Sephadex-bound histamine in the catalysis of ester hydrolysis

Ester hydrolysis by Sephadex-bound catalysts was studied in a flow-through system. Three different immobilized preparations were synthesized and used: histamine-, coimmobilized histamine-octylamine-, and octylamine-Sephadex; octylamine-Sephadex was used as a reference. Immobilization was carried out using water-soluble carbodiimide, which gave amide linkages between carboxymethyl Sephadex and the groups attached. It was found that the coimmobilized histamine-octylamine preparation was three times more efficient than immobilized histamine alone in the hydrolysis of the esterp-nitrophenylcaproate, whereas hardly any difference was found in the hydrolysis of the less hydrophobic substratep-nitrophenylacetate. We attribute this enhancement of the hydrolysis ofp-nitrophenylcaproate to local enrichment of the substrate on the histamine-octylamine matrix caused by the presence of hydrophobic octyl groups.     

Study of modified Sephadex-bound insulin in animal experiments

Preparations of insulin covalently coupled to Sephadex were obtained using modified aldehyde-containing Sephadex carriers. In all cases carriers used had the same structure but differed in molecular weight. Depending on the reaction conditions insulin might or might not be split from the carrier. In animal experiments it was shown that in the homogeneous state all preparations retain high biological activity independent of molecular weight of the carriers, which varied from 10,000 to 150,000. The binding of insulin with polymeric carriers prolonged the time of its action from 5-6 to 14-16 h.     

Separation of tin from some metals by extraction chromatography

Much attention has been devoted to Sn (IV) strongly retained on the TBP-Daiflon column from 2M HCl in extraction chromatography. The separations of Sn?Cu, Zn, As, Cd, Sb, Pb,¹¹³Sn-¹²⁵Sb,¹¹³Sn-^113mIn (^113mIn milking) and Sn?Hg?Fe were successfully achieved without any contamination. In the separations, except for the last, only tin was retained separately on the column upon passing the mixed solution. The daughter indium was eluted with 0.5M HCl. In the last separation, iron was eluted with 0.5 M HCl, tin with 0.1M HCl and mercury with 2M HNO₃, for these metals retained on the column. Radioactive tracers for tin, iron, mercury and antimony were used.     

Isolation of nonprotamine proteins from the nuclei of the gonad cells of the Russian sturgeonAcipenser güldenstadti

Eight nonprotamine proteins have extracted from the nuclei of the cells of the gonads of the Russian sturgeon with 0.35 M sodium chloride solution followed by fractionation of carboxymethyl-Sephadex G-25 and desalting on Bio-Gel P-2, and their amino acid compositions have been determined.     

Phosphatase activities of fractions of the venom of Renard's viperVipera ursini renardi

The separation of the venom of Renard's viper by filtration through Sephadex G-75 gel has been performed and phosphatase activities have been determined in the fractions and in the course of separation. The bulk of the activity of the phosphatases investigated was concentrated in the high-molecular-weight fractions I and II. A correlation has been found in the distribution of the DNase and ATP-pyrophosphatase activities with the phosphodiesterase activity in the venom fractions. There is no correlation in the distribution of the RNase and phosphodiesterase activity.     

Extraction and chemical investigation of jute (Corchorus olitorius,Linn.) seed protein

Studies were made on the protein solubility of deoiled jute (Corchorus olitorius, Linn.) seed in aqueous solution over various pHs and in different concentrations of NaCl at pH 8.0. Chemical analysis of the seed protein showed 16 amino acids, of which 9 were essential. Gel filtration on Sephadex G-200 revealed the presence of four components, and their molecular weights were determined by two standard methods. Extractable jute seed proteins in salt solution were separated into six fractions electrophoretically (SDS-PAGE). The molecular weights of the six fractions were found to be 118,000; 103,000; 96,000; 67,500; 48,500; and 15,000.     

Purification of nerve growth factor from the venom of the Central Asian cobraNaja oxiana

A highly purified electrophoreticaly homogeneous protein with a NGF activity of 10·10⁵ BU/mg of protein have been isolated from the venom of the Central Asian cobra by gel-filtration and ion-exchange chromatography followed by preparative isolectric focusing in a thin layer of Sephadex. It has been shown that the NGF isolated is characterized by a molecular weight in the range of 20–30 kD and a pI value of about 7.0.     

