DISTRIBUTIONS OF FREE RADICALS AMONG AMINO ACIDS FROM LYOPHILIZED ULTRAVIOLET-IRRADIATED PROTEINS

Abstract— The effects of u.v.-irradiation at 254 nm upon lyophilized ribonuclease, lysozyme, insulin, and chymotrypsinogen have been investigated by electron spin resonance (ESR). enzymatic assay, and labeling of free radical sites with tritiated hydrogen sulfide (HST). The ESR signal of the irradiated protein diminishes on exposure to HST, and tritium becomes covalently bound to carbon. The distribution of tritium among the amino acids of each protein. studied as an indicator of the carbon free radical distribution, differs markedly from those observed previously to result from exposure to gamma radiation, electrical discharge. or hydrogen atoms. However, the earlier observation that the tritium distribution is influenced by protein conformation holds true as well for u.v.-irradiation. Moreover, the distributions of tritium among the amino acids of u.v.-irradiated proteins indicate a broad scattering of free radicals. Tyrosine and phenylalanine, residues that absorb light energy in the region of the wavelength employed, are not particularly important as radical carriers. Thus, for ribonuclease, these residues incorporated 3.8 and 1.5 per cent of the total tritium, but they absorb 51 and 12 per cent of the light, respectively. These results, together with the observed low recoveries of methionine, an amino acid that does not absorb at 254 nm, add weight to the concept that a migration of energy ensues after the initial absorption of light energy and that photolytic damage may thus be due to destruction of amino acids other than those initially absorbing the u.v.-radiation.     

LA PHOTOOXYDATION DU GROUPE PEPTIDE—I. PROTÉINES FIBREUSES*

Abstract— The determination of the α-ketoacids, glyoxylic and pyruvic in partial hydrolysates of fibrous proteins (keratin and fibroin) after exposition to u.v. light has allowed us to suggest a photooxydation mechanism for the α-aminoacid residues of glycine and alanine in proteins. The primary reaction is believed to be the formation of the peptide radical -NH-CR-CO-, and the breakdown of polypeptide chains is explained by the formation of α-ketoacyl peptide linkages.     

ELECTRON SPIN RESONANCE STUDIES OF ULTRAVIOLET IRRADIATED KERATIN AND RELATED PROTEINS

Abstract— The formation of free radicals in keratin and related proteins by exposure to u.v. light has been examined by electron spin resonance spectroscopy. From a comparison of the spectra obtained and the effects of different wavelengths and the stabilities of the radicals produced, it has been shown that several species of free radicals are produced by the u.v. irradiation of keratin. However it has been possible to identify only those free radicals associated with the cystine residues. The free radicals produced in keratin by exposure to shorter wavelength irradiation (below 3250 Å) were found to be quite different to those produced at longer wavelengths.     

Study by electron paramagnetic resonance and spectrophotometry of absorption of the primary processes in photochemistry of frozen aqueous solutions at 77 degree K of aromatic amino acids and polypeptides

Abstract— Neutral, acidic or basic frozen aqueous solutions of aromatic amino acids undergo photoionisation under u.v. irradiation, at 77°K. In neutral or basic solutions, photo-ejected electrons are trapped in the solvent matrix and exhibit a characteristic absorption band in the visible region. In acidic solutions electrons are trapped by protons and ESR signal spectrum of hydrogen atoms may be observed. Hydrogen atoms are also produced in low yield in neutral or basic frozen aqueous solutions, u.v. irradiated at 77°K. In basic media the ESR spectrum of 0^- radical ions is observed. Kinetic studies as a function of light intensity show that photoionisation takes place after absorption of a second photon by the phosphorescent molecule in its lowest triplet state. Recombination of trapped electrons in neutral or basic solutions may be induced using secondary excitation with visible light. In all instances we could record the absorption spectrum of photolytic products of aromatic amino acids and polypeptides which are u.v. irradiated at 77°K.     

RETARDATION OF ULTRAVIOLET LIGHT ACCELERATED LEAF SENESCENCE BY A CYTOKININ: N6 BENZYLADENINE

O ne clear evidence of u.v. injury is the manifestation of chlorosis after leaf irradiation, and such chlorosis can be photorepaired by blue light[1]. We found that such u.v.-accelerated chlorosis could be prevented in the dark by applying N⁶ benzyladenine (BA), a synthetic cytokinin, either before or after u.v. irradiation. Cytokinins are known to retard natural senescence of detached leaves kept in the dark [2]. The present finding suggests that u.v. damage may accelerate the natural senescence process of detached leaves kept in the dark. N. glutinosa leaves were used in this experiment, because chlorosis develops quickly (within one week) when the mature leaves are detached and incubated in the dark. The plants were grown in a glasshouse without white-wash. The light intensity at noon often reached 9000 ft-c., and the temperature ranged from 19–35°C. The leaves shown in Fig. 1 were from mature plants, i.e. plants with the terminal growing point developed into a flower bud. The floral buds were cut off several days before the leaves were detached for exposure to u.v. light.     

