Twisted protein aggregates and disease: the stability of sickle hemoglobin fibers

We describe how twist could play an essential role in stabilizing 20 nm diameter sickle hemoglobin fibers. Our theory successfully reproduces the observed variation of helical pitch length with fiber diameter. With no remaining adjustable parameters it also yields a prediction for the torsional rigidity of sickle hemoglobin fibers that is in good agreement with experiment and hence retains the striking feature that such fibers can be highly mechanically anisotropic, even with a ratio of bending to torsional rigidity of about 50. We discuss how our study might be relevant to the development of treatment strategies.     

Optical wavelength selection for portable hemoglobin determination by near-infrared spectroscopy method

Hemoglobin concentration is commonly used in clinical medicine to diagnose anemia, identify bleeding, and manage red blood cell transfusions. The golden standard method for determining hemoglobin concentration in blood requires reagent. Spectral methods were advantageous at fast and non-reagent measurement. However, model calibration with full spectrum is time-consuming. Moreover, it is necessary to use a few variables considering size and cost of instrumentation, especially for a portable biomedical instrument. This study presents different wavelength selection methods for optical wavelengths for total hemoglobin concentration determination in whole blood. The results showed that modelling using only two wavelengths combination (1143 nm, 1298 nm) can keep on the fine predictability with full spectrum. It appears that the proper selection of optical wavelengths can be more effective than using the whole spectra for determination hemoglobin in whole blood. We also discussed the influence of water absorptivity on the wavelength selection. This research provides valuable references for designing portable NIR instruments determining hemoglobin concentration, and may provide some experience for noninvasive hemoglobin measurement by NIR methods.     

Endogenous contrast blood flow imaging in embryonic hearts using hemoglobin contrast subtraction angiography

The genetic basis of congenital heart disease is yet to be defined, and the interactions between the malformed heart and biomechanical cardiac performance remain poorly understood. Functional optical imaging enables detailed biomechanical phenotyping of cardiac dysfunction in small animal models, which in turn enables specific gene-phenotype relationship. We have developed a new microangiography technique based on flow imaging using endogenous hemoglobin contrast enabling in vivo assessment and biomechanical phenotyping of Xenopus tropicalis embryonic heart. We demonstrated that hemoglobin contrast angiography can be used to quantify physiological response to treatment with well-established cardioactive drugs.     

Spectral studies of photomodification of blood by therapeutic doses of optical radiation at different wavelengths

We analyze changes in electronic and IR absorption spectra of samples of blood and its components, in the fluorescence spectra of plasma, as well as in the gas composition of blood, the hemoglobin concentration, and acid-base balance indices, upon the irradiation of blood by therapeutic doses of optical radiation at 254, 632.8, 670, and 806 nm. We show that the irradiation of blood by radiation at these wavelengths initiates similar molecular changes in blood and its components and that monochromatic incoherent light acts equally as efficiently as laser radiation. We find that, if the blood irradiation wavelength is in the range of the absorption bands of hemoglobin, the hemoglobin acts as a primary photoacceptor and that the dissociation of hemoglobin complexes with ligands directly in erythrocytes is a primary photoprocess. We conclude that the photomodification of blood should be attributed to therapeutic methods capable of controlling the balance between the production of active forms of oxygen and their inhibition by antioxidant systems of the organism.     

Characteristics of blood fluorescence spectra using low-level, 457.9-nm excitation from Ar+ laser

We measured the fluorescence spectra of the whole blood, the red blood cell (RBC) and the hemoglobin using 457.9-nm Ar+ laser excitation. It was found that the fluorescence spectra of the whole blood and the RBC have much similarities in the intensity, the emission peaks and the emitting region, and abundant peaks can be found. But for the hemoglobin, fluorescence could only be found in the wavelength range 580-650 nm. It was concluded that in the wavelength range of 650-850 nm, the fluorescence spectra were emitted by the new fluorophores generated by the breakdown of some weak bonds on the RBC membrane,such as the C-C bond and the C-N bond. In the wavelength range of 590 - 650 nm, the fluorescence spectra are mainly emitted by the hemoglobin, but the hemoglobin solution of cracked RBC has a strong quencher effect on the fluorescence spectrum. The experimental result and the theoretical analysis are meaningful for the medical diagnostics and the therapy.     

