Modification of radiation resistance of Bacillus spores by water

Spores of Bacillus subtilis, containing plasmid (B. subtilis BD 366(pUB110, Trp C2 thr 5)), were prepared free of additives and the effect of water on their radiation sensitivity under aerobic condition was investigated. The moisture content of spores was controlled by (i) drying over CaCl₂ (Aw = 0.0), (ii) equilibrating with the vapor of saturated aqueous NaCl solution (Aw = 0.7), or (iii) suspending in water (Aw = 1.0). The resistance of spores to radiation increased with increasing moisture content. The analysis of spore DNA by agarose gel electrophoresis showed that DNA damage (strand breaks) decreased with increasing water activity of the spores, a finding concomitant with the observed survival rates.     

The liquid-liquid extraction using hydrophilic solvents occluded to an oil-adsorbent as an organic phase: extraction of 8-quinolinolato complexes of alkaline earth metals into tetrahydrofuran.

In order to use a hydrophilic solvent as an organic phase in liquid-liquid extraction by retaining it to a solid support, the occlusion and desorption of nine hydrophilic solvents, such as methanol, to three kinds of commercially available oil-adsorbent were examined. The combination of tetrahydrofuran and WOSEP made by fibrous polypropylene was finally selected, and was subjected to the extraction of alkaline Earth metals with 8-quinolinol. It was proved that Mg2+, Ca2+, and Sr2+ ions could be successfully extracted into the tetrahydrofuran phase in the form of 8-quinolinolato complexes, which could not be extracted into CHCl3.     

Enzyme immunoelectrode for insulin incorporating a membrane partially treated with water vapour plasma

An enzyme immunoelectrode for the amperometric determination of serum insulin is described. The device consists of an immunoreactive membrane combined with a hydrogen peroxide electrode. The surface of a microporous hydrophobic polypropylene membrane is modified by water vapour plasma treatment to make it partially hydrophilic. Subsequent treatment with octamethylenediamine and glutaraldehyde enables the surface of this membrane to interact with various proteins. Anchoring of the antibody to Protein A immobilized on the membrane was effective for immunoreactivity.     

Exopeptidase degradation for the analysis of phosphorylation site in a mono-phosphorylated peptide with matrix-assisted laser desorption/ionization mass spectrometry.

The utility of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) coupled with a peptide ladder sequencing method employing exopeptidase degradation for the analysis of phosphorylation site in a mono-phosphorylated peptide is investigated. MALDI-TOFMS analysis of time-dependent exopeptidase digestion using carboxypeptidase W and aminopeptidase M of the mono-phosphorylated 33-48 fragment isolated from a beta-casein tryptic digestion mixture allowed for the sequencing analysis from both the C-terminus and N-terminus. Negative ion detection MALDI-TOFMS made it possible to clearly measure the peptide ladder of mono-phosphorylated peptide by the strong negative charge localized at the phosphoric acid group. Since exopeptidase activity was suppressed by the existence of a phosphorylated amino acid residue, the termination exopeptidase degradation therefore suggested the existence of a phosphorylated amino acid residue at that site. This peptide ladder sequencing method using exopeptidases was effective for the identification of the site of a phosphorylated amino acid residue by a simple MALDI-TOFMS analysis in the negative ion detection mode.     

Parametrization of calcium binding site in proteins and molecular dynamics simulation on phospholipase A2

The empirical energy parameters for a calcium ion and its ligands in proteins were determined within a pairwise additive framework. The interaction energies of Ca²⁺-water, Ca²⁺-peptide group and Ca²⁺-carboxyl group systems were calculated using the ab initio molecular orbital method with basis sets of double zeta quality including polarization or diffuse functions. The resulting potential energy surfaces served as references for the determination of the nonbonded parameters in the empirical energy function. The nonadditive corrections for the Ca²⁺-ligand pair potentials are incorporated implicitly in the nonbonded paremeters by treating three-body (1:2 complex) or seven-body (1:6 complex) systems in reference calculations. Ligand polarizations induced by Ca²⁺ are estimated from the partial atomic charges of two-body (1:1 complex) systems. The charge sets were determined by scaling so as to reproduce the reference potential energy surfaces. The newly determined parameter set was used in a stochastic boundary molecular dynamics simulation of phospholipase A₂. The solvated structure of the Ca²⁺-binding site obtained from an X-ray crystallographic study is well reproduced by the parameter set.     

