Improved signal to noise in high-resolution magic angle spinning total correlation spectroscopy studies of prostate tissues using rotor-synchronized adiabatic pulses.

A rotor-synchronized WURST-8 adiabatic pulse scheme was compared to the conventional MLEV-17 hard pulse scheme for isotropic mixing in total correlation spectroscopy (TOCSY) studies of intact human prostate tissues under high-resolution magic angle spinning (HR-MAS) conditions. Both mixing schemes were extremely sensitive to the rotational resonance condition and dramatic reductions in signal to noise were observed when pulse durations deviated from 1/(spin rate). A significant increase in cross-peak intensities was observed using rotor-synchronized WURST-8 adiabatic pulses versus those observed using the rotor-synchronized MLEV-17 hard pulse scheme in both solution and tissue. In tissue, absolute signal intensities ranged from 1.5x to 10.5x greater (average: 4.75x) when WURST-8 was used in place of MLEV-17. Moreover, the difference was so dramatic that several metabolite cross peaks observed using WURST-8 pulses were not observed using MLEV-17 pulses, including cross peaks corresponding to many of the choline- and ethanolamine-containing metabolites. Due to the complex modulation of TOCSY cross peaks for multiply coupled spins and the shorter T(2) relaxation times of tissue metabolites, maximum cross-peak intensities occurred at shorter mixing times than predicted by theory. In summary, a WURST-8 adiabatic mixing scheme produced significantly greater absolute cross-peak signal intensities than MLEV-17 hard pulse mixing, and maximum cross-peak intensity versus mixing time must be established for specific spin systems and T(2) relaxation times.     

High-resolution magic angle spinning proton NMR analysis of human prostate tissue with slow spinning rates.

The development of high-resolution magic angle spinning (HR-MAS) NMR spectroscopy for intact tissue analysis and the correlations between the measured tissue metabolites and disease pathologies have inspired investigations of slow-spinning methodologies to maximize the protection of tissue pathology structures from HR-MAS centrifuging damage. Spinning sidebands produced by slow-rate spinning must be suppressed to prevent their complicating the spectral region of metabolites. Twenty-two human prostatectomy samples were analyzed on a 14.1T spectrometer, with HR-MAS spinning rates of 600 Hz, 700 Hz, and 3.0 kHz, a repetition time of 5 sec, and employing various rotor-synchronized suppression methods, including DANTE, WATERGATE, TOSS, and PASS pulse sequences. Among them, DANTE, as the simplest scheme, has shown the most potential in suppression of tissue water signals and spinning sidebands, as well as in quantifying metabolic concentrations.     

Assessment of intervertebral disc degeneration with magnetic resonance single‐voxel spectroscopy

This study examined the feasibility of using short‐echo water‐suppressed point‐resolved spectroscopy (PRESS) on a clinical 3T magnetic resonance (MR) scanner for evaluating biochemical changes in degenerated bovine and cadaveric human intervertebral discs. In bovine discs (N = 17), degeneration was induced with papain injections. Degeneration of human cadaveric discs (N = 27) was assessed using the Pfirrmann grading on T₂‐weighted images. Chemicals in the carbohydrate region (Carb), the choline head group (Cho), the N‐acetyl region (N‐acetyl), and the lipid and lactate region (Lac+Lip) were quantified using ¹H PRESS, and were compared between specimens with different degrees of degeneration. The correlation between the spectroscopic findings and glycosaminoglycan (GAG) quantification using biochemical assays was determined. Significant differences were found between the ratios (N‐acetyl/Cho, N‐acetyl/Lac+Lip) acquired before and after papain injection in bovine discs. For human cadaveric discs, significant differences in the ratios (N‐acetyl/Carb, N‐acetyl/Lac+Lip) were found between discs having high and low Pfirrmann scores. Significant correlations were found between N‐acetyl/Lac+Lip and GAG content in bovine discs (R = 0.77, P = 0.0007) and cadaveric discs (R = 0.83, P < 0.0001). Significant correlation between N‐acetyl/Cho and GAG content was also found in cadaver discs (R = 0.64, P = 0.0039). This study demonstrates for the first time that short‐echo PRESS on a clinical 3T MR scanner can be used to noninvasively and can reproducibly quantify metabolic changes associated with degeneration of intervertebral discs. Magn Reson Med, 2009. © 2009 Wiley‐Liss, Inc.     

