Photodynamic activity of a nickel diimine complex and its interaction with DNA

Employing 1,3-diphenylisobenzofuran as a probe, the photodynamic activity of a nickel diimine complex, bis(o-diiminobenzosemiquinonato)nickel(II), in red light has been studied by fluorescence spectra. These results show that the nickel complex can generate singlet oxygen efficiently after irradiation in red light. The interaction of the metal complex with DNA has also been studied by electronic absorption spectra, fluorescence spectra and viscosity measurements. The electronic spectra of the metal complex exhibit dramatic hypochromism on interaction with DNA. Scatchard plot analyses indicate that the metal complex can competitively inhibit the binding between DNA and ethidium bromide. Viscosity experiments show that the binding of the metal complex increases the relative viscosity of DNA. These results suggest that the photoactive nickel diimine complex may interact with DNA by intercalation binding mode. Potential applications of the complex in photodynamic therapy are discussed.     

褐藻糖胶的提取纯化及其抗凝血活性的研究

从海带中提取出的水溶性提取物，经超滤，醇析获得褐藻糖胶粗多糖，以DEAE-Sephrose Fast Flow离子交换层析分离纯化，得到了F1，F2和F3三个级分，这三个级分具有明显的抗凝血作用。尤其是F3级分，能显地延长激活部分凝血活酶时间和凝血酶时间。     

槲皮素-铝配合物的制备及其与BSA的结合作用

在甲醇溶液中合成了槲皮素-铝配合物(Que-Al),并用紫外-可见吸收光谱和红外光谱进行了表征;运用荧光光谱探讨了Que-Al与牛血清白蛋白(BSA)的相互作用;求得了结合常数KA和热力学参数△H、△G、和△S.结果表明,Que-Al对BSA具有荧光猝灭作用,其猝灭方式为动态猝灭;Que-Al与BSA之间的作用力主要为疏水作用力.     

Voltammetric study of the interaction of the ofloxacin–copper complex with DNA, and its analytical application

The electrochemical behavior of the ofloxacin–copper complex, Cu(II)L₂, at a mercury electrode, and the interaction of DNA with the complex have been investigated. The experiments indicate that the electrode reaction of Cu(II)L₂ is an irreversible surface electrochemical reaction and that the reactant is of adsorbed character. In the presence of DNA, the formation of the electrochemically non-active complexes Cu(II)L₂-DNA, results in the decrease of the peak current of Cu(II)L₂. Based on the electrochemical behavior of the Cu(II)L₂ with DNA, binding by electrostatic interaction is suggested and a new method for determining nucleic acid is proposed. Under the optimum conditions, the decrease of the peak current is in proportional to the concentration of nucleic acids in the range from 3 × 10⁻⁸ to 3 × 10⁻⁶ g · mL⁻¹ for calf thymus DNA, from 1.6 × 10⁻⁸ to 9.0 × 10⁻⁷ g · mL⁻¹ for fish sperm DNA, and from 3.3 × 10⁻⁸ to 5.5 × 10⁻⁷ g · mL⁻¹ for yeast RNA. The detection limits are 3.3 × 10⁻⁹, 6.7 × 10⁻⁹ and 8.0 × 10⁻⁹ g · mL⁻¹, respectively. The method exhibits good recovery and high sensitivity in synthetic samples and in real samples.     

Interaction of fibrinogen with n-alkylagaroses and its purification by critical hydrophobicity hydrophobic interaction chromatograpy

A rational application of hydrophobic interaction chromatography (HIC) to the purification of proteins has remained an enigma in spite of over 30 years of research. The critical hydrophobicity parameter, which can be determined from a concentration series of n-alkyl Sepharose 4B (Seph-Cn) offers the possibility of adapting the HIC gel to the needs of purification. To this end a library of HIC gels (Seph-C4 to Seph-C6) of different immobilized alkyl residue concentrations was synthesized and tested with purified bovine fibrinogen. Binding of fibrinogen to such a concentration series resulted in sigmoidal binding curves. Analysis of the Seph-C5 data according to the lattices-site binding model yielded adsorption coefficients (nS) between 5 and 10 indicating that 5-10 lattice-sites (alkyl residues) interact multivalently with a fibrinogen molecule for adsorption at low ionic strength. The apparent lattice-site half-saturation constant of dissociation lies between 21 and 25 micromol/ml packed gel. For each alkyl chain length a critical hydrophobicity could be determined. For fibrinogen purification the critical hydrohobicity gel, Seph-C5 (13 micromol/ml packed gel), was selected. With the help of the cosolvents NaCl or glycine a fully reversible adsorption of fibrinogen could be facilitated on the critical hydrophobicity gel. Application of the method to human and bovine blood plasma resulted in a single step purification of fibrinogen in high yields. A comparison of the classical purification of fibrinogen with the critical hydrophobicity HIC (CHIC) method demonstrates a reduction in preparation time from several days to ca. 1 h. The subunit structure of HIC-purified human fibrinogen is identical to the classically purified protein. In the case of bovine fibrinogen however HIC-purified fibrinogen displayed a different subunit structure in that the Aalpha chain of fibrinogen had a ca. 5 kDa higher molecular mass. This may be due to the rapidity of the new one-step method and an avoidance of proteolysis.     