Rapid determination of fumagillin residues in honey by liquid chromatography-tandem mass spectrometry using the QuEChERS method

A new, rapid, and efficient method for determining the fumagillin residues in honey was developed. The samples extracted were analyzed using LC/MS/MS. Chromatographic separation of fumagillin was performed in gradient mode on a C8 column (100 x 2.0 mm, 5 microm) at 40 degrees C. The mobile phase consisted of a mixture of 2 mM ammonium formate-0.01% formic acid solution and methanol; the flow rate was set to 0.2 mL/min. Under these conditions, it was possible to measure fumagillin and its isomers as a single peak. The sample preparation procedure used is based on the QuEChERS (quick, easy, cheap, effective, rugged, and safe) method, which is fast (approximately 30 min) and uses less organic solvent. The fumagillin was extracted with acetonitrile containing 0.1% formic acid, then purified using a solid-phase extraction method with an Oasis mixed-mode weak anion-exchange cartridge. The overall recovery of fumagillin ranged from 88.1 to 99.4%; the intra- and interassay CVs were <4.5% and <4.9%, respectively. The LOQ was 0.1 microg/kg. LC/MS/MS coupled with the QuEChERS method showed strong potential as a method for determining fumagillin residues in honey.     

Thermal analysis graphite intercalation compounds with dichlorides of some d-elements: A.S. Skoropanov,G.S. Petrov,A.A. Vecher,Yu.N. Novikov and M.E. Vol'pin,Thermochim. Acta, 90 (1985) 339-342

Kinetic and thermodynamic aspects of the axial base dissociation of solid Co(salen) (X-py) complexes, X = H (1), 3-Me (2), 4-Me (3), 3,4-Me₂ (4), 3,5-Me₂ (5), 3-NH₂ (6), 3-Cl (7), 3-CN (8), 4-CN (9), have been investigated by means of TG-DSC and isothermal weight-loss measurements. These adducts endothermically dissociate the axial base giving rise to the oxygen-active Co(salen) complex. The axial base dissociation reactions fit the contracting disc equation and the kinetic compensation effect is observed for all the adducts excepting Adducts 4–6. For the remaining adducts the kinetic and thermodynamic stabilities of the Co-(X-py) bond are found to increase linearly with increasing Hammett's substitution constants of X except for Adducts 3 and 9. These results are discussed in terms of the σ and π interactions between cobalt(II) and substituted pyridine. Factors dominating the kinetic bond stability are briefly discussed.     

Determination of bithionol, bromophen, nitroxynil, oxyclozanide, and tribromsalan in milk with liquid chromatography coupled with tandem mass spectrometry

LC/MS/MS was developed to determine the residues of bithionol (BTN), bromofen (BMF), nitroxynil (NTX), oxyclozanide (OCZ), and tribromsalan (TBS) in milk. Samples were extracted with ethyl acetate and cleaned up by liquid-liquid separation with acetonitrile and n-hexane. The compounds were determined by RP-LC using a C18 column with 0.1% formic acid-methanol. Mass spectral acquisition was performed in the negative mode by applying selected-reaction monitoring. The method was validated in milk spiked with these compounds at 5-600 microg/kg; average recoveries were in the range 83.8-97.1%, with RSD values of 1.4-8.0%. The interassay RSDs were less than 11%. The LODs of these compounds in milk were 0.1 microg/kg. The method was applied to 24 raw milk samples. The concentration of these compounds in all samples was lower than the Japanese maximum residue limits. The method is rapid, sensitive, and specific for monitoring residues of BTN, BMF, NTX, OCZ, and TBS in milk.     

Photoinduced ionic polymerization. III. Cationic polymerization and copolymerization of cyclohexene oxide and α-methylstyrene in the presence of pyromellitic dianhydride

The photoinduced ionic polymerization of cyclohexene oxide was studied in the presence of pyromellitic dianhydride. The polymerization is initiated by the excited chargetransfer complex between cyclohexene oxide and the electron-acceptor and proceeds by a cationic mechanism. Photoinduced cationic polymerization of α-methylstyrene was also observed in the presence of pyromellitic dianhydride. The initiation mechanism of the polymerization was elucidated by means of electron spin resonance measurements. The concentration of pyromellitic dianhydride anion-radicals measured in this way was found to be proportional to the rate of polymerization. This result shows clearly that the photopolymerization is initiated by cation-radicals formed from photoexcited donoracceptor complexes. The attempted photocopolymerization of cyclohexene oxide and α-methylstyrene gave a mixture of homopolymers. The composition of the product depends on the wavelength of the light used.