Feasibility study of in vivo fingernail dosimetry by electron paramagnetic resonance / 中华放射医学与防护杂志

Objective To explore the feasibility of EPR in vivo fingernail dosimetry to address the difficulty in separating mechanically induced signals from fingernail EPR dosimetry for need of nuclear medical emergency aid.Methods Using the specially designed EPR in vivo measurement system,uncut fingernails were measured to obtain the characteristics of EPR signal without mechanically induced signals.The in vivo fingernail experiment was carried out to evaluate the impact of in vivo condition on the spectra.Actual in vivo measurement experiment was conducted to evaluate the interference of the in vivo condition on EPR spectra.Results The background signal distribution of uncut fingernails was obtained and background signals had no significant difference between male and female(P＞0.05).The dose response curve in the range of 2-10 Gy was established,and the half-life of the fingernail radiation-induced signal was approximately 5 d.The water treatment combined with temperature-changing was established for restoring the background signal.EPR signal obtained after restoring treatment has no significant difference with background signal (P＞ 0.05).The EPR spectra of in vivo fingernails were obtained.Conclusions The EPR spectra without mechanically induced signals can be acquired by this method.The feasibility of the in vivo fingernail EPR dosimetry is preliminarily verified.

Radiolytic studies of cefozopran hydrochloride in the solid state

Background: The radiation sterilization is one of the best methods for sterilizing vulnerable degradation drugs like cefozopran hydrochloride. Results: Chemical stability of radiosterylized cefozopran hydrochloride, was confirmed by spectrophotometric and chromatographic methods. EPR studies showed that radiation has created some radical defects whose concentration was no more than several dozen ppm. The antibacterial activity of cefozopran hydrochloride irradiated with a dose of 25 kGy was unaltered for Gram-positive bacteria but changed for two Gram-negative strains. The radiation sterilized cefozopran hydrochloride was not in vitro cytotoxic against human CCD39Lu normal lung fibroblast cell line. Conclusions: Cefozopran hydrochloride in solid state is not resistant to radiation sterilization and this method cannot be used for sterilization of this compound.

Scheduling of chemotherapy based on direct monitoring ofpO2 in tumor microenvironment by EPR oximetry / 中国癌症杂志

Background and purpose:Tumor microenvironment plays an important role in the introduction of foreign factors that mediate tumor acquired resistance. The antitumor effects of many chemotherapeutic agents depend on the level of oxygen pressure (pO2) in tumor microenvironment. This study aimed to evaluate electron paramagnetic reso-nance (EPR)-based monitoring on an oxygen-enriched tumor microenvironment to increase chemotherapeutic sensitivity. Methods:MCF-7 cells were used to establish human breast cancer in nude mice. EPR was used to directly measure pO2 levelin vivo. Tumor tissues were collected, and mitochondrial activity was assayed on the basis of the kinetics of enzyme-catalyzed reactions. A laser Doppler monitor was used to detect regional blood flow. Tumor apoptotic rate was analyzed by flow cytometry.Results:The tumor volume decreased more evidently in the chemotherapy group with oxy-gen-enriched environment than that in the conventional chemotherapy group after the treatment was administered (P<0.01). After chemotherapy was completed, the apoptotic rate of tumor cells was significantly higher in the chemotherapy group with oxygen-enriched environment than that in the conventional chemotherapy group (P<0.001). This study examined the mechanism ofpO2 changes in tumor microenvironment: This was related to the change of the balance between the oxygen consumption and the regional blood flow in the tumor tissues after chemotherapy.Conclusion:Based on the characteristics ofpO2 changes in the tumor microenvironment after chemotherapy was completed, the selection of chemotherapy mode forthe treatment inpO2 peak time window improves the sensitivity of chemotherapy, which provides a new idea for individual-ized chemotherapy in clinical applications.

Accuracy of dose planning for prostate radiotherapy in the presence of metallic implants evaluated by electron spin resonance dosimetry

Radiotherapy is one of the main approaches to cure prostate cancer, and its success depends on the accuracy of dose planning. A complicating factor is the presence of a metallic prosthesis in the femur and pelvis, which is becoming more common in elderly populations. The goal of this work was to perform dose measurements to check the accuracy of radiotherapy treatment planning under these complicated conditions. To accomplish this, a scale phantom of an adult pelvic region was used with alanine dosimeters inserted in the prostate region. This phantom was irradiated according to the planned treatment under the following three conditions: with two metallic prostheses in the region of the femur head, with only one prosthesis, and without any prostheses. The combined relative standard uncertainty of dose measurement by electron spin resonance (ESR)/alanine was 5.05%, whereas the combined relative standard uncertainty of the applied dose was 3.35%, resulting in a combined relative standard uncertainty of the whole process of 6.06%. The ESR dosimetry indicated that there was no difference (P>0.05, ANOVA) in dosage between the planned dose and treatments. The results are in the range of the planned dose, within the combined relative uncertainty, demonstrating that the treatment-planning system compensates for the effects caused by the presence of femur and hip metal prostheses.

