Interaction of Low-Intensity Laser Radiation with Blood and Its Components

The response of the blood of rabbits to the intravenous irradiation by a He-Ne laser radiation (λ = 632.8 nm) has been investigated by the UV-visible and IR absorption spectra of the whole blood, plasma, and erythrocyte mass. It has been established that hemoglobin is a primary photoacceptor absorbing low-intensity He-Ne-laser radiation. The exposure of blood to this radiation causes clearly defined changes in the IR and visible absorption spectra of the blood and erythrocytes. These spectral changes arise as a result of partial photodissociation of hemoglobin-ligand complexes in the process of absorption of laser radiation. It is suggested that photodissociation is a primary reaction that arises in blood exposed to a low-intensity laser radiation.__________Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 72, No. 2, pp. 230–235, March–April, 2005.     

A new method for quantitation of spin concentration by EPR spectroscopy: application to methemoglobin and metmyoglobin

A new method of EPR spectral analysis is developed to quantitate overlapping signals. The method requires double integration of a number of spectra containing the signals in different proportions and the subsequent solution of a system of linear equations. The result gives the double integral values of the individual lines, which can then be further used to find the concentrations of all the paramagnetic species present. There is no requirement to deconvolute the whole spectrum into its individual components. The method is employed to quantify different heme species in methemoglobin and metmyoglobin preparations. A significantly greater intensity of the high-spin signal in metmyoglobin, compared to methemoglobin at the same heme concentration, is shown to be due to larger amounts of low-spin forms in methemoglobin. Three low-spin types in methemoglobin and two in metmyoglobin are present in these samples. When their calculated concentrations are added to those of the high-spin forms, the results correspond to the total heme concentrations obtained by optical spectroscopy.     

Estimation of Absorbing Components in a Local Layer Embedded in the Turbid Media on the Basis of Visible to Near- Infrared (VIS-NIR) Reflectance Spectra

A new method is studied for estimating the hemoglobin concentration and oxygenation of a local blood layer in a human skin tissue model on the basis of visible and near infrared reflectance spectra. This method uses the iterative Monte Carlo technique for a multi-layered skin tissue model. The numerical simulation was performed to investigate the effects of estimation errors for epidermis and dermis layers on the results for the local blood layer. Experiments with skin tissue phantoms were performed to verify the possibility of this method.     

血红蛋白生物催化合成导电聚苯胺

利用血红蛋白在十二烷基磺酸钠阴离子表面活性剂胶束体系中生物催化合成水溶性导电聚苯胺/十二烷基磺酸复合物(PANI/SDS), 讨论了不同反应体系及溶液pH值对聚合反应产物的影响. 结果表明该反应具有明显的pH值依赖性, pH (1.0～4.0)是合成导电聚苯胺所必需的, 其最适pH值为3.0, 聚苯胺由导电的翠绿亚胺盐转变为本征态发生在pH 10.4. 用元素分析法、紫外-可见分光光度法、FT-IR、循环伏安法、粘度测试、电导率测试、热重分析法等对PANI/SDS复合物表征, 结果表明该复合物具有较好的热稳定性和可逆的电化学活性.     

竹红菌甲素与血红蛋白相互作用光谱

竹红菌甲素与血红蛋白相互作用光谱;马心血红蛋白;竹红菌甲素;相互作用;UV-Vis吸收光谱;荧光光谱;荧光猝灭动力学常数     

开环棉籽糖分子内交联血红蛋白及其戊二醛的二次交联

用高碘酸钠氧化制备开环棉籽糖(O-Raffinose),将其作为交联剂对脱氧态无基质血红蛋白(Stroma-freehemoglobinSFHb)进行修饰.十二烷基磺酸钠-聚丙烯酰胺不连续电泳(SDS-PAGE)结果显示,开环棉籽糖交联血红蛋白(O-Ra-Hb)与SFHb相比抗解聚性有明显提高;琼脂糖(Sepharose)凝胶层析结果显示,SFHb和O-Ra-Hb出峰时间和峰形大小基本相同,说明交联反应主要发生在分子内的亚基间,基本没有分子间交联产物.用戊二醛(GDA)在优化条件下对O-Ra-Hb进行分子间二次交联,SDS-PAGE结果显示,二次交联产物稳定性比O-Ra-Hb有了进一步提高;Sepharose凝胶层析结果表明,产物中分子量为65000的未聚合的O-Ra-Hb组分为22.6%,分子量为130000～600000的组分为76.9%,分子量为600000以上的组分仅为0.5%,达到了预期目的.     

