High resolution electron paramagnetic resonance imaging of biological samples with a single line paramagnetic label

The application of electron paramagnetic resonance imaging (EPRI) to obtain information from biological samples has been limited by the lack of ideal single line radical labels. The commonly used nitroxides exhibit multiple lines causing either hyperfine-based limitations in the maximum obtainable image resolution or hyperfine-based artifacts in the reconstructed image. The use of a novel single-line triarylmethyl paramagnetic label that enables marked enhancement in image quality and resolution is reported. This label exhibits a single line EPR spectrum that is sharp (linewidth ～60 mG in the absence of oxygen) and relatively stable in tissues. The potential of this label in enabling high resolution EPR imaging of biological samples was demonstrated in a series of phantoms and isolated biological organs such as the rat kidney. The images demonstrate that resolutions better than 100 μm could be obtained at L-band on samples of up to 20 mm in size.     

Carbon blacks as EPR sensors for localized measurements of tissue oxygenation.

New electron paramagnetic resonance (EPR) oximetry probes were identified in the class of carbon black materials. These compounds exhibit very high oxygen sensitivity and favorable EPR characteristics for biological applications. At low pO(2), the linewidth is particularly sensitive to changes in oxygen tension (sensitivity of 750 mG/mmHg). The application of the probes for oximetry was demonstrated in vivo: the pO(2) was measured in muscle in which the blood flow was temporarily restricted as well as in tumor-bearing mice during a carbogen breathing challenge. The responsiveness to pO(2) was stable in muscle for at least 3 months. No toxicity was observed using these materials in cellular experiments and in histological studies performed 2, 7, and 28 days after implantation. In view of their EPR characteristics (high sensitivity) as well as the well-characterized production procedure that make them available on a large scale, these probes can be considered as very promising tools for future developments in EPR oximetry.     

Use of epr oximetry with india ink to measure the po2 in the liver in vivo in mice

The partial pressure of oxygen (pO₂) of the liver in vivo in unanesthetized mice was determined using electron paramagnetic resonance (EPR) oximetry with India ink. The EPR spectra were obtained using a low-frequency (1.2 GHz) EPR spectrometer with a loop gap cavity resonator. The line width of the India ink used in this experiment was reversibly broadened by oxygen and was particularly sensitive to pO₂ below 30 torr. After the administration of India ink into the tail vein, the India ink particles were taken up mainly by Kupffer cells in the liver and in part by phagocytes in the spleen. The pO₂ measured in the normal liver was about 14 torr and was constant for the 2-week experimental period. The pO₂ decreased when measured at 1, 2, and 6 days after treatment with a hepatotoxin (carbon tetrachloride (CCI₄)); within 2 weeks, it returned almost to the initial level. Measurements by EPR at sacrifice of controls and CCI₄-treated mice indicated that more than 90% of the India ink went to the liver; the spleen contained 4.7% of total amount in control mice and 8.8% in CCI₄-treated mice when measured 2 weeks after the treatment. These data indicate the usefulness of India ink for measuring the pO₂ of the liver in vivo and that the pO₂ in the Kupffer cells is decreased when the liver is damaged by CCI₄.     

基于 C/S与B/S混合模式的重症肌无力管理系统的设计与研制

重症肌无力临床医生在就诊过程中收集了大量的临床资料,但是面对庞大而又杂乱的临床资料,人工对其进行管理、查询、统计分析等显然不可取,费时费力且更不用说从中“挖掘”出有用的信息以辅助诊断;此外,对于病人服药情况、疗效的随访等也难于开展,更不用说在医院各科室内部、Internet上共享临床资料。基于此,本文开发了以C/S和B/S混合模式的重症肌无力信息系统,以期为临床和科研做出一定的贡献。     

大鼠腹部岛状皮瓣氧自由基产生的实验研究

目的 :为了研究大鼠腹部皮瓣在缺血再灌注时氧自由基的变化 ;方法 :应用电子顺磁共振 (EPR)技术 ,对缺血再灌注后的SD大鼠腹壁岛状皮瓣进行了直接检测 ;结果 :大鼠腹壁皮瓣在缺血再灌注以后产生大量的超氧阴离子自由基 (O ·2 ) ;而且O ·2 变化十分活跃 ;结论 :大鼠腹壁岛状皮瓣是EPR研究氧自由基较为理想的模型     

