Fetal mouse selenophosphate synthetase 2 (SPS2): biological activities of mutant forms in Escherichia coli.

A novel gene, sps2, detected in mouse embryo at the early stages of development has been identified as an analog of the E. coli selenophosphate synthetase gene. Unlike the E. coli enzyme, the presence of selenocysteine in the mouse enzyme is indicated by a TGA codon in the open reading frame of the cDNA. Using an N-FLAG monoclonal antibody, it was shown that the full length N-FLAG-sps2 gene product was expressed in COS-7 cells. To investigate the biological activity of the sps2 gene product in vivo, the mutated sps2 gene, which contains cysteine in the place of the TGA encoded selenocysteine in the wild type, was expressed in the E. coli selD deficient mutant, MB08. Like the E. coli wild type selD gene, the mutant sps2 gene complemented the selD mutation. However, replacement of Cys with either Ala, Ser, or Thr resulted in a loss of ability to complement the selD mutation. The SPS2-CYS protein expressed in E. coli was purified and its catalytic activity was determined. The Km value for ATP was 0.75 mM and Vmax was 9.23 nmole/min/mg protein. These results confirm that the mouse embryonic sps2 gene encodes an eukaryotic selenophosphate synthetase, and that availability of selenophosphate as a selenium donor compound is widespread.     

Osteogenic and adipogenic differentiation potential of an immortalized fibroblast-like cell line derived from porcine peripheral blood

Primary fibroblast-like cells isolated from the peripheral blood of a healthy pig were immortalized by transduction of cells with a replication-defective retrovirus vector expressing the E6/E7 proteins of human papillomavirus type 16 (pLXSN-16E6E7). The immortalized cells grow rapidly in cell culture and exhibit a distinct cell surface phenotype that was positive for CD90, CD44, collagen I, and vimentin and negative for CD14 and MHC II. Additionally, these immortalized blood derived-fibroblast-like cells had the potential to differentiate into osteoblasts and adipocytes in vitro as evidenced by the deposition of calcium, increased alkaline phosphatase activity, upregulated osteogenic and adipogenic marker gene expression, and accumulation of fat droplets in cells when osteogenic (dexamethasone, ascorbic acid, and β-glycerophosphate) or adipogenic supplements (dexamethasone, 3-isobutyl-1-methylxanthine, indomethacin, and insulin) were added to the culture. Overall, the results suggest that the immortalized blood-derived fibroblast-like cells exhibit some of the features of mesenchymal precursor cells, which may have implications in tissue repair and remodeling process.     

Distinguishing Drift and Selection Empirically: “The Great Snail Debate” of the 1950s

Biologists and philosophers have been extremely pessimistic about the possibility of demonstrating random drift in nature, particularly when it comes to distinguishing random drift from natural selection. However, examination of a historical case – Maxime Lamotte’s study of natural populations of the land snail, Cepaea nemoralis in the 1950s – shows that while some pessimism is warranted, it has been overstated. Indeed, by describing a unique signature for drift and showing that this signature obtained in the populations under study, Lamotte was able to make a good case for a significant role for␣drift. It may be difficult to disentangle the causes of drift and selection acting in a population, but it is not (always) impossible.     

The effects of manipulation of sedimentary iron and organic matter on sediment biogeochemistry and seagrasses in a subtropical carbonate environment

The microbial metabolism of organic matter (OM) in seagrass beds can create sulfidic conditions detrimental to seagrass growth; iron (Fe) potentially has ameliorating effects through titration of the sulfides and the precipitation of iron-sulfide minerals into the sediment. In this study, the biogeochemical effects of Fe availability and its interplay with sulfur and OM on sulfide toxicity, phosphorous (P) availability, seagrass growth and community structure were tested. The availability of Fe and OM was manipulated in a 2 × 2 factorial experiment arranged in a Latin square, with four replicates per treatment. The treatments included the addition of Fe, the addition of OM, the addition of both Fe and OM as well as no addition. The experiment was conducted in an oligotrophic, iron-deficient seagrass bed. Fe had an 84.5% retention efficiency in the sediments with the concentration of Fe increasing in the seagrass leaves over the course of the experiment. Porewater chemistry was significantly altered with a dramatic decrease in sulfide levels in Fe addition plots while sulfide levels increased in the OM addition treatments. Phosphorus increased in seagrass leaves collected in the Fe addition plots. Decreased sulfide stress was evidenced by heavier δ³⁴S in leaves and rhizomes from plots to which Fe was added. The OM addition negatively affected seagrass growth but increased P availability; the reduced sulfide stress in Fe added plots resulted in elevated productivity. Fe availability may be an important determinant of the impact that OM has on seagrass vitality in carbonate sediments vegetated with seagrasses.     

Engineering Hybrid Guided Modes in Subwavelength Uniaxial Metamaterial Waveguides

We report anomalous dispersion properties of hybrid guided modes (HGMs) and their group velocity in a subwavelength uniaxial metamaterial waveguide with metal cladding. We derive exact dispersion relations and modal fields of HGMs by solving eigenvalue equation based on basic electromagnetic field theory in detail. Numerical results show that two fundamental HGMs and two types of high-order HGMs can be excited, and their exciting conditions are clarified. In addition, such HGMs can be engineered to belong to normal dispersion or anomalous dispersion. Importantly, the HGMs may be controlled to be forward or backward, and their group velocities may be very small in a certain frequency band. These properties make such metamaterial waveguides have many potential applications in integrated optics, information storage and biosensing.