Spectral priors improve near-infrared diffuse tomography more than spatial priors

We compare the benefits of spatial and spectral priors in near-infrared diffuse tomography image reconstruction. Although previous studies that incorporated anatomical spatial priors have shown improvement in algorithm convergence and resolution, our results indicate that functional parameter quantification by this approach can be suboptimal. The incorporation of a priori spectral information significantly improves the accuracy observed in recovered images. Specifically, phantom results show that the maximum total hemoglobin concentration ([Hb(T)]) in a region of heterogeneity reached 91% of the true value compared to 63% using spatial priors. The combination of both priors produced results accurate to 98% of the true [Hb(T)]. When both spatial and spectral priors were applied in a healthy volunteer, glandular tissue showed a higher [Hb(T)], water fraction, and scattering power compared to adipose tissue.     

Investigation on the interaction of glipizide with bovine hemoglobin by spectroscopy and molecular docking

This study aims to investigate the interaction between glipizide and bovine hemoglobin using fluorescence quenching, circular dichroism spectroscopy in various temperatures (293, 303, and 310?K) and molecular docking methods. The results demonstrated that glipizide could cause strong fluorescence quenching of bovine hemoglobin by a dynamic quenching mechanism, during which the hydrophobic interaction played a dominant role in this system. The order of magnitude of binding constant is 10⁴, and the number of binding site in the system was close to 1. It also showed that tyrosine residues and tryptophan residues were both involved in the binding of glipizide with bovine hemoglobin, and was closer to the later. Circular dichroism spectra revealed that the conformation of bovine hemoglobin was changed during the binding reaction. The interaction of the system was studied by both spectroscopic method and molecular docking simulation, and the conclusions are consistent.     

Fluorescent analysis of interaction of flavonols with hemoglobin and bovine serum albumin

We have studied the fluorescent properties of flavonols (quercetin, fisetin, morin, rutin) with the aim of studying possible interaction with hemoglobin and bovine serum albumin (BSA). We observed an increase in the intensity of intrinsic fluorescence for all the flavonols except rutin in the presence of BSA. From the changes in the fluorescence spectra, we concluded that tautomeric forms are formed on interaction with hemoglobin. We determined the interconnection between the structure of related flavonols and their fluorescent properties on interaction with proteins, and we determined the binding constants for binding with BSA and hemoglobin. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 74, No. 5, pp. 659–664, September–October, 2007.     

Fluorescence intensity and anisotropy decays of the intrinsic tryptophan emission of hemoglobin measured with a 10-GHz fluorometer using front-face geometry on a free liquid surface

We measured the intensity and anisotropy decays of the intrinsic tryptophan emission from hemoglobin solutions obtained using a 10-GHz frequency-domain fluorometer and a specially designed cuvette which allows front-face excitation on a free liquid surface. The cuvette eliminates reflections and stray emissions, which become significant for low-intensity fluorescence such as in hemoglobin. Three lifetimes are detectable in the subnanosecond range. The average lifetime of hemoglobin emission is ligand dependent. The measured values of average lifetimes are 91, 174, and 184 ps for deoxy-, oxy-, and carboxyhemoglobin, respectively. Fluorescence anisotropy decays of oxy-, deoxy-, and carbonmonoxyhemoglobin can be fitted with up to three correlation times. When three components are used, the floating initial anisotropyr _o is, in each case, higher than the steady-state anisotropy of tryptophan in vitrified solution. For deoxy hemoglobin it is close to 0.4. The data are consistent with an initial loss of anisotropy from 0.4 to about 0.3 occurring in the first 2 ps.     