Skin oxygenation after topical application of liposome-entrapped benzyl nicotinate as measured by EPR oximetry in vivo: Influence of composition and size

New and improved drug delivery systems are the important subject of much scientific research. The development of formulations that increase skin oxygenation and of methods for measuring oxygen levels in skin are important for dealing with healing processes affected by the level of oxygen. We have use EPR oximetry in vivo to compare the influence of liposomal formulations of different size and composition with that of hydrogel with respect to the action of the entrapped benzyl nicotinate (BN). Following the topical application of BN onto the skin of mice, pO₂ increase was measured by low-frequency EPR as a function of time. The effect of BN was evaluated by 3 different parameters: lag-time, time needed for maximum pO₂ increase, and overall effectiveness expressed by the area under the response-time curve. An increase in skin oxygenation was observed after BN application. The results show that the effect of BN incorporated in liposomes is achieved more rapidly than the effect from hydrophilic gel. The composition of the liposomes significantly affects the time at which BN starts to act and, to a lesser extent, the maximum increase of pO₂ in skin and the effectiveness of BN action. However, the size of the liposomes influences both the effectiveness of BN action and the time at which BN starts to act. After repeated application of liposomes, the pO₂ baseline increased and the response of the skin tissue was faster. Our results demonstrate that EPR oximetry is a useful method for evaluating oxygen changes after drug application and for following the time course of their action.     

EPR技术研究枸杞多糖清除·OH自由基作用

目的:探讨枸杞多糖清除自由基的作用.方法:采用电子顺磁共振技术(EPR)检测不同浓度枸杞多糖对·OH自由基的清除作用.结果:枸杞多糖清除·OH自由基的能力在0.25 mg/ml为18.64%,在1.0 mg/ml为87.29%,再增大浓度时清除能力呈下降趋势.结论:枸杞多糖低浓度时具显著清除·OH自由基作用,但高浓度时其清除能力却下降.     

甘草渣中总黄酮清除·OH自由基能力的研究

目的研究甘草渣中的总黄酮体外对羟自由基(·OH)的清除作用。方法将甘草渣中的总黄酮提取物制成不同浓度溶液,利用电子顺磁共振(EPR)技术检测甘草渣中总黄酮对体外产生羟基自由基(·OH)的清除率。结果甘草渣中的总黄酮在0.25~25 mg/m L范围内对·OH自由基清除率随浓度增加呈上升趋势,分别为50.53%、59.89%、65.64%、69.30%、80.06%。结论甘草渣中的总黄酮有明显的清除羟自由基的作用,EPR技术和试验结果可用于甘草渣中黄酮类物质有效活性成分的初步筛选。     

Mechanistic analysis of PLGA/HPMC-based in-situ forming implants for periodontitis treatment

In-situ forming implant formulations based on poly(lactic-co-glycolic acid) (PLGA), acetyltributyl citrate (ATBC), minocycline HCl, N-methyl pyrrolidone (NMP) and optionally hydroxypropyl methylcellulose (HPMC) were prepared and thoroughly characterized in vitro. This includes electron paramagnetic resonance (EPR), nuclear magnetic resonance (¹H NMR), mass change and drug release measurements under different conditions, optical microscopy, size exclusion chromatography (SEC) as well as antibacterial activity tests using gingival crevicular fluid samples from periodontal pockets of periodontitis patients. Based on these results, deeper insight into the physico-chemical phenomena involved in implant formation and the control of drug release could be gained. For instance, the effects of adding HPMC to the formulations, resulting in improved implant adherence and reduced swelling, could be explained. Importantly, the in-situ formed implants effectively hindered the growth of bacteria present in the patients’ periodontal pockets. Interestingly, the systems were more effectively hindering the growth of pathogenic bacterial strains (e.g., Fusobacterium nucleatum) than that of strains with a lower pathogenic potential (e.g., Streptococcus salivarius). In vivo, such a preferential action against the pathogenic bacteria can be expected to give a chance to the healthy flora to re-colonize the periodontal pockets.