Generation of Mouse Small Intestinal Epithelial Cell Lines That Allow the Analysis of Specific Innate Immune Functions

Cell lines derived from the small intestine that reflect authentic properties of the originating intestinal epithelium are of high value for studies on mucosal immunology and host microbial homeostasis. A novel immortalization procedure was applied to generate continuously proliferating cell lines from murine E19 embryonic small intestinal tissue. The obtained cell lines form a tight and polarized epithelial cell layer, display characteristic tight junction, microvilli and surface protein expression and generate increasing transepithelial electrical resistance during in vitro culture. Significant up-regulation of Cxcl2 and Cxcl5 chemokine expression upon exposure to defined microbial innate immune stimuli and endogenous cytokines is observed. Cell lines were also generated from a transgenic interferon reporter (Mx2-Luciferase) mouse, allowing reporter technology-based quantification of the cellular response to type I and III interferon. Thus, the newly created cell lines mimic properties of the natural epithelium and can be used for diverse studies including testing of the absorption of drug candidates. The reproducibility of the method to create such cell lines from wild type and transgenic mice provides a new tool to study molecular and cellular processes of the epithelial barrier.     

The effect of the 4B chromosomes of hexaploid wheat on the growth and regeneration of callus cultures

Summary Calli were initiated from immature embryos of nine lines of hexaploid wheat (Triticum aestivum L. em. Thell). These were the euploid lines Chinese Spring and Cappelle-Desprez, a line of Chinese Spring ditelocentric for the long arm of 4B, four substitution lines of Chinese Spring in which chromosome 4B has been replaced by its homologues from different wheat varieties and substituted into Chinese Spring and a substitution line of Besostaya I 4B into Cappelle-Desprez. The calli from these lines were found to differ in their growth rates and morphogenic and regenerative activities. The substitution of different 4B chromosomes into Chinese Spring significantly increased morphogenesis and shoot regeneration from callus. The potential for developing wheat lines with improved culture characteristics is discussed.     

Water, temperature and life

Cold is the fiercest and most widespread enemy of life on earth. Natural cold adaptation and survival are discussed in terms of physicochemical and biochemical water management mechanisms, relying on thermodynamic or kinetic stabilization. Distinctions are drawn between general effects of low temperature (chill) and specific effects of freezing. Freeze tolerance is a misnomer because tolerance does not extend to the cell fluids. Freezing is confined to the extracellular spaces where it acts as a means of protecting the cytoplasm against freezing injury. Freeze resistance depends on the phenomenon of undercooling, a survival mechanism that relies on the long-term maintenance of a thermodynamically highly unstable state. Correct water management involves many factors, among them the control of membrane composition and transmembrane osmotic equilibrium, the biosynthesis of compounds able to afford protection against injury through freeze desiccation and the availability (or inactivation) of biogenic ice nucleation catalysts.     

Accelerating the convergence of the back-propagation method

The utility of the back-propagation method in establishing suitable weights in a distributed adaptive network has been demonstrated repeatedly. Unfortunately, in many applications, the number of iterations required before convergence can be large. Modifications to the back-propagation algorithm described by Rumelhart et al. (1986) can greatly accelerate convergence. The modifications consist of three changes:1) instead of updating the network weights after each pattern is presented to the network, the network is updated only after the entire repertoire of patterns to be learned has been presented to the network, at which time the algebraic sums of all the weight changes are applied:2) instead of keeping , the learning rate (i.e., the multiplier on the step size) constant, it is varied dynamically so that the algorithm utilizes a near-optimum , as determined by the local optimization topography; and3) the momentum factor is set to zero when, as signified by a failure of a step to reduce the total error, the information inherent in prior steps is more likely to be misleading than beneficial. Only after the network takes a useful step, i.e., one that reduces the total error, does again assume a non-zero value. Considering the selection of weights in neural nets as a problem in classical nonlinear optimization theory, the rationale for algorithms seeking only those weights that produce the globally minimum error is reviewed and rejected.     

Copper deficiency depresses rat aortae superoxide dismutase activity and prostacyclin synthesis

Prostaglandin synthesis shows dependence on lipid hydroperoxides and resultant oxygen derived radical formation. In view of the importance of dietary copper in cytosolic copper dependent superoxide dismutase (Cu/Zn SOD) activity and the role of SOD in oxygen radical formation, the influence of dietary copper on prostacylin (PGI2) synthesis and SOD activity in rat aorta was examined. Copper deficient (0.5 micrograms Cu/g diet) rats showed a significant 47% reduction in PGI2 synthesis rates by aortic ring incubations in comparison to copper adequate (6.0 micrograms Cu/g diet) animals. Aortic SOD activity was reduced by 46% in copper deficiency in comparison to copper adequate animals. Marginal dietary copper (1.6 micrograms Cu/g diet) significantly reduced aortic SOD activity by 32% but was without effect on aortic ring incubation PGI2 synthesis. These results indicate that dietary copper deficiency, and the resultant decrease in SOD activity, depresses aortic PGI2 synthesis.