Mechanisms of antioxidant action: Autosynergistic polymer-bound antioxidants as heat and light stabilizers for ABS

An antioxidant (3,5-di-tert-butyl-4-hydroxybenzyl mercaptan) and a u.v. stabilizer (4-benzoyl-3-hydroxyphenyl-O-ethylthioglycollate) containing the thiol group have been reacted with ABS latex to give polymers with exceptionally high heat and light stability. Both chemical and physical phenomena contribute to the high activity of the bound antioxidants. The concentration of the molecularly dispersed antioxidants in the most oxidatively vulnerable part of the polyblend (the polybutadiene segment) has an important role in determining stabilizer efficiency and, particularly under high temperature conditions, the immobilization of the antioxidants by chemical bonding leads to much lower rates of physical loss. The autosynergistic hindered phenol is primarily responsible for the thermal stabilizing activity of the combination but it also powerfully augments u.v. stabilizing effectiveness of the u.v. stabilizer.     

THERMOLUMINESCENCE OF NUCLEIC ACID CONSTITUENTS IN SOLUTION AFTER U.V.-IRRADIATION

Abstract— The thermoluminescence of purines and pyrimidines has been studied in ethylene glycol (EG)/water (2:1) glass matrices after ultraviolet (u.v.) irradiation at 77°K. An investigation of the mechanism of thennoluminescence has been made for adenine. The thermoluminescence involved biphotonic ionisation and recombination. The thermoluminescence depended on the square of the excitation light intensity, and the observed lifetime of the second photonabsorbing intermediate was the same as the phosphorescence lifetime. The emission spectrum lay in the same wavelength region as the phosphorescence spectrum. It was possible to bleach the thennoluminescence with visible light.     

藉L-半胱氨酸包覆的ZnS纳米粒子的共振光散射定量分析蛋白质

合成和表征了功能性L-半胱氨酸包覆的ZnS纳米粒子。在pH5．12的NaAc-HAc溶液介质中,L-半胱氨酸包覆ZnS纳米粒子于波长308．0nm处出现共振光散射峰。一定量蛋白质的加入能明显增强体系的共振光散射,且峰强度增加值与蛋白质浓度间存在良好线性关系,据此建立了一种灵敏的测定微量蛋白质的方法。用L-半胱氨酸包覆ZnS纳米粒子作为探针,不仅克服了有机染料可能出现的光漂白等缺点,而且本身不具毒性。该法用于人血清试样中总蛋白的测定,其结果与临床数据一致。     

Change in the concentration of free radicals on the photolysis of lignin

It has been shown that under the action of UV light on solid preparations of lignin the generation of electron paramagnetic centers (EPCs) takes place, their concentration rising with a fall in the wavelength of the exciting light. In a dark stage there is a slow death of EPCs with time. The reactivities of individual model compounds of lignin in radical reactions in solutions were followed with the aid of a stable nitrogen radical used as a spin label. The results show that the efficiency of quenching differs considerably for individual chromophores and decreases in the sequence benzoquinone, vanillin, quinone methide, Björkman lignin, and soda lignin. The change in the concentration of EPCs of the stable radical with time is described by a first-order equation.Siberian Pulp and Paper Scientific-Research Institute, Bratsk. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 236–239, March–April, 1980.     

Chemistry of dithiocarbamate derivatives of amino acids. Part II. Complexes of nickel(II) with derivatives of branched and cyclic amino acids

Summary Barium salts of the dithiocarbamate derivates of the amino acids: DL-valine, L-valine, DL-leucine, DL-isoleucine and L-proline have been synthesized. The anions have been used to obtain the corresponding nickel(II) complexes in acid form. The complexes are diamagnetic, and coordination takes place in a near-square planar geometry around the nickel(II) ion through the sulphur atoms of the dithiocarbamate moiety, the structure being confirmed by i.r.,¹H n.m.r., u.v.-vis spectros copies and by chemical analysis.No significant difference has been found between the physicochemical properties (i.r. and 'H n.m.r. spectra and magnetic properties) of the complexes prepared from the dithiocarbamate derivatives of DL- and L-valine, but d-d bands in the u.v.-vis spectra show a slight different intensity.     

