Free Radical Formation from the Antineoplastic Agent VP 16-213

Free radical formation from VP 16-213 was studied by ESR spectroscopy. Incubation of VP 16-213 with the one-electron oxidators persulphate-ferrous, myeloperoxidase (MPO)/hydrogen peroxide and horseradish peroxidase (HRP)/hydrogen peroxide readily led to the formation of a free radical. The ESR spectra obtained in the last two cases, were in perfect accord with that of a product obtained by electrochemical oxidation of VP 16-213 at +550 mV. The half-life of the free radical in 1 mM Tris (pH 7.4), 0.1 MNaClat 20°C, was 257 ± 4 s. The signal recorded on incubation with HRP/H₂O₂ or MPO/H₂O₂ did not disappear on addition of 0.3 - 1.2 mg/ml microsomal protein. From incubations with rat liver microsomes in the presence of NADPH, no ESR signals were obtained.     

Free Radical Formation from the Antineoplastic Agent VP 16-213

Free radical formation from VP 16-213 was studied by ESR spectroscopy. Incubation of VP 16-213 with the one-electron oxidators persulphate-ferrous, myeloperoxidase (MPO)/hydrogen peroxide and horseradish peroxidase (HRP)/hydrogen peroxide readily led to the formation of a free radical. The ESR spectra obtained in the last two cases, were in perfect accord with that of a product obtained by electrochemical oxidation of VP 16-213 at +550 mV. The half-life of the free radical in 1 mM Tris (pH 7.4), 0.1 MNaClat 20°C, was 257 ± 4 s. The signal recorded on incubation with HRP/H₂O₂ or MPO/H₂O₂ did not disappear on addition of 0.3 - 1.2 mg/ml microsomal protein. From incubations with rat liver microsomes in the presence of NADPH, no ESR signals were obtained.     

Precision parameters from spin-probe studies of membranes using a partitioning technique. Application to two model membrane vesicles

A new version of the ESR spin probe partitioning method is developed and applied to the study of hydration properties of dimyristoyl-phosphatidylglycerol (DMPG) and dimyristoyl-phosphatidylcholine (DMPC) vesicles as functions of salt concentration and temperature above the lipid phase transition. The small spin probe di-tert-butyl nitroxide (DTBN) is used in order to achieve motionally narrowed Electron Spin Resonance (ESR) spectra which may be analyzed with high precision. The new method relies on the use of the second harmonic display of the ESR spectrum followed by spectral line fitting. Spectral fitting yields precise ESR parameters giving detailed information on the surroundings of the spin probe in both phospholipid and aqueous phases. The nitrogen hyperfine coupling constant of DTBN arising from those probes occupying the vesicles is used to study the hydration of the vesicle surface. The hydration properties of the negatively charged vesicle surface of DMPG vesicles are affected by the addition of salt at all temperatures. In contrast, the hydration of DMPC vesicles does not change with salt concentration at the low temperatures. However, at higher temperatures the hydration properties of DMPC vesicle are affected by salt which is interpreted to be due to the faster motion of the phospholipid molecules. The partitioning of the spin probe increases with salt concentration for both DMPG and DMPC vesicles, while water penetration decreases simultaneously. The spin probe in the phospholipid bilayer exhibits anisotropic motion and the extent of the anisotropy is increased at the higher salt concentrations.     

大荔人所在层位贝壳的电子自旋共振年龄

大荔人属早期智人中较古老的类型 ,在人类起源研究中具有重要意义。采用电子自旋共振 (ESR)测年技术直接测定了采自大荔人所在层位的 3个贝壳样品的年龄 ,同时 ,进行了铀系测年分析 ,根据贝壳ESR的年龄结果 ,大荔人可能老于距今 250ka。     

Real-time MRI and articulatory coordination in speech

This paper describes the real-time MRI technique and its use for the study of speech production. The two major problems, (i) the simultaneous recording of the MR images and the speech signal and (ii) the synchronisation of the images and of the speech signal, are addressed. Measurement accuracy on real-time images is evaluated by comparison with similar measurements on static MR images.     