Amylase inhibitors ofActiniae of the Caribbean Sea amylase inhibitor of protein nature fromStoichactis helianthus

Natural amylase inhibitors have been found in actiniae of the Caribbean Sea. From the actiniaS. helianthus an inhibitor has been isolated by gel filtration on Sephadex G-50 and ion-exchange chromatography on CM-cellulose that is highly specific in relation to the amylases of marine mollusks and is inactive in relation of the amylases from other sources.     

Coupling of effector molecules to solid supports

In view of the limited stability of the isourea bond, formed in ligand coupling to CNBractivated polysaccharides, an alternative to this current activation method has been developed. 2,4,6-Trifluoro-5-chloropyrimidine (FCP), known as a reactive group in reactive dyes, was used to activate Sepharose. Under appropriate conditions a thermally stable product with unimpaired beaded structure was obtained, which was reactive toward amines and mercaptans. Coupling with hexamethylenediamine, aniline, and ethanethiol, respectively, yielded an incorporation of 0.2-2.7, 0.9-1.7, and 1.1 mmol ligand/g dry agarose. The stability of immobilized ligands based on FCP-Sepharose between pH 4 and 8 was about 200 times higher as compared to products originating from CNBr-Sepharose; ligand leakage was only 0.5 x 10^p-3%/h. The possibility of obtaining a high degree of substitution is a further advantage of the FCP activation. In addition, the FCP-activated Sepharose can be stored in the wet state at 4°C without substantial decrease in coupling capacity. The FCP analogs 2,4,5,6-tetrachloro- and 2,4,5,6-tetrafluoropyrimidine, and other polymers (cellulose, Sephadex, aminomethylpolystyrene) appeared to be applicable also.     

Flavin-protein interaction in bound glucose oxidase

Glucose oxidase was bound to Sepharose, Sephadex, gelatin, and dextran, yielding immobilized soluble and insoluble derivatives of the enzyme. The soluble preparations possessed higher enzymic activity than the analogous insoluble ones. The reversible dissociation process of the bound enzyme into apoenzyme and flavin adenine dinucleotide (FAD) was studied with the soluble and insoluble glucose oxidase in relation to enzymic activity and conformational changes as measured by circular dichroism and fluorescence methods. Bound apoenzyme was found to be more stable than the apoenzyme obtained from the unmodified glucose oxidase. The binding constant of FAD in bound glucose oxidase (K_diss≈10^-8M) calculated from fluorescent studies was lower than that of FAD in the native enzyme (K_diss10^-10M). The circular dichroism measurements indicated that dextran-bound glucose oxidase has a conformation similar to that of the native enzyme.     

Solid-phase microextraction/gas chromatography–mass spectrometry method optimization for characterization of surface adsorption forces of nanoparticles

A complete characterization of the different physico-chemical properties of nanoparticles (NPs) is necessary for the evaluation of their impact on health and environment. Among these properties, the surface characterization of the nanomaterial is the least developed and in many cases limited to the measurement of surface composition and zetapotential. The biological surface adsorption index approach (BSAI) for characterization of surface adsorption properties of NPs has recently been introduced (Xia et al. Nat Nanotechnol 5:671–675, 2010; Xia et al. ACS Nano 5(11):9074–9081, 2011). The BSAI approach offers in principle the possibility to characterize the different interaction forces exerted between a NP's surface and an organic—and by extension biological—entity. The present work further develops the BSAI approach and optimizes a solid-phase microextraction gas chromatography–mass spectrometry (SPME/GC-MS) method which, as an outcome, gives a better-defined quantification of the adsorption properties on NPs. We investigated the various aspects of the SPME/GC-MS method, including kinetics of adsorption of probe compounds on SPME fiber, kinetic of adsorption of probe compounds on NP's surface, and optimization of NP's concentration. The optimized conditions were then tested on 33 probe compounds and on Au NPs (15 nm) and SiO₂ NPs (50 nm). The procedure allowed the identification of three compounds adsorbed by silica NPs and nine compounds by Au NPs, with equilibrium times which varied between 30 min and 12 h. Adsorption coefficients of 4.66?±?0.23 and 4.44?±?0.26 were calculated for 1-methylnaphtalene and biphenyl, compared to literature values of 4.89 and 5.18, respectively. The results demonstrated that the detailed optimization of the SPME/GC-MS method under various conditions is a critical factor and a prerequisite to the application of the BSAI approach as a tool to characterize surface adsorption properties of NPs and therefore to draw any further conclusions on their potential impact on health. Graphical Abstract

