Lipid metabolism of ripening apples

Little change was observed in the concentration of sitosterol, the principal free sterol of apple, during ripening of the fruit in air at 12°. Phospholipid increased by ca 10% during the first 15–18 days and thereafter showed little change. Phosphatidylcholine increased during ripening whilst phosphatidylethanolamine exhibited a transitory increase in the first 7–18 days. Incorporation of [¹⁴C]acetate into free sterol by apple cortical discs showed little change during ripening but incorporation into phospholipids increased substantially between days 1 and 15 with a 27-fold increase in incorporation into phosphatidylcholine and an 8-fold increase into phosphatidylethanolamine and phosphatidylinositol.     

Induced pluripotent stem cells derived from the developing striatum as a potential donor source for cell replacement therapy for Huntington disease

BackgroundCell replacement therapy (CRT) for Huntington disease (HD) requires a source of striatal (STR) progenitors capable of restoring the function lost due to STR degeneration. Authentic STR progenitors can be collected from the fetal putative striatum, or whole ganglionic eminence (WGE), but these tissues remain impractical for widespread clinical application, and alternative donor sources are required. Here we begin exploring the possibility that induced pluripotent stem cells (iPSC) derived from WGE may retain an epigenetic memory of their tissue of origin, which could enhance their ability to differentiate into STR cells.ResultsWe generate four iPSC lines from human WGE (hWGE) and establish that they have a capacity similar to human embryonic stem cells with regard to their ability to differentiate toward an STR phenotype, as measured by expression and demethylation of key STR genes, while maintaining an overall different methylome. Finally, we demonstrate that these STR-differentiated hWGE iPSCs share characteristics with hWGE (i.e., authentic STR tissues) both in vitro and following transplantation into an HD model. Overall, iPSCs derived from human WGE show promise as a donor source for CRT for HD.     

Can Zipf's law be adapted to normalize microarrays?

Background  

Normalization is the process of removing non-biological sources of variation between array experiments. Recent investigations of data in gene expression databases for varying organisms and tissues have shown that the majority of expressed genes exhibit a power-law distribution with an exponent close to -1 (i.e. obey Zipf's law). Based on the observation that our single channel and two channel microarray data sets also followed a power-law distribution, we were motivated to develop a normalization method based on this law, and examine how it compares with existing published techniques. A computationally simple and intuitively appealing technique based on this observation is presented.     

The larval feeding inhibition assay for the diagnosis of nematode anthelmintic resistance

A larval feeding assay for detection of nematode anthelmintic resistance to macrocyclic lactones and imidazothiazoles is described. The estimated concentration of anthelmintic required to inhibit larval feeding in 50% of L1's (IC50) that were resistant to either macrocyclic lactones or imidazothiazoles were significantly higher (P < or = 0.05) than those of susceptible isolates. Some variations in IC50 values were observed during the patent period of infection in all strains, although the pattern of the IC50 followed the same course. IC50 values varied in larvae developing from eggs shed throughout the patent period, with low values in the earliest larvae followed by higher values as the infection progressed, before decreasing at 70-90 days post-infection, although the low values of the first part of the patent period were not reached. However, the IC50 differences between all resistant and susceptible strains were significant throughout the whole patent period for ivermectin and levamisole. These results suggest that this technique may provide an alternative in vitro to detect anthelmintic resistance.     

Structure of MurF from Streptococcus pneumoniae co-crystallized with a small molecule inhibitor exhibits interdomain closure

In a broad genomics analysis to find novel protein targets for antibiotic discovery, MurF was identified as an essential gene product for Streptococcus pneumonia that catalyzes a critical reaction in the biosynthesis of the peptidoglycan in the formation of the cell wall. Lacking close relatives in mammalian biology, MurF presents attractive characteristics as a potential drug target. Initial screening of the Abbott small-molecule compound collection identified several compounds for further validation as pharmaceutical leads. Here we report the integrated efforts of NMR and X-ray crystallography, which reveal the multidomain structure of a MurF-inhibitor complex in a compact conformation that differs dramatically from related structures. The lead molecule is bound in the substrate-binding region and induces domain closure, suggestive of the domain arrangement for the as yet unobserved transition state conformation for MurF enzymes. The results form a basis for directed optimization of the compound lead by structure-based design to explore the suitability of MurF as a pharmaceutical target.     

The effect of inhibitors on the formation of phosphoenolpyruvate by rat liver mitochondria

1. Rat liver mitochondria oxidizing malate produce PEP (phosphoenolpyruvate) without the addition of ATP or other nucleotides. 2. The addition of oligomycin in the presence of 2,4-dinitrophenol did not abolish PEP formation and in some instances stimulated its formation. 3. Formation of PEP was inhibited by arsenate. 4. Arsenite decreased PEP formation and caused accumulation of pyruvate. 5. Added GTP and ITP had no effect on PEP formation. 6. PEP formed from malate in the presence of GTP and labelled P(i) had a specific radioactivity approximately the same as the P(i) with no contribution from the phosphate of the added GTP. 7. There was no parallelism between the effects of inhibitors on PEP formation from malate and their effects on the assayed activity of PEP carboxykinase. 8. In a direct comparison it was shown that the PEP carboxykinase content of mitochondria was insufficient to account for the PEP formation from malate. 9. Consideration of the kinetic characteristics of PEP carboxykinase and mitochondrial content of oxaloacetate and GTP show that this enzyme cannot account for the PEP formed from malate by mitochondria.