Constraining CP violation in neutral meson mixing with theory input

There has been a lot of recent interest in the experimental hints of CP violation in B_{d,s}^0 mixing, which would be a clear signal of beyond the standard model physics (with higher significance). We derive a new relation for the mixing parameters, which allows clearer interpretation of the data in models in which new physics enters in M_12 and/or \Gamma_12. Our results imply that the central value of the D\O\ measurement of the semileptonic CP asymmetry in B_{d,s}^0 decay is not only in conflict with the standard model, but in a stronger tension with data on \Delta\Gamma_s than previously appreciated. This result can be used to improve the constraint on \Delta\Gamma or A_SL, whichever is less precisely measured.Comment: 5 pages, 2 figures, informed of prior derivation of eq. (21), title modifie

Higher Order Power Corrections in Inclusive B Decays

We discuss order 1/m_b^4 and 1/m_b^5 corrections in inclusive semileptonic decay of a $B$ meson. We identify relevant hadronic matrix elements of dimension seven and eight and estimate them using the ground-state saturation approximation. Within this approach the effects on the integrated rate and on kinematic moments are estimated. The overall relative shift in V_{cb} turns out about +0.4% as applied to the existing fits. Similar estimates are presented for B -> X_s+\gamma decays.Comment: 30 pages, 16 figure

Proceedings of the 2nd Workshop on Flavor Symmetries and Consequences in Accelerators and Cosmology (FLASY12)

These are the proceedings of the 2nd Workshop on Flavor Symmetries and Consequences in Accelerators and Cosmology, held 30 June 2012 - 4 July 2012, Dortmund, Germany.Comment: Order 400 pages, several figures including the group picture v2: corrected author list and contributio

Highly multiplexed subcellular RNA sequencing in situ

Understanding the spatial organization of gene expression with single-nucleotide resolution requires localizing the sequences of expressed RNA transcripts within a cell in situ. Here, we describe fluorescent in situ RNA sequencing (FISSEQ), in which stably cross-linked complementary DNA (cDNA) amplicons are sequenced within a biological sample. Using 30-base reads from 8102 genes in situ, we examined RNA expression and localization in human primary fibroblasts with a simulated wound-healing assay. FISSEQ is compatible with tissue sections and whole-mount embryos and reduces the limitations of optical resolution and noisy signals on single-molecule detection. Our platform enables massively parallel detection of genetic elements, including gene transcripts and molecular barcodes, and can be used to investigate cellular phenotype, gene regulation, and environment in situ

Taming the higher power corrections in semileptonic B decays

AbstractWe study the effect of dimension 7 and 8 operators on inclusive semileptonic B decays and the extraction of |Vcb|. Using moments of semileptonic B decay spectra and information based on the Lowest-Lying State saturation Approximation (LLSA) we perform a global fit of the non-perturbative parameters of the heavy quark expansion including for the first time the O(1/mb4,5) contributions. Higher power corrections appear to have a very small effect on the extraction of |Vcb|, independently of the weight we attribute to the LLSA

A novel in vitro assay for assessing efficacy and toxicity of antifungals using human leukemic cells infected with Candida albicans

Aims This study describes a novel in vitro assay that simultaneously determines antifungal efficiency and host cell toxicity using suspensions of human leukemic cells (HL-60) infected with Candida albicans. Methods and Results The effect of Candida infection on host cell viability was evaluated by microscopy of trypan blue stained cells and lactate dehydrogenase (LDH) activity. The in vitro ‘drug potency assay’ utilized the Cell Counting Kit-8 and measured post antifungal treatment viability of Candida-infected HL-60 cells and ability of the antifungal to prevent infection. LDH activity showed that 42% ± 4.0 and 85.3% ± 7.40 of HL-60 cells were killed following Candida infection at multiplicity of infection (MOI) of 1:1 and 1:5, respectively. Using the assay, the antifungal nystatin (0.78-25 μmol l−1) was found to inhibit C. albicans infection as seen by significantly increased viability of HL-60 cells. Using the assay, cytotoxicity of nystatin towards infected HL-60 cells was evident at higher concentrations and this was also confirmed by propidium iodide staining. Conclusions An assay using undisturbed cell suspension conditions was successfully developed for assessing selectivity of antifungal therapy in the host-Candida environment. Significance and Impact of the Study The assay employing Candida infection of host cell suspensions represents a promising method for testing interactions of antifungal compounds with both fungal and host cells