Magnetization transfer ratio mapping of intervertebral disc degeneration

The magnetization transfer ratio of the lumbar discs was spatially quantified from age‐matched subjects and the nucleus pulposus magnetization transfer ratio was correlated with T2‐weighted Pfirrmann grades. A moderate and significant linear correlation between magnetization transfer ratio and Pfirrmann grades was observed, suggesting that nucleus pulposus collagen relative density increases with degeneration. High‐resolution axial magnetization transfer ratio maps revealed elevated magnetization transfer ratio in the nucleus pulposa of injured and heavily degenerated discs. In the injured disc, significant elevation in nucleus pulposa magnetization transfer ratio was not accompanied by significant decrease in disc height. This observation may suggest a possible increase in absolute collagen content, in addition to increased collagen relative density. In summary, magnetization transfer MRI of the disc may serve as a noninvasive diagnostic tool for disc degeneration, in addition to other MRI techniques specific to proteoglycan content. Magn Reson Med, 2010. © 2010 Wiley‐Liss, Inc.     

腰椎失稳致大鼠椎间盘蜕变的组织学和影像学观察

目的:建立大鼠椎间盘蜕变的动物模型,并对其组织学和影像学改变进行观察。方法:采用SD大鼠30只,手术对照组15只,手术模型组15只。后正中切口,剥离骶棘肌并向两侧分开,显露L1~6棘突,切除,暴露双侧椎板、关节突,咬除L1~6的小关节的外1/2部分,切断椎板间韧带;对照组仅行皮肤切开。X线检查大鼠腰椎的影像学改变,收集大鼠腰椎间盘组织,行HE染色观察组织学的表现。结果:手术模型组于术后3个月时出现椎间隙的楔形改变,于术后6个月时出现终板软骨的钙化和椎间隙的狭窄,术后9个月时则更为明显,同手术对照组相比有显著性差异(P<0.05);组织学改变于术后3个月时即可见髓核的皱缩,终板的轻度钙化,纤维环的排列不规则,术后6个月和9个月时可见髓核的纤维化,终板钙化明显纤维环板层状结构消失。结论:腰椎失稳可以引起大鼠的椎间盘的蜕变,为不直接损伤椎间盘组织建立椎间盘蜕变动物模型的好方法。     

Biochemical correlates of thiazolidinedione-induced adipocyte differentiation by high-resolution magic angle spinning NMR spectroscopy.

Thiazolidinediones, a class of synthetic ligands to the peroxisome proliferator-activated receptor-gamma, induce terminal adipocyte differentiation of 3T3 F442A cells, and have already been used as alternative therapeutic agents for the treatment of liposarcoma in clinical trials. The biochemical changes occurring in the 3T3 F442A cell line and well-differentiated liposarcoma following induction of adipocyte differentiation with the thiazolidinedione troglitazone were measured using high-resolution magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. 3T3 F442A cell differentiation was characterized by a large accumulation of intracellular triglyceride and withdrawal from the cell cycle. Phosphatidylcholine (PTC), phosphocholine (PC), myo-inositol, and glycerol were found to be possible biochemical markers for adipocyte differentiation induced by thiazolidenediones. The molar ratio of PTC to PC increased fourfold in differentiated 3T3 F442A cells compared to undifferentiated cells, suggesting a substantial increase in CTP:phosphocholine cytidylyltransferase activity with differentiation. A 2.8-fold increase in the PTC:PC ratio was observed in the lipoma-like well-differentiated liposarcoma of three patients who were treated with troglitazone when compared to liposarcoma from patients not treated with this drug. Thus, this ratio may be an NMR-detectable marker of troglitazone efficacy and response to differentiation therapy for liposarcoma.     

486例腰椎间盘退变CT分析

目的 探讨腰椎间盘退变的CT表现及诊断价值。方法 通过对486例腰椎间盘退变的临床表现及CT所见进行回顾性分析并总结。结果 根据CT表现，把腰椎间盘退变分为椎间盘突出321例，椎间盘膨出165例。结论 CT是诊断腰椎间盘退变的重要检查方法。     

2H double quantum filtered (DQF) NMR spectroscopy of the nucleus pulposus tissues of the intervertebral disc.

Deuterium (2H) double-quantum filtered (DQF) NMR spectroscopy of nucleus pulposus (NP) tissues from human intervertebral discs is reported. The DQF spectral intensities, DQ build-up rates, and DQF-detected rotating-frame spin-lattice relaxation times are sensitive to the degree of hydration of the NP tissue, and display a monotonous correlation with age between 15 and 80 years. The implications of this work are that the changes in water dynamics as detected via DQF NMR spectroscopy may be used as a probe of tissue degeneration in NP, particularly in the early stages of degeneration to which most standard NMR methods are not sensitive.     

Reduction of spinning sidebands in proton NMR of human prostate tissue with slow high-resolution magic angle spinning.