温度对单个活态人红细胞携氧能力的即时影响

利用快速显微多道分光光度技术,在无损、在位、实时的情况下,测定了不同环境温度下单个活态人红细胞内血红蛋白吸收光谱的即时变化情况。发现人红细胞内血红蛋白吸收峰的高度和位置均随环境温度的变化而变化,表明人红细胞的携氧能力与温度密切相关。初步揭示了人红细胞内血红蛋白分子结构、浓度与功能之间的密切关系,并且探讨了温度变化对人红细胞生理功能即时影响的分子机制。     

拉曼光谱在血红蛋白结构及功能研究中的应用进展

血红蛋白发挥多种重要生理功能,但对其结构及功能的认识尚不能满足疾病诊治的需求。拉曼光谱在血红蛋白结构及功能研究中具有很大应用潜力,不但可以检测血红蛋白中血红素及其周围分子结构变化,还可以反映血红蛋白反应动力学具体过程。同时,血红蛋白拉曼光谱在疾病状态下异常血红蛋白检测,血氧饱和度定量测定及血液代用品的高铁血红蛋白含量检测中突显优势。本文综述了拉曼光谱在血红蛋白结构及功能领域中的研究,简述了拉曼光谱在一些病变血红蛋白诊断中的研究进展,分析了影响血红蛋白拉曼光谱检测的因素,以促进拉曼光谱技术在血红蛋白结构和功能研究中的应用。     

利用近红外光谱监测皮肤血氧输运

利用近红外光谱无创监测移植手术的皮瓣成活状况，是近红外组织氧检测技术的最新应用之一，本文介绍子我们设计的利用光纤传导的近外皮肤血氧监测系统。该系统可以灵活调整光谱－探测器间距以适应皮层检测。利用该系统进行的前臂皮皮肤阻断试验中，静脉阻断时皮肤内有显著的血液充盈现现象，全阻断时氧含量明显下降，实验表明，利用近红外光谱法监测瓣血氧输运情况是非常有效的，特别是可以比较好地监测脉回流情况。     

Change in the absorption spectra of blood exposed to a low-frequency magnetic field

We have used the absorption spectra of whole blood in the UV-visible and IR regions of the spectrum to study changes in the structure of the molecular components of blood when exposed to a low-frequency pulsed magnetic field used to treat ischemic heart disease. We show that pronounced changes in the spectra when the blood is directly exposed in vivo to a magnetic field may be due to breaking of the bond between the heme group and the protein of the hemoglobin, as a consequence of changes in the intermolecular interactions in the polypeptide chains of the hemoglobin and also the spin states of the paramagnetic heme components. Exposure to a magnetic field results in changes in the conformations of the polypeptide chains of hemoglobin and the rate of dissociation of oxyhemoglobin. The structural changes in the hemoglobin molecule are considered as one of the possible primary mechanisms of action on blood in vivo for a low-frequency pulsed magnetic field. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 74, No. 2, pp. 199–204, March–April, 2007.     

基于光谱法研究盐酸胍诱导的人血红蛋白去折叠过程

借助于紫外-可见吸收光谱法、荧光光谱法以及停流-荧光光谱法研究了盐酸胍(GdmHcl)诱导人血红蛋白的去折叠过程。实验发现,盐酸胍诱导的血红蛋白去折叠有两个不同的过程,即随着GdmHcl浓度增加到1.0 mol·L-1左右时,血红蛋白亚基发生解聚,形成中间态;持续增加其浓度时,各亚基发生内部去折叠,最终导致血红素发生崩解。加入还原剂(β-巯基乙醇)对血红蛋白亚基解聚、血红素崩解有协同作用且直接引起亚基和全分子同步变构。血红蛋白去折叠过程从“三态模型”转变为“二态模型”。     

Effect of pigment packaging on diffuse reflectance spectroscopy of samples containing red blood cells

We present the results of diffuse reflectance measurements made on the surface of a tissue-simulating phantom containing intact human erythrocytes. These measurements indicate that the absorption spectrum of hemoglobin in its natural environment is significantly different from that measured in homogeneous fluid solution, especially in the spectral regions of highest absorption. We show that this difference can be explained by the pigment packaging theory developed by Duysens [Biochim. Biophys. Acta 19, 1 (1956)] and that the adoption of basis spectra that take this effect into account improves the accuracy of fitting diffuse reflectance spectra.     

