Mechanism of action of VP1-001 in cryAB(R120G)-associated and age-related cataracts

PurposeWe previously identified an oxysterol, VP1-001 (also known as compound 29), that partially restores the transparency of lenses with cataracts. To understand the mechanism of VP1-001, we tested the ability of its enantiomer, ent-VP1-001, to bind and stabilize αB-crystallin (cryAB) in vitro and to produce a similar therapeutic effect in cryAB(R120G) mutant and aged wild-type mice with cataracts. VP1-001 and ent-VP1-001 have identical physicochemical properties. These experiments are designed to critically evaluate whether stereoselective binding to cryAB is required for activity.MethodsWe compared the binding of VP1-001 and ent-VP1-001 to cryAB using in silico docking, differential scanning fluorimetry (DSF), and microscale thermophoresis (MST). Compounds were delivered by six topical administrations to mouse eyes over 2 weeks, and the effects on cataracts and lens refractive measures in vivo were examined. Additionally, lens epithelial and fiber cell morphologies were assessed via transmission electron microscopy.ResultsDocking studies suggested greater binding of VP1-001 into a deep groove in the cryAB dimer compared with ent-VP1-001. Consistent with this prediction, DSF and MST experiments showed that VP1-001 bound cryAB, whereas ent-VP1-001 did not. Accordingly, topical treatment of lenses with ent-VP1-001 had no effect, whereas VP1-001 produced a statistically significant improvement in lens clarity and favorable changes in lens morphology.ConclusionsThe ability of VP1-001 to bind native cryAB dimers is important for its ability to reverse lens opacity in mouse models of cataracts

Deconvolving mutational patterns of poliovirus outbreaks reveals its intrinsic fitness landscape.

Vaccination has essentially eradicated poliovirus. Yet, its mutation rate is higher than that of viruses like HIV, for which no effective vaccine exists. To investigate this, we infer a fitness model for the poliovirus viral protein 1 (vp1), which successfully predicts in vitro fitness measurements. This is achieved by first developing a probabilistic model for the prevalence of vp1 sequences that enables us to isolate and remove data that are subject to strong vaccine-derived biases. The intrinsic fitness constraints derived for vp1, a capsid protein subject to antibody responses, are compared with those of analogous HIV proteins. We find that vp1 evolution is subject to tighter constraints, limiting its ability to evade vaccine-induced immune responses. Our analysis also indicates that circulating poliovirus strains in unimmunized populations serve as a reservoir that can seed outbreaks in spatio-temporally localized sub-optimally immunized populations

A partition-free approach to transient and steady-state charge currents

We construct a non-equilibrium steady state and calculate the corresponding current for a mesoscopic Fermi system in the partition-free setting. To this end we study a small sample coupled to a finite number of semi-infinite leads. Initially, the whole system of quasi-free fermions is in a grand canonical equilibrium state. At t = 0 we turn on a potential bias on the leads and let the system evolve. We study how the charge current behaves in time and how it stabilizes itself around a steady state value, which is given by a Landauer-type formula.Comment: 14 pages, submitte

Three-term polynomial progressions in subsets of finite fields

Bourgain and Chang recently showed that any subset of $\mathbb{F}_p$ of density $\gg p^{-1/15}$ contains a nontrivial progression $x,x+y,x+y^2$. We answer a question of theirs by proving that if $P_1,P_2\in\mathbb{Z}[y]$ are linearly independent and satisfy $P_1(0)=P_2(0)=0$, then any subset of $\mathbb{F}_p$ of density $\gg_{P_1,P_2}p^{-1/24}$ contains a nontrivial polynomial progression $x,x+P_1(y),x+P_2(y)$.Comment: 21 pages; v2: minor exposition changes based on referee comment

Identification of hepatitis a virus mimotopes by phage display, antigenicity and immunogenicity

A phage-displayed peptide approach was used to identify ligands mimicking antigenic determinants of hepatitis A virus (HAV) for the first time. Bacteriophages displaying HAV mimotopes were isolated from a phage-display peptide library by affinity selection on serum antibodies from hepatitis A patients. Selected phage-peptides were screened for reactivity with sera from HAV infected patients and healthy controls. Four cloned peptides with different sequences were identified as mimotopes of HAV; three of them showed similarity in their amino acid sequences with at least one of the VP3 and VP1 antigenic proteins of HAV. One clone was recognised by 92% of the positive sera. The phagotopes competed effectively with HAV for absorption of anti-HAV-specific antibodies in human sera, as determined by ELISA. The four phage clones induced neutralising anti-HAV antibodies in immunised mice. These results demonstrate the potential of this method to elucidate the disease related epitopes of HAV and to use these mimotopes in diagnostic applications or in the development of a mimotope-based hepatitis A vaccine without the necessity of manipulation of the virus

Pseudospin-Resolved Transport Spectroscopy of the Kondo Effect in a Double Quantum Dot

We report measurements of the Kondo effect in a double quantum dot (DQD), where the orbital states act as pseudospin states whose degeneracy contributes to Kondo screening. Standard transport spectroscopy as a function of the bias voltage on both dots shows a zero-bias peak in conductance, analogous to that observed for spin Kondo in single dots. Breaking the orbital degeneracy splits the Kondo resonance in the tunneling density of states above and below the Fermi energy of the leads, with the resonances having different pseudospin character. Using pseudospin-resolved spectroscopy, we demonstrate the pseudospin character by observing a Kondo peak at only one sign of the bias voltage. We show that even when the pseudospin states have very different tunnel rates to the leads, a Kondo temperature can be consistently defined for the DQD system.Comment: Text and supplementary information. Text: 4 pages, 5 figures. Supplementary information: 4 pages, 4 figure

The Minimum Spectral Radius of Graphs with the Independence Number

In this paper, we investigate some properties of the Perron vector of connected graphs. These results are used to characterize that all extremal connected graphs with having the minimum (maximum) spectra radius among all connected graphs of order $n=k\alpha$ with the independence number $\alpha$, respectively.Comment: 28 pages, 3 figure

Genetic characterization of human coxsackievirus A6 variants associated with atypical hand, foot and mouth disease: a potential role of recombination in emergence and pathogenicity

Human coxsackievirus A6 (CVA6) is an enterically transmitted enterovirus. Until recently, CVA6 infections were considered as being of minor clinical significance, and only rarely aetiologically linked with hand, foot and mouth disease (HFMD) associated with other species A enteroviruses (particularly EV71 and CVA16). From 2008 onwards, however, CVA6 infections have been associated with several outbreaks worldwide of atypical HFMD (aHFMD) accompanied by a varicelliform rash. We recently reported CVA6-associated eczema herpeticum occurring predominantly in children and young adults in Edinburgh in January and February 2014. To investigate genetic determinants of novel clinical phenotypes of CVA6, we genetically characterized and analysed CVA6 variants associated with eczema herpeticum in Edinburgh in 2014 and those with aHFMD in CAV isolates collected from 2008. A total of eight recombinant forms (RFs) have circulated worldwide over the past 10 years, with the particularly recent appearance of RF-H associated with eczema herpeticum cases in Edinburgh in 2014. Comparison of phylogenies and divergence of complete genome sequences of CVA6 identified recombination breakpoints in 2A-2C, within VP3, and between 5' untranslated region and VP1. A Bayesian temporal reconstruction of CVA6 evolution since 2004 provided estimates of dates and the actual recombination events that generated more recently appearing recombination groups (RF-E, -F, -G and -H). Associations were observed between recombination groups and clinical presentations of herpangina, aHFMD and eczema herpeticum, but not with VP1 or other structural genes. These observations provided evidence that NS gene regions may potentially contribute to clinical phenotypes and outcomes of CVA6 infection