Antioxidant effect of 4-nerolidylcatechol and -tocopherol in erythrocyte ghost membranes and phospholipid bilayers

4-Nerolidylcatechol (4-NC) is found in Pothomorphe umbellata root extracts and is reported to have a topical protective effect against UVB radiation-induced skin damage, toxicity in melanoma cell lines, and antimalarial activity. We report a comparative study of the antioxidant activity of 4-NC and α-tocopherol against lipid peroxidation initiated by two free radical-generating systems: 2,2′-azobis(2-aminopropane) hydrochloride (AAPH) and FeSO4/H2O2, in red blood cell ghost membranes and in egg phosphatidylcholine (PC) vesicles. Lipid peroxidation was monitored by membrane fluidity changes assessed by electron paramagnetic resonance spectroscopy of a spin-labeled lipid and by the formation of thiobarbituric acid-reactive substances. When lipoperoxidation was initiated by the hydroxyl radical in erythrocyte ghost membranes, both 4-NC and α-tocopherol acted in a very efficient manner. However, lower activities were observed when lipoperoxidation was initiated by the peroxyl radical; and, in this case, the protective effect of α-tocopherol was lower than that of 4-NC. In egg PC vesicles, malondialdehyde formation indicated that 4-NC was effective against lipoperoxidation initiated by both AAPH and FeSO4/H2O2, whereas α-tocopherol was less efficient in protecting against lipoperoxidation by AAPH, and behaved as a pro-oxidant for FeSO4/H2O2. The DPPH (2,2-diphenyl-1-picrylhydrazyl) free-radical assay indicated that two free radicals were scavenged per 4-NC molecule, and one free radical was scavenged per α-tocopherol molecule. These data provide new insights into the antioxidant capacity of 4-NC, which may have therapeutic applications for formulations designed to protect the skin from sunlight irradiation.

Avaliação por ressonância paramagnética eletrônica da concentração de radicais livres gerados em um cimento resinoso dual utilizado na cimentação de um pino de fibra de vidro / Evaluation by electron paramagnetic resonance of the concentration of free radicals generated in a dual resin cement used on the cementing of translucent glass fiber post

Foi analisado por espectro de Ressonância Paramagnética Eletrônica (RPE), a concentração de radicais livres (RL) formados quando da cimentação de pino de fibra de vidro (PFV), com cimento resinoso dual (CRD), bem como a influência da forma anatômica do PFV (cônico ou cilíndrico). A primeira etapa foi composta de 3 grupos, de acordo com o método de cimentação do PFV numa matriz de silicone: G1- CRD fotopolimerizado por 40 segundos, com aparelho fotopolimerizador de diodo emissor de luz (LED), com potência de 1.500mW/cm2, encostado à superfície externa do pino; G2- CRD não recebeu ativação física; G3- tratamento semelhante ao G1, mas com bloqueio da interface pino/silicone. Na segunda etapa, os PFV foram seccionados de forma a obter dois pinos: PFVCl, cilíndrico e PFVCn cônico, de 9,5mm de comprimento cada, cimentadas semelhantemente ao G1. A concentração de radicais livres foi avaliada em: T0-10 minutos após espatulação do cimento e T1-24 horas após, em secções obtidas em diferentes profundidades do PFV. A análise do espectro das ressonâncias indicou que G1 e G3, apresentaram maior CRL. A translucidez do pino não influenciou na concentração de RL em profundidades acima de 12mm. Os pinos cilíndricos apresentaram maior concentração de RL. A concentração de radicais livres é influenciada pelo método de polimerização, bem como pela forma anatômica do PFV; a capacidade de condução da luz pelo PFV não interfere no grau de conversão do CRD nas regiões de maior profundidade.