Spin Label Studies of the Hemoglobin–Membrane Interaction During Sickle Hemoglobin Polymerization

An enhanced hemoglobin–membrane association has been previously documented in sickle cell anemia. However, it is not known how this interaction is modified during the hemoglobin S polymerization process. In this work, we use a model of reconstituted erythrocytes from ghost membranes whose cytoskeleton proteins had been previously labeled with the 4-maleimido Tempo spin label, and that were subsequently resealed with hemoglobin S or A solutions. Using electron paramagnetic resonance spectroscopy, we studied the time dependence of the spectral W/S parameter, indicative of the conformational state of cytoskeleton proteins (mainly spectrin) under spontaneous deoxygenation, with the aim of detecting the eventual effects due to hemoglobin S polymerization. The differences observed in the temporal behavior of W/S in erythrocytes reconstituted with both hemoglobins were considered as experimental evidence of an increment in hemoglobin S–membrane interaction as a result of the polymerization process of hemoglobin S under spontaneous deoxygenation.     

Oxyhemoglobin saturation measurements by green spectral shift

From an analysis of new hemoglobin solution transmission spectra at various oxygen saturations (SO2), path lengths, and pH, we find the determination of SO2 by using the classical oximetry technique to be poorly calibrated. We used this data set to develop a proposed method for SO2 determination based on the spectral shift of the hemoglobin transmission minimum between 475 and 510 nm. The method does not require accurate knowledge of hemoglobin extinction coefficients and is linear in relation to SO2 despite changes in path length, pH, or hemoglobin concentration.     

Mechanism of the toxicological interactions of decabrominated diphenyl ether with hemoglobin

Polybrominated diphenyl ethers are vital flame retardants in production and human life and they are widespread global organic pollutants in the environment and likewise in food and feed, causing a potential health concern. Hemoglobin is the main protein in the blood, which is a carrier of oxygen in red blood cells. This work aimed at investigating the toxic interactions of polybrominated diphenyl ethers with hemoglobin, by molecular modeling and spectroscopic analysis methods. Decabrominated diphenyl ether and bovine hemoglobin were selected as representatives for polybrominated diphenyl ethers and hemoglobin, respectively. The experimental results indicated that decabrominated diphenyl ether changed the frames’ conformation and the microenvironment of bovine hemoglobin, which can affect the physiological function of the protein. Decabrominated diphenyl ether combined with bovine hemoglobin with the average number of binding sites (0.7) to form bovine hemoglobin–decabrominated diphenyl ether complex. The binding constant was 860.95?L mol^?1. In addition, the molecular docking data revealed that the van der Waals forces played the primary role in the interaction between bovine hemoglobin and decabrominated diphenyl ether. The study provides insight into the molecular toxicity mechanism of decabrominated diphenyl ether during the blood transportation in vivo.     

Relationship between wavelength combination and signal-to-noise ratio in measuring hemoglobin concentrations using visible or near-infrared light

The signal-to-noise ratio (SNR), when using visible or near-infrared light to measure the change in hemoglobin concentration length (the product of hemoglobin concentration and optical path length in this study), depends on the wavelength combination and the analysis method. Although the SNRs increase when detected or incident optical power increases, the optical power should be limited because of safety standards. Considering these safety standards, we assumed that the total optical power was constant by using the relationship between optical power and measurement error. We investigated the theoretical estimation errors of the changes in hemoglobin concentration length using two, three, and four different wavelengths. The SNRs of the changes in hemoglobin concentration length were high when fewer wavelengths were used. These SNRs decreased when the redox state change in cytochrome oxidase was included in the analysis.     

Fourier Self-Deconvolution Analysis of β-Sheet Contents in the Amide I Region of Hemoglobin Aqueous Solutions under Exposure to 900 MHz Microwaves and Bioprotective Effectiveness of Sugar and Salt Solutions

Spectroscopic Fourier self-deconvolution analysis was used to investigate β-sheet features in the secondary structure of hemoglobin under mobile phone microwaves at 900 MHz. To this end, four samples of hemoglobin in bidistilled water, sucrose, trehalose, and sodium chloride aqueous solutions were exposed for up to 4 hr to 900 MHz microwaves at an average H-field intensity of 42 mA/m. Quantitative spectral analyses highlighted significant increases in β-sheet contents in the Amide I region of hemoglobin samples in bidistilled water solution, but no appreciable change was observed in hemoglobin samples in sucrose, trehalose, and sodium chloride solutions. These results led us to conclude that mobile phone microwaves can denaturate hemoglobin in bidistilled water solution whereas sucrose, trehalose, and sodium chloride solutions produce a protective effect against microwaves, preserving the protein from unfolding.