Development of a Sensitive Phospho-p70 S6 Kinase ELISA to Quantify mTOR Proliferation Signal Inhibition

Background: Drug blood levels can only serve as a surrogate because of the lack of information on the drug's direct pharmacological effects in the individual patient. Measurement of the mammalian target of rapamycin (mTOR) activity dependent on the phosphorylation status of p70 S6 kinase (p70 S6K) offers a practical way for monitoring pharmacodynamic drug activity, with the potential to better assess the state of immunosuppression in individual patients. Material and Methods: Here, we established a novel in vitro model system by treating Jurkat cells and peripheral blood mononuclear cells with different concentrations of sirolimus after stimulation with phorbol 12-myristate 13-acetate. Results: A dose-dependent reduction of the p70 S6K phosphorylation status was demonstrated by Western blot and a newly established enzyme-linked immunosorbent assay (ELISA). Relative phospho-p70 S6K values from ELISA and relative densities from Western blot analysis in peripheral blood mononuclear cells revealed a strong correlation (Spearman correlation coefficient r(s) = 0.7, P = 0.01). Finally, parallel assays confirmed a sirolimus dose-dependent reduction of cytokine production and cell proliferation in the in vitro model. Conclusions: Pharmacodynamic monitoring of mTOR inhibition with a p70 S6K ELISA could guide mTOR inhibitor immunosuppression therapy toward a more individualized therapy. The usage of this technique now has to be evaluated in a clinical series of patients

Atomic and Molecular Interstellar Absorption Lines toward the High Galactic Latitude Stars HD 141569 and HD 157841 at Ultra-High Resolution

We present ultra-high-resolution (0.32 km s-1) spectra obtained with the 3.9 m Anglo-Australian Telescope (AAT) and Ultra-High-Resolution Facility (UHRF) of interstellar Na I D1, Na I D2, Ca II K, K I, and CH absorption toward two high Galactic latitude stars HD 141569 and HD 157841. We have compared our data with 21 cm observations obtained from the Leiden/Dwingeloo H I survey. We derive the velocity structure and column densities of the clouds represented by the various components and identify the clouds with ISM structures seen in the region at other wavelengths. We further derive abundances, linear depletions, and H2 fractional abundances for these clouds wherever possible. Both stars are located in regions of IRAS 100 μm emission associated with high Galactic latitude molecular clouds (HLCs): HD 141569 lies, in projection, close to MBM 37 and the Lynds dark cloud L134N, whereas HD 157841 is in the vicinity of the MBM 151. Toward HD 141569 we detect two components in our UHRF spectra: a weak, broad b = 4.5 km s-1 component at -15 km s-1, seen only in Ca II K absorption, and another component at 0 km s-1, seen in Na I D1, Na I D2, Ca II K, K I, and CH absorption. The cloud represented by the -15 km s-1 component is warm and may be located in a region close to the star. The cloud represented by the 0 km s-1 component has a Ca linear depletion δ(Ca) = 1.4 × 10-4 and shows evidence for the presence of dust, consistent with strong 100 μm emission seen in this region. The H2 fractional abundance f(H2) derived for this cloud is 0.4, which is typically what is observed toward HLCs. We conclude that this 0 km s-1 cloud is associated with MBM 37 and L134N based on the presence of dust and molecular gas (CH) and good velocity agreement with CO emission from these two clouds. This places HD 141569 beyond MBM 37 and L134N, which are estimated to be at ≈ 110 pc. In the case of the HD 157841 sight line, a total of six components are seen on our UHRF spectra in Na I D1, Na I D2, Ca II K, K I, and CH absorption. Two of these six components are seen only in a single species. The cloud represented by the components at 1.85 km s-1 has a Ca linear depletion δ(Ca) = 2.8 × 10-4, indicating the presence of dust. The f(H2) derived for this cloud is 0.45, and there is good velocity agreement with CO emission from MBM 151. To the best of our knowledge, this 1.85 km s-1 component toward HD 157841 is the first one found to have relative line widths that are consistent with pure thermal broadening only. We associate the 1.85 km s-1 cloud seen in our UHRF spectra with MBM 151 and conclude that HD 157841 must lie beyond ~200 pc, the estimated distance to MBM 151