Phosphorylation of RSK2 at Tyr529 by FGFR2-p38 enhances human mammary epithelial cells migration

AbstractThe members of p90 ribosomal S6 kinase (RSK) family of Ser/Thr kinases are downstream effectors of MAPK/ERK pathway that regulate diverse cellular processes including cell growth, proliferation and survival. In carcinogenesis, RSKs are thought to modulate cell motility, invasion and metastasis. Herein, we have studied an involvement of RSKs in FGF2/FGFR2-driven behaviours of mammary epithelial and breast cancer cells. We found that both silencing and inhibiting of FGFR2 attenuated phosphorylation of RSKs, whereas FGFR2 overexpression and/or its stimulation with FGF2 enhanced RSKs activity. Moreover, treatment with ERK, Src and p38 inhibitors revealed that p38 kinase acts as an upstream RSK2 regulator. We demonstrate for the first time that in FGF2/FGFR2 signalling, p38 but not MEK/ERK, indirectly activated RSK2 at Tyr529, which facilitated phosphorylation of its other residues (Thr359/Ser363, Thr573 and Ser380). In contrast to FGF2-triggered signalling, inhibition of p38 in the EGF pathway affected only RSK2-Tyr529, without any impact on the remaining RSK phosphorylation sites. p38-mediated phosphorylation of RSK2-Tyr529 was crucial for the transactivation of residues located at kinase C-terminal domain and linker-region, specifically, in the FGF2/FGFR2 signalling pathway. Furthermore, we show that FGF2 promoted anchorage-independent cell proliferation, formation of focal adhesions and cell migration, which was effectively abolished by treatment with RSKs inhibitor (FMK). These indicate that RSK2 activity is indispensable for FGF2/FGFR2-mediated cellular effects. Our findings identified a new FGF2/FGFR2-p38-RSK2 pathway, which may play a significant role in the pathogenesis and progression of breast cancer and, hence, may present a novel therapeutic target in the treatment of FGFR2-expressing tumours

Precise Cas9 targeting enables genomic mutation prevention

Here, we present a generalized method of guide RNA “tuning” that enables Cas9 to discriminate between two target sites that differ by a single-nucleotide polymorphism. We employ our methodology to generate an in vivo mutation prevention system in which Cas9 actively restricts the occurrence of undesired gain-of-function mutations within a population of engineered organisms. We further demonstrate that the system is scalable to a multitude of targets and that the general tuning and prevention concepts are portable across engineered Cas9 variants and Cas9 orthologs. Finally, we show that the mutation prevention system maintains robust activity even when placed within the complex environment of the mouse gastrointestinal tract.National Human Genome Research Institute (U.S.) ( Grant P50 HG005550)Wyss Institute for Biologically Inspired EngineeringUnited States. Defense Threat Reduction Agency (Grant HDTRA1-15-1-0051)Paul G. Allen Frontiers Grou

Precise Cas9 targeting enables genomic mutation prevention

ABSTRACTHere we present a generalized method of guide RNA “tuning” that enables Cas9 to discriminate between two target sites that differ by a single nucleotide polymorphism. We employ our methodology to generate a novelin vivomutation prevention system in which Cas9 actively restricts the occurrence of undesired gain-of-function mutations within a population of engineered organisms. We further demonstrate that the system is scalable to a multitude of targets and that the general tuning and prevention concepts are portable across engineered Cas9 variants and Cas9 orthologs. Finally, we show that the designed mutation prevention system maintains robust activity even when placed within the complex environment of the mouse gastrointestinal tract.</jats:p