THE PHOTOCHEMICAL INTERACTION OF DEOXYRIBONUCLEIC ACID AND PROTEIN IN VIVO AND ITS BIOLOGICAL IMPORTANCE*,†

Summary When bacteria are irradiated with u.v. light there is a dose dependent decrease in the amount of DNA that can subsequently be extracted free of protein with detergent. This appears to be due to the crosslinking of the DNA with protein and the precipitation of the linked DNA when the denatured proteins are precipitated in the procedure used for the isolation of the DNA. The type of linkage between the DNA and the protein is unknown except that it resists the sequential attack of 2% sodium lauryl sulfate and 0.5 M KCI or 55% CsCI. The main evidence that the loss of DNA in vivo is due to the crosslinking of DNA and protein is that the crosslinking of DNA and protein can be demonstrated in vitro . X-rays do not crosslink DNA and protein in vivo , but acridine orange and visible light cause the crosslinking of DNA and protein both in vivo and in vitro .
By pulse labeling the DNA of bacteria with tritated thymine it can be shown that newly synthesized DNA is most sensitive to crosslinking and that this sensitivity shows a cyclic response keyed to the generation time of the bacteria. Under conditions of thymine starvation where the intrinsic sensitivity of the cells to killing by u.v. is markedly increased, there is a parallel increase in the sensitivity of the DNA of these cells to be crosslinked to protein. The similarity in the time sequence of these two events strongly suggests that the crosslinking may play an important role in the loss of viability following u.v. irradiation.     

FLASH PHOTOLYSIS OF SEVERAL AROMATICS BY ULTRAVIOLET LIGHT AND VISIBLE LIGHT IN THE PRESENCE OF EOSIN Y*

Abstract— A flash photolysis investigation was made of the photo-oxidation of aqueous aniline, resorcinol, βnaphthol, p-sulfanilic acid, and p-bromophenol induced by ultraviolet and visible light irradiation in the presence of eosin Y. The transient spectra show that u.v. irradiation generates the hydrated electron (except in p-bromophenol) and the radical products of one-electron oxidation. The initial products of the eosin-sensitized oxidations are the dye semi-quinone and aromatic radicals which coincide with the u.v. photolysis products in at least several cases. The investigation of the reaction kinetics by rapid spectrophotometry with analog computer analysis shows that the aromatics quench the triplet state of eosin and also react with it in a slower electron-transfer process, in competition with ‘dye-dye’ quenching and electron-transfer reactions. The u.v. and dye-sensitized oxidations are discussed in terms of their energetics.     

Synthesis,crystal structure and properties of the nickel(II) complex of a new dioxocyclam bearing 2-methylfuran pendants

A new dioxocyclam ligand, bearing two 2-methylfuran pendants, has been synthesized and its nickel(II) complex isolated as a single crystal. The crystal structure of the complex has been determined by X-ray diffraction analysis. The complex adopts an unusual cis-configuration. The square-planar N4 coordination of the four basal nitrogen atoms with Ni II is evident, with the dioxocyclam moiety composed of two tertiary amine Ns and two deprotonated amide N; s; the latter nitrogens have stronger coordination bonds than those in the tertiary amines. There is a hydrogen bond between O(1) and a water molecule [O(1) H2O = 2.698]. The solution behaviour of NiL (1) has been further studied by u.v.-vis. and cyclic voltammetric techniques. A remarkable red-shift was observed for the maxima absorption band of the electronic spectra of NiL (1) in comparison to that of NiL 0 (H2L0, dioxocyclam = 1,4,8,11-tetraazacyclo-tetradecane-5,7-dione).     

THE LETHAL EFFECTS OF RADIATION ON SIX SPECIES OF PROTOZOANS

Abstract. The lethal response of six strains of protozoa to both ionizing radiation and u.v. light has been established. Values have been determined by observation of isolated animals and death was considered to be failure of irradiated individuals to produce unlimited progeny regardless of the actual time of death following irradiation. The strains were selected to give a large variety of size. Lethal doses of u.v. radiation have been determined in the same units used for ionizing radiation (the rad). The results reported here have been supplimented by values from the literature and plotted in the form of a graph of LD-50 value versus individual organism size.     

SOME STABLE FLUORESCENT CONVERTER SOLUTIONS ENHANCING OPTICAL PUMPING IN THE VISIBLE REGION

Abstract A series of fluorescent converter solutions is described, increasing the radiation intensity of flash lamps in the region 4000–6000 Å with a factor R_Λ1.5˜3. The converter solutions are composed for use at an optical path of 1 cm, are easy to prepare, and are stable under high intensity irradiation with u. v. and visible light.     

