[11C]flumazenil Binding Is Increased in a Dose-Dependent Manner with Tiagabine-Induced Elevations in GABA Levels

Evidence indicates that synchronization of cortical activity at gamma-band frequencies, mediated through GABA-A receptors, is important for perceptual/cognitive processes. To study GABA signaling in vivo, we recently used a novel positron emission tomography (PET) paradigm measuring the change in binding of the benzodiazepine (BDZ) site radiotracer [11C]flumazenil associated with increases in extracellular GABA induced via GABA membrane transporter (GAT1) blockade with tiagabine. GAT1 blockade resulted in significant increases in [11C]flumazenil binding potential (BPND) over baseline in the major functional domains of the cortex, consistent with preclinical studies showing that increased GABA levels enhance the affinity of GABA-A receptors for BDZ ligands. In the current study we sought to replicate our previous results and to further validate this approach by demonstrating that the magnitude of increase in [11C]flumazenil binding observed with PET is directly correlated with tiagabine dose. [11C]flumazenil distribution volume (VT) was measured in 18 healthy volunteers before and after GAT1 blockade with tiagabine. Two dose groups were studied (n = 9 per group; Group I: tiagabine 0.15 mg/kg; Group II: tiagabine 0.25 mg/kg). GAT1 blockade resulted in increases in mean (± SD) [11C]flumazenil VT in Group II in association cortices (6.8±0.8 mL g−1 vs. 7.3±0.4 mL g−1;p = 0.03), sensory cortices (6.7±0.8 mL g−1 vs. 7.3±0.5 mL g−1;p = 0.02) and limbic regions (5.2±0.6 mL g−1 vs. 5.7±0.3 mL g−1;p = 0.03). No change was observed at the low dose (Group I). Increased orbital frontal cortex binding of [11C]flumazenil in Group II correlated with the ability to entrain cortical networks (r = 0.67, p = 0.05) measured via EEG during a cognitive control task. These data provide a replication of our previous study demonstrating the ability to measure in vivo, with PET, acute shifts in extracellular GABA

Constitutive activation of T cells by γ2-herpesviral GPCR through the interaction with cellular CXCR4

Members of the herpesviral family use multiple strategies to hijack infected host cells and exploit cellular signaling for their pathogenesis and latent infection. Among the most intriguing weapons in the arsenal of pathogenic herpesviruses are the constitutively active virally-encoded G protein-coupled receptors (vGPCRs). Even though vGPCRs contribute to viral pathogenesis such as immune evasion and proliferative disorders, the molecular details of how vGPCRs continuously activate cellular signaling are largely unknown. Here, we report that the vGPCR of Herpesvirus saimiri (HVS), an oncogenic gamma 2-herpesvirus, constitutively activates T cells via a heteromeric interaction with cellular CXCR4. Constitutive T cell activation also occurs with expression of the vGPCR of Kaposi's sarcoma-associated herpesvirus (KSHV), but not the vGPCR of Epstein-Barr virus. Expression of HVS vGPCR down-regulated the surface expression of CXCR4 but did not induce the degradation of the chemokine receptor, suggesting that vGPCR/CXCR4 signaling continues in cytosolic compartments. The physical association of vGPCR with CXCR4 was demonstrated by proximity ligation assay as well as immunoprecipitation. Interestingly, the constitutive activation of T cells by HVS vGPCR is independent of proximal T cell receptor (TCR) signaling molecules, such as TcR beta, Lck, and ZAP70, whereas CXCR4 silencing by shRNA abolished T cell activation by vGPCRs of HVS and KSHV. Furthermore, previously identified inactive vGPCR mutants failed to interact with CXCR4. These findings on the positive cooperativity of vGPCR with cellular CXCR4 in T cell activation extend our current understanding of the molecular mechanisms of vGPCR function and highlight the importance of heteromerization for GPCR activity. (C) 2016 Elsevier B.V. All rights reserved.1111Ysciescopu

An Extension for Direct Gauge Mediation of Metastable Supersymmetry Breaking

We study the direct mediation of metastable supersymmetry breaking by a \Phi^2-deformation to the ISS model and extend it by splitting both Tr\Phi and Tr\Phi^2 terms in the superpotential and gauging the flavor symmetry. We find that with such an extension the enough long-lived metastable vacua can be obtained and the proper gaugino masses can be generated. Also, this allows for constructing a kind of models which can avoid the Landau pole problem. Especially, in our metastable vacua there exist a large region for the parameter m_3 which can satisfy the phenomenology requirements and allow for a low SUSY breaking scale (\sim 100 TeV).Comment: version in Europhys. Let

