Pattern Recognition Analysis of Proton Nuclear Magnetic Resonance Spectra of Brain Tissue Extracts from Rats Anesthetized with Propofol or Isoflurane

Background

General anesthesia is routinely used as a surgical procedure and its safety has been endorsed by clinical outcomes; however, its effects at the molecular level have not been elucidated. General anesthetics influence glucose metabolism in the brain. However, the effects of anesthetics on brain metabolites other than those related to glucose have not been well characterized. We used a pattern recognition analysis of proton nuclear magnetic resonance spectra to visualize the changes in holistic brain metabolic phenotypes in response to the widely used intravenous anesthetic propofol and the volatile anesthetic isoflurane.

Methodology/Principal Findings

Rats were randomized into five groups (n = 7 each group). Propofol and isoflurane were administered to two groups each, for 2 or 6 h. The control group received no anesthesia. Brains were removed directly after anesthesia. Hydrophilic compounds were extracted from excised whole brains and measured by proton nuclear magnetic resonance spectroscopy. All spectral data were processed and analyzed by principal component analysis for comparison of the metabolite profiles. Data were visualized by plotting principal component (PC) scores. In the plots, each point represents an individual sample. The propofol and isoflurane groups were clustered separately on the plots, and this separation was especially pronounced when comparing the 6-h groups. The PC scores of the propofol group were clearly distinct from those of the control group, particularly in the 6-h group, whereas the difference in PC scores was more subtle in the isoflurane group and control groups.

Conclusions/Significance

The results of the present study showed that propofol and isoflurane exerted differential effects on holistic brain metabolism under anesthesia.     

Recent Advances in Computational Methods for Nuclear Magnetic Resonance Data Processing

Although three-dimensional protein structure determination using nuclear magnetic resonance (NMR) spectroscopy is a computationally costly and tedious process that would benefit from advanced computational techniques, it has not garnered much research attention from specialists in bioinformatics and computational biology. In this paper, we review recent advances in computational methods for NMR protein structure determination. We summarize the advantages of and bottlenecks in the existing methods and outline some open problems in the field. We also discuss current trends in NMR technology development and suggest directions for research on future computational methods for NMR.     

A Guide to the Metabolic Pathways and Function of Metabolites Observed in Human Brain 1H Magnetic Resonance Spectra

The current knowledge of the normal biochemistry of compounds that give rise to resonances in human brain proton magnetic resonance spectra measureable at readily available field strengths (i.e. ≤3 T) is reviewed. Molecules covered include myo- and scyllo-inositol, glycerophospho- and phospho-choline and choline, creatine and phosphocreatine, N-acetylaspartate, N-acetylaspartylglutamate, glutamate, glutamine, γ-aminobutyrate, glucose, glutathione and lactate. The factors which influence changes in the levels of these compounds are discussed. As most proton resonances in the brain at low field are derived from a combination of moieties whose biochemistry is complex and interrelated, an understanding of the mechanisms underlying why these species change is crucial to meaningful interpretation of human brain spectra.     

基于fMRI观察正常人脑中枢对针刺得气感反应的初步研究

目的:运用功能性磁共振成像技术观察正常人脑中枢对针刺得气感的反应特点,探索针刺得气的物质基础。方法:按照纳入排除标准收集男、女健康志愿者各35例,随机分为2组,针刺组40人,假针刺组30人,分别以外关穴针刺与假针刺作为刺激因素。采用Block方法设计刺激程序,运用功能性磁共振成像技术收集脑中枢反应信号。扫描完毕随即使用针刺感觉量表评价受试对象的感觉,依据感觉量化结果将图像数据分为针刺得气感组与无感觉对照组,以P≤0.0001,uncorrected,K10为统计检验显著性标准,以对照无感觉组为基线,在Matlab平台上采用SPM2.0软件包对图像数据进行处理和分析。结果:(1)得气效应:针刺组29人、假针刺组1人以酸麻胀重等得气指征为主,VAS评分4.62±3.05;针刺组2人轻微刺痛感,假针刺组21人无感觉、5人轻微刺痛感。(2)脑中枢激活效应:针刺得气激活的脑功能区有双侧额上回(BA6)、额下回(BA47)、缘上回(BA40),左侧额中回(BA47)、中央前回(BA10)、颞中回(BA21),右侧额中回(BA10)、顶下小叶(BA40)、中央前回(BA6)、颞上回(BA10)、颞中回(BA39)、颞下回(BA20)。结论:针刺得气感能特异性的激活正常人脑功能区,针刺得气效应与脑中枢密切相关。     

13C Solid-State Cross Polarization/Magic-Angle-Spinning Nuclear Magnetic Resonance Spectra of Natural and Synthetic Melanins

The availability of NMR spectrometers operating in cross polarization/magic angle spinning (CP-MAS) has provided a powerful tool for the structural elucidation of insoluble materials. In this ¹³C NMR study of eumelanins we report the first direct evidence of the presence of different chemical functionalities in synthetic and natural eumelanins. These spectra contain useful information for the characterization of melanins from different sources.     

