激光拉曼光谱技术在生物分子DNA研究中的应用和进展

激光技术的兴起使拉曼光谱成为激光分析中最活跃的研究领域之一,已被广泛地用于物质成分的分析和分子结构的鉴定。本文综述了拉曼技术在DNA研究中近年来的最新进展,包括：DNA的常规拉曼光谱分析;DNA的激光共振拉曼光谱分析;DNA在金属表面或电极上吸附行为的表面增强拉曼光谱研究;DNA的傅立叶变换拉曼光谱研究等。并对拉曼光谱技术在DNA等生物大分子领域中的研究前景做了进一步的展望。     

激光拉曼光谱在蛋白质构象研究中的应用和进展

激光拉曼光谱法被公认为是研究生物大分子的结构、动力学和功能的有效方法。近年来拉曼光谱在蛋白质构象研究中的最新进展,涉及到拉曼光谱在非折叠蛋白质、蛋白质装配的特征描述,拉曼晶体学在实时监控蛋白质单晶中化学变化等方面的应用。另外,介绍了蛋白质拉曼光谱分析在生物技术中的应用现状。并对拉曼光谱技术在蛋白质等生物大分子领域中的研究前景做了进一步的展望。     

Fabrication of layer-by-layer deposited multilayer films containing DNA and its interaction with methyl green.

Multilayer films were fabricated by layer-by-layer electrostatic deposition techniques between poly(diallyldimethylammonium chloride) (PDDA) and calf thymus DNA (CT DNA) on glassy carbon and quartz substrates. Electrochemical impedance spectroscopy (EIS), Fourier transform infrared (FTIR) spectroscopy and UV-vis spectroscopy demonstrated the uniform assembly of PDDA/DNA multilayer films, and X-ray photoelectron spectroscopy confirmed the elemental composition of the films. Moreover, the interaction of DNA in PDDA/DNA films with methyl green was investigated by UV-vis spectroscopy and circular dichroism (CD).     

功能红细胞研究的拉曼光谱技术与进展

红细胞在人体新陈代谢和物质运输方面起着非常重要的作用。作为一种无损、快速而灵敏的光谱学检测方法,拉曼光谱技术在生物医学领域的应用日渐广泛。本文综述了近年来拉曼光谱技术在红细胞及血红蛋白研究中的应用进展,详尽评述当前研究功能红细胞中较有优势的拉曼技术—显微拉曼光谱技术和光镊拉曼光谱技术,并对其技术的改进做出总结,以期为利用激光拉曼光谱研究红细胞提供较系统的参考。     

The structural and topological analysis of membrane-associated polypeptides by oriented solid-state NMR spectroscopy: established concepts and novel developments

Solid-state NMR spectroscopy is a powerful technique for the investigation of membrane-associated peptides and proteins as well as their interactions with lipids, and a variety of conceptually different approaches have been developed for their study. The technique is unique in allowing for the high-resolution investigation of liquid disordered lipid bilayers representing well the characteristics of natural membranes. Whereas magic angle solid-state NMR spectroscopy follows approaches that are related to those developed for solution NMR spectroscopy the use of static uniaxially oriented samples results in angular constraints which also provide information for the detailed analysis of polypeptide structures. This review introduces this latter concept theoretically and provides a number of examples. Furthermore, ongoing developments combining solid-state NMR spectroscopy with information from solution NMR spectroscopy and molecular modelling as well as exploratory studies using dynamic nuclear polarization solid-state NMR will be presented.     

Circular dichroism and guanine quadruplexes

Circular dichroism (CD) is remarkably sensitive to the conformational states of nucleic acids; therefore, CD spectroscopy has been used to study most features of DNA and RNA structures. Quadruplexes are among the significant noncanonical nucleic acids architectures that have received special attentions recently. This article presents examples on the contribution of CD spectroscopy to our knowledge of quadruplex structures and their polymorphism. The examples were selected to demonstrate the potential of this simple method in the quadruplex field. As CD spectroscopy detects only the global feature of a macromolecule, it should preferably be used in combination with other techniques. On the other hand, CD spectroscopy, often as a pioneering approach, can reveal the formation of particular structural arrangements, to search for the conditions stabilizing the structures, to follow the transitions between various structural states, to explore kinetics of their appearance, to determine thermodynamic parameters and also detect formation of higher order structures. This article aims to show that CD spectroscopy is an important complementary technique to NMR spectroscopy and X-ray diffraction in quadruplex studies.     