The basic principle of SPME/GC-MS method for characterization of nanoparticles surface adsorption forces     

Penning ionization electron spectrometry of hydrogen chloride

An electron spectrometric study has been performed on HCl using metastable helium and neon atoms as well as neon resonance photons. High resolution electron spectra were obtained with two different beam apparatuses for a mixed He(2¹ S, 2³ S) beam, a pure He(2³ S) beam, and, for the first time, state-selected pure Ne(3s ³ P ₂) and pure Ne(3s ³ P ₀) beams, and for NeI resonance photons. For the system He(2³ S)+HCl the vibrational populationsP(υ′) of the formed HCl⁺ (X ²∏ _i , υ′) and HCl⁺ (A ²Ω⁺, υ′) ions are found to differ from the Franck-Condon factors for unperturbed potentials, indicating slight bond stretching in HCl upon He(2³ S) approach. For He(2¹ S)+HCl the vibrational peak shapes and vibrational populations are substantially different from the He(2³ S) case, pointing to an additional, charge exchanged interaction (He⁺+HCl^?) in the entrance channel of the former system. For the first time, we have detected the electrons in both the He(2¹ S)+HCl and He(2³ S)+HCl spectra associated with the major mechanism for the formation of Cl⁺ ions: energy transfer to repulsive HCl** Rydberg states, dissociating toH(1s) and autoionizing Cl**(¹ D ₂ nl) atoms. For both Ne(³ P ₂)+HCl and Ne(³ P ₀)+HCl, the populationsP(υ′) of both final molecular states HCl⁺ (X, A) agree closely with the Franck-Condon factors at the average relative collision energyē _coll=55 meV and, for HCl⁺ (A ²Ω⁺), also atē _coll=130 meV.     

Seed proteins of Ricinus communis. II. Isolation and comparative characterization of the acid form of ricin

An acid form of ricin — ricin T — with a molecular weight of 58 kDa, an isoelectric point, pI, of 7.0–7.1, and a sedimentation coefficient of 4.60 S has been isolated from the seeds of the Central Asian castor-oil plantRicinus communis by gel filtration on Sephadex G-75 followed by ion exchange chromatography on DEAE- and CM-celluloses. The amino acid composition and the N-terminal amino acid residues of ricin T have been determined.     

Heat of reaction and curing of epoxy resin

The heat of reaction and kinetics of curing of diglycidyl ether of bisphenol-A (DGEBA) type of epoxy resin with catalytic amounts of ethylmethylimidazole (EMI) have been studied by differential power-compensated calorimetry as a part of the program for the study of process monitoring for composite materials. The results were compared with those from 1∶1 and 1∶2 molar mixtures of DGEBA and EMI. A method of determination of heat of reaction from dynamic thermoanalytical instruments was given according to basic thermodynamic principles. The complicated mechanism, possibly involving initial ionic formation, has also been observed in other measurements, such as by time-domain dielectric spectroscopy. The behavior of commercially available DGEBA resin versus purified monomeric DGEBA were compared. The melting point of purified monomeric DGEBA crystals is 41.4 °C with a heat of fusion of 81 J/g. The melt of DGEBA is difficult to crystallize upon cooling. The glass transition of purified DGEBA monomer occurs around ?22 °C with aΔC _p of 0.60 J/K/g.     

Cell-free protein synthesis by posterior silk gland polyribosome

Polyribosome was prepared from the posterior gland of silkworms, where silkfibroin is specifically synthesized. Coupled with the corresponding enzymes, amino acids, ATP and GTP, silkfibroin was produced in a cell-free system. The production of silkfibroin was monitored by SDS polyacrylamide gel electrophoresis of the dansylated polypeptides. The polysome fraction was adsorbed on DEAE Sephadex. Using the immobilized polysomes, a genetic information-transducing bioreactor for producing a specified protein was assembled. The promising features of this bioreactor are discussed.     