Fingerprinting Analysis of Saposhnikovia divaricata using 1H Nuclear Magnetic Resonance Spectroscopy and High Performance Liquid Chromatography

The 1H nuclear magnetic resonance (1H NMR) fingerprints of fractionated non-polar and polar extracts (control substance for plant drug [CSPD] A and B) from the roots of 12 specimens of Saposhnikovia divaricata (Turcz.) Schischk were achieved with Fourier Transform (FT)-NMR spectrometer and assigned by comparison to each other and to the 1H NMR spectra of the isolated individual compounds. These fingerprints were found to be uniform in terms of the specificity for the implication of all 12 specimens being systematically of the same origin. The uniformity was further affirmed by high performance liquid chromatography (HPLC), which also revealed exactly identical specificity for the identified S. divaricata species with the 1H NMR appearances of corresponding CSPD on the part of the composition of characteristic constituents when comparing to corresponding individual compounds. This investigation unambiguously shows that the specific signals from the chemotaxonomically significant compounds of chromones and coumarins in S. divaricata are exhibited distinctively in the composite features of both 1H NMR fingerprints and HPLC profiles. The 1H NMR and HPLC profiles established can successfully be used as reference for the authentication of the origin of S. divaricata species as well as for chemotaxonomic studies.     

Nuclear Magnetic Resonance of Sodium-23 Linoleate-Water: Basis for an Alternative Interpretation of Sodium-23 Spectra within Cells

The ²³Na spectrum from liquid crystals of sodium linoleate in water has been studied by nuclear magnetic resonance (NMR) techniques. The integrated intensity of the visible central spectral line was 34-39% of the intensity of a reference sample containing an equal quantity and concentration of ²³Na nuclei. Since satellite signals were clearly demonstrable, the effect reflected a nuclear quadrupolar interaction rather than a splitting of the ²³Na into two populations of bound and free nuclei. It is proposed that a similar quadrupolar effect may be the basis for the apparent binding of the ²³Na observed in biological systems.     

Fingerprinting Analysis of Rhizoma Chuanxiong of Commercial Types using 1H Nuclear Magnetic Resonance Spectroscopy and High Performance Liquid Chromatography Method

The 1H nuclear magnetic resonance (1H NMR) fingerprints of fractionated non-polar extracts (control substance for a plant drug (CSPD) A) from Rhizoma chuanxiong, the rhizomes of Ligusticum chuanxiong Hort., of seven specimens from different sources were measured on Fourier Transform (FT)-NMR spectrometer and assigned by comparing them with the 1H NMR spectra of the isolated pure compounds. The 1H NMR fingerprints showed exclusively characteristic resonance signals of the major special constituents of the plant. Although the differences in the relative intensity of the 1H NMR signals due to a discrepancy in the ratio of the major constituents among these samples could be confirmed by high performance liquid chromatography analysis, the general features of the 1H NMR fingerprint established for an authentic sample of the rhizomes of L. chuanxiong exhibited exclusive data from those special compounds and can be used for authenticating L. Chuanxiong species.     

激光极化技术制备核磁共振探针的方法和应用原理

通过激光增强核自旋极化技术,可以得到很高的非热平衡极化度和长驰豫时间的惰性气体（129Xe和3He),其作为核磁共振探针,具有广泛的生物医学应用价值,该文探讨了激光增强129Xe和3He核自旋极化的磁共振成像和波谱学原理,介绍了激光极化惰性气体的设备和方法,以及极化气体的收集贮存和输入过程,最后对氢核及极化核的磁共振成像,极化气体129Xe和3He的应用进行了比较性总结。     

研究报告: 小动物高分辨扩散磁共振成像数据分析方法

扩散磁共振成像(dMRI)是一种非侵入性的、能提供生物体内水分子扩散运动相关信息的成像技术,可用于检测神经纤维微观结构的变化．dMRI的出现为大脑结构与功能研究提供了全新的检测手段．过去的20年中,dMRI在实验方法和临床应用上均取得了重大进展．然而dMRI应用在基于动物模型的临床前脑成像研究中却并不常见．本文针对dMRI在临床前脑成像研究中的应用,建立了系列针对小动物高分辨dMRI数据的分析方法：a．构建了大鼠高分辨dMRI图像模板;b．实现了适用于小动物研究的基于体素的统计分析(VBA)方法与基于纤维束的空间统计分析(TBSS)方法;c．实现了小动物脑白质纤维束的确定性与概率性跟踪．这些方法的实现不仅能为小动物脑dMRI研究提供统一的图像模板与完善的计算方法,还将大大促进dMRI技术在小动物脑成像研究中的应用．     