Identification of candidate downstream genes for the homeodomain transcription factor Labial in Drosophila through oligonucleotide-array transcript imaging

BACKGROUND: Homeotic genes are key developmental regulators that are highly conserved throughout evolution. Their encoded homeoproteins function as transcription factors to control a wide range of developmental processes. Although much is known about homeodomain-DNA interactions, only a small number of genes acting downstream of homeoproteins have been identified. Here we use a functional genomic approach to identify candidate target genes of the Drosophila homeodomain transcription factor Labial. RESULTS: High-density oligonucleotide arrays with probe sets representing 1,513 identified and sequenced genes were used to analyze differential gene expression following labial overexpression in Drosophila embryos. We find significant expression level changes for 96 genes belonging to all functional classes represented on the array. In accordance with our experimental procedure, we expect that these genes are either direct or indirect targets of labial gene action. Among these genes, 48 were upregulated and 48 were downregulated following labial overexpression. This corresponds to 6.3% of the genes represented on the array. For a selection of these genes, we show that the data obtained with the oligonucleotide arrays are consistent with data obtained using quantitative RT-PCR. CONCLUSIONS: Our results identify a number of novel candidate downstream target genes for Labial, suggesting that this homeoprotein differentially regulates a limited and distinct set of embryonically expressed Drosophila genes

Impact of appropriate pharmaceutical therapy for chronic conditions on direct medical costs and workplace productivity: A review of the literature

This paper presents the findings of a literature review investigating the economic impact of appropriate pharmaceutical therapy in treating four prevalent chronic conditions - asthma, diabetes, heart failure, and migraine. The goal of the review was to identify high-quality studies examining the extent to which appropriate pharmaceutical therapy impacts overall medical expenditure (direct costs) and workplace productivity (indirect costs). The working hypothesis in conducting the review was that the costs of pharmaceuticals for the selected chronic conditions are offset by savings in direct and indirect costs in other areas. The literature provides evidence that appropriate drug therapy improves the health status and quality of life of individuals with chronic illnesses while reducing costs associated with utilization of emergency room, inpatient, and other medical services. A growing body of evidence also suggests that workers whose chronic conditions are effectively controlled with medications are more productive. For employers, the evidence translates into potential direct and indirect cost savings. The findings also confirm the importance of pharmaceutical management as a cornerstone of disease management

Atomic and molecular interstellar absorption lines toward the high galactic latitude stars HD~141569 and HD~157841 at ultra-high resolution

We present ultra-high resolution (0.32 km/s) spectra obtained with the 3.9m Anglo-Australian Telescope (AAT) and Ultra-High-Resolution Facility (UHRF), of interstellar NaI D1, D2, Ca II K, K I and CH absorption toward two high galactic latitude stars HD141569 and HD157841. We have compared our data with 21-cm observations obtained from the Leiden/Dwingeloo HI survey. We derive the velocity structure, column densities of the clouds represented by the various components and identify the clouds with ISM structures seen in the region at other wavelengths. We further derive abundances, linear depletions and H2 fractional abundances for these clouds, wherever possible. Toward HD141569, we detect two components in our UHRF spectra : a weak, broad component at - 15 km/s, seen only in CaII K absorption and another component at 0 km/s, seen in NaI D1, D2, Ca II K, KI and CH absorption. In the case of the HD157841 sightline, a total of 6 components are seen on our UHRF spectra in NaI D1, D2 Ca II K, K I and CH absorption. 2 of these 6 components are seen only in a single species.Comment: 16 pages, Latex, 4 figures, ps files Astrophysical Journal (in press

Star formation in bright-rimmed clouds and cluster associated with W5 E H{\sc ii} region

