Structure-based Discovery of Novel Small Molecule Wnt Signaling Inhibitors by Targeting the Cysteine-rich Domain of Frizzled.

Frizzled is the earliest discovered glycosylated Wnt protein receptor and is critical for the initiation of Wnt signaling. Antagonizing Frizzled is effective in inhibiting the growth of multiple tumor types. The extracellular N terminus of Frizzled contains a conserved cysteine-rich domain that directly interacts with Wnt ligands. Structure-based virtual screening and cell-based assays were used to identify five small molecules that can inhibit canonical Wnt signaling and have low IC50 values in the micromolar range. NMR experiments confirmed that these compounds specifically bind to the Wnt binding site on the Frizzled8 cysteine-rich domain with submicromolar dissociation constants. Our study confirms the feasibility of targeting the Frizzled cysteine-rich domain as an effective way of regulating canonical Wnt signaling. These small molecules can be further optimized into more potent therapeutic agents for regulating abnormal Wnt signaling by targeting Frizzled

Characterization of the Adeno-Associated Virus 1 and 6 Sialic Acid Binding Site

ABSTRACT The adeno-associated viruses (AAVs), which are being developed as gene delivery vectors, display differential cell surface glycan binding and subsequent tissue tropisms. For AAV serotype 1 (AAV1), the first viral vector approved as a gene therapy treatment, and its closely related AAV6, sialic acid (SIA) serves as their primary cellular surface receptor. Toward characterizing the SIA binding site(s), the structure of the AAV1-SIA complex was determined by X-ray crystallography to 3.0 Å. Density consistent with SIA was observed in a pocket located at the base of capsid protrusions surrounding icosahedral 3-fold axes. Site-directed mutagenesis substitution of the amino acids forming this pocket with structurally equivalent residues from AAV2, a heparan sulfate binding serotype, followed by cell binding and transduction assays, further mapped the critical residues conferring SIA binding to AAV1 and AAV6. For both viruses five of the six binding pocket residues mutated (N447S, V473D, N500E, T502S, and W503A) abolished SIA binding, whereas S472R increased binding. All six mutations abolished or decreased transduction by at least 50% in AAV1. Surprisingly, the T502S substitution did not affect transduction efficiency of wild-type AAV6. Furthermore, three of the AAV1 SIA binding site mutants—S472R, V473D, and N500E—escaped recognition by the anti-AAV1 capsid antibody ADK1a. These observations demonstrate that common key capsid surface residues dictate both virus binding and entry processes, as well as antigenic reactivity. This study identifies an important functional capsid surface “hot spot” dictating receptor attachment, transduction efficiency, and antigenicity which could prove useful for vector engineering. IMPORTANCE The adeno-associated virus (AAV) vector gene delivery system has shown promise in several clinical trials and an AAV1-based vector has been approved as the first gene therapy treatment. However, limitations still exist with respect to transduction efficiency and the detrimental effects of preexisting host antibodies. This study aimed to identify key capsid regions which can be engineered to overcome these limitations. A sialic glycan receptor recognition pocket was identified in AAV1 and its closely related AAV6, using X-ray crystallography. The site was confirmed by mutagenesis followed by cell binding and transduction assays. Significantly, residues controlling gene expression efficiency, as well as antibody escape variants, were also identified. This study thus provides, at the amino acid level, information for rational structural engineering of AAV vectors with improved therapeutic efficacy

CFHTLenS: A Weak Lensing Shear Analysis of the 3D-Matched-Filter Galaxy Clusters

We present the cluster mass-richness scaling relation calibrated by a weak lensing analysis of >18000 galaxy cluster candidates in the Canada-France-Hawaii Telescope Lensing Survey (CFHTLenS). Detected using the 3D-Matched-Filter cluster-finder of Milkeraitis et al., these cluster candidates span a wide range of masses, from the small group scale up to $\sim10^{15} M_{\odot}$, and redshifts 0.2 $\lesssim z\lesssim$ 0.9. The total significance of the stacked shear measurement amounts to 54$\sigma$. We compare cluster masses determined using weak lensing shear and magnification, finding the measurements in individual richness bins to yield 1$\sigma$ compatibility, but with magnification estimates biased low. This first direct mass comparison yields important insights for improving the systematics handling of future lensing magnification work. In addition, we confirm analyses that suggest cluster miscentring has an important effect on the observed 3D-MF halo profiles, and we quantify this by fitting for projected cluster centroid offsets, which are typically $\sim$ 0.4 arcmin. We bin the cluster candidates as a function of redshift, finding similar cluster masses and richness across the full range up to $z \sim$ 0.9. We measure the 3D-MF mass-richness scaling relation $M_{200} = M_0 (N_{200} / 20)^\beta$. We find a normalization $M_0 \sim (2.7^{+0.5}_{-0.4}) \times 10^{13} M_{\odot}$, and a logarithmic slope of $\beta \sim 1.4 \pm 0.1$, both of which are in 1$\sigma$ agreement with results from the magnification analysis. We find no evidence for a redshift-dependence of the normalization. The CFHTLenS 3D-MF cluster catalogue is now available at cfhtlens.org.Comment: 3D-MF cluster catalog is NOW AVAILABLE at cfhtlens.org. Magnification-shear mass comparison in Figure 10. 19 pages, 10 figures. Accepted to MNRA

Ex Situ Characterization of 1T/2H MoS2 and Their Carbon Composites for Energy Applications, a Review

The growing interest in the development of next-generation net zero energy systems has led to the expansion of molybdenum disulfide (MoS2) research in this area. This activity has resulted in a wide range of manufacturing/synthesis methods, controllable morphologies, diverse carbonaceous composite structures, a multitude of applicable characterization techniques, and multiple energy applications for MoS2. To assess the literature trends, 37,347 MoS2 research articles from Web of Science were text scanned to classify articles according to energy application research and characterization techniques employed. Within the review, characterization techniques are grouped under the following categories: morphology, crystal structure, composition, and chemistry. The most common characterization techniques identified through text scanning are recommended as the base fingerprint for MoS2 samples. These include: scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy. Similarly, XPS and Raman spectroscopy are suggested for 2H or 1T MoS2 phase confirmation. We provide guidance on the collection and presentation of MoS2 characterization data. This includes how to effectively combine multiple characterization techniques, considering the sample area probed by each technique and their statistical significance, and the benefit of using reference samples. For ease of access for future experimental comparison, key numeric MoS2 characterization values are tabulated and major literature discrepancies or currently debated characterization disputes are highlighted

Human phenotypes caused by PIEZO1 mutations; one gene, two overlapping phenotypes?

PIEZO1 is a large mechanosensitive ion channel protein. Diseases associated with PIEZO1 include autosomal recessive Generalised Lymphatic Dysplasia of Fotiou (GLDF) and autosomal dominant Dehydrated Hereditary Stomatocytosis with or without pseudohyperkalemia and/or perinatal oedema (DHS). The two disorders show overlapping features, fetal hydrops/perinatal oedema have been reported in both. Electrophysiological studies suggest opposite mechanisms of action, the mutations identified in GLDF patients cause a loss-of-function mechanism of disease and mutations in DHS patients cause gain-of-function. This raises the question, is the pathogenic disease mechanism behind the fetal oedema the same in the two phenotypes? In this symposium review, we will discuss the two conditions and highlight key questions that remain to be answered. For instance, the perinatal oedema often resolves soon after birth and we are still at a loss to understand why. Are there any mechanisms which could compensate for the faulty PIEZO1 in these patients? Are there physiological changes at birth that are less reliant on the function of PIEZO1? Thus, there is a clear need for further studies into the two disorders, in order to fully understand the role of PIEZO1 in health and disease

Consistent cosmic shear in the face of systematics: a B-mode analysis of KiDS-450, DES-SV and CFHTLenS

We analyse three public cosmic shear surveys; the Kilo-Degree Survey (KiDS-450), the Dark Energy Survey (DES-SV) and the Canada France Hawaii Telescope Lensing Survey (CFHTLenS). Adopting the “COSEBIs” statistic to cleanly and completely separate the lensing E-modes from the non-lensing B-modes, we detect B-modes in KiDS-450 and CFHTLenS at the level of ∼2.7σ. For DES-SV we detect B-modes at the level of 2.8σ in a non-tomographic analysis, increasing to a 5.5σB-mode detection in a tomographic analysis. In order to understand the origin of these detected B-modes we measure the B-mode signature of a range of different simulated systematics including PSF leakage, random but correlated PSF modelling errors, camera-based additive shear bias and photometric redshift selection bias. We show that any correlation between photometric-noise and the relative orientation of the galaxy to the point-spread-function leads to an ellipticity selection bias in tomographic analyses. This work therefore introduces a new systematic for future lensing surveys to consider. We find that the B-modes in DES-SV appear similar to a superposition of the B-mode signatures from all of the systematics simulated. The KiDS-450 and CFHTLenS B-mode measurements show features that are consistent with a repeating additive shear bias

The Type IIb Supernova 2013df and its Cool Supergiant Progenitor

We have obtained early-time photometry and spectroscopy of supernova (SN) 2013df in NGC 4414. The SN is clearly of Type IIb, with notable similarities to SN 1993J. From its luminosity at secondary maximum light, it appears that less 56Ni (≲0.06 M_☉) was synthesized in the SN 2013df explosion than was the case for the SNe IIb 1993J, 2008ax, and 2011dh. Based on a comparison of the light curves, the SN 2013df progenitor must have been more extended in radius prior to explosion than the progenitor of SN 1993J. The total extinction for SN 2013df is estimated to be AV = 0.30 mag. The metallicity at the SN location is likely to be solar. We have conducted Hubble Space Telescope (HST) Target of Opportunity observations of the SN with the Wide Field Camera 3, and from a precise comparison of these new observations to archival HST observations of the host galaxy obtained 14 yr prior to explosion, we have identified the progenitor of SN 2013df to be a yellow supergiant, somewhat hotter than a red supergiant progenitor for a normal Type II-Plateau SN. From its observed spectral energy distribution, assuming that the light is dominated by one star, the progenitor had effective temperature Teff = 4250 ± 100 K and a bolometric luminosity L_bol = 10^4.94 ± 0.06)L_☉. This leads to an effective radius R_eff = 545 ± 65 R_☉. The star likely had an initial mass in the range of 13–17 M_☉; however, if it was a member of an interacting binary system, detailed modeling of the system is required to estimate this mass more accurately. The progenitor star of SN 2013df appears to have been relatively similar to the progenitor of SN 1993J

Variability of Voriconazole Trough Levels in Haematological Patients: Influence of Comedications with cytochrome P450(CYP) Inhibitors and/or with CYP Inhibitors plus CYP Inducers

Voriconazole plasma exposure greatly varies among haematological patients. The purpose of this study was to identify the magnitude of influence of comedications with CYP inhibitors and/or with CYP inhibitors plus CYP inducers on voriconazole trough level (Cmin). Voriconazole Cmin was retrospectively assessed among haematological patients who underwent therapeutic drug monitoring (TDM). Univariate and multivariate linear mixed-effect regression analyses were performed to identify the independent predictors of normalized Cmin. Of the 83 included patients, 35 had comedications with CYP inhibitors (omeprazole or pantoprazole) and 21 with CYP inhibitors (omeprazole or pantoprazole) plus CYP inducers (methylprednisolone, dexamethasone, phenobarbital, rifampin or carbamazepine). Median Cmin value (n = 199) was 2.4 mg/L with a wide range of distribution (<0.2-13.5 mg/L). Median (IQR) normalized voriconazole Cmin value was significantly higher in the presence of CYP inhibitors (4.20 mg/L, 3.23-5.51 mg/L) than either in the absence of interacting cotreatments (2.55 mg/L, 1.54-3.47 mg/L) or in the presence of CYP inhibitors plus CYP inducers (2.16 mg/L, 1.19-3.09 mg/L). The presence of CYP inhibitors was highly significantly associated with Cmin >5.5 mg/L (OR: 23.22, 95% CI: 3.01-179.09, p = 0.003). No significant association emerged when CYP inhibitors were coadministered with CYP inducers (OR: 3.53, 95% CI: 0.36-34.95, p = 0.280). The amount of expected Cmin increase was significantly influenced by both the type and the dose of the administered proton pump inhibitor. The study highlights that the benefit from TDM of voriconazole may be maximal in those patients who are cotreated with CYP inhibitors and/or with CYP inhibitors plus CYP inducers, especially when receiving proton pump inhibitors (PPIs) at very high dosages intravenously

CFHTLenS: a weak lensing shear analysis of the 3D-Matched-Filter galaxy clusters

We present the cluster mass-richness scaling relation calibrated by a weak lensing analysis of ≳ 18000 galaxy cluster candidates in the Canada-France-Hawaii Telescope Lensing Survey (CFHTLenS). Detected using the 3D-Matched-Filter (MF) cluster-finder of Milkeraitis etal., these cluster candidates span a wide range of masses, from the small group scale up to∼1015 M⊙, and redshifts 0.2≲z≲0.9. The total significance of the stacked shear measurement amounts to 54σ. We compare cluster masses determined using weak lensing shear and magnification, finding the measurements in individual richness bins to yield 1σ compatibility, but with magnification estimates biased low. This first direct mass comparison yields important insights for improving the systematics handling of future lensing magnification work. In addition, we confirm analyses that suggest cluster miscentring has an important effect on the observed 3D-MF halo profiles, and we quantify this by fitting for projected cluster centroid offsets, which are typically∼0.4arcmin. We bin the cluster candidates as a function of redshift, finding similar cluster masses and richness across the full range up to z∼0.9. We measure the 3D-MF mass-richness scaling relation M200=M0(N200/20)β. We find a normalization $M_0 \sim (2.7^{+0.5}_{-0.4}) \times 10^{13} \,\mathrm{M}_{{\odot }}$, and a logarithmic slope of β∼1.4±0.1, both of which are in 1σ agreement with results from the magnification analysis. We find no evidence for a redshift dependence of the normalization. The CFHTLenS 3D-MF cluster catalogue is now available at cfhtlens.or