Fingerprinting Analysis of Rhizoma Chuanxiong of Commercial Types using ^1 H Nuclear Magnetic Resonance Spectroscopy and High Performance Liquid Chromatography Method

The ^1 H nuclear magnetic resonance （^1 H NMR） fingerprints of fractionated non-polar extracts （control substance for a plant drug （CSPD） A） from Rhizoma chuanxiong, the rhizomes of Ligusticum chuanxiong Hort., of seven specimens from different sources were measured on Fourier Transform （FT）-NMR spectrometer and assigned by comparing them with the ^1 H NMR spectra of the isolated pure compounds. The ^1 H NMR fingerprints showed exclusively characteristic resonance signals of the major special constituents of the plant. Although the differences in the relative intensity of the ^1H NMR signals due to a discrepancy in the ratio of the major constituents among these samples could be confirmed by high performance liquid chromatography analysis, the general features of the ^1H NMR fingerprint established for an authentic sample of the rhizomes of L. chuanxiong e. hibited exclusive data from those special compounds and can be used for authenticating L. Chuanxiong species.     

Nuclear Magnetic Resonance Structure and Dynamics of the Response Regulator Sma0114 from Sinorhizobium meliloti

Receiver domains control intracellular responses triggered by signal transduction in bacterial two-component systems. Here, we report the solution nuclear magnetic resonance structure and dynamics of Sma0114 from the bacterium Sinorhizobium meliloti, the first such characterization of a receiver domain from the HWE-kinase family of two-component systems. The structure of Sma0114 adopts a prototypical α(5)/β(5) Rossman fold but has features that set it apart from other receiver domains. The fourth β-strand of Sma0114 houses a PFxFATGY sequence motif, common to many HWE-kinase-associated receiver domains. This sequence motif in Sma0114 may substitute for the conserved Y-T coupling mechanism, which propagates conformational transitions in the 455 (α4-β5-α5) faces of receiver domains, to prime them for binding downstream effectors once they become activated by phosphorylation. In addition, the fourth α-helix of the consensus 455 face in Sma0114 is replaced with a segment that shows high flexibility on the pico- to nanosecond time scale by (15)N relaxation data. Secondary structure prediction analysis suggests that the absence of helix α4 may be a conserved property of the HWE-kinase-associated family of receiver domains to which Sma0114 belongs. In spite of these differences, Sma0114 has a conserved active site, binds divalent metal ions such as Mg(2+) and Ca(2+) that are required for phosphorylation, and exhibits micro- to millisecond active-site dynamics similar to those of other receiver domains. Taken together, our results suggest that Sma0114 has a conserved active site but differs from typical receiver domains in the structure of the 455 face that is used to effect signal transduction following activation.     

13C and 1H Nuclear Magnetic Resonance Study of Glycogen Futile Cycling in Strains of the Genus Fibrobacter

We investigated the carbon metabolism of three strains of Fibrobacter succinogenes and one strain of Fibrobacter intestinalis. The four strains produced the same amounts of the metabolites succinate, acetate, and formate in approximately the same ratio (3.7/1/0.3). The four strains similarly stored glycogen during all growth phases, and the glycogen-to-protein ratio was close to 0.6 during the exponential growth phase. ¹³C nuclear magnetic resonance (NMR) analysis of [1-¹³C]glucose utilization by resting cells of the four strains revealed a reversal of glycolysis at the triose phosphate level and the same metabolic pathways. Glycogen futile cycling was demonstrated by ¹³C NMR by following the simultaneous metabolism of labeled [¹³C]glycogen and exogenous unlabeled glucose. The isotopic dilutions of the CH₂ of succinate and the CH₃ of acetate when the resting cells were metabolizing [1-¹³C]glucose and unlabeled glycogen were precisely quantified by using ¹³C-filtered spin-echo difference ¹H NMR spectroscopy. The measured isotopic dilutions were not the same for succinate and acetate; in the case of succinate, the dilutions reflected only the contribution of glycogen futile cycling, while in the case of acetate, another mechanism was also involved. Results obtained in complementary experiments are consistent with reversal of the succinate synthesis pathway. Our results indicated that for all of the strains, from 12 to 16% of the glucose entering the metabolic pathway originated from prestored glycogen. Although genetically diverse, the four Fibrobacter strains studied had very similar carbon metabolism characteristics.     

Integrating Functional and Diffusion Magnetic Resonance Imaging for Analysis of Structure-Function Relationship in the Human Language Network

Background

The capabilities of magnetic resonance imaging (MRI) to measure structural and functional connectivity in the human brain have motivated growing interest in characterizing the relationship between these measures in the distributed neural networks of the brain. In this study, we attempted an integration of structural and functional analyses of the human language circuits, including Wernicke''s (WA), Broca''s (BA) and supplementary motor area (SMA), using a combination of blood oxygen level dependent (BOLD) and diffusion tensor MRI.