Study on the interaction of a palladium complex with DNA

The complex [Pd(bipy)Cl₂] (1) (bipy = 2,2′-bipyridyl) has been synthesized and characterized by NMR spectroscopy, elemental analysis and X-ray diffraction method. The first step hydrolysis reaction kinetics for the complex was studied by UV-absorption spectroscopy; the speed constant (k ₁) was found to be 3.0×10^?4 s^?1. The fluorescence spectra have been collected to investigate the interaction of complex (1) with fish sperm DNA (FS-DNA) and the results indicate that the complex (1) has an effective intercalation within DNA. The reaction of complex (1) with adenine in ethanol/water results in the compound [Pd₂(bipy)₂(ade)₂]Cl₂·3H₂O (2) (ade = adenine) whose crystal structure was determined by X-ray diffraction method. The structure is orthorhombic, Pmmn, a = 12.993(4) Å, b = 14.512(5) Å, c = 9.837(3) Å, V = 1854.8(11) ų, Z = 2 (C₃₀H₃₀Cl₂N₁₄O₃Pd₂), final R ₁ = 0.0675. The palladium complex is a binuclear cation, where two ade ligands bridge two Pd(II) centers, while each Pd(II) is also chelated by one bipy ligand.     

Isolation,purification, and properties of thioredoxin fromAnkistrodesmus braunii

Two forms of thioredoxin possessing the capacity for reactivating glutamine synthetase have been isolated from the cells of a green alga. Thioredoxins I and II are heat-stable proteins with molecular masses of 12 and 24 kDa, respectively. Analysis of the amino acid compositions of (I) and (II) have shown that they each contain two cysteine residues participating in the reduction of the oxidized form of glutamine synthetase.Institute of Microbiology, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 79–83, January–February, 1990.     

Isolation and purification of digalactosyldiacylglycerols

Summary A convenient method for the isolation of digalactosyldiacylglycerols from plant material has been developed using solid phase extraction. Commerical cartridges were utilized to prepare 100 mg fractions of galactolipids containing more than 97% of digalactosyldiacylglycerol. The purity of the fractions was monitored by an isocratic normal phase analytical HPLC procedure, which was optimized using factorial design. The optimized system was successfully scaled up to a semi-preparative HPLC system with a capacity of about 10 mg/injection which was used to further purify the digalactosyldiacylglycerol fraction.     

Isolation and purification of alkaloids

A literature review is given of methods of isolating and purifying alkaloids. Material is collected on the influence of the nature of the solvent on the isolation of alkaloids and on the interaction of alkaloids with acids and alkalis.All-Union Scientific-Research Institute of Medicinal Plants, Moscow. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 415–429, July–August, 1983.     

Isolation,purification, and characterization of ferritin from human liver

Ferritin from human liver has been isolated and purified by thermal denaturation and salt precipitation in combination with the methods of ion-exchange and gel permeation chromatography. The partial physicochemical characterization of this protein has been performed. It is proposed to use it in the creation of a radio-immunological method of determining the concentration of ferritin in human blood serum.Institute of Hematology and Blood Transfusion, Ministry of Health of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 96–99, January–February, 1986.     

In vitro antitumor activity of titanocene alanine complex and its interaction with nucleic acids

Titanocene alanine complex, Cp2Ti(O2CCH(CH3)NH3+Cl-)2, has been found to have in vitro antitumor activity against Ehrlich ascites tumor cells. Study of the interaction between Cp2Ti(O2CCH(CH3)NH+Cl-)2 and DNA by UV-VIS spectra, fluorescence spectra, agarose gel elec-trophoresis reveals that Cp2Ti(O2CCH(CH3)NH3 Cl-)2 forms a stable compound with DNA in molar ratio 1:1 and reduces the migration rate of ccc-DNA band and oc-DNA band. However, unlike cis-platin, the complex does not induce nicking of DNA even after a long incubation at a high ratio of the complex to DNA. The results suggest that titanocene-based antitumor agents have different mechanism of action from cisplatin-like antitumor agents.     

从猪血中分离纯化高纯度的猪血红蛋白

为了从猪血中分离纯化高纯度的猪血红蛋白,建立了通过超滤、DEAE-Sepharose Fast Flow离子交换色谱和Sephadex G-75凝胶 排阻色谱三步法制备高纯度猪血红蛋白的方法,并通过十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、高效凝胶排阻色谱和反相高 效液相色谱方法,对纯化后的猪血红蛋白进行了鉴定。经三步分离纯化后,猪血红蛋白的纯度大于99%,含量为1.328 g/L。     

Simple method for the isolation and purification of hemoglobin components.

A simple method for the isolation and purification of hemoglobin components from starch gel by electrophoresis is described. An inverted bottle with a cut off bottom is used. Pieces of gel containing the hemoglobin are embedded in potato starch, wetted with buffer, and placed in the neck of the inverted bottle, the mouth of which has been closed with filter paper and tape. Connection with the negative tank buffer is achieved by inserting a layer of starch gel between the potato starch and the negative tank buffer. By electrophoresis, the hemoglobin migrates and accumulates in a dialysing tube tied around the mouth of the bottle and hanging into the positive tank buffer.     

Immobilized hemoglobin in the purification of hemoglobin-based oxygen carriers.

Chemically modified hemoglobins can be used as oxygen carriers in cell-free fluids provided that they have a low oxygen affinity and are stable towards dissociation into subunits. The latter species are undesirable because they are filtered rapidly through the kidneys, have renal toxicity and are characterized by a high oxygen affinity. A most important step in the preparation of hemoglobin-based oxygen carriers is therefore their purification from any dissociable material. Hemoglobin immobilized as alpha beta dimers on Sepharose lends itself naturally to this purpose as it is able to interact in a specific and reversible way with soluble alpha beta dimers. Hemoglobin affinity columns are very effective in the purification of cross-linked and pseudo-cross-linked human and bovine hemoglobin. The applicability of the technique is enhanced by the ease with which alpha beta dimers from different species cross-interact to yield hybrid alpha 2 beta 2 tetramers. It is shown that hemoglobin affinity columns may provide analytical information on the cross-linking reaction itself.