A paradigm for axonal transport

Axonal transport has been extensively studied for a period of 20–30 years, but there is still no general consensus concerning the mechanism by which this transport process operates. An important development in this regard is the recent studies in the physical biochemistry group in the Department of Biochemistry at Monash University where it has been demonstrated that ordered flows may be generated spontaneously in polymer systems under non-equilibeium conditions. The new phenomenon exhibits many novel features, particularly with respect to polymer transport, which bear marked similarity to the behaviour of components in axonal transport. This article sets out to essentiallybring to the attention of those in the neurosciences some of the properties of ordered structured flows in polymer solutions. These properties may generate a different view in the understanding of the mechanism of axonal transport.     

Altered processing of integrin receptors during keratinocyte activation

We used monoclonal antibodies against specific integrin subunits to examine the role of integrin receptors in keratinocyte activation. We found that before activation, beta 1 subunits in keratinocytes showed a diffuse distribution, whereas after activation, keratinocytes organized beta 1 receptors into marginal adhesion plaques. In immunoprecipitation experiments with antibodies against beta 1 integrin subunits, we found mostly immature subunits synthesized in keratinocytes freshly harvested from skin. Moreover, integrin receptor complexes immunoprecipitated from these cells by monoclonal antibodies against alpha 2, alpha 3, or alpha 5 subunits contained only immature beta 1 subunits. With keratinocytes cultured 4-7 days, anti-beta 1 antibodies immunoprecipitated mostly mature beta 1 subunits, and integrin complexes immunoprecipitated from cultured cells by anti-alpha subunit antibodies contained mostly mature beta 1 subunits. Antibodies directed against beta 1 subunits also inhibited keratinocyte migration. Based on these results, we suggest that up-regulation of migration by activated keratinocytes depends on changes in processing of pre-beta 1 subunits to mature beta 1 subunits. We also studied the distribution of integrin subunits in skin and on keratinocytes migrating out of skin explants. Whereas beta 1, alpha 2, and alpha 3 subunits were detected in keratinocytes in skin and migrating out of explants, alpha 5 subunits were observed only in migrating cells.     

The inheritance of restriction fragment length polymorphisms in the flax rust Melampsora lini

Summary Random cDNA sequences synthesized from poly A⁺ RNA extracted from germinated urediospores of the flax rust fungus, Melampsora lini, were used as probes to detect restriction fragment length polymorphisms (RFLPs) in three races of M. lini originating from cultivated flax, Linum usitatissimum, and one race originating from Australian native flax, L. marginale. Fourteen out of 22 probes tested detected RFLPs in the three races from cultivated flax while 19 of the probes detected polymorphisms between these three races and the race from L. marginale. The segregation of seven RFLPs was determined in a family of 19 F₂ progeny derived from a cross between two of the rust races. With six of these the inheritance was consistent, in each case, with the segregation of alleles at a single locus. Inheritance of the seventh was unusual and an explanation involving two loci with null alleles at each was proposed. No linkage was detected between any of the RFLP loci and nine unlinked loci specifying avirulence.     

Quantitative PCR: Procedures and precisions

We develop a multitype branching-process model for the Polymerase Chain Reaction (PCR). We apply the model to a comparison of three methods for estimating the initial number of molecules of target present in a PCR. These three methods are: one which uses a coamplified, internal control; one which uses an external control series; and one which uses simple extrapolation of log outputvs time (no control). We identify assumptions for each method which permit mathematical analysis of bias and precision. All three methods perform well if: (1) replication efficiencies are stable among reactions; (2) other method-specific conditions on efficiencies are met; and (3) product accumulates exponentially throughout the range where it is observed. When replication efficiencies vary among reactions but other optimal conditions for each method hold, the no-control and external-control methods lose precision relative to the internal control method, but they may still perform satisfactorily for many applications. The internal control method continues to perform well even if accumulation of product plateaus. This method depends, however, on a condition we call equivalence of replication efficiencies, the attainability of which in practice remains to be proven.     

Anhydrobiosis in Nematodes II: Carbohydrate and Lipid Analysis in Undesiccated and Desiccate Nematodes

Glycogen, trehalose, glucose, and total lipid contents of six nematode species were studied. Anhydrobiotic Anguina tritici and Ditylencbus dipsaci stored trehalose in preference to glycogen and only small amounts of glucose were detected. Glycogen content was also reduced in anhydrobiotic Aphelenchus avenae. Conversely, Panagrellus redivivus and Turbatrix aceti contained large amounts of glycogen, appreciable amounts of glucose, and minimal amounts of trehalose. Ditylenchus myceliophagous "curds" contained low amounts of glycogen and very little trehalose; total lipid was 60% of that in fresh samples. The lipid contents of fresh samples of P. redivivus, T. aceti, and A. avenae were high (23.1, 21.9, and 36.7% dry weight, respectively), but in anhydrobiotic A. avenae larvae the level was reduced by over 60%. In contrast, lipid levels remained high in anhydrobiotic A. tritici and D. dipsaci larvae (40.6 and 38.3%, respectively). Analysis of lipid composition in anhydrobiotic A. tritici and A. avenae did not indicate any specific metabolic adaptations to desiccation survival.     

Cell-free synthesis of glial fibrillary acidic protein

Glial fibrillary acidic (GFA) protein has been synthesized in an RNA-dependent cell-free system derived from rabbit reticulocytes. The cell-free synthesized product appears to have the same size as GFA protein isolated from bovine spinal cord, thus showing that GFA protein does not undergo detectable proteolytic processing.