The free radical concentration (FR) formed during GFP cementation, using dual cure resin cement (DCRC), as well as the influence of the anatomical shape of the GFP (conical or cylindrical), were analyzed by Electron Paramagnetic Resonance (EPR) spectrum. The first stage consisted of 3 groups, according to the GFP cementation method in a silicone matrix: G1- DCRC light cured for 40 seconds, with light emitting diode (LED), with intensity of 1.500mW/cm2 , leaning against the outer surface of the post; G2 - DCRC without light activation; G3- treatment similar to G1, but blocking the post/matrix interface. In the second stage, GFP were sectioned to obtain two sections: PFVCl, cylindrical and PFVCn, conical, 9,5mm in length each, cemented similarly to G1. The FRC was evaluated in: T0-10 minutes after cement's spatulation e T1-24 hours after, in sections obtained from different GFP depths. The resonance spectrum analysis indicated that G1 and G3 had a greater FR concentration. Post translucency did not influenced the FRC at depths greater than 12mm. Cylindrical posts presented greater FRC. The FR concentration is influenced by the polymerization method, as well as the GFP anatomical shape; GFP capacity to conduct light does not interferes in the degree of conversion on DCRC in regions of greater depth.

Electron paramagnetic resonance study of lipid and protein membrane components of erythrocytes oxidized with hydrogen peroxide

Electron paramagnetic resonance (EPR) spectroscopy of spin labels was used to monitor membrane dynamic changes in erythrocytes subjected to oxidative stress with hydrogen peroxide (H2O2). The lipid spin label, 5-doxyl stearic acid, responded to dramatic reductions in membrane fluidity, which was correlated with increases in the protein content of the membrane. Membrane rigidity, associated with the binding of hemoglobin (Hb) to the erythrocyte membrane, was also indicated by a spin-labeled maleimide, 5-MSL, covalently bound to the sulfhydryl groups of membrane proteins. At 2% hematocrit, these alterations in membrane occurred at very low concentrations of H2O2 (50 µM) after only 5 min of incubation at 37°C in azide phosphate buffer, pH 7.4. Lipid peroxidation, suggested by oxidative hemolysis and malondialdehyde formation, started at 300 µM H2O2 (for incubation of 3 h), which is a concentration about six times higher than those detected with the probes. Ascorbic acid and α-tocopherol protected the membrane against lipoperoxidation, but did not prevent the binding of proteins to the erythrocyte membrane. Moreover, the antioxidant (+)-catechin, which also failed to prevent the cross-linking of cytoskeletal proteins with Hb, was very effective in protecting erythrocyte ghosts from lipid peroxidation induced by the Fenton reaction. This study also showed that EPR spectroscopy can be useful to assess the molecular dynamics of red blood cell membranes in both the lipid and protein domains and examine oxidation processes in a system that is so vulnerable to oxidation.

Plasma lipid pattern and red cell membrane structure in β-thalassemia patients in Jakarta.

Background: Over the last 10 years, we have investigated thalassemia patients in Jakarta to obtain a comprehensive picture of iron overload, oxidative stress, and cell damage. Methods: In blood samples from 15 transfusion-dependent patients (group T), 5 non-transfused patients (group N) and 10 controls (group C), plasma lipids and lipoproteins, lipid-soluble vitamin E, malondialdehyde (MDA) and thiol status were measured. Isolated eryhtrocyte membranes were investigated with electron paramagnetic resonance (EPR) spectroscopy using doxyl-stearic acid and maleimido-proxyl spin lables. Data were analyzed statistically with ANOVA. Results: Plasma triglycerides were higher and cholesterol levels were lower in thalassemic patients compared to controls. Vitamin E, group C: 21.8 vs T: 6.2 μmol/L) and reactive thiols (C: 144 vs. T: 61 μmol/L) were considerably lower in transfused patients, who exert clear signs of oxidative stress (MDA, C: 1.96 vs T: 9.2 μmol/L) and of tissue cell damage, i.e., high transaminases plasma levels. Non-transfused thalassemia patients have slight signs of oxidative stress, but no signifi cant indication of cell damage. Erythrocyte membrane parameters from EPR spectroscopy differ considerably between all groups. In transfusion-dependent patients the structure of the erythrocyte membrane and the gradients of polarity and fl uidity are destroyed in lipid domains; binding capacity of protein thiols in the membrane is lower and immobilized. Conclusion: In tranfusion-dependent thalassemic patients, plasma lipid pattern and oxidative stress are associated with structural damage of isolated erythrocyte membranes as measured by EPR spectroscopy with lipid and proteinthiol spin labels.

