Functional Fluorescence Labeling of Carbohydrates and Its Use for Preparation of Neoglycoconjugates

Abstract

New bifunctional reagents, 2-amino-6-carboxyethylpyridine and 2-amino-6-cyanoethylpyridine, were designed and synthesized in order to provide a novel procedure for preparation of neoglycoconjugates from fluorescence-labeled and purified sugar chains. Labeling of model sugar chains with these reagents was effected by reductive amination using BH₃.Me₂NH to give corresponding 6-carboxyethylpyridylaminated (CEPA-) and 6-cyanoethylpyridylaminated (CNEPA-) derivatives, which were readily purified by reversed phase HPLC. The reagent parts of the labeled products possess the functional groups which then serve as linkers for coupling with matrices such as proteins and polymers. A CEPA-derivative of glucose was directly coupled with the ε-amino group of a Lys derivative to give a neoglycoprotein model. CNEPA-sugars were hydrogenated to give 6-aminopropylpyridylaminated (APPA-) derivatives, which can then be readily used for the preparation of various types of neoglycoconjugates by use of appropriate spacers as exemplified by the coupling of APPA-maltotriose with bovine serum albumin (BSA), biotin, and acrylic acid. The reaction of APPA-maltotriose with succinimidyl 3-(3-nitro-2-pyridyldithio)propionate gave the corresponding amide to be used for a disulfide formation with BSA. Condensation with biotin was effected by use of N-hydroxysuccinimidobiotin. The conjugation of APPA-maltotriose with acrylic acid was carried out by use of 4-acryloyloxyphenyldimethylsulfonium methylsulfate to give the corresponding acrylamide, which can be used for the preparation of sugar-acrylamide copolymers.     

Spatially variant regularization of lateral displacement measurement using variance

The purpose of this work is to confirm the effectiveness of our proposed spatially variant displacement component-dependent regularization for our previously developed ultrasonic two-dimensional (2D) displacement vector measurement methods, i.e., 2D cross-spectrum phase gradient method (CSPGM), 2D autocorrelation method (AM), and 2D Doppler method (DM). Generally, the measurement accuracy of lateral displacement spatially varies and the accuracy is lower than that of axial displacement that is accurate enough. This inaccurate measurement causes an instability in a 2D shear modulus reconstruction. Thus, the spatially variant lateral displacement regularization using the lateral displacement variance will be effective in obtaining an accurate lateral strain measurement and a stable shear modulus reconstruction than a conventional spatially uniform regularization. The effectiveness is verified through agar phantom experiments. The agar phantom [60 mm (height) × 100 mm (lateral width) × 40 mm (elevational width)] that has, at a depth of 10 mm, a circular cylindrical inclusion (dia. = 10 mm) of a higher shear modulus (2.95 and 1.43 × 10⁶ N/m², i.e., relative shear modulus, 2.06) is compressed in the axial direction from the upper surface of the phantom using a commercial linear array type transducer that has a nominal frequency of 7.5-MHz. Because a contrast-to-noise ratio (CNR) expresses the detectability of the inhomogeneous region in the lateral strain image and further has almost the same sense as that of signal-to-noise ratio (SNR) for strain measurement, the obtained results show that the proposed spatially variant lateral displacement regularization yields a more accurate lateral strain measurement as well as a higher detectability in the lateral strain image (e.g., CNRs and SNRs for 2D CSPGM, 2.36 vs 2.27 and 1.74 vs 1.71, respectively). Furthermore, the spatially variant lateral displacement regularization yields a more stable and more accurate 2D shear modulus reconstruction than the uniform regularization (however, for the regularized relative shear modulus reconstructions, slightly accurate, e.g., for 2D CSPGM, 1.51 vs 1.50). These results indicate that the spatially variant displacement component-dependent regularization will enable the 2D shear modulus reconstruction to be used as practical diagnostic and monitoring tools for the effectiveness of various noninvasive therapy techniques of soft tissue diseases (e.g., breast, liver cancers). Application of the regularization to the elevational displacement will also increase the stability of a three-dimensional (3D) reconstruction.     

Affine geometric crystals and limit of perfect crystals

For every non-exceptional affine Lie algebra, we explicitly construct a positive geometric crystal associated with a fundamental representation. We also show that its ultra-discretization is isomorphic to the limit of certain perfect crystals of the Langlands dual affine Lie algebra.