Tissue oxygenation and perfusion in inferior pedicle reduction mammaplasty by near-infrared reflection spectroscopy and color-coded duplex sonography

Near-infrared reflection spectroscopy has been used in various experimental and clinical settings to investigate tissue perfusion and oxygenation noninvasively. Its application in plastic surgery has only recently been reported. The current study used near-infrared reflection spectroscopy to monitor cutaneous microcirculation in breast skin flaps after inferior pedicle reduction mammaplasty. Thirty patients underwent bilateral reduction mammaplasty by a modified Robbins technique. Near-infrared reflection spectroscopy measurements were performed preoperatively and postoperatively at several defined positions of the breast. The reflection spectroscopy system was capable of detecting absolute values of total hemoglobin in milligrams per milliliter of tissue and tissue hemoglobin oxygen saturation in percent. Color-coded duplex sonography was used to visualize nutrient vessels of the inferior dermoglandular pedicle and to measure systolic peak flow in the arteries supplying the nipple-areola complex. Reflection spectroscopy values were examined for changes during the postoperative course. Reflection spectroscopy and duplex sonography values were analyzed for differences between patients with normal and compromised skin flap perfusion and wound healing, which was assessed clinically and by ultrasound. Preoperative reflection spectroscopy values demonstrated local, regional, and interindividual variations. Postoperatively, characteristic changes of tissue hemoglobin oxygen saturation and total hemoglobin were observed in all patients during the 2-week follow-up. Reflection spectroscopy values differed significantly between breast and nipple-areola skin. Tissue hemoglobin oxygen saturation was significantly lower, and total hemoglobin significantly higher, in patients with impaired wound healing compared with patients having normal wound healing. However, systolic peak flow in arteries of the inferior dermoglandular pedicle did not reveal differences between patients with impaired or normal wound healing of the nipple-areola complex. Near-infrared reflection spectroscopy allows the detection of hemoglobin content and oxygenation in skin flaps. Changes in tissue hemoglobin oxygen saturation and total hemoglobin reflect hemodynamic changes in skin flaps during normal and pathological wound healing. Because of considerable intraindividual and interindividual variations, trend values seem to be superior to single measurements. Although in this study, near-infrared reflection spectroscopy was capable of distinguishing between normal and impaired perfusion in skin flaps in a clinical model, its future implication may be the early detection of vascular compromise in free flaps.     

Observation volumes and {gamma}-factors in two-photon fluorescence fluctuation spectroscopy

Fluorescence fluctuation spectroscopy has become an important measurement tool for investigating molecular dynamics, molecular interactions, and chemical kinetics in biological systems. Although the basic theory of fluctuation spectroscopy is well established, it is not widely recognized that saturation of the fluorescence excitation can dramatically alter the size and profile of the fluorescence observation volume from which fluorescence fluctuations are measured, even at relatively modest excitation levels. A precise model for these changes is needed for accurate analysis and interpretation of fluctuation spectroscopy data. We here introduce a combined analytical and computational approach to characterize the observation volume under saturating conditions and demonstrate how the variation in the volume is important in two-photon fluorescence correlation spectroscopy. We introduce a simple approach for analysis of fluorescence correlation spectroscopy data that can fully account for the effects of saturation, and demonstrate its success for characterizing the observed changes in both the amplitude and relaxation timescale of measured correlation curves. We also discuss how a quantitative model for the observed phenomena may be of broader importance in fluorescence fluctuation spectroscopy.     

Isolation and characterization of a fluorescent material in bovine achilles tendon collagen

A fluorescent material in bovine achilles tendon collagen was isolated and characterized by ultraviolet spectroscopy, fluorescence spectroscopy and nmr spectroscopy. The data suggest that the compound is a 3-hydroxypyridinium derivative with three amino acid side chains. The name “pyridinoline” is proposed. Pyridinoline is a novel type crosslink of collagen.     