High-resolution magic angle spinning (HRMAS) NMR spectroscopy has proven useful for analyzing intact tissue and permitting correlations to be made between tissue metabolites and disease pathologies. Extending these studies to slow-spinning methodologies helps protect tissue pathological structures from HRMAS centrifuging damage and may permit the study of larger objects. Spinning sidebands (SSBs), which are produced by slow spinning, must be suppressed to prevent the complication of metabolic spectral regions. In this study human prostate tissues, as well as gel samples of a metabolite mixture solution, were measured with continuous-wave (CW) water presaturation on a 14.1T spectrometer, with HRMAS spinning rates of 250, 300, 350, 600, and 700 Hz, and 3.0 kHz. Editing the spectra by means of a simple minimum function (Min(A, B, ..., N) for N spectra acquired at different but close spinning rates) produced SSB-free spectra. Statistically significant linear correlations were observed for metabolite concentrations quantified from the Min(A, B, ..., N)-edited spectra generated at low spinning rates, with concentrations measured from the 3 kHz spectra, and also with quantitative pathology. These results indicate the empirical utility of this scheme for analyzing intact tissue, which also may be used as an adjunct tool in pathology for diagnosing disease.     

Water exchange across the erythrocyte plasma membrane studied by HR-MAS NMR spectroscopy.

Water exchange across the plasma membrane of erythrocytes (red blood cells (RBCs)) was studied by means of high-resolution magic angle spinning (HR-MAS) NMR spectroscopy. Under HR-MAS conditions, the centrifugal force causes the splitting of RBC suspensions into a two-phase system composed of a central core of cell free water and an outer layer of tightly packed cells. Water belonging to each of these phases gives rise to two separated resonances. Chemical exchange between them is not detectable on the chemical shift or saturation transfer (ST) NMR time scale because of the physical separation between the phases. When the RBCs are dispersed and immobilized within a matrix made of cross-linked albumin, the splitting into a two-phase system is prevented and a single exchange-averaged peak for water is detected in (1)H HR-MAS NMR spectra. The lineshape of this peak is dependent on transmembrane exchange kinetics, since MAS averages out all the anisotropic magnetic interactions that are responsible for additional line-broadening under conventional liquid conditions. Line-shape analysis according to a two-site exchange model yielded a residence lifetime on the order of about 10 ms (at 37 degrees C) for a water molecule within the intracellular compartment, which is not too far from the generally accepted value of 9.6-14.8 ms.     

DTI及T2 mapping对腰椎间盘退变的诊断价值

目的：探讨磁共振扩散张量成像(DTI)表观扩散系数(ADC)、各向异性分数(FA)以及 T2 mapping T2值定量评估腰椎间盘退变程度的价值。方法健康志愿者12例,男8例,女4例;慢性下腰痛患者28例,男15例,女13例;年龄19~70岁。行腰椎MRI矢状位T2 WI、DTI、T2 mapping成像。对 L1~S1椎间盘进行Pfirrminn退变分级,测量各腰椎间盘髓核 ADC 值、FA 值以及T2值并分析其与不同腰椎间盘退变分级的相关性,比较 ADC值、FA值以及T2值对腰椎间盘退变的诊断价值。结果腰椎间盘的平均ADC值、T2值与Pfirrminn分级呈显著负相关关系,相关系数分别为r=-0.779,r=-0.708(P<0.001);FA值与Pfirrminn分级呈正相关关系,但是相关关系不密切(相关系数r=0.474,P<0.001)。结论 DTI和T2 mapping可以定量评估腰椎间盘的退变程度,其诊断价值为 ADC 值最高,T2值次之,FA 值较差。     

High-resolution (1)H NMR and magic angle spinning NMR spectroscopic investigation of the biochemical effects of 2-bromoethanamine in intact renal and hepatic tissue.