     

Synthesis of 2-azabicyclo[2.1.0]pentanes by the intramolecular nucleophilic substitution of cyclopropylmagnesium carbenoids with magnesium anilide

A variety of 2-azabicyclo[2.1.0]pentanes were synthesized by the intramolecular nucleophilic substitution of cyclopropylmagnesium carbenoids with magnesium anilide. The 1-chlorocyclopropyl p-tolyl sulfoxides possessing an N-aryl-substituted aminomethyl group were prepared from dichloromethyl p-tolyl sulfoxide, α,β-unsaturated carboxylic acid esters, and anilines in four steps. The deprotonation of the amine with t-BuMgCl followed by sulfoxide/magnesium exchange of the sulfoxides with i-PrMgCl led to the generation of the cyclopropylmagnesium carbenoids possessing a magnesium anilide moiety. Subsequent intramolecular nucleophilic substitution of the cyclopropylmagnesium carbenoids occurred in a 4-exo-tet manner to give the 2-azabicyclo[2.1.0]pentanes. The optically active 2-azabicyclo[2.1.0]pentane was synthesized using a p-tolylsulfinyl group as a chiral auxiliary.     

Addition of ArSSAr to carbon-carbon multiple bonds using electrochemistry

ArS(ArSSAr)⁺ (arylbis(arylthio)sulfonium ions), which were generated and accumulated by the electrochemical oxidation of diaryl disulfides (ArSSAr) in CH₂Cl₂ at −78 °C, reacted with alkenes to give the corresponding diarylthio-substituted compounds in a stereospecific manner in good yields, when the reaction was quenched with a soft nucleophile, such as allylsilanes, ketene silyl acetals, and triethylamine. A mechanism involving the initial formation of an episulfonium ion followed by ring-opening by the attack of ArSSAr has been suggested. The reactions of ArS(ArSSAr)⁺ with alkynes also took place to give 1,2-diorganothio-substitued alkenes stereoselectively under similar conditions.     

Unveiling residual molecular binding in triply charged hydrogen bromide

We present an experimental and theoretical study of triply charged hydrogen bromide ions formed by photoionization of the inner 3d shell of Br. The experimental results, obtained by detecting the 3d photoelectron in coincidence with the two subsequent Auger electrons, are analyzed using calculated potential energy curves of HBr3+. The competition between the short-range chemical binding potential and the Coulomb repulsion in the dissociative process is shown. Two different mechanisms are observed for double Auger decay: one, a direct process with simultaneous ejection of two Auger electrons to final HBr3+ ionic states and the other, a cascade process involving double Auger decay characterized by the autoionization of Br*+ ion subsequent to the HBr2+ fragmentation.     

Fredholm operators,evolution semigroups,and periodic solutions of nonlinear periodic systems

Let X be a Banach space and L the generator of the evolution semigroup associated with the τ  -periodic evolutionary process _{{U(t,s)}t≥s}${\{U(t,s)\}}_{t\ge s}$ on the space _Pτ(X)$P_{\tau }(X)$ of all τ-periodic continuous X  -valued functions. We give criteria for the existence of periodic solutions to nonlinear systems of the form Lp=−?F(p,?)$Lp=-?F(p,?)$ under the condition that 1 is a normal eigenvalue of the monodromy operator U(τ,0)$U(\tau ,0)$. The proof is based on a new decomposition of the space _Pτ(X)$P_{\tau }(X)$ by constructing a right inverse of L.     

A novel approach for LC-MS/MS-based chiral metabolomics fingerprinting and chiral metabolomics extraction using a pair of enantiomers of chiral derivatization reagents

Chiral metabolites are found in a wide variety of living organisms and some of them are understood to be physiologically active compounds and biomarkers. However, the overall analysis of chiral metabolomics is quite difficult due to the high number of metabolites, the significant diversity in their physicochemical properties, and concentration range from metabolite-to-metabolite. To solve this difficulty, we developed a novel approach for chiral metabolomics fingerprinting and chiral metabolomics extraction, which is based on the labeling of a pair of enantiomers of chiral derivatization reagents (i.e., DMT-(S,R)-Pro-OSu and DMT-3(S,R)-Apy) and precursor ion scan chromatography of the derivatives. The multivariate statistics is also required for this strategy. The proposed procedures were evaluated by the detection of a diagnostic marker (i.e., d-lactic acid) using the saliva of diabetic patients. This method was used for the determination of biomarker candidates of chiral amines and carboxyls in Alzheimer's disease (AD) brain homogenates. As the results, l-phenylalanine (L-Phe) and l-lactic acid (L-LA) were identified as the decreased and increased biomarker candidates in the AD brain, respectively. Therefore, the proposed approach seems to be helpful for the determination of non-target chiral metabolomics possessing amines and carboxyls.