Biochemical characterization of metastatic lymph nodes of breast cancer patients by in vitro 1H magnetic resonance spectroscopy: a pilot study

Using one-dimensional (1D) and two-dimensional (2D) proton nuclear magnetic resonance (NMR) methods, the perchloric acid extract of involved (n = 11) and noninvolved (n = 12) axillary lymph nodes (ALN) of breast cancer patients was investigated. Resonances from 40 metabolites such as lactate (Lac), glucose, several amino acids (alanine, lