Universal and high-fidelity DNA single nucleotide polymorphism detection based on a CRISPR/Cas12a biochip

Single nucleotide polymorphisms (SNPs) are associated with many human diseases, so accurate and efficient SNP detection is of great significance for early diagnosis and clinical prognosis. This report proposes a universal and high-fidelity genotyping method in microfluidic point-of-care equipment based on the clustered regularly interspaced short palindromic repeat (CRISPR) system. Briefly, by systematically inserting the protospacer-adjacent-motif (PAM) sequence, we improved the universality of the CRISPR/Cas12a based SNP detection; by removing the complementary ssDNA and introducing an additional nucleotide mismatch, we improved the sensitivity and specificity. We preloaded the CRISPR/Cas12a reagents into the point-of-care biochip for automating the process, increasing the stability and long-term storage. This biochip enables us to rapidly and conveniently detect the genotypes within 20 min. In a practical application, the CRISPR/Cas12a biochip successfully distinguished three genotypes (homozygous wild type; the homozygous mutant type; and the heterozygous mutant type) of the CYP1A1*2 (A4889G, rs1048943), CYP2C19*2 (G681A, rs4244285), CYP2C9*3 (A1075C, rs1057910), and CYP2C19*3 (G636A, rs4986893) genes related to multiple cancers from 17 clinical blood samples. This CRISPR/Cas12a-based SNP genotyping method, being universal, accurate, and sensitive, will have broad applications in molecular diagnostics and clinical research.

A universal and high-fidelity genotyping method based on the clustered regularly interspaced short palindromic repeat (CRISPR) system was performed on the microfluidic point-of-care equipment.     

Synthesis, Structure, and Properties of a Model for Galactose Oxidase

An active-site analog of the radical copper enzyme galactose oxidase has been prepared from a synthetic tripod chelate ((2-pyridylmethyl)[(2-hydroxy-3,5-dimethylphenyl)methyl][(2-hydroxy-5-methyl-3-(methylthio)phenyl)methyl]amine, duncamine (dnc)) that binds a single Cu(II) ion through phenolate, thioether-substituted phenolate, and pyridylamine arms. The Cu complex crystallizes as a dinucleated dimer bridged by phenolate oxygens, and the structure has been determined by X-ray crystallography. Addition of pyridine (or other coordinating bases) dissociates the complex into a monomeric derivative that has been characterized spectroscopically (optical absorption and EPR) and electrochemically. The model provides insight into the properties of a mutant form of galactose oxidase which retains the same copper ligand complement as the wild type protein but lacks catalytic activity.     

Multi-stage portfolio selection problem with dynamic stochastic dominance constraints

We study the multi-stage portfolio selection problem where the utility function of an investor is ambiguous. The ambiguity is characterized by dynamic stochastic dominance constraints, which are able to capture the dynamics of the random return sequence during the investment process. We propose a multi-stage dynamic stochastic dominance constrained portfolio selection model, and use a mixed normal distribution with time-varying weights and the K-means clustering technique to generate a scenario tree for the transformation of the proposed model. Based on the scenario tree representation, we derive two linear programming approximation problems, using the sampling approach or the duality theory, which provide an upper bound approximation and a lower bound approximation for the original nonconvex problem. The upper bound is asymptotically tight with infinitely many samples. Numerical results illustrate the practicality and efficiency of the proposed new model and solution techniques.

     

The Greatest Lower Bound of a Boros–Moll Sequence

Mathematical Notes - The Boros–Moll polynomials $$P_m(a)$$ arise in the evaluation of a quartic integral. In the past few years, there has been some remarkable research on the properties of...     

Global energy spectra of bands and density of states in a class of two-tile systems

Electronic properties of a general class of one-dimensional two-tile systems are calculated exactly. The systems containing periodic crystals, generalized Fibonacci quasicrystals, generalized Thue-Morse aperiodic lattices and even other two-tile aperiodic lattices, can be divided into two different families which are constructed by the inflation rules: {A, B}{A ^m11 B ^m12,A ^m21 B ^m22} and {A, B}{A ⁿ¹¹ B ⁿ¹²,B ⁿ²¹ A ⁿ²²}, respectively. As typical examples, global spectra of bands and density of states in some two-tile aperiodic systems are numerically calculated. Some interesting properties are obtained.     

Aspects of nucleon Compton scattering

We consider the spin-averaged nucleon forward Compton scattering amplitude in heavy baryon chiral perturbation theory including all terms to order . The chiral prediction for the spin-averaged forward Compton scattering amplitude is in good agreement with the data for photon energies110 MeV. We also evaluate the nucleon electric and magnetic Compton polarizabilities to this order and discuss the uncertainties of the various counter terms entering the chiral expansion of these quantities.     

Gas-discharge XeF* (B→X) laser with high specific output energy

The discharge characteristics of the XeF* (BX) laser are investigated. The NF₃ and Xe partial pressure of the laser gas mixture and the total gas pressure have been varied. A highest specific output energy of 4.7 J/l with an efficiency of 0.5% was obtained from a X-ray preionized Ne/Xe/NF₃ gas mixture at 6 bar with single-pulse excitation through a multichannel spark gap.     

黑腔靶中超热电子特性研究

近几年来,在“神光”装置上进行了1.053μm激光与平面靶及一系列柱形黑腔靶相互作用实验。用一台多道滤波—荧光X光能谱仪(FFS)测得各种靶发射的超热X射线谱,由谱推导超热电子温度T_h和超热电子总能量E_h当照射靶单束激光能量E_(tar)为400～670J、脉宽τ=650～1150ps时,发现黑腔内明显存在两群服从Maxwell分布高能电子(T_h=35～45keV;T_(hh)=150～350kev),而且E_(he)占E_(tar)的份额为10％～12％。实验还表明:腔内的E_(he)与非线性过程特征量(SRS)有较好的线性关系,因此推断出腔内超热电子产生的主要机制是受激Raman散射。在相同照射条件下,黑腔靶产生的超热电子比平面靶严重。     

Effect of Low Energy Electron Exposure on Ion Scattering Spectroscopy

The effect of low energy electron (LEE) exposure on ISS, including on the sputter peak and scatter peak as well as on ion neutralization, has been investigated for different samples. Some new results are discussed.     

Intermolecular packing and alignment in an ordered beta-hairpin antimicrobial peptide aggregate from 2D solid-state NMR

The aggregation and packing of a membrane-disruptive beta-hairpin antimicrobial peptide, protegrin-1 (PG-1), in the solid state are investigated to understand its oligomerization and hydrogen-bonding propensity. Incubation of PG-1 in phosphate buffer saline produced well-ordered nanometer-scale aggregates, as indicated by 13C and 15N NMR line widths, chemical shifts, and electron microscopy. Two-dimensional 13C and 1H spin diffusion experiments using C-terminus strand and N-terminus strand labeled peptides indicate that the beta-hairpin molecules in these ordered aggregates are oriented parallel to each other with like strands lining the intermolecular interface. In comparison, disordered and lyophilized peptide samples are randomly packed with both parallel and antiparallel alignments. The PG-1 aggregates show significant immobilization of the Phe ring near the beta-turn, further supporting the structural ordering. The intermolecular packing of PG-1 found in the solid state is consistent with its oligomerization in lipid bilayers. This solid-state aggregation approach may be useful for determining the quaternary structure of peptides in general and for gaining insights into the oligomerization of antimicrobial peptides in lipid bilayers in particular.