The aim of this paper is to present the results of photometric investigations of the central cluster of the W5 E region as well as a follow-up study of the triggered star formation in and around bright-rimmed clouds (BRCs). We have carried out wide field $UBVI_c$ and deep $VI_c$ photometry of the W5 E H{\sc ii} region. A distance of $\sim$2.1 kpc and a mean age of $\sim$1.3 Myr have been obtained for the central cluster. The young stellar objects (YSOs) associated with the region are identified on the basis of near-infrared and mid-infrared observations. We confirmed our earlier results that the average age of the YSOs lying on/inside the rim are younger than those lying outside the rim. The global distribution of the YSOs shows an aligned distribution from the ionising source to the BRCs. These facts indicate that a series of radiation driven implosion processes proceeded from near the central ionising source towards the periphery of the W5 E H{\sc ii} region. We found that, in general, the age distributions of the Class II and Class III sources are the same. This result is apparently in contradiction with the conclusion by Bertout, Siess & Cabrit (2007) and Chauhan et al. (2009) that classical T Tauri stars evolve to weak-line T Tauri stars. The initial mass function of the central cluster region in the mass range $0.4 \le M/M_\odot \le 30$ can be represented by $\Gamma = -1.29 \pm 0.03$. The cumulative mass functions indicate that in the mass range $0.2 \le M/M_\odot \le 0.8$, the cluster region and BRC NW have more low mass YSOs in comparison to BRCs 13 and 14.Comment: 43 pages, 17 figures, accepted for publication in MNRA

Choosing celebrity endorsers for advertising campaigns in cosmopolitan China: Does their ethnicity matter?

This study asks if endorsers function as surrogates for country-of-origin, and if the effectiveness of the tactic varies with the perceived ethnicity (Western versus Chinese) of the celebrities. It finds no significant influence on the country-oforigin perception, brand attitudes or product quality evaluations in the reactions of 797 consumers in Shanghai to magazine advertisements featuring celebrities of Chinese vs. Western ethnicity. The lack of any significant effect of the ethnicity of celebrities on a brand’s perceived country of origin precludes country image having any effect on those variables for the case of the cosmopolitan Chinese consumer

Elucidating how the saprophytic fungus Aspergillus nidulans uses the plant polyester suberin as carbon source

Lipid polymers in plant cell walls, such as cutin and suberin, build recalcitrant hydrophobic protective barriers. Their degradation is of foremost importance for both plant pathogenic and saprophytic fungi. Regardless of numerous reports on fungal degradation of emulsified fatty acids or cutin, and on fungi-plant interactions, the pathways involved in the degradation and utilisation of suberin remain largely overlooked. As a structural component of the plant cell wall, suberin isolation, in general, uses harsh depolymerisation methods that destroy its macromolecular structure. We recently overcame this limitation isolating suberin macromolecules in a near-native state.work partially supported by a grant from Iceland, Liechtenstein and Norway through the EEA financial mechanism (Project PT015), FCT: grant (PEst-OE/EQB/LA0004/2013) and fellowships (SFRH/BD/38378/2007, SFRH/BD/66396/2009, SFRH/BD/66030/2009, SFRH/BD/48286/2008), Fundação Calouste Gulbenkian fellowship (21-95587-B)

Gate-controlled Guiding of Electrons in Graphene

Ballistic semiconductor structures have allowed the realization of optics-like phenomena in electronics, including magnetic focusing and lensing. An extension that appears unique to graphene is to use both n and p carrier types to create electronic analogs of optical devices having both positive and negative indices of refraction. Here, we use gate-controlled density with both p and n carrier types to demonstrate the analog of the fiber-optic guiding in graphene. Two basic effects are investigated: (1) bipolar p-n junction guiding, based on the principle of angle-selective transmission though the graphene p-n interface, and (2) unipolar fiber-optic guiding, using total internal reflection controlled by carrier density. Modulation of guiding efficiency through gating is demonstrated and compared to numerical simulations, which indicates that interface roughness limits guiding performance, with few-nanometer effective roughness extracted. The development of p-n and fiber-optic guiding in graphene may lead to electrically reconfigurable wiring in high-mobility devices.Comment: supplementary materal at http://marcuslab.harvard.edu/papers/OG_SI.pd