Anomeric and other substrate specificity studies with myo-inositol-1-P synthase

L-myo-Inositol-1-phosphate synthase has been found to have at least a 5-fold preference for the beta-anomer of its natural substrate D-Glc-6-P. The alpha-anomer appears to be an inhibitor of the reaction and may be converted to product as well. As well as showing an enzymatic preference for the equatorial C-1 hydroxyl of D-Glc-6-P, our results suggest that it is the pyranose form of D-Glc-6-P that binds to the enzyme and that ring-opening is an enzymatic step. We have also found D-2-dGlc-6-P, D-2-F-2-dGlc-6-P, and D-Man-6-P each to be both competitive inhibitors and substrates that are converted to inositol phosphates by the synthase. D-Allose-6-P is a weak inhibitor of the enzyme, but not a substrate. D-Gal-6-P is neither substrate nor inhibitor. Thus the specificity of the synthase with respect to single position epimers of D-Glc-6-P increases in the order C1 less than C2 much less than C3 less than C4.     

Ultrastructure of interstitial cells of Cajal at the gastro-oesophageal junction of the monkey (Macaca fascicularis)

The ultrastructure of the interstitial cells of Cajal (ICC) in the oesophagus of the monkey resembled that described in the oesophagus of other mammalian species but differed in their paucity and almost lack of smooth endoplasmic reticulum, caveolae and filaments. The plasmalemma of the ICC was in close contact (20- to 30-nm gaps) with that of smooth muscle cells. This may occasionally take the form of a desmosome, but gap junctions have not been observed. Vesiculated axon profiles, containing large granular or agranular vesicles were in close contact (20- to 30-nm gaps) with the plasmalemma of ICC. In a few vesiculated profiles a presynaptic density could be recognized. The intercalation of the ICC between the vesiculated axon profiles and the smooth muscle cells suggest a role in oesophageal motility. Between 3 and 21 days following bilateral vagotomy some ICC showed regressive changes such as increased electron density and shrinkage of the cytoplasm, crowding of the organelles and dissolution of the nuclear chromatin material. Axon profiles in the vicinity of the affected ICC contained glycogen granules suggesting injury. In late stages, the number of ICC and smooth muscle contacts was reduced. The results suggest that the vagus nerves exert a trophic influence on the ICC and that the intercellular relationships between ICC and smooth muscle cells possess a degree of plasticity. It is tentatively suggested that these vagal effects may be mediated via the oesophageal myenteric ganglia.     

Ultrastructure of the epithelial cells of the ventral prostate from the hopping mouse Notomys alexis

The large ventral prostate of the hopping mouse has abundant secretory units whose epithelial cells vary in height and which often have nuclei in the apical region of the cell. TEM observations indicated two epithelial cell types in which some unusual features occurred. Type A cells had granular endoplasmetic reticulum (GER) whose membranes often formed intracytoplasmic confronting cisternae. Type B cells had more fragmented and vesiculated GER with sparse ribosomes and less frequently also intracytoplasmic confronting confronting cisternae. In the latter cells, two types of granules were found, one of which was derived from the Golgi and the other possibly directly from the GER. Type A cells only had one type of granule present. A highly convoluted membrane was also found at the basal region in many of the cells. The significance of these unusual ultrastructural features has yet to be ascertained.     

Monitoring signal transduction in cancer: cDNA microarray for semiquantitative analysis.

This study targeted the development of a novel microarray tool to allow rapid determination of the expression levels of 58 different tyrosine kinase (tk) genes in small tumor samples. The goals were to define a reference probe for multi-sample comparison and to investigate the variability and reproducibility of the image acquisition and RT-PCR procedures. The small number of tk genes on our arrays enabled us to define a reference probe by artificially mixing all genes on the arrays. Such a probe provided contrast reference for comparative hybridization of control and sample DNA and enabled cross-comparison of more than two samples against one another. Comparison of signals generated from multiple scanning eliminated the concern of photo bleaching and scanner intrinsic noise. Tests performed with breast, thyroid, and prostate cancer samples yielded distinctive patterns and suggest the feasibility of our approach. Repeated experiments indicated reproducibility of such arrays. Up- or downregulated genes identified by this rapid screening are now being investigated with techniques such as in situ hybridization.     

Autumnal moth – why autumnal?

1. As for some other spring‐feeding moths, adult flight of Epirrita autumnata (Lepidoptera: Geometridae) occurs in late autumn. Late‐season flight is a result of a prolonged pupal period. Potential evolutionary explanations for this phenological pattern are evaluated. 2. In a laboratory rearing, there was a weak correlation between pupation date and the time of adult emergence. A substantial genetic difference in pupal period was found between two geographic populations. Adaptive evolution of eclosion time can thus be expected. 3. Metabolic costs of a prolonged pupal period were found to be moderate but still of some ecological significance. Pupal mortality is likely to form the main cost of the prolonged pupal period. 4. Mortality rates of adults, exposed in the field, showed a declining temporal trend from late summer to normal eclosion time in autumn. Lower predation pressure on adults may constitute the decisive selective advantage of late‐season flight. It is suggested that ants, not birds, were the main predators responsible for the temporal trend. 5. Egg mortality was estimated to be low; it is thus unlikely that the late adult period is selected for to reduce the time during which eggs are exposed to predators. 6. In a laboratory experiment, oviposition success was maximal at the time of actual flight peak of E. autumnata, however penalties resulting from sub‐optimal timing of oviposition remained limited.