Photosensitization by hematoporphyrin: ESP evidence for free radical induction in unsaturated fatty acids and for singlet oxygen production.

Hematoporphyrin ability to photoreact by type I and type II mechanisms was investigated in some model systems. At room temperature, visible irradiation of hematoporphyrin-unsaturated fatty acids and hematoporphyrin-cholesterol systems resulted in the Electron Spin Resonance (ESR) spectrum of the hematoporphyrin free radical. Triplet state hematoporphyrin is shown to be involved in the electron transfer from the lipid moiety. Moreover an ESR method to monitor the singlet oxygen production by hematoporphyrin was used. β-carotene effect on both mechanisms (type I and type II) was tested.     

Nucleotidic cofactors and the origin of the genetic code

In this article it is suggested that the first coding system started from specific interactions between the nucleotide part of nucleotidic cofactors and enzymes. These interactions generated a first primitive code of four words of one letter each for the four primittive amino acids (phenylalanine, lysine, glycine and proline); when the triplet code (which allowed the integration of 20 amino acids into proteins) progressively appeared, it must have been modulated by the existence of this first coding system.     

超氧化物歧化酶的自旋标记研究

本文应用对-SH基特异性结合的自旋标记物对Cu_2Zn_2-SOD进行自旋标记研究,进一步观察了电离辐射对-SH基部位的影响,实验结果:(1)自旋标记后的Cu_2Zn_2-SOD表现出典型的弱固定化的ESR波谱信号特征。标记对Cu_2Zn_2-SOD的UV谱无明显影响。这表明该游离-SH基是位于酶蛋白分子的相对表层而不是包埋在疏水内部。(2)标记保温后立即取样及保温后17小时取样测酶活力,发现与未标记酶的活力相同。这表明该-SH基与酶的催化活性无直接关系。(3)在10—100Krad γ-射线照射下,酶活性及ESR信号幅度均随照射剂量增加而下降,两者之间有一定的相关性。     

Electron spin resonance-spin trapping. Detection of superoxide formation during aerobic microsomal reduction of nitro-compounds.

ESR measurements demonstrate that in aerobic systems reduction of aromatic nitro-compounds by rat liver microsomes leads to the production of superoxide which may be detected as its adduct with nitrone spin traps.     

The Lipid-binding Domain of Wild Type and Mutant α-Synuclein: COMPACTNESS AND INTERCONVERSION BETWEEN THE BROKEN AND EXTENDED HELIX FORMS*

α-Synuclein (αS) is linked to Parkinson disease through its deposition in an amyloid fibril form within Lewy Body deposits, and by the existence of three αS point mutations that lead to early onset autosomal dominant Parkinsonism. The normal function of αS is thought to be linked to the ability of the protein to bind to the surface of synaptic vesicles. Upon binding to vesicles, αS undergoes a structural reorganization from a dynamic and disordered ensemble to a conformation consisting of a long extended helix. In the presence of small spheroidal detergent micelles, however, this extended helix conformation can convert into a broken helix state, in which a region near the middle of the helix unwinds to form a linker between the two resulting separated helices. Membrane-bound conformations of αS likely mediate the function of the protein, but may also play a role in the aggregation and toxicity of the protein. Here we have undertaken a study of the effects of the three known PD-linked mutations on the detergent- and membrane-bound conformations of αS, as well as factors that govern the transition of the protein between the extended helix and broken helix states. Using pulsed dipolar ESR measurements of distances up to 8.7 nm, we show that all three PD-linked αS mutants retain the ability to transition from the broken helix to the extended helix conformation. In addition, we find that the ratio of protein to detergent, rather than just the absolute detergent concentration, determines whether the protein adopts the broken or extended helix conformation.     

