Characterization and de novo sequencing of Atlantic salmon vitellogenin protein by electrospray tandem and matrix-assisted laser desorption/ionization mass spectrometry

Vitellogenin (VTG) is a protein produced by the liver of oviparous animals in response to circulating estrogens. In the plasma of males and immature females, VTG is undetectable. VTG has been used as a biomarker for exposure to endocrine disruptors in many species. In the present study, characterization of intact Atlantic salmon VTG was effected using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI ToF MS). Tryptic digest peptides were analyzed by MALDI ToF MS to obtain a peptide mass fingerprint. De novo sequencing of the tryptic peptides used low-energy collisionally-induced dissociation (CID) in an electrospray ionization quadrupole-ToF orthogonal hybrid mass spectrometer (ESI Q-ToF MS/MS). The interpretation of the product-ion spectra obtained from the ESI Q-ToF MS/MS was done by Lutefisk, a computer-based software algorithm. The molecular mass of the intact protein was found to be 187335 Da. A total of 14 tryptic peptides were sequenced and compared with the complete rainbow trout VTG and the partial Atlantic salmon VTG sequences found in the Swiss-Prot database. De novo sequencing by CID MS/MS of 11 Atlantic salmon tryptic digest peptides with selected precursor ions at m/z 788.24, 700.20, 794.75, 834.31, 889.28, 819.79, 865.27, 843.81, 572.20, 573.66 and 561.68 showed high homology with the known sequence of rainbow trout VTG. The last two precursor peptide ions, found at m/z 573.66 and m/z 561.68, also specifically matched the known portion of the Atlantic salmon VTG sequence. Finally, three tryptic precursor peptide ions found at m/z 795.18, 893.28 and 791.05, provided product-ion spectra, which were exclusive to the unsequenced portion of the Atlantic salmon VTG.     

Homogeneous liquid-liquid extraction method for the selective separation and preconcentration of ultra trace molybdenum

A new simple and efficient homogeneous liquid-liquid extraction method for the selective separation and preconcentration of molybdenyl ions was developed. α-Benzoin oxime (ABO) was investigated as a complexing ligand, and perfluorooctanoate ion (PFOA⁻) was applied as a phase-separator agent under strongly acidic conditions. Under the optimal conditions ([ABO] = 2.1 × 10⁻³ M, [PFOA⁻] = 1.8 × 10⁻² M, [HNO₃] = 1.7 M, [acetone] = 11.8% (v/v)), 10 μg of molybdenum in 5 ml aqueous phase could be extracted quantitatively into 40 μl of the sedimented phase. The maximum concentration factor was 125-fold. Thiocyanate was applied as a chromogenic reagent for the direct spectrophotometric determination of molybdenum in the sedimented phase. The reproducibility of the proposed method is at the most 2.4%.The influence of the type and concentration of acid solution, the concentration of ABO, the type and volume of the water-miscible organic solvent, the concentration of PFOA⁻, and the effect of different diverse ions on the extraction and determination of molybdenum(VI) were investigated. The proposed method was applied to the extraction and determination of molybdenum(VI) in natural water, Spinach, and Lucerne samples. A satisfactory agreement exists between the results obtained by the proposed method and those reported by GF-AAS.     

Dielectric relaxation of ball-like labels in polyisoprene

Ball-like molecules with strong dipoles (labels) were mixed with synthetic polyisoprene (IR305) in low concentrations (<1%) and measured dielectrically in the frequency range 10^–2–10⁷ Hz and the temperature range –70–0°C (glass relaxation region). Calorimetric measurements showed that this type of label has a plasticizing effect on the polymeric matrix. The dielectric measurements showed that these ball-like molecules relax through cooperative rotations with the polymeric segments and at the same relaxation frequency. In addition, the label molecules showed a high-frequency local relaxation process. The relaxation strength ratio of the local process (X _local) to the total relaxation strength of the label was found to be dependent on the volume as well as on the shape of the label. A comparison between the relaxation behaviors of the ball-and rod-like molecules, having the same volume, showed that the length of the label is also an important parameter for the determination of the local contribution as well as of the cooperative relaxation mechanism of the label. The label relaxation process is discussed in relation to the molecular packing of the host polymer.     

