Effects of As(III) binding on beta-hairpin structure

While arsenic(III) compounds can exert profound toxicological and pharmacological effects, their modes of action and, in particular, the structural consequences of their binding to cysteinyl side chains in proteins, remain poorly understood. To gain an understanding of how arsenic binding influences beta-structure, pairs of cysteines were introduced into a model monomeric beta-hairpin to yield a family of peptides such that coordination occurs either across the strands or within the same strand of the beta-hairpin. Circular dichroism, NMR, UV-vis spectroscopy, and rapid-reaction studies were used to characterize the binding of monomethylarsonous acid or p-succinylamidephenyl arsenoxide (PSAO) to these peptides. Placement of cysteines at non-hydrogen bond (NHB) positions across the beta-hairpin, such that they occupy the same face of the sheet, was found to enhance the structure as assessed by CD. Cross-strand cysteine residues that project on opposite faces close to the termini of the hairpin can still bind arsenic tightly and show modestly increased beta-sheet content. NMR and modeling studies suggest that arsenic can be accommodated at this locus without disrupting the core interactions stabilizing the turn. However, As(III) binding to nonopposed cysteines, or to cysteines at HB and NHB positions along one strand of the hairpin, caused loss of structure. UV-vis titrations show that all these hairpin peptides bind PSAO stoichiometrically with K(d) values from 13 to 106 nM. Further, binding is moderately rapid, with second-order rate constants for association of 10,000-22,000 M(-1) s(-)1 irrespective of the placement of the cysteines within the hairpin and the consequent extent of structural reorganization required as a result of binding. These studies complement recent work with alpha-helices and further demonstrate that capture of a pair of thiols by As(III) may result in significant changes in local secondary structure in the protein targets of these potent bioactive agents.     

Studies with azoles and benzoazoles: A novel simple approach for synthesis of 3‐functionally substituted 3‐acylindoles

3‐Substituted acylindoles 8 are obtained via refluxing carboxylic acids with indole in acetic anhydride solutions. The formed 3‐substituted acylindole 8a is readily converted into 4‐aminopyrazol‐3‐ylindoles 20 , and into 22 . Indole reacts with chloroacetyl chloride to yield: 3‐chloroacetylindole 9 which could also be utilized for synthesis of a number of 3‐substituted indoles.     

Eu(III) extraction by bis(2-ethylhexyl)phosphoric acid and 8-hydroxyquinoline in dodecane from perchlorate medium

Europium(III) was extracted by bis(2-ethylhexyl)phosphoric acid (HDEHP) and 8-hydroxyquinoline (HQ) in dodecane from aqueous perchlorate media of constant ionic strength (0.1M; H⁺, NaClO₄). Slope analysis of the data indicate that three molecules of HDEHP or HQ are attached to Eu³⁺. Extraction constants were obtained at different temperatures. The data were used to calculate the thermodynamic parameters (G, H and S) for the extraction process in the two systems. When using mixtures of crown ethers with HDEHP no synergism was observed.     

Enzyme immunoassays for the analysis of streptomycin in milk, serum and water: development and assessment of a polyclonal antiserum and assay procedures using novel streptomycin derivatives

A polyclonal antiserum to streptomycin was generated in sheep using a streptomycin-bovine serum albumin conjugate as immunogen. Streptomycin was linked to the carrier protein with cyanuric chloride using a new two-step conjugation method. Plate coating antigen conjugates of streptomycin and gelatine were prepared using either cyanuric chloride (homologous bridge) or 1,4-butanediol diglycidyl ether to provide an heterologous complex. The reagents enabled the generation of a specific antiserum with a titre of 1/40,000 and the development of a sensitive ELISA method suitable for the measurement of streptomycin sulfate in milk, serum and water samples. A minimum detection value of 1 ng mL(-1) and a dynamic range of 1 to 200 ng mL(-1) were demonstrated in the three matrices. No detectable cross reactivity with any of the common aminoglycosides was found except the related dihydrostreptomycin which gave a 75% cross reaction value. The details of the preparation of the hapten-protein conjugates, characterisation of the antiserum and assay construction and assessment methods are presented. The introduction of new coupling methods and antibody assessment provide improved basic methodologies necessary for the advancement of immuno-analysis of streptomycin, one of the most widely used antimicrobial substances.     

Bis (trimethylsilyl) thioketone

Tris (trimethylsilyl) methylsulphenylbromide undergoes facile Me₃ SiBr elimination to give bis (trimethylsilyl) thioketone.     

Regioselective synthesis of 5-aminopyrazoles from reactions of amidrazones with activated nitriles: NMR investigation and X-ray structural analysis

N-Arylbenzamidrazones and ethyl 2-cyano-3-ethoxybut-2-enoate reacted together in ethanol and catalyzed by triethylamine (Et₃N) to give 5-amino-3-methyl-1-(aryl(phenylimino)methyl)-1H-pyrazole derivatives. Reaction of the target amidrazones with bis-(methylthio)methylidene)malononitrile in EtOH/Et₃N/DMF mixture proceeded to give the corresponding 5-aminopyrazoles. The structure of the obtained products was proved by IR, mass, and NMR spectra and elemental analyses. Two-dimensional NMR spectroscopy and X-ray structural analyses were used to differentiate the assigned structures from other possible ring systems and regioisomers. The reaction mechanism is discussed.     

A numerical method based on hybrid orthonormal Bernstein and improved block-pulse functions for solving Volterra–Fredholm integral equations

This paper deals with the numerical solution of the integral equations of linear second kind Volterra–Fredholm. These integral equations are commonly used in engineering and mathematical physics to solve many of the problems. A hybrid of Bernstein and improved block-pulse functions method is introduced and used where the key point is to transform linear second-type Volterra–Fredholm integral equations into an algebraic equation structure that can be solved using classical methods. Numeric examples are given which demonstrate the related features of the process.     

Solar thermochemical reactions Ⅱ: Synthesis of 2-aminothiophenes via Gewald reaction induced by solar thermal energy

Green conditions have been developed for the synthesis of substituted 2-aminothiopbenes employing multicomponent reactions of a ketone with active methylene nitrile and elemental sulphur induced by free solar thermal energy.     

Determination of GR-IN optical fibre parameters from transverse interferograms considering the refraction of the incident ray by the fibre

The core–cladding refractive index difference Δn and the index gradient profile parameter of the graded index (GR-IN) optical fibre has been determined. The curve representing the real path, in the core region, due to refraction of the beam and the change in the exit wavefront are considered. Multiple-beam Fizeau fringes and two-beam interference Pluta polarizing microscope, are used for this investigation. A new method, based on a derived mathematical expression, is used with a prepared computer program to estimate the fringe shift inside GR-IN of the fibre core. The estimated and experimental values of the fringe shift, along the core radius, are used to obtain the refractive index profile of the optical fibre. A comparison between the new method and a previous model considering the refraction has been shown. Microinterferograms are given for illustrations.