Preparation,Characterization of CeO2-ZrO2 Composite Hollow Microspheres and Their Application as Electrocatalysis Materials for Hemoglobin in Biosensor

Composite hollow microspheres of CeO₂-ZrO₂ were prepared by using anionic PSA latex particles as template and then calcining the PSA core particles at 600°C. The samples were characterized by XRD, TEM, TG, SEM, EDS, and FT-IR spectra. The effects of the proportion of CeO₂ on the structure of the microspheres were also studied. Electrochemical assay showed that the CeO₂-ZrO₂ composite hollow spheres hydrided with chitosan could be used as a novel biosensing material to detect the direct electron transfer of hemoglobin.     

基于近红外光谱的SG-MSC-MC-UVE-PLS算法在全血血红蛋白浓度检测中的应用

为提高全血血红蛋白浓度预测模型的预测精度,基于近红外光谱分析,首先对原始全血透射光谱数据分别进行均值中心化、标准化、标准正态变量变换(SNV)、多元散射校正(MSC)以及Savitzky-Golay（SG）卷积平滑结合MSC的预处理操作,最终选择预处理效果最好的SG-MSC方法作为数据预处理方法,其最大相关系数达到0.944 1。对SG平滑的平滑窗口宽度进行讨论,找出平滑效果最好的窗口宽度为27。数据预处理消除了全血吸收光谱的基线失真,提高了全血吸收光谱数据的信噪比。将190个样本（190个血红蛋白浓度对应的透射光谱数据）分为具有相近血红蛋白浓度分布的校正集和测试集,其中校正集为143个样本（对应血红蛋白浓度分布为10.6～17.3 g·dL-1）,测试集为47个样本（对应血红蛋白浓度分布为10.3～17.3 g·dL-1）,确保建立模型的适用性。对校正集数据预处理后利用蒙特卡洛无信息变量消除（MC-UVE）方法对其进行波长变量选择,剔除含信息量少的波长点,提高含信息量多的波长占比。设置蒙特卡洛迭代次数为1 000,最终从全血吸收光谱的700个波长变量中筛选出191个波长变量用于建立全血血红蛋白浓度偏最小二乘（PLS）回归模型。对比分析原始全血透射光谱全谱PLS模型、原始全血吸收光谱全谱PLS模型、预处理全血吸收光谱全谱PLS模型、SG-MSC-MC-UVE-PLS模型以及已有二阶导数PLS模型的模型效果,表明基于SG-MSC-MC-UVE-PLS算法的全血血红蛋白浓度预测模型效果较其他模型效果更优,预测相关系数由0.676 3提高到0.979 1,预测集均方根误差由0.898 1减小到0.220 3,最大绝对误差由2.426 1减小到0.411 2。同时,利用MC-UVE方法进行波长变量选择,在保证预测精度的前提下,筛选出建模的波长个数更少,有利于提高模型计算效率。研究结果表明,SG-MSC-MC-UVE-PLS方法能够提高全血吸收光谱信号的信噪比,简化模型结构,提高模型的预测精度和计算效率,对推动血红蛋白浓度检测技术的发展具有进步意义。     

Direct electrochemistry and electrocatalysis of hemoglobin immobilized by methacrylic acid

The direct electrochemistry of hemoglobin (Hb) incorporated in methacrylic acid (MAA) film on a paraffin-impregnated graphite electrode (PIGE) was described. A pair of well-defined and quasi-reversible cyclic voltametric peaks are obtained. The formal potentials (E ^0′) linearly depend on the pH of solution, indicating that the electron transfer was proton-coupled. Ultraviolet-visible (UV-Vis) spectra showed that the secondary structure of Hb in the MAA film was similar to individual Hb. The immobilized Hb retained its biological activity well and exhibited a nice response to the reduction of both NO₂⁻, and H₂O₂, on the basis of which a new biosensor has been developed. Published in Russian in Elektrokhimiya, 2008, Vol. 44, No. 9, pp. 1079–1086. The text was submitted by the authors in English.