The prosthetic group of methanol dehydrogenase from Hyphomicrobium X: electron spin resonance evidence for a quinone structure.

Partial reduction of the isolated prosthetic group of methanol dehydrogenase yields a free radical with the same characteristics as the one contained in the enzyme. The Electron Spin Resonance spectrum in alkaline aqueous solution displays hyperfine structure and is interpreted in terms of an isotropic g-value, hyperfine coupling constants and nuclear spins. The magnitudes of these parameters indicate that the prosthetic group is a quinone containing two nitrogen atoms.     

Heterogeneite des variations spectrales photoinduites vers 700 nm observees sur les membranes chlorophylliennes et les complexes chlorophylle-proteines isoles de divers organismes photosynthetiques

A heterogeneity of photoinduced spectral variations near 700 nm is observed at −196°C, by comparison of difference spectra obtained from several materials (two blue-green algae, Spirulina and Fremyella; a red alga Porphyridium and a higher plant, Nicotiana tabacum). The maximum of complexity is seen with Spirulina chlorophyllous membranes where three bands are detected at 718, 705 and 695 nm. These bands are recovered in relatively different amounts in two chlorophyll-protein complexes prepared after sodium dodecyl sulfate electrophoresis of Spirulina membranes.

The photoinduced changes near 700 nm with Spirulina membranes are reversible and seem to be synchronous and related to the bleaching of three different pigments. This spectral diversity for photosensizable pigments is comparable with the spectral heterogeneity of the major forms of the bulk chlorophyll. It is assumed that these pigments may play the same functional role. This assumption may be related to a possible heterogeneity of Photosystem I centres recently detected by Breton using linear dichroism method (Breton, J. (1977) Biochim. Biophys. Acta 459, 66–70).     

Étude polarographique de la mobilisation du fer de la ferritine

Polarographic study of the mobilization of ferritin ironPolarographic study allows to propose a model for mobilization of ferritin iron: an equilibrium exists between iron core and small quantities of iron outside the protein.These iron atoms would be lying on electron acceptor sites including SH groups. The number of sites is dependent on iron content of ferritin.Therefore, the iron could be removed by the action of reducing agents such as xanthine oxidase or ascorbic acid, and then chelated by a complexing agent.     

Electron spin resonance dating in paleoanthropology

Many archeological and paleoanthropological sites cannot be dated by well established and common dating techniques such as uranium series (U-series) or argon-argon (⁴⁰Ar/³⁹Ar) because of the lack of materials that are suitable for these techniques. Most sites, however, contain bones and teeth, and the latter can be used to obtain electron spin resonance (ESR) age estimates. The theoretical age range of ESR dating accuracy lies between a few thousand and more than a million years. In practice, continuing uranium accumulation increases the uncertainty of ESR age assessments in such a way that most age assignments beyond 300,000 years are very uncertain.     

用ESR能级电子饱和法早期诊断高发肿瘤研究

用顺磁共振能级电子饱和法的饱和功率点分别对健康人、肺癌、食道癌及与肺癌相关的肺结核及矽肺5.19例的人发进行测定,并与临床诊断相比较.测定结果表明,其诊断率分别为1.66,85.24%,90.48%,27.27%,及20.00%.其中肺癌、食道癌与健康人相比较,经统计处理差异显著(P<0.001).肺癌和食道癌经治疗后其饱和功率点后移,增高明显,介于10.0—24.0mW及10.0—19.9mW者分别占71.43%、71.23%.从肺癌和食道癌治疗后的典型病例所见,饱和功率点越高、疗效越好.对食道癌高发区56例调查结果表明,饱和功率点测定结果与临床诊断一致者为39.29%,排除食道癌者为60.71%.     