Single cell Raman spectroscopy for cell sorting and imaging

Single cell Raman spectroscopy (SCRS) is a non-invasive and label-free technology, allowing in vivo and multiple parameter analysis of individual living cells. A single cell Raman spectrum usually contains more than 1000 Raman bands which provide rich and intrinsic information of the cell (e.g. nucleic acids, protein, carbohydrates and lipids), reflecting cellular genotypes, phenotypes and physiological states. A Raman spectrum serves as a molecular 'fingerprint' of a single cell, making it possible to differentiate various cells including bacterial, protistan and animal cells without prior knowledge of the cells. However, a key drawback of SCRS is the fact that spontaneous Raman signals are naturally weak; this review discusses recent research progress in significantly enhancing and improving the signal of spontaneous Raman spectroscopy, including resonance Raman spectroscopy (RRS), coherent anti-Stokes Raman spectroscopy (CARS), stimulated Raman spectroscopy (SRS) and surface enhanced Raman scattering (SERS). This review focuses on the biotechnological development and the associated applications of SCRS, including Raman activated cell sorting (RACS) and Raman imaging and mapping.     

The application of spectroscopy techniques for diagnosis of malaria parasites and arboviruses and surveillance of mosquito vectors: A systematic review and critical appraisal of evidence

Spectroscopy-based techniques are emerging diagnostic and surveillance tools for mosquito-borne diseases. This review has consolidated and summarised recent research in the application of Raman and infrared spectroscopy techniques including near- and mid-infrared spectroscopy for malaria and arboviruses, identified knowledge gaps, and recommended future research directions. Full-length peer-reviewed journal articles related to the application of Raman and infrared (near- and mid-infrared) spectroscopy for malaria and arboviruses were systematically searched in PUBMED, MEDILINE, and Web of Science databases using the PRISMA guidelines. In text review of identified studies included the methodology of spectroscopy technique used, data analysis applied, wavelengths used, and key findings for diagnosis of malaria and arboviruses and surveillance of mosquito vectors. A total of 58 studies met the inclusion criteria for our systematic literature search. Although there was an increased application of Raman and infrared spectroscopy-based techniques in the last 10 years, our review indicates that Raman spectroscopy (RS) technique has been applied exclusively for the diagnosis of malaria and arboviruses. The mid-infrared spectroscopy (MIRS) technique has been assessed for the diagnosis of malaria parasites in human blood and as a surveillance tool for malaria vectors, whereas the near-infrared spectroscopy (NIRS) technique has almost exclusively been applied as a surveillance tool for malaria and arbovirus vectors.Conclusions/SignificanceThe potential of RS as a surveillance tool for malaria and arbovirus vectors and MIRS for the diagnosis and surveillance of arboviruses is yet to be assessed. NIRS capacity as a surveillance tool for malaria and arbovirus vectors should be validated under field conditions, and its potential as a diagnostic tool for malaria and arboviruses needs to be evaluated. It is recommended that all 3 techniques evaluated simultaneously using multiple machine learning techniques in multiple epidemiological settings to determine the most accurate technique for each application. Prior to their field application, a standardised protocol for spectra collection and data analysis should be developed. This will harmonise their application in multiple field settings allowing easy and faster integration into existing disease control platforms. Ultimately, development of rapid and cost-effective point-of-care diagnostic tools for malaria and arboviruses based on spectroscopy techniques may help combat current and future outbreaks of these infectious diseases.     

Surface analysis and materials characterization for the study of biodeterioration and weathering effects on cultural property

Several methods for material characterization and surface analysis such as scanning electron microscopy (SEM), energy dispersion X-ray analysis (EDX), environmental scanning electron microscopy (ESEM), petrographic analyses, Mössbauer spectroscopy (MS), conventional X-ray diffraction (XRD), grazing incidence diffraction (GID), Raman spectroscopy (RS), other spectroscopic techniques like X-ray photoelectron spectroscopy (XPS), reflection electron energy-loss spectroscopy (REELS) and advanced combined applications of synchrotron based μ-X-ray diffraction/μ-X-ray fluorescence (SR-μXRD/μXRF) can be used for assessing weathering and biodeterioration effects on materials (such as stone buildings, metallic artefacts, pigments, mixtures, and processes) of cultural property. Molecular biology techniques to identify the microbial components of biofilms are also described. Different examples of the use of these methods in the field of cultural property preservation are presented.     