Enamel dose calculation by electron paramagnetic resonance spectral simulation technique / 中华放射医学与防护杂志

Objective To optimize the enamel electron paramagnetic resonance (EPR) spectral processing by using the EPR spectral simulation method to improve the accuracy of enamel EPR dosimetry and reduce artificial error.Methods The multi-component superimposed EPR powder spectral simulation software was developed to simulate EPR spectrum models of the background signal(BS) and the radiation- induced signal (RS) of irradiated enamel respectively.RS was extracted from the multi-component superimposed spectrum of irradiated enamel and its amplitude was calculated.The dose-response curve was then established for calculating the doses of a group of enamel samples.The result of estimated dose was compared with that calculated by traditional method.Results BS was simulated as a powder spectrum of gaussian line shape with the following spectrum parameters:g =2.00 35 and Hpp =0.65 - 1.1 mT,RS signal was also simulated as a powder spectrum but with axi-symmetric spectrum characteristics.The spectrum parameters of RS were:g(1) =2.0018,g (11) =1.996 5,Hpp =0.335 - 0.4 mT.The amplitude of RS had a linear response to radiation dose with the regression equation as y =240.74x + 76 724 ( R2 =0.9947 ).The expectation of relative error of dose estimation was 0.13.Conclusions EPR simulation method has improved somehow the accuracy and reliability of enamel EPR dose estimation.

Study of nitric oxide during ischemia-reperfusion of kidney transplantation in rats using electron paramagnetic resonance / 中华泌尿外科杂志

Objective To monitor the production of nitric oxide(NO)and to study its role during ischemia-reperfusion of kidney transplantation in rats using electron paramagnetic resonance(EPR). Methods LEW male rats (age of 8 to 10 months,body weight of 200 to 230 g) were included.Of them 30 were assigned to donor group whose kidney grafts were stored at 0℃ for 24 hours.The remaining 45 rats were equally divided into 3 groups (15 in each group).Group 1 served as controls.In Group 2,allotransplantations of kidneys were performed,and both original kidneys were removed during reperfusions.In Group 3,2 hours prior to the operation,the donors and the recipients were both treated with L-NAME at the dose of 30 mg/kg.Reperfusions were done to the transplanted kidneys and both original kidneys were removed.The production of NO was measured with ERP at each time point before and after blood flow recovery.The creatinine level,GFR and the tissue carbonyl content were recorded on the first and fifth days after operation. Results In Group 2,the production of NO remarkably increased after 15 minutes' reperfusion and kept increasing for 120 minutes. Then the level fell to normal after 210 minutes.In Group 3,the recorded creatinine levels were higher than those in Group 2 at the 24th hour ( P

Antioxidant Effects of Munziq and Mushil of Abnormal Savda by Electron Paramagnetic Resonance / 中药新药与临床药理

To explore the antioxidant mechanism of munziq and mushil of Abnormal Savda. Scavenging effects of munziq and mushil of Abnormal Savda on superoxide anion radical(O -? 2) and hydroxyl radical (OH ?) were measured by electron paramagnetic resonance(EPR). Munziq and mushil possessed obvious scavenging effects on O -? 2 and OH ? and the effect of mushil was superior to that of the munziq.[Conclusion] The EPR is a reliable, effective, accurate, sensitive method for measuring the antioxidant properties of herbal medicines. Munziq and mushil are two kinds of effective antioxidants, and the antioxidant effect may be one of the therapeutic mechanisms of munziq and mushil.

THE SCAVENGING ROLE OF OXYGEN FREE RADICALS BY SALVIA MILTIORRHIZA / 中国药理学通报

Using Xanthine-Xanthine Oxidase system to generate superoxide ani-on, using H2O2 -Fe2 + system to generate hydroxyl radical, the role of scavenging oxygen free radical by Miltiorrhiza was studied by electron paramagnetic resonance and spin trapping. The result suggested that the scavenging rate of hydroxyl radical by Salvia Miltiorrhiza injection is 65%, and that of superoxide anion is 100%. This result suggests that scavenging cytotoxic oxygen free radicals may be one of important pharmacological mechanism of Salvia Miltiorrhiza.

Effects of Several Natural Medicines on the Superoxide Anion Radicals / 第二军医大学学报

The superoxide anion radical (Of) produced in the alkaline dimethyl sulfoxide (DMSO) system was directly measured by electron paramagnetic resonance (EPR) at low temperature. This reaction was used as a model system for generating O2-. Astragalus membranaceus, Codonopsis pilousla, Paeonia laetiflora, Angelica sincnsis and Sophora flavescens, traditional natural medicines which have the effect of delaying anility, were added into the model system to observe their intensifying or inhibiting effect on EPR signals of O2. These results suggest that these traditional natural medicines have the mostly eliminating effect on Of produced in the alkaline DMSO system.