Protein Expression, Characterization and Activity Comparisons of Wild Type and Mutant DUSP5 Proteins

Background The mitogen-activated protein kinases (MAPKs) pathway is critical for cellular signaling, and proteins such as phosphatases that regulate this pathway are important for normal tissue development. Based on our previous work on dual specificity phosphatase-5 (DUSP5), and its role in embryonic vascular development and disease, we hypothesized that mutations in DUSP5 will affect its function. Results In this study, we tested this hypothesis by generating full-length glutathione-S-transferase-tagged DUSP5 and serine 147 proline mutant (S147P) proteins from bacteria. Light scattering analysis, circular dichroism, enzymatic assays and molecular modeling approaches have been performed to extensively characterize the protein form and function. We demonstrate that both proteins are active and, interestingly, the S147P protein is hypoactive as compared to the DUSP5 WT protein in two distinct biochemical substrate assays. Furthermore, due to the novel positioning of the S147P mutation, we utilize computational modeling to reconstruct full-length DUSP5 and S147P to predict a possible mechanism for the reduced activity of S147P. Conclusion Taken together, this is the first evidence of the generation and characterization of an active, full-length, mutant DUSP5 protein which will facilitate future structure-function and drug development-based studies

Thermodynamic properties of sea air

Very accurate thermodynamic potential functions are available for fluid water, ice, seawater and humid air covering wide ranges of temperature and pressure conditions. They permit the consistent computation of all equilibrium properties as, for example, required for coupled atmosphere-ocean models or the analysis of observational or experimental data. With the exception of humid air, these potential functions are already formulated as international standards released by the International Association for the Properties of Water and Steam (IAPWS), and have been adopted in 2009 for oceanography by IOC/UNESCO. <br><br> In this paper, we derive a collection of formulas for important quantities expressed in terms of the thermodynamic potentials, valid for typical phase transitions and composite systems of humid air and water/ice/seawater. Particular attention is given to equilibria between seawater and humid air, referred to as "sea air" here. In a related initiative, these formulas will soon be implemented in a source-code library for easy practical use. The library is primarily aimed at oceanographic applications but will be relevant to air-sea interaction and meteorology as well. <br><br> The formulas provided are valid for any consistent set of suitable thermodynamic potential functions. Here we adopt potential functions from previous publications in which they are constructed from theoretical laws and empirical data; they are briefly summarized in the appendix. The formulas make use of the full accuracy of these thermodynamic potentials, without additional approximations or empirical coefficients. They are expressed in the temperature scale ITS-90 and the 2008 Reference-Composition Salinity Scale

Universality in the Crossover between Edge Channel and Bulk Transport in the Quantum Hall Regime

We present a new theoretical approach for the integer quantum Hall effect, which is able to describe the inter-plateau transitions as well as the transition to the Hall insulator. We find two regimes (metallic and insulator like) of the top Landau level, in which the dissipative bulk current appears in different directions. The regimes are separated by a temperature invariant point.Comment: 4 page, 2 eps figures included, submitte

Relationship between apoptotic markers in semen from fertile men and demographic, hormonal and seminal characteristics

Apoptosis in the testis has two putative roles during normal spermatogenesis; limitation of the germ cell population to numbers that can be supported by the Sertoli cells, and, possibly, selective depletion of meiotic and postmeiotic abnormal germ cells. We investigated the demographic and biological correlates of the pro-apoptotic marker Fas and the anti-apoptotic marker Bcl-xL in sperm cells of fertile men. Six hundred and four men from Greenland, Poland and Ukraine were consecutively enrolled during their pregnant wife's antenatal visits. Semen analysis was performed as recommended by the World Health Organization. Immunofluorescence coupled to flow cytometry was utilized for detection of apoptotic markers in the sperm cell. DNA damage was assessed by flow cytometry using both the sperm chromatin structure assay (SCSA) and the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. The percentage of Fas-positive sperm cells was higher in men with high total sperm count (P<0.01), more motile sperms (P=0.04) and fewer sperm head defects (P=0.05). These associations were consistent within and across study regions. Furthermore, testosterone, follicle-stimulating hormone (FSH) and sexual hormone-binding globulin (SHBG) were significantly negatively correlated with Fas within and across regions as well. The data indicated no association between the anti-apoptotic Bcl-xL marker and semen or personal characteristics. The finding of Fas-positive sperm cells associated with better semen quality in a cohort of spouses of pregnant women seems different from previous data obtained in infertile men and warrants further investigation to clarify the biological significance of sperm apoptotic markers

Recent distribution of Red-legged Partridge Alectoris rufa in Piedmont (North Western Italy): signs of recent spreading.

The red-legged partridge Alectoris rufa in Piedmont (NW Italy) is historically limited to the hilly and low-mountain areas of the southern half of the region, in particular lying in the Langhe and the Apennines on the Ligurian border, in the districts of Cuneo, Asti and Alessandria. In the Cuneo district, on the Maritime Alps, hybrid populations with Alectoris graeca were also known. The distribution of the species has been mapped in some occasions with the \u201cAtlases\u201d projects of the GPSO (Gruppo Piemontese Studi Ornitologici) and special enquiries. On this occasion we tried to update the geographical distribution of the species using data conferred by bird-watchers on www.regione.piemonte.it/aves/, managed by the GPSO, enquiries among hunters\u2019 organizations and specific field researches. The resulting distribution is shown on a 10 km square grid. The species shows signs of distribution dynamism, with the occupation of some lowland areas, where it was totally absent until the \u201890s. A little range expansion is noticeable also in the Western Monferrato. The situation seems to be more stable (or even declining) in the Langhe area and the species is now disappeared from the alpine border. In general we are seeing an areal contraction or density reduction on hills and mountains and an extension on lowlands. The observed changes seem to be attributable mainly to environmental and climate changes