Role of Dicer Enzyme in the Regulation of Store Operated Calcium Entry (SOCE) in CD4+ T Cells

Background/Aims: Activation of T cell receptors (TCRs) in CD4+ T cells leads to a cascade of signalling reactions including increase of intracellular calcium (Ca2+) levels with subsequent Ca2+ dependent stimulation of gene expression, proliferation, cell motility and cytokine release. The increase of cytosolic Ca2+ results from intracellular Ca2+ release with subsequent activation of store-operated Ca2+ entry (SOCE). Previous studies suggested miRNAs are required for the development and functions of CD4+ T cells. An enzyme called Dicer is required during the process of manufacturing mature miRNAs from the precursor miRNAs. In this study, we explored whether loss of Dicer in CD4+ T cells affects SOCE and thus Ca2+ dependent regulation of cellular functions. Methods: We tested the expression of Orai1 by q-RT-PCR and flow cytometry. Further, we measured SOCE by an inverted phase-contrast microscope with the Incident-light fluorescence illumination system using Fura-2. Intracellular Ca2+ was also measured by flow cytometry using Ca2+ sensitive dye Fluo-4. Results: We found that in Dicer deficient (DicerΔ/Δ) mice Orai1 was downregulated at mRNA and protein level in CD4+ T cells. Further, SOCE was significantly smaller in DicerΔ/Δ CD4+ T cells than in CD4+ T cells isolated from wild-type (Dicerfl/fl) mice. Conclusion: Our data suggest that miRNAs are required for adequate Ca2+ entry into CD4+ T cells and thus triggering of Ca2+ sensitive immune functions

Heterotrimeric G-protein subunit Gαi2 contributes to agonist-sensitive apoptosis and degranulation in murine platelets

Gαi2, a heterotrimeric G-protein subunit, regulates various cell functions including ion channel activity, cell differentiation, proliferation and apoptosis. Platelet-expressed Gαi2 is decisive for the extent of tissue injury following ischemia/reperfusion. However, it is not known whether Gαi2 plays a role in the regulation of platelet apoptosis, which is characterized by caspase activation, cell shrinkage and cell membrane scrambling with phosphatidylserine (PS) translocation to the platelet surface. Stimulators of platelet apoptosis include thrombin and collagen-related peptide (CoRP), which are further known to enhance degranulation and activation of αII bβ3-integrin and caspases. Using FACS analysis, we examined the impact of agonist treatment on activation and apoptosis in platelets drawn from mice lacking Gαi2 and their wild-type (WT) littermates. As a result, treatment with either thrombin (0.01 U/mL) or CoRP (2 μg/mL or 5 μg/mL) significantly upregulated PS-exposure and significantly decreased forward scatter, reflecting cell size, in both genotypes. Exposure to CoRP triggered a significant increase in active caspase 3, ceramide formation, surface P-selectin, and αII bβ3-integrin activation. These molecular alterations were significantly less pronounced in Gαi2-deficient platelets as compared to WT platelets. In conclusion, our data highlight a previously unreported role of Gαi2 signaling in governing platelet activation and apoptosis.Fil: Cao, Hang. Universität Tübingen; AlemaniaFil: Qadri, Syed M.. Canadian Blood Services; Canadá. McMaster University; CanadáFil: Lang, Elisabeth. Heinrich-heine-universität Düsseldorf; AlemaniaFil: Pelzl, Lisann. Universität Tübingen; AlemaniaFil: Umbach, Anja T.. Universität Tübingen; AlemaniaFil: Leiss, Veronika. Universität Tübingen; AlemaniaFil: Birnbaumer, Lutz. National Institutes of Health; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Nürnberg, Bernd. Universität Tübingen; AlemaniaFil: Pieske, Burkert. Berlin Institute of Health; Alemania. Universitätsmedizin Berlin; AlemaniaFil: Voelkl, Jakob. Berlin Institute of Health; Alemania. Universitätsmedizin Berlin; AlemaniaFil: Gawaz, Meinrad. Universität Tübingen; AlemaniaFil: Bissinger, Rosi. Universität Tübingen; AlemaniaFil: Lang, Florian. Universität Tübingen; Alemania. Heinrich-heine-universität Düsseldorf; Alemani