Solution and solid-state NMR structural studies of antimicrobial peptides LPcin-I and LPcin-II

Lactophoricin (LPcin-I) is an antimicrobial, amphiphatic, cationic peptide with 23-amino acid residues isolated from bovine milk. Its analogous peptide, LPcin-II, lacks six N-terminal amino acids compared to LPcin-I. Interestingly, LPcin-II does not display any antimicrobial activity, whereas LPcin-I inhibits the growth of both Gram-negative and Gram-positive bacteria without exhibiting any hemolytic activity. Uniformly ¹⁵N-labeled LPcin peptides were prepared by the recombinant expression of fusion proteins in Escherichia coli, and their properties were characterized by electrospray ionization mass spectrometry, circular dichroism spectroscopy, and antimicrobial activity tests. To understand the structure-activity relationship of these two peptides, they were studied in model membrane environments by a combination of solution and solid-state NMR spectroscopy. We determined the tertiary structure of LPcin-I and LPcin-II in the presence of dodecylphosphorylcholine micelles by solution NMR spectroscopy. Magnetically aligned unflipped bicelle samples were used to investigate the structure and topology of LPcin-I and LPcin-II by solid-state NMR spectroscopy.     

The complex formation of copper(II) with GHL and HSA

The formation constants for complexes of copper(II) with GHL have been determined by means of pH titrations and ESR spectroscopy in aqueous solutions. GHL has an extremely high affinity for copper(II) and forms very stable 1:1 complexes and a comparatively weak 1:2 complex. The ? amino group of GHL seems not to be involved in complex formation as can be deducted from both equilibrium constants and ESR spectroscopy. The ternary system copper(II)-GHL-HSA was investigated by ESR spectroscopy and optical absorption spectroscopy in aqueous solution at physiological pH (7.4). At equimolar concentrations, copper(II), HSA and GHL form a ternary complex.     

Investigation of TRAM flap oxygenation and perfusion by near-infrared reflection spectroscopy and color-coded duplex sonography

Near-infrared reflection spectroscopy, used experimentally for investigation of tissue hemoglobin content and oxygenation in various flaps, was tested in the pedicled transverse rectus abdominis musculocutaneous (TRAM) flap, chosen as a simple clinical model because of its well-known vascular anatomy and clinical relevance. The study intended to answer the following questions: Does the near-infrared reflection spectroscopy system used in this study measure tissue hemoglobin content and oxygenation in the superficial skin layers only, as proposed by the manufacturer? Is near-infrared reflection spectroscopy able to detect differences of tissue hemoglobin content and oxygenation in distinct zones of the TRAM flap skin before, early, and late after surgery? Does tissue hemoglobin content and oxygenation correspond to blood flow in the supplying superior epigastric artery and to clinical signs of TRAM flap perfusion and viability? In 11 patients, tissue hemoglobin content and oxygenation in the lower abdomen/TRAM flap, mastectomy skin flap, and contralateral breast were measured by a new near-infrared reflection spectroscopy system preoperatively, early postoperatively, and late postoperatively. Simultaneously, systolic peak flow in the ipsilateral superior epigastric artery was obtained by color-coded duplex sonography. Routine clinical monitoring was performed throughout the early postoperative period. Tissue hemoglobin content and oxygenation in the lower abdomen, mastectomy skin flap, and contralateral breast were similar before surgery but varied considerably between different patients. There were no significant differences among preoperative, early postoperative, and late postoperative values of tissue hemoglobin content and oxygenation in the mastectomy skin flap and contralateral breast. However, near-infrared reflection spectroscopy measurements of the TRAM flap revealed significant differences between preoperative and early postoperative values of tissue hemoglobin content and oxygenation and among zones I, II, and III early after surgery. Tissue hemoglobin content in the TRAM flap skin increased and oxygenation decreased early after surgery. Near-infrared reflection spectroscopy values corresponded to clinical signs of venous congestion predominantly in zone III. Late postoperative return of hemoglobin content and oxygenation in the TRAM flap toward preoperative values can be attributed to improved venous return by reversed flow across regurgitant valves and development of collateral circulation. Finally, there was a significant increase of systolic peak flow in the ipsilateral superior epigastric artery early after surgery. This could be related to the opening of small-caliber choke arteries between the superior and deep inferior epigastric arteries following ligation of the dominant deep inferior epigastric artery and TRAM flap transfer to the chest. Systolic peak flow returned to preoperative values late after surgery. The near-infrared reflection spectroscopy system used in this study appeared to measure hemoglobin content and oxygenation in the superficial skin layers only. Near-infrared reflection spectroscopy was able to detect differences of tissue hemoglobin content and oxygenation in the TRAM flap between preoperative and postoperative measurements and between distinct zones of the TRAM flap early postoperatively. Postoperative changes in near-infrared reflection spectroscopy values corresponded to clinical observations and blood flow in the superior epigastric artery measured by color-coded duplex sonography. Further experience is needed before near-infrared reflection spectroscopy can be advocated for routine clinical flap monitoring.     