Screening of protein crystallization conditions on a microfluidic chip using nanoliter-size droplets

Protein crystallization is a major bottleneck in determining tertiary protein structures from genomic sequence data. This paper describes a microfluidic system for screening hundreds of protein crystallization conditions using less than 4 nL of protein solution for each crystallization droplet. The droplets are formed by mixing protein, precipitant, and additive stock solutions in variable ratios in a flow of water-immiscible fluids inside microchannels. Each droplet represents a discrete trial testing different conditions. The system has been validated by crystallization of several water-soluble proteins.     

芳香叔胺引发烯类单体聚合研究——单体结构与聚合的关系

不同烯类单体在芳香叔胺存在下的聚合机构不一样。甲基丙烯酸甲酯等有α-甲基的烯类单体在不照光的条件下即可被芳香叔胺引发聚合,其聚合机构认为是首先α—甲基被胺-氧复合物氧化,生成单体过氧化物。再与胺形成氧化还原体系,分解产生自由基。 丙烯酸酯,丙烯腈等没有α-甲基的单体,只有光照时才被芳香叔胺氧化聚合,不光照时完全不聚合。这是因为这些单体不被胺-氧复合物氧化。光照下聚合的机构认为是因光的激发,这些单体与芳香叔胺形成电子转移激发络合物,再分解产生自由基。 研究了单体结构,胺结构对光聚合速度的影响。不同单体的活性次序是: AN>MA>VA>St 不同芳香叔胺的活性次序是: DMT>DMA>DMB>DNA 表明单体双键电子云密度越小,芳香叔胺氮原子上电子云密度越大,越容易形成激发态电子转移络合物,从而越容易聚合。     

Mechanisms of antioxidant action: Antioxidant behaviour of nickel complex u.v. stabilisers

The antioxidant mechanisms involved in the u.v. stabilising activity of two nickel complexes, nickel dibutyl dithiocarbamate (NiDBC) and nickel acetophenone oxime (NiOx), have been investigated both by oxygen absorption measurements and in peroxide decomposition studies. NiDBC gives rise to a powerful catalyst for non-radical hydroperoxide decomposition; the rate of the reaction is faster in the presence than in the absence of light although the overall mechanisms appear to be very similar. NiOx is decomposed by hydroperoxides in a series of stoichiometric reactions which involve the consumption of at least six molecules of hydroperoxide per molecule of NiOx. Again the reaction is catalysed by light but the mechanism, which does not involve free-radical formation, is the same in the presence and absence of light. NiOx also appears to have weak radical trapping properties. Attempts to show that NiDBC and NiOx might interact sacrificially with photo-excited states of molecules (triplet carbonyl and singlet oxygen) were unsuccessful; the rates of photo-destruction of both complexes were unaffected by the presence of ketones both in the presence and absence of oxygen. This work confirms earlier conclusions that the nickel complex u.v. stabilisers function by auto-synergistic mechanisms involving u.v. screening and conventional chain-breaking and preventive antioxidant processes which operate during thermal processing operations as well as during environmental exposure. Although excited state quenching processes may occur, they appear to be less important in the overall scheme than the well established antioxidant mechanism.     

PHOTOCHEMISTRY OF THYMIDINE AS A THIN SOLID FILM*

Abstract— –Photochemistry of thymidine in the solid state has been investigated. Four isomers of the cyclobutane-type thymidine dimer, two bimolecular addition products with u.v. absorbance maxima above 300 nm, (the formation of which involves the C=C and C=O groups of adjacent thymine residues) and 5-thyminyl-5.6 dihydrothymine (TDHT) (formed by the addition of a thyminyl radical at the C₅ position of a thymyl radical) were isolated. TDHT was found to be the major photoproduct. The results are compared with those for thymidine irradiated in the frozen state.     

Assignment in the near-threshold absorption spectrum of N2

The absorption spectrum of N(2), just above the first ionization limit, in the region between 125,750 and 126,600 cm(-1), is dominated by two broad features which have the appearance of the towers of a "cathedral." Recently, this energy region has been measured by Sommavilla et al. [J. Phys. B 35, 3901 (2002)] with a resolution of 0.04 cm(-1) and an assignment for one of these two bands has been suggested. In order to discuss this assignment, we have solved the coupled-channel Schr?dinger equation with the parameters of Spelsberg and Meyer above the v=28 level of the b(') (1)Sigma(u)(+) state for determining the high levels of b('). Furthermore, we evaluate the autoionization widths and the rotational structure of the different possible assignments. Finally, we propose as assignment for the second tower of the cathedral the Rydberg state (A (2)Pi(u)v(+)=1)3(')d(')sigma(g) (1)Pi(u).