Amplified detection of nucleic acids and proteins using an isothermal proximity CRISPR Cas12a assay

Herein, we describe an isothermal proximity CRISPR Cas12a assay that harnesses the target-induced indiscrimitive single-stranded DNase activity of Cas12a for the quantitative profiling of gene expression at the mRNA level and detection of proteins with high sensitivity and specificity. The target recognition is achieved through proximity binding rather than recognition by CRISPR RNA (crRNA), which allows for flexible assay design. A binding-induced primer extension reaction is used to generate a predesigned CRISPR-targetable sequence as a barcode for further signal amplification. Through this dual amplification protocol, we were able to detect as low as 1 fM target nucleic acid and 100 fM target protein isothermally. The practical applicability of this assay was successfully demonstrated for the temporal profiling of interleukin-6 gene expression during allergen-mediated mast cell activation.

Herein, we develop an isothermal proximity CRISPR Cas12a assay that harnesses the target-induced collateral cleavage activity of Cas12a for the quantitative profiling of gene expression and detection of proteins with high sensitivity and specificity.     

A new design of the elimination and choice translating reality method for multi-criteria group decision-making in an intuitionistic fuzzy environment

The purpose of this paper is to design a new extension of the ELECTRE, known as the elimination and choice translating reality method, for multi-criteria group decision-making problems based on intuitionistic fuzzy sets. This method is widely utilized when a set of alternatives should be identified and evaluated with respect to a set of conflicting criteria by reflecting decision makers’ (DMs’) preferences. However, handling the exact data and numerical measure is difficult to be precisely focused because the DMs’ judgments are often vague in real-life decision problems and applications. A more realistic and practical approach can be to use linguistic variables expressed in intuitionistic fuzzy numbers instead of numerical data to model DMs’ judgments and to describe the inputs in the ELECTRE method. The proposed intuitionsitic fuzzy ELECTRE utilizes the truth-membership function and non-truth-membership function to indicate the degrees of satisfiability and non-satisfiability of each alternative with respect to each criterion and the relative importance of each criterion, respectively. Then, a new discordance intuitionistic index is introduced, which is extended from the concept of the fuzzy distance measure. Outranking relations are defined by pairwise comparisons and a decision graph is depicted to determine which alternative is preferable, incomparable or indifferent in the intuitionistic fuzzy environment. Finally, a comprehensive sensitivity analysis is employed to further study regarding the impact of threshold values on the final evaluation, and a comparative analysis is demonstrated with an application example in flexible manufacturing systems between the proposed ELECTRE method and the existing intuitionistic fuzzy technique for order preference by similarity to ideal solution (IF-TOPSIS) method.     

Chromium(VI) Based Oxidants III.1,2 Ferric Dichromate,Polyvinylpyridine Supported Zinc,and Ferric Dichromates as New Oxidizing Agents

Ferric dichromate, polyvinylpyridine supported zinc and ferric dichromates complexes are prepared and used as stable, mild, and efficient oxidizing reagents for the oxidation of different organic compounds. Ferric dichromate complex is more efficient.     

Silver nanoparticles immobilized onto poly(4‐vinylpyridine)‐functionalized magnetic nanoparticles: A robust magnetically recyclable catalyst for oxidant‐free alcohol dehydrogenation

A heterogeneous and recyclable catalyst with a high loading of silver nanoparticles was synthesized via the silver nanoparticles being supported onto the surface of magnetic nanoparticles coated with poly(4‐vinylpyridine). The synthesized catalyst was used in the dehydrogenation of alcohols to corresponding carbonyl compounds. A broad diversity of alcohols was converted into their corresponding carbonyl compounds in excellent yields. The catalyst was easily recovered by applying an external magnetic field and reused for seven reaction cycles without considerable loss of activity. The catalyst was fully characterized using various techniques.