1H and 31P NMR characterisation of a double breast coil for spectroscopic measurements and imaging.

For the first time a double turn breast coil has been described which can be used for 1H imaging, 1H spectroscopy and 31P spectroscopy. The paper describes basic technical features of the coil, coil design, B1 field/excitation field distribution for 1H and 31P, sensitivity, and feasibility for 31P spectroscopic in vivo studies. The main advantage of the double frequency tuneable coil is that 1H imaging for tumor localization and 31P spectroscopy for response control can be done without an additional repositioning of the patient.     

Discrimination of geographical origin and adulteration of radix astragali using fourier transform infrared spectroscopy and chemometric methods

Introduction – Radix Astragali, one of most widely used and important traditional Chinese medicines, is cultivated in different geographical regions. Because of varying growing conditions, the qualities of Radix Astragali vary, which can give rise to differences in clinical therapy. Detecting adulteration is a routine requirement in pharmaceutical practice. Objective – To develop a simple and accurate approach to discriminate the geographical origin and potential adulteration of Radix Astragali, derived from the root of Astragalus membranaceus (Fischer) Bunge var. mongholicus (Bunge) Hsiao, using Fourier transform infrared (FT‐IR) spectroscopy and chemometric methods. Methodology – To obtain characteristic IR spectra for accurate discrimination, a one‐solvent extraction method was utilised following a novel evaluation method for selecting appropriate solvents. Samples of Radix Astragali from different geographical origins were discriminated using FT‐IR spectroscopy and discriminant partial least squares (DPLS) methods. FT‐IR spectroscopy combined with Mahalanobis distance was employed to detect adulteration of Radix Astragali. Results – In comparison with other solvents, butanone was more effective at extracting samples. Radix Astragali samples were accurately assigned to their corresponding geographical origins by using FT‐IR spectroscopy and DPLS method. Most adulterated samples were detected accurately by application of FT‐IR spectroscopy combined with Mahalanobis distance. Conclusion – FT‐IR spectroscopy combined with chemometric method was developed and demonstrated to be a useful tool to discriminate geographical origin and adulteration of Radix Astragali. Copyright © 2010 John Wiley & Sons, Ltd.     

Study of a low-selenium environment in China by INAA and Mössbauer spectrometry

Biological Trace Element Research - The neutron activation analysis, γ coincidence spectroscopy, nondispersive hydrogen flame atomic fluorescence spectroscopy, and Mössbauer spectrometry...     

Hydrogen bonding-assisted interaction between amitriptyline hydrochloride and hemoglobin: spectroscopic and molecular dynamics studies

Herein, we have explored the interaction between amitriptyline hydrochloride (AMT) and hemoglobin (Hb), using steady-state and time-resolved fluorescence spectroscopy, UV–visible spectroscopy, and circular dichroism spectroscopy, in combination with molecular docking and molecular dynamic (MD) simulation methods. The steady-state fluorescence reveals the static quenching mechanism in the interaction system, which was further confirmed by UV–visible and time-resolved fluorescence spectroscopy. The binding constant, number of binding sites, and thermodynamic parameters viz. ΔG, ΔH, ΔS are also considered; result confirms that the binding of the AMT with Hb is a spontaneous process, involving hydrogen bonding and van der Waals interactions with a single binding site, as also confirmed by molecular docking study. Synchronous fluorescence, CD data, and MD simulation results contribute toward understanding the effect of AMT on Hb to interpret the conformational change in Hb upon binding in aqueous solution.