Methodology/Principal Findings

Functional connectivity was measured by low frequency inter-regional correlations of BOLD MRI signals acquired in a resting steady-state, and structural connectivity was measured by using adaptive fiber tracking with diffusion tensor MRI data. The results showed that different language pathways exhibited different structural and functional connectivity, indicating varying levels of inter-dependence in processing across regions. Along the path between BA and SMA, the fibers tracked generally formed a single bundle and the mean radius of the bundle was positively correlated with functional connectivity. However, fractional anisotropy was found not to be correlated with functional connectivity along paths connecting either BA and SMA or BA and WA.

Conclusions/Significance

These findings suggest that structure-function relations in the human language circuits may involve a number of confounding factors that need to be addressed. Nevertheless, the insights gained from this work offers a useful guidance for continued studies that may provide a non-invasive means to evaluate brain network integrity in vivo for use in diagnosing and determining disease progression and recovery.     

1H Nuclear Magnetic Resonance Spectroscopy-Based Studies of the Metabolism of Food-Borne Carcinogen 2-Amino-3-Methylimidazo[4,5-f]Quinoline by Human Intestinal Microbiota

2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) is a mutagenic/carcinogenic compound formed from meat and fish during cooking. Following ingestion, IQ is metabolized mainly by liver xenobiotic-metabolizing enzymes, but intestinal bacteria may also contribute to its biotransformation. The aim of this study was to investigate the metabolism of IQ by the human intestinal microbiota. Following incubation of IQ (200 μM) under anoxic conditions with 100-fold dilutions of stools freshly collected from three healthy volunteers, we quantified residual IQ by high-pressure liquid chromatography (HPLC) analysis and characterized the production of IQ metabolites by in situ ¹H nuclear magnetic resonance (¹H-NMR) spectroscopic analysis of crude incubation media. In addition, we looked for IQ-degrading bacteria by screening collection strains and by isolating new strains from the cecal contents of human-microbiota-associated rats gavaged with IQ on a regular basis. HPLC and ¹H-NMR analyses showed that the three human microbiota degraded IQ with different efficiencies (range, 50 to 91% after 72 h of incubation) and converted it into a unique derivative, namely, 7-hydroxy-IQ. We found 10 bacterial strains that were able to perform this reaction: Bacteroides thetaiotaomicron (n = 2), Clostridium clostridiiforme (n = 3), Clostridium perfringens (n = 1), and Escherichia coli (n = 4). On the whole, our results indicate that bacteria belonging to the predominant communities of the human intestine are able to produce 7-hydroxy-IQ from IQ. They also suggest interindividual differences in the ability to perform this reaction. Whether it is a metabolic activation is still a matter of debate, since 7-hydroxy-IQ has been shown to be a direct-acting mutagen in the Ames assay but not carcinogenic in laboratory rodents.     

高分辨魔角旋转核磁共振和主成分分析研究人类低级星形细胞瘤和脑膜瘤的代谢组特征

采用高分辨魔角旋转核磁共振（HRMAS ^1H NMR）技术结合主成分分析（PCA）方法研究了39例人体脑肿瘤组织的代谢组特征．39例肿瘤样本分别来自39个脑肿瘤患者,包括15例低级星形细胞瘤,13例纤维型脑膜瘤和11例过渡型脑膜瘤．核磁共振波谱分析结果表明,脑肿瘤组织的代谢组中丰要含有脂肪酸、乳酸、胆碱代谢物（如胆碱、磷酸胆碱和甘油磷酸胆碱）、氯基酸（如丙氨酸、谷氨酸、谷氮酰胺、牛磺酸）、N-乙酰天门冬氨酸（NAA）和谷胱甘肽等代谢物．通过对核磁共振谱进行主成分分析（PCA）,发现低级星形细胞瘤和脑膜瘤的代谢组之间具有明显的差异,而在过渡型和纤维型两个亚类脑膜瘤之间该差别相对较小．与脑膜瘤相比,低级星形细胞瘤中甘油磷酸胆碱、磷酸胆碱、肌醇与肌酸的含量较高,而丙氨酸、谷氨酸、谷氨酰胺、谷胱甘肽和牛磺酸的含量较低．NAA的含量在低级星形细胞瘤中尽管较低但能观察到,而脑膜瘤中却未发现NAA的信号．结果衷明,HRMAS ^1H NMR和多变量统计分析相结合的组织代谢组学方法,不仅能有效区分不同类型的脑肿瘤,而且还可以为脑肿瘤提供丰富的代谢组信息,这些信息对研究肿瘤发生发展的机制具有潜在的意义．