 14–16. The two major fractions from aSBB and SBB were examined, and a great similarity was found between the absorption spectra of the respective fractions of aSBB and SBB. The major fraction of aSBB was investigated by mass spectroscopy and found to have a similar molecular weight to that expected of SBB. This demonstrates that aSBB is not in fact acetylated, and that the components of aSBB are chemically no different from the corresponding components of SBB.     

PATTERNS OF VARIATION IN AQUATIC AMBYSTOMATID SALAMANDERS: KINEMATICS OF THE FEEDING MECHANISM

Patterns of variation in the feeding mechanism of three species of ambystomatid salamanders (Ambystoma dumerilii, A. mexicanum, and A. ordinarium) were studied to provide insight into the nature of variation in kinematic parameters of the jaw mechanism associated with prey capture. A nested analysis of variance design provided an assessment of the amount of variation in six kinematic variables (measured from 200 frames/sec films of feeding behavior) both among species and among individuals within species. For all six variables, a highly significant proportion of the variance was explained at the intraspecific level. Among species, the most robust discriminators were variables associated with movement of the hyoid. The variables reflecting gape and lifting of the head provided no significant discrimination among species and had large error variances. The hyoid apparatus is the most phylogenetically conservative component of the feeding mechanism in lower vertebrates and was the most stereotyped component of feeding behavior within the salamander species studied here.     

Changes in odor quality discrimination following recovery from olfactory nerve transection

Following recovery from olfactory nerve transection, animals regain their ability to discriminate between odors. Odor discrimination is restored after new neurons establish connections with the olfactory bulb. However, it is not known if the new connections alter odor quality perception. To address this question, 20 adult hamsters were first trained to discriminate between cinnamon and strawberry odors. After reaching criterion (> or = 90% correct response), half of the animals received a bilateral nerve transection (BTX) and half a surgical sham procedure. Animals were not tested again until day 40, a point in recovery when connections are re-established with the bulb. When BTX animals were tested without food reinforcement, they could not perform the odor discrimination task. Sham animals, however, could discriminate, demonstrating that the behavioral response had not been extinguished during the 40 day period. When reinforcement was resumed, BTX animals were able to discriminate between cinnamon and strawberry after four test sessions. In addition, their ability to discriminate between these two familiar odors was no different than that of BTX and sham animals tested with two novel odors, baby powder and coffee. These findings suggest that, after recovery from nerve transection, there are alterations in sensory perception and that restoration of odor quality discrimination requires that the animal must again learn to associate individual odor sensations with a behavioral response.      

How well does Turing's theory of morphogenesis work?

In 1952 Turing published a paper which showed how under restricted conditions a class of chemical reactions could give biological patterns in diffusion-coupled cells. Although this theory has been much discussed, little has been learnt about the range and type of pattern it can generate. In order to do this and to see how stable the patterns are, we have examined the system in detail and written a computer program to simulate Turing's kinetics for two morphogens over various assemblies of cells. We find that on one-dimensional lines of cells, patterns can indeed be produced and that the chemical wavelengths follow all of Turing's predictions. The results show that stable repeating peaks of chemical concentration of periodicity 2–20 cells can be obtained in embryos in periods of time of less than an hour. We do find however that these patterns are not reliable: small variations in initial conditions give small but significant changes in the number and positions of observed peaks. Similar results are observed in two-dimensional assemblies of cells. On rectangles, random blotches are observed whose position cannot be reliably predicted. On cylinders whose circumference is less than the chemical wavelength, annular stripes are produced. For larger cylinders, blotches that lie very approximately on helices are generated; again sharp prediction of the detailed pattern is impossible.The significance of these results for the developing embryo is discussed. We conclude that Turing kinetics, at least in the simple cases that we have studied, are too unreliable to serve as the generating mechanism for features such as digits which are characterized by a consistent number of units. The theory is however more than adequate by these criteria to specify less well-defined developing patterns such as those of hair follicles or leaf organization. It is emphasized however that the Turing theory is quite unable to generate regulative systems, only mosaicpatterns can be produced.     

Biomechanical comparison of unilateral and bilateral power snatch lifts

Biomechanical characteristics of the one-handed dumbbell power snatch (DBPS) were examined to determine whether significant differences existed between unilateral and bilateral weightlifting movements. Kinetic and kinematic movement data were recorded from 10 male weightlifters (mean +/- SD: age: 30.2 +/- 10.2 years; height: 174.2 +/- 4.4 cm; body mass: 81.5 +/- 14.6 kg) during one-handed dumbbell (DB) and traditional barbell (BBPS) power snatch performance with loads of approximately 80% of respective lift one repetition maximums (1RM) with the use of 2 synchronized Kistler force plates and high-speed 3-dimensional video. Results highlighted asymmetry in the ground reaction force and kinematic profile of the DBPS, which deviated from the observed patterns of the bilateral movement. This study found that the nonlifting side (the side corresponding with the hand that did not hold the DB) tended to generate a greater pull phase peak vertical ground reaction forces significantly faster (p = 0.001) than the lifting side (the side corresponding with the hand that held the DB) during the DBPS. In addition, the DBPS nonlifting side catch phase loading rate was approximately double that of the lifting side loading rate (p < 0.05). These results quantify symmetrical deviations in the movement patterns of the unilateral power snatch movement both during the concentric muscular contraction of load vertical displacement, and the loading implications of unilateral landing. This asymmetry supports the contention that unilateral variations of weightlifting movements may provide a different training stimulus to athletes.     

Therapeutic dendritic cell vaccine preparation using tumor RNA transfection: A promising approach for the treatment of prostate cancer

Background

Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery.

Methods

Enhanced electrically-mediated delivery, and less extensively, liposome complexed delivery, of a plasmid encoding the reporter luciferase was tested in rodent skin. Expression kinetics and tissue damage were explored as well as testing in a second rodent model.

Results

Experiments confirm that electroporation alone is more effective in enhancing reporter gene expression than plasmid injection alone, plasmid conjugation with liposomes followed by injection, or than the combination of liposomes and electroporation. However, with two time courses of multiple electrically-mediated plasmid deliveries, neither the levels nor duration of transgene expression are significantly increased. Tissue damage may increase following a second treatment, no further damage is observed after a third treatment. When electroporation conditions utilized in a mouse model are tested in thicker rat skin, only higher field strengths or longer pulses were as effective in plasmid delivery.

Conclusion

Electroporation enhances reporter plasmid delivery to the skin to a greater extent than the liposome conjugation method tested. Multiple deliveries do not necessarily result in higher or longer term expression. In addition, some impact on tissue integrity with respect to surface damage is observed. Pulsing conditions should be optimized for the model and for the expression profile desired.