Thermally Stable Gel Polymer Electrolytes

To prepare miscible polyethylene glycol diacrylate/polyvinylidene fluoride (PEGDA/PVdF) blend gel polymer electrolytes, low molecular weight (M = 742) liquid PEGDA oligomer was mixed with PVdF-HFP dissolved in ethylene carbonate/dimethyl carbonate/LiPF6 liquid electrolytes, and then cured under ultraviolet irradiation. Room temperature conductivity of PEGDA/PVdF blend films was found to be comparable to that of PVdF-HFP gel polymer electrolytes, and they were electrochemically stable up to 4.6 V vs. Li/Li+. Scanning electron micrographs revealed that PEGDA/PVdF blend electrolytes have pore size intermediate between dense PEGDA and highly porous PVdF-HFP. It was confirmed by weight change measurement that liquid electrolyte was likely to evaporate through large pores in PVdF-HFP at 80°C, while PEGDA/PVdF blend showed better liquid electrolyte retention ability. This result was in good agreement with more stable interfacial properties of PEGDA/PVdF blend at 80°C in ac impedance analysis. Consequently, both PVdF-HFP and PEGDA/PVdF gel polymer electrolytes delivered similar discharge capacity at room temperature, but PEGDA/PVdF blend gel polymer electrolyte showed much better cycle performance than pure PVdF-HFP at 80°C

Correlation between Discharged Worms and Fecal Egg Counts in Human Clonorchiasis

Clonorchiasis is a major neglected disease in East Asia. Worm data in infected humans are very limited, and only egg counts roughly estimate infection burden of the worms. In endemic areas, we recruited infected cases and tried to collect the adult worms from them. They were treated with 3 doses of praziquantel, and purged next day under fasting. Adult worms of C. sinensis were recovered from their diarrheal feces. The worms discharged from humans after treatment are minimum confirmed numbers. The worm recovery rate noticeably increased in subjects with higher egg counts. The number of collected worms was well-correlated with the egg counts. Worm collection by praziquantel medication and purgation is a safe non-invasive method to get worm information from human. The present study confirms that at least 110 worms are infected in a human body with egg counts per gram of feces around 3,000, and egg productivity of a worm per day is around 4,000

The vacuum bubbles in de Sitter background and black hole pair creation

We study the possible types of the nucleation of vacuum bubbles. We classify vacuum bubbles in de Sitter background and present some numerical solutions. The thin-wall approximation is employed to obtain the nucleation rate and the radius of vacuum bubbles. With careful analysis we confirm that Parke's formula is also applicable to the large true vacuum bubbles. The nucleation of the false vacuum bubble in de Sitter background is also evaluated. The tunneling process in the potential with degenerate vacua is analyzed as the limiting cases of the large true vacuum bubble and false vacuum bubble. Next, we consider the pair creation of black holes in the background of bubble solutions. We obtain static bubble wall solutions of junction equation with black hole pair. The masses of created black holes are uniquely determined by the cosmological constant and surface tension on the wall. Finally, we obtain the rate of pair creation of black holes.Comment: 3 figures, minor including errors and typos corrected, and refs. adde

Reprogramming human T cell function and specificity with non-viral genome targeting.

Decades of work have aimed to genetically reprogram T cells for therapeutic purposes1,2 using recombinant viral vectors, which do not target transgenes to specific genomic sites3,4. The need for viral vectors has slowed down research and clinical use as their manufacturing and testing is lengthy and expensive. Genome editing brought the promise of specific and efficient insertion of large transgenes into target cells using homology-directed repair5,6. Here we developed a CRISPR-Cas9 genome-targeting system that does not require viral vectors, allowing rapid and efficient insertion of large DNA sequences (greater than one kilobase) at specific sites in the genomes of primary human T cells, while preserving cell viability and function. This permits individual or multiplexed modification of endogenous genes. First, we applied this strategy to correct a pathogenic IL2RA mutation in cells from patients with monogenic autoimmune disease, and demonstrate improved signalling function. Second, we replaced the endogenous T cell receptor (TCR) locus with a new TCR that redirected T cells to a cancer antigen. The resulting TCR-engineered T cells specifically recognized tumour antigens and mounted productive anti-tumour cell responses in vitro and in vivo. Together, these studies provide preclinical evidence that non-viral genome targeting can enable rapid and flexible experimental manipulation and therapeutic engineering of primary human immune cells