HD 17156b: A Transiting Planet with a 21.2 Day Period and an Eccentric Orbit

We report the detection of transits by the 3.1 M_Jup companion to the V=8.17 G0V star HD 17156. The transit was observed by three independant observers on Sept. 9/10, 2007 (two in central Italy and one in the Canary Islands), who obtained detections at confidence levels of 3.0 sigma, 5.3 sigma, and 7.9 sigma, respectively. The observations were carried out under the auspices of the Transitsearch.org network, which organizes follow-up photometric transit searches of known planet-bearing stars during the time intervals when transits are expected to possibly occur. Analyses of the 7.9 sigma data set indicates a transit depth d=0.0062+/-0.0004, and a transit duration t=186+/-5 min. These values are consistent with the transit of a Jupiter-sized planet with an impact parameter b=a*cos(i)/R_star ~ 0.8. This planet occupies a unique regime among known transiting extrasolar planets, both as a result of its large orbital eccentricity (e=0.67) and long orbital period (P=21.2 d). The planet receives a 26-fold variation in insolation during the course of its orbit, which will make it a useful object for characterization of exoplanetary atmospheric dynamics.Comment: Accepted for publication to A&A, 4 pages, 2 figure

Mevalonate-Farnesal Biosynthesis in Ticks: Comparative Synganglion Transcriptomics and a New Perspective

Juvenile hormone (JH) controls the growth, development, metamorphosis, and reproduction of insects. For many years, the general assumption has been that JH regulates tick and other acarine development and reproduction the same as in insects. Although researchers have not been able to find the common insect JHs in hard and soft tick species and JH applications appear to have no effect on tick development, it is difficult to prove the negative or to determine whether precursors to JH are made in ticks. The tick synganglion contains regions which are homologous to the corpora allata, the biosynthetic source for JH in insects. Next-gen sequencing of the tick synganglion transcriptome was conducted separately in adults of the American dog tick, Dermacentor variabilis, the deer tick, Ixodes scapularis, and the relapsing fever tick, Ornithodoros turicata as a new approach to determine whether ticks can make JH or a JH precursor. All of the enzymes that make up the mevalonate pathway from acetyl-CoA to farnesyl diphosphate (acetoacetyl-CoA thiolase, HMG-S, HMG-R, mevalonate kinase, phosphomevalonate kinase, diphosphomevalonate decarboxylase, and farnesyl diphosphate synthase) were found in at least one of the ticks studied but most were found in all three species. Sequence analysis of the last enzyme in the mevalonate pathway, farnesyl diphosphate synthase, demonstrated conservation of the seven prenyltransferase regions and the aspartate rich motifs within those regions typical of this enzyme. In the JH branch from farnesyl diphosphate to JH III, we found a putative farnesol oxidase used for the conversion of farnesol to farnesal in the synganglion transcriptome of I. scapularis and D. variabilis. Methyltransferases (MTs) that add a methyl group to farnesoic acid to make methyl farnesoate were present in all of the ticks studied with similarities as high as 36% at the amino acid level to insect JH acid methyltransferase (JHAMT). However, when the tick MTs were compared to the known insect JHAMTs from several insect species at the amino acid level, the former lacked the farnesoic acid binding motif typical in insects. The P450s shown in insects to add the C10,11 epoxide to methyl farnesoate, are in the CYP15 family; this family was absent in our tick transcriptomes and in the I. scapularis genome, the only tick genome available. These data suggest that ticks do not synthesize JH III but have the mevalonate pathway and may produce a JH III precursor

In Situ Quench Reactions of Enantioenriched Secondary Alkyllithium Reagents in Batch and Continuous Flow Using an I/Li-Exchange

We report a practical in situ quench (ISQ) procedure involving the generation of chiral secondary alkyllithiums from secondary alkyl iodides (including functionalized iodides bearing an ester or a nitrile) in the presence of various electrophiles such as aldehydes, ketones, Weinreb amides, isocyanates, sulfides, or boronates. This ISQ-reaction allowed the preparation of a broad range of optically enriched ketones, alcohols, amides, sulfides and boronic acid esters in typically 90-98 % ee. Remarkably, these reactions were performed at -78 degrees C or -40 degrees C in batch. A continuous flow set-up permitted reaction temperatures between -20 degrees C and 0 degrees C and allowed a scale-up up to a 40-fold without further optimization

Call Me Caitlyn: Making and making over the 'authentic' transgender body in Anglo-American popular culture

