Gas exchange of a desert shrub (Zygophyllum dumosum Boiss.) under different soil moisture regimes during summer drought

The effects of soil water potential on photosynthesis and transpiration of whole Zygophyllum dumosum Boiss. shrubs were examined with a field IRGA system during a rainless summer. Daily photosynthesis and transpiration activities were not notably different on a unit phyllode area basis among shrubs at naturally differing soil water potentials. Irrigation of shrubs caused phyllodes to increase significantly in water content and new leaflets to appear. Leaflets had three times as many stomata per unit area (23000 stomata cm^-2) as phyllodes (7100 stomata cm^-2) but photosynthesis and transpiration rates were not measurably different between irrigated and non-irrigated shrubs on a unit area basis. This finding suggests that sufficient soil moisture will lead to increased carbon uptake of the entire shrub simply because the total area of photosynthesizing tissue increases. Gas exchange rates appear to be controlled solely by atmospheric conditions under the stresses of summer.     

An Extracellular Disulfide Bond Forming Protein (DsbF) from Mycobacterium tuberculosis: Structural, Biochemical, and Gene Expression Analysis

Disulfide bond forming (Dsb) proteins ensure correct folding and disulfide bond formation of secreted proteins. Previously, we showed that Mycobacterium tuberculosis DsbE (Mtb DsbE, Rv2878c) aids in vitro oxidative folding of proteins. Here, we present structural, biochemical, and gene expression analyses of another putative Mtb secreted disulfide bond isomerase protein homologous to Mtb DsbE, Mtb DsbF (Rv1677). The X-ray crystal structure of Mtb DsbF reveals a conserved thioredoxin fold although the active-site cysteines may be modeled in both oxidized and reduced forms, in contrast to the solely reduced form in Mtb DsbE. Furthermore, the shorter loop region in Mtb DsbF results in a more solvent-exposed active site. Biochemical analyses show that, similar to Mtb DsbE, Mtb DsbF can oxidatively refold reduced, unfolded hirudin and has a comparable pK_a for the active-site solvent-exposed cysteine. However, contrary to Mtb DsbE, the Mtb DsbF redox potential is more oxidizing and its reduced state is more stable. From computational genomics analysis of the M. tuberculosis genome, we identified a potential Mtb DsbF interaction partner, Rv1676, a predicted peroxiredoxin. Complex formation is supported by protein coexpression studies and inferred by gene expression profiles, whereby Mtb DsbF and Rv1676 are upregulated under similar environments. Additionally, comparison of Mtb DsbF and Mtb DsbE gene expression data indicates anticorrelated gene expression patterns, suggesting that these two proteins and their functionally linked partners constitute analogous pathways that may function under different conditions.     

New embedding medium for sectioning undecalcified bone

Methylmethacrylate (MMA) is the most commonly used embedding medium for sectioning undecalcified bone; however, a number of problems exist with its use in a research laboratory. MMA requires a long infiltration time and temperature control, and it reacts with many polymers. We used Kleer Set resin™ as an alternative embedding medium for sectioning undecalcified bone specimens. Fluorochrome labeled bone specimens were sectioned transversely using a ground section technique and longitudinally on a sledge macrotome. The slides were viewed using both transmitted light and epifluorescence microscopy. High quality sections were obtained using Kleer Set resin™ for both sectioning techniques. We have shown that this new embedding medium is simpler, safer, quicker to use and does not interfere with visualization of fluorochromes.     

On the relative importance of marker heterozygosity and intermarker distance in gene mapping.

Molecular biologists are often confronted with the problem of whether they should try to generate large numbers of very closely linked markers of low heterozygosity or smaller numbers of less closely linked markers of high heterozygosity. In other words, What is more important for gene mapping, high marker heterozygosity or dense marker spacing? We investigated that problem by analytically computing the expected lod score per meiosis in which the new locus is informative and phase known. We also looked at the length of the 1-unit-of-lod-score support interval for the expected lod score from 100 such meioses. We found that while both quantities have an influence on the number of meioses needed to find linkage, the length of the support interval is almost entirely dependent on the intermarker distance, for heterozygosities between 20 and 100%. However, the probability of any given meiosis being phase known and the ability to develop an accurate map of the markers are functions of marker heterozygosity, further complicating the issue.