Établissement d'une nouvelle lignée cellulaire (IAFEs-1) a partir d'ovarioles d'Euxoa scandens [Lep.: Noctuidae]

Résumé Une nouvelle lignée cellulaire stable (IAFEs-1) des ovarioles de chrysalides du lepidoptèreEuxoa scandens Riley a été obtenue. La forme prédominante des cellules est sphérique et le doublement de la population cellulaire s'effectue en 18 et 24 h en utilisant respectivement 20 et 10% de sérum de veau foetal. Cette lignée est sensible à divers virus des polyédroses nucléaires et cytoplasmiques.

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Summary A new continuous cell line designated IAFEs-1 was obtained from ovaries ofEuxoa scandens Riley in order to study the replication and the productionin vitro of viruses infecting this cutworm. Predominant cell morphology is spherical and cell population doubling time is 18 and 24 h using respectively 20 and 10% fetal calf serum. Susceptiblity of the cell line to several nuclear and cytoplasmic polyhedrosis viruses is demonstrated.

     

A simple framework to describe the regulation of gene expression in prokaryotes

Based on the bimolecular mass action law and the derived mass conservation laws, we propose a mathematical framework in order to describe the regulation of gene expression in prokaryotes. It is shown that the derived models have all the qualitative properties of the activation and inhibition regulatory mechanisms observed in experiments. The basic construction considers genes as templates for protein production, where regulation processes result from activators or repressors connecting to DNA binding sites. All the parameters in the models have a straightforward biological meaning. After describing the general properties of the basic mechanisms of positive and negative gene regulation, we apply this framework to the self-regulation of the trp operon and to the genetic switch involved in the regulation of the lac operon. One of the consequences of this approach is the existence of conserved quantities depending on the initial conditions that tune bifurcations of fixed points. This leads naturally to a simple explanation of threshold effects as observed in some experiments.     

On the Origin of Large Flexibility of P-glycoprotein in the Inward-facing State

P-glycoprotein (Pgp) is one of the most biomedically relevant transporters in the ATP binding cassette (ABC) superfamily due to its involvement in developing multidrug resistance in cancer cells. Employing molecular dynamics simulations and double electron-electron resonance spectroscopy, we have investigated the structural dynamics of membrane-bound Pgp in the inward-facing state and found that Pgp adopts an unexpectedly wide range of conformations, highlighted by the degree of separation between the two nucleotide-binding domains (NBDs). The distance between the two NBDs in the equilibrium simulations covers a range of at least 20 Å, including, both, more open and more closed NBD configurations than the crystal structure. The double electron-electron resonance measurements on spin-labeled Pgp mutants also show wide distributions covering both longer and shorter distances than those observed in the crystal structure. Based on structural and sequence analyses, we propose that the transmembrane domains of Pgp might be more flexible than other structurally known ABC exporters. The structural flexibility of Pgp demonstrated here is not only in close agreement with, but also helps rationalize, the reported high NBD fluctuations in several ABC exporters and possibly represents a fundamental difference in the transport mechanism between ABC exporters and ABC importers. In addition, during the simulations we have captured partial entrance of a lipid molecule from the bilayer into the lumen of Pgp, reaching the putative drug binding site. The location of the protruding lipid suggests a putative pathway for direct drug recruitment from the membrane.     

Electron Movement Through Proteins and DNA

Nature utilizes the phenomenon of single electron transfer very widely, especially in metallo-proteins. In systems when the metal donor (D) is well separated from the acceptor (A) by polypeptide chains, the transferring electron is presumed to be bonded, in part, by these chains, which may influence the pathway taken. This situation can be probed by radiolytic injection of electrons into proteins at low temperatures. One aim of this brief review is to consider how information derived from such radiolysis studies, and followed by ESR spectroscopy, may possibly impinge on studies of D-A systems. Electrons can also be injected into duplex DNA in this way, and the results are compared with those for proteins. They are also considered in the light of recent studies of D-A electron-transfer via a polynucleotide strand. It seems that such transfers are very efficient, and it is tentatively suggested that Nature may also use this conductivity in some as yet undiscovered systems. © 1997 Elsevier Science Inc.