A conception of transgender identity as an ‘authentic’ gendered core ‘trapped’ within a mismatched corporeality, and made tangible through corporeal transformations, has attained unprecedented legibility in contemporary Anglo-American media. Whilst pop-cultural articulations of this discourse have received some scholarly attention, the question of why this 'wrong body' paradigm has solidified as the normative explanation for gender transition within the popular media remains underexplored. This paper argues that this discourse has attained cultural pre-eminence through its convergence with a broader media and commercial zeitgeist, in which corporeal alteration and maintenance are perceived as means of accessing one’s ‘authentic’ self. I analyse the media representations of two transgender celebrities: Caitlyn Jenner and Nadia Almada, alongside the reality TV show TRANSform Me, exploring how these women’s gender transitions have been discursively aligned with a cultural imperative for all women, cisgender or trans, to display their authentic femininity through bodily work. This demonstrates how established tropes of authenticity-via-bodily transformation, have enabled transgender to become culturally legible through the wrong body trope. Problematically, I argue, this process has worked to demarcate ideals of ‘acceptable’ transgender subjectivity: self-sufficient, normatively feminine, and eager to embrace the possibilities for happiness and social integration provided by the commercial domain

XO-3b: A Massive Planet in an Eccentric Orbit Transiting an F5V Star

We report the discovery of a massive (Mpsini = 13.02 +/- 0.64 Mjup; total mass 13.25 +/- 0.64 Mjup), large (1.95 +/- 0.16 Rjup) planet in a transiting, eccentric orbit (e = 0.260 +/- 0.017) around a 10th magnitude F5V star in the constellation Camelopardalis. We designate the planet XO-3b, and the star XO-3, also known as GSC 03727-01064. The orbital period of XO-3b is 3.1915426 +/- 0.00014 days. XO-3 lacks a trigonometric distance; we estimate its distance to be 260 +/- 23 pc. The radius of XO-3 is 2.13 +/- 0.21 Rsun, its mass is 1.41 +/- 0.08 Msun, its vsini = 18.54 +/- 0.17 km/s, and its metallicity is [Fe/H] = -0.177 +/- 0.027. This system is unusual for a number of reasons. XO-3b is one of the most massive planets discovered around any star for which the orbital period is less than 10 days. The mass is near the deuterium burning limit of 13 Mjup, which is a proposed boundary between planets and brown dwarfs. Although Burrows et al. (2001) propose that formation in a disk or formation in the interstellar medium in a manner similar to stars is a more logical way to differentiate planets and brown dwarfs, our current observations are not adequate to address this distinction. XO-3b is also unusual in that its eccentricity is large given its relatively short orbital period. Both the planetary radius and the inclination are functions of the spectroscopically determined stellar radius. Analysis of the transit light curve of XO-3b suggests that the spectroscopically derived parameters may be over estimated. Though relatively noisy, the light curves favor a smaller radius in order to better match the steepness of the ingress and egress. The light curve fits imply a planetary radius of 1.25 +/- 0.15 Rjup, which would correspond to a mass of 12.03 +/- 0.46 Mjup.Comment: 26 pages, 10 figures. Accepted by ApJ. Current version has several small corrections as a result of a bug in the fitting softwar

A Transiting Planet of a Sun-like Star

A planet transits an 11th magnitude, G1V star in the constellation Corona Borealis. We designate the planet XO-1b, and the star, XO-1, also known as GSC 02041-01657. XO-1 lacks a trigonometric distance; we estimate it to be 200+-20 pc. Of the ten stars currently known to host extrasolar transiting planets, the star XO-1 is the most similar to the Sun in its physical characteristics: its radius is 1.0+-0.08 R_Sun, its mass is 1.0+-0.03 M_Sun, V sini < 3 km/s, and its metallicity [Fe/H] is 0.015+-0.04. The orbital period of the planet XO-1b is 3.941534+-0.000027 days, one of the longer ones known. The planetary mass is 0.90+-0.07 M_Jupiter, which is marginally larger than that of other transiting planets with periods between 3 and 4 days. Both the planetary radius and the inclination are functions of the spectroscopically determined stellar radius. If the stellar radius is 1.0+-0.08 R_Sun, then the planetary radius is 1.30+-0.11 R_Jupiter and the inclination of the orbit is 87.7+-1.2 degrees. We have demonstrated a productive international collaboration between professional and amateur astronomers that was important to distinguishing this planet from many other similar candidates.Comment: 31 pages, 9 figures, accepted for part 1 of Ap