The metabolic consequences of xenobiotic-induced toxicity were investigated using high-resolution magic angle spinning (MAS) NMR spectroscopy of intact tissue. Renal papillary necrosis (RPN) was induced in Sprague-Dawley rats (n = 12) via a single i.p. dose of 250 mg/kg 2-bromoethanamine (BEA) hydrobromide. At 2, 4, 6, and 24 h after treatment with BEA, three animals were killed and tissue samples were obtained from liver, renal cortex, and renal medulla. Tissue samples were also removed at 2 and 24 h from matched controls (n = 6). (1)H MAS NMR spectroscopic techniques were used to analyze samples of intact tissue ( approximately 10 mg). Decreased levels of nonperturbing renal osmolytes (glycerophosphocholine, betaine, and myo-inositol) were observed in the renal papilla of BEA-treated animals at 6 and 24 h postdose (p.d.), concomitant with a relative increase in the tissue concentration of creatine. Increased levels of glutaric acid were found in all tissues studied in BEA-treated animals at 4 and 6 h p.d., indicating the inhibition of mitochondrial fatty acyl CoA dehydrogenases and mitochondrial dysfunction. Increased levels of trimethylamine-N-oxide occurred in the renal cortex at 6 h p.d. Changes in the metabolite profile of liver included an increase in the relative concentrations of triglycerides, lysine, and leucine. The novel application of (1)H MAS NMR to the biochemical analysis of intact tissues following a toxic insult highlights the potential of this technique as a toxicological probe in providing a direct link between urinary biomarkers of toxicity and histopathological evaluation of toxicological lesions.     

腰椎间盘镜下髓核摘除术治疗腰椎间盘突出症疗效分析

目的 探讨显微内镜椎间盘摘除术(microendoscopic discectomy,MED)治疗腰椎间盘突出症的疗效.方法 回顾性分析32例经MED治疗腰椎间盘突出症效果.结果 32例中27例完成镜下操作,随访4～54个月,优24例,良3例;术中出血30～400ml,平均120ml,切口长度平均2cm.中转开放手术5例,马尾神经损伤1例,硬膜损伤1例,损伤椎管内静脉丛3例.结论 MED具有手术切口小、创伤小、出血少、神经根减压彻底、不影响脊柱稳定性、术后恢复快等优点,但易出现较多的并发症.     

Proton high-resolution magic angle spinning NMR analysis of fresh and previously frozen tissue of human prostate.

The previously observed improvement in spectral resolution of tissue proton NMR with high-resolution magic angle spinning (HRMAS) was speculated to be due largely to freeze-thawing artifacts resulting from tissue storage. In this study, 12 human prostate samples were analyzed on a 14.1T spectrometer at 3 degrees C, with HRMAS rates of 600 and 700 Hz. These samples were measured fresh and after they were frozen for 12-16 hr prior to thawing. The spectral linewidths measured from fresh and previously frozen samples were identical for all metabolites except citrate and acetate. The metabolite intensities of fresh and freeze-thawed samples depend on the quantification procedures used; however, in this experiment the differences of means were <30%. As expected, it was found that tissue storage impacts tissue quality for pathological analysis, and HRMAS conditions alone are not sufficiently destructive to impair pathological evaluation. Furthermore, although storage conditions affect absolute metabolite concentrations in NMR analysis, relative metabolite concentrations are less affected.     

Localized in vivo isotropic-anisotropic correlation 1H NMR spectroscopy using ultraslow magic angle spinning.

In a previous work (1), the susceptibility broadening in the (1)H NMR metabolite spectrum obtained in a live mouse was separated from the isotropic information, which significantly increased the spectral resolution. This was achieved using ultraslow magic angle spinning (MAS) of the animal combined with a modified phase-corrected magic angle turning (PHORMAT) pulse sequence. However, PHORMAT cannot be used for spatially selective spectroscopy. This article introduces a modified sequence called localized magic angle turning (LOCMAT) that makes this possible. Proton LOCMAT spectra were obtained from the liver and heart of a live mouse while the animal was spun at a speed of 4 Hz in a 2 Tesla field. It was found that even in this relatively low field, LOCMAT provided isotropic line widths that were a factor of 4-10 times smaller than those obtained in a stationary animal. Furthermore, the susceptibility broadening of the heart metabolites showed unusual features that are not observed in dead animals. The limitations of LOCMAT and possible ways to improve the technique are discussed. It is concluded that in vivo LOCMAT can significantly enhance the utility of NMR spectroscopy for biomedical research.     

High-resolution (1)H NMR spectroscopy in organs and tissues using slow magic angle spinning.

It is shown that high-resolution (1)H NMR spectra of intact excised tissues and organs can be obtained by rotating the sample slowly about an axis at the magic angle of 54 degrees 44' with the external magnetic field. In this way tissue and cellular damage invoked by standard magic angle spinning (MAS) experiments, where spinning speeds of several kHz are typically employed, are minimized. Special RF pulse sequences, developed originally in solid state NMR, can be used to produce a spinning sideband-free isotropic spectrum. In this article the first results are shown of the brain, heart, liver, gluteus muscle, and kidney excised from mice using the 2D-phase-altered spinning sidebands (PASS) technique and employing MAS spinning speeds of 43-125 Hz. It was found that with slow sample spinning similar, and in some cases even better, spectral resolutions are obtained as compared with fast MAS.