Structural and thermal properties of HDPE/n-AlN polymer nanocomposites

High-density polyethylene (HDPE) containing various volume fractions (0–20 vol%) of aluminum nitride nanoparticles (n-AlN) is prepared by melt mixing. Structural and morphological characterizations of the prepared composites are carried out by X-ray diffraction (XRD), high-resolution transmission electron microscopy (HR-TEM), and atomic force microscopy (AFM). Thermal stability and degradation kinetics of HDPE/AlN (nano) composites are investigated by Thermogravimetric analysis (TG). HR-TEM micrographs confirm fairly uniform dispersion of AlN nanoparticles, as well as the existence of long interconnected chain-like aggregates. AFM images also confirm homogeneous dispersion of n-AlN in the polymer matrix. Roughness analysis from the AFM data indicates the presence of substantial undulation from the mean surface level. Thermogravimetric data indicate small improvement in the thermal stability of the composites. Kinetic parameters, viz., the activation energy (E _a), frequency factor (A), and reaction order (n) are estimated using the isoconversional methods of Kissinger, Flynn–Wall–Ozawa (FWO), KAS, and Friedman. Activation energies (E _a) calculated by the above four models display nearly similar features and are enhanced by the presence of AlN nanoparticles. Kinetics of degradation of HDPE-AlN (nano) composites follows a first-order reaction.     

Adsorption chromatographic separation of radioiodine labelled iodothyronines

3,3′-diiodothyronine, 3,3′,5-triiodothyronine, 3,3′,5′-triiodothyronine and thyroxine was labelled with¹²⁵I and/or¹³¹I by the use of the chloramine T method.¹.² The labelled products were separated by adsorption chromatography using Sephadex LH-20 dextrangel as adsorbent and aqueous solution of ethanol as eluent.     

Gel-assisted synthesis of anatase TiO2 nanoparticles and application for electrochemical determination of L-tryptophan

Anatase titanium dioxide nanoparticles (TiO₂-NPs) were synthesized with and without gelatin via the sol-gel method. The TiO₂-NPs were characterized by a number of techniques, such as thermogravimetric analysis (TGA), X-ray diffraction analysis (XRD), transmission electron microscopy (TEM), fourier transform infrared spectroscopy (FT-IR) and ultraviolet visible spectroscopy (UV-Vis). The particle sizes of the TiO₂-NPs prepared with and without gelatin were ～13 and ～17 nm, respectively. The main advantage of using gelatin as a stabilizing agent is that it provides long-term stability for nanoparticles by preventing particles agglomeration. The results indicated that gelatin was a reliable green stabilizer, which can be used as a polymerization agent in the sol-gel method for synthesis of tiny size TiO₂-NPs. Moreover, the composite film was prepared by synthesized TiO₂-NPs nanoparticles and multi wall carbon nanotube (MWNT) on glassy carbon electrode (TiO₂-MWNT/GCE). The TiO₂-MWNT/GCE responded linearly to L-tryptophan (L-Trp) in the concentration of 1.0 × 10^?6 to 1.5 × 10^?4 M with detection limit of 5.2 × 10^?7 M at 3 using amperometry. The studied sensor exhibited good reproducibility and long-term stability.     

SDS 800 — The new data system for quantitative depth profiling of inhomogeneous samples by SIMS

More than 13 years of SIMS application field experience of numerous users of the ATOMIKA Ionmicroprobes have been the basis for the new SIMS Data System SDS 800. The hardware and software concept of the SDS 800, therefore, pays special attention to the following requirements:

1. Convenient set-up, modification and re-use of the measuring parameter sets for easy, time-saving operation.
2. Individual parameter selection from the very broad range of SIMS measuring parameters for optimum SIMS data quality.
3. Multitasking operation for simultaneous handling of SIMS measurement, data processing, data output and of auxiliary techniques.
4. Simultaneous depth profile/ion image acquisition and processing to enhance data quality and to validate data interpretation.
5. User-friendly data processing and output.