Human SULT1A genes: Cloning and activity assays of the SULT1A promoters

The three human SULT1A sulfotransferase enzymes are closely related in amino acid sequence (>90%), yet differ in their substrate preference and tissue distribution. SULT1A1 has a broad tissue distribution and metabolizes a range of xenobiotics as well as endogenous substrates such as estrogens and iodothyronines. While the localization of SULT1A2 is poorly understood, it has been shown to metabolize a number of aromatic amines. SULT1A3 is the major catecholamine sulfonating form, which is consistent with it being expressed principally in the gastrointestinal tract. SULT1A proteins are encoded by three separate genes, located in close proximity to each other on chromosome 16. The presence of differential 5′-untranslated regions identified upon cloning of the SULT1A cDNAs suggested the utilization of differential transcriptional start sites and/or differential splicing. This chapter describes the methods utilized by our laboratory to clone and assay the activity of the promoters flanking these different untranslated regions found on SULT1A genes. These techniques will assist investigators in further elucidating the differential mechanisms that control regulation of the human SULT1A genes. They will also help reveal how different cellular environments and polymorphisms affect the activity of SULT1A gene promoters

A major cellular substrate for protein kinases, annexin II, is a DNA-binding protein

AbstractWe have screened a human cDNA expression library in λgt11 for clones encoding Alu-binding proteins using direct binding of labeled Alu DNA to recombinant phage lysates fixed on a membrane, and isolated a clone 98% identical in sequence to the well-known substrate of protein kinases, annexin II, which was suggested earlier to play a role in transduction of mitogenic signals and DNA replication. A diagnostic property of annexins is their binding to phospholipids in the presence of calcium ions, and we have found that the interaction of proteins of human nuclear extracts with Alu subsequences is suppressed by Ca/phosphatidylserine liposomes, suggesting overlapping of Ca/phospholipid- and DNA-binding domains in annexin II

Proton NMR measurements of the local magnetic field in the paramagnetic metal and antiferromagnetic insulator phases of λ\lambda-(BETS)2_{2}FeCl4_{4}

Measurements of the $^{1}$H-NMR spectrum of a small ($\sim$ 4 $\mu$g) single crystal of the organic conductor $\lambda$-(BETS)$_{2}$FeCl$_{4}$ are reported with an applied magnetic field $\bf{B}$$_{0}$ = 9 T parallel to the a-axis in the $ac$-plane over a temperature $(T)$ range 2.0 $-$ 180 K. They provide the distribution of the static local magnetic field at the proton sites in the paramagnetic metal (PM) and antiferromagnetic insulator (AFI) phases, along with the changes that occur at the PM$-$AFI phase transition. The spectra have six main peaks that are significantly broadened and shifted at low $T$. The origin of these features is attributed to the large dipolar field from the 3d Fe$^{3+}$ ion moments (spin $S_{\rm{d}}$ = 5/2). Their amplitude and $T-$dependence are modeled using a modified Brillouin function that includes a mean field approximation for the total exchange interaction ($J_{0}$) between one Fe$^{3+}$ ion and its two nearest neighbors. A good fit is obtained using $J_{0}$ = $-$ 1.7 K. At temperatures below the PM$-$AFI transition temperature $T_{MI}$ = 3.5 K, an extra peak appears on the high frequency side of the spectrum and the details of the spectrum become smeared. Also, the rms linewidth and the frequency shift of the spectral distribution are discontinuous, consistent with the transition being first-order. These measurements verify that the dominant local magnetic field contribution is from the Fe$^{3+}$ ions and indicate that there is a significant change in the static local magnetic field distribution at the proton sites on traversing the PM to AFI phase transition.Comment: 11 pages, 7 figures. Revised version of cond-mat/0605044 resubmitted to Phys. Rev. B in response to comments of Editor and reviewer

Laboratory study on heterogeneous decomposition of methyl chloroform on various standard aluminosilica clay minerals as a potential tropospheric sink

International audienceMethyl chloroform (1,1,1-trichloroethane, CH3CCl3) was found to decompose heterogeneously on seven types of standard clay minerals (23 materials) in dry air at 313 K in the laboratory. All reactions proceeded through the elimination of HCl; CH3CCl3 was converted quantitatively to CH2=CCl2. The activities of the clay minerals were compared via their pseudo-first-order reaction rate constants (k1). A positive correlation was observed between the k1 value and the specific surface area (S) of clay minerals, where the S value was determined by means of the general Brunauer-Emmett-Teller (BET) equation. The k1 value was anti-correlated with the value of n, which was a parameter of the general BET equation and related to the average pore size of the clay minerals, and correlated with the water content that can be removed easily from the clay minerals. The reaction required no special pretreatment of clay minerals, such as heating at high temperatures; hence, the reaction can be expected to occur in the environment. Photoillumination by wavelengths present in the troposphere did not accelerate the decomposition of CH3CCl3, but it induced heterogeneous photodecomposition of CH2=CCl2. The temperature dependence of k1, the adsorption equilibrium coefficient of CH3CCl3 and CH2=CCl2, and the surface reaction rate constant of CH3CCl3 were determined for an illite sample. The k1 value increased with increasing temperature. The amount of CH3CCl3 adsorbed on the illite during the reaction was proportional to the partial pressure of CH3CCl3. The reaction was sensitive to relative humidity and the k1 value decreased with increasing relative humidity. However, the reaction was found to proceed at a relative humidity of 22% at 313 K, although the k1 value was about one-twentieth of the value in non-humidified air. The conditions required for the reaction may be present in major desert regions of the world. A simple estimation indicates that the possible heterogeneous decomposition of CH3CCl3 on the ground surface in arid regions is worth taking into consideration when inferring the tropospheric lifetime of CH3CCl3 and global OH concentration from the global budget concentration of CH3CCl3

Catchment geology preconditions spatio-temporal heterogeneity of ecosystem functioning in forested headwater streams

Catchment geology can affect water chemistry and groundwater influence, eventually affecting macroinvertebrate communities, but its effects on stream functions such as leaf decomposition have been scarcely investigated. To understand the effects of geology on leaf decomposition, we conducted leaf litter experiments in streams with volcanic and non-volcanic substrata using fine and coarse mesh bags. Volcanic spring-fed streams showed lower temperature in summer and higher temperature in winter (with temperature difference being more pronounced later in incubation) than non-volcanic streams. Macroinvertebrate communities captured inside coarse litter bags differed in the two stream types in both seasons, mainly because of shredder communities. Shredder abundance and biomass were higher in volcanic streams in both seasons. Geology-dependent temperature influenced microbe-mediated decomposition in both seasons, with total phosphorus as an additional driver in winter. Summer temperature was associated with an overall positive effect on the abundance of shredders, which affected invertebrate-mediated decomposition, but this was not evident in winter. Shredder activity in volcanic streams compensated for temperature-dependent microbial activity resulting in an overall balance in leaf decomposition. Spring-fed systems are valuable ecosystems, particularly for cold-adapted species. Thus, understanding these understudied ecosystems will significantly aid in their appropriate conservation

Application of the Hilbert-Huang transform for analyzing SASI induced gravitational waves in a core-collapse supernova

Through numerical simulations, it is predicted that the gravitational waves (GWs) reflect the characteristics of the core-collapse supernova (CCSN) explosion mechanism. There are multiple GW excitation processes that occur inside a star before its explosion, and it is suggested that the GWs originating from the CCSN have a mode for each excitation process in terms of time-frequency representation. Therefore, we propose an application of the Hilbert-Huang Transform (HHT), which is a high-resolution time-frequency analysis method, to analyze these GW modes for theoretically probing and increasing our understanding of the explosion mechanism. The HHT defines frequency as a function of time, and is not bound by the trade-off between time and frequency resolutions. In this study, we analyze a gravitational waveform obtained from a three-dimensional general-relativistic CCSN model that showed a vigorous activity of the standing-accretion-shock-instability (SASI). We succeed in extracting the SASI induced GWs with high resolution on a time-frequency representation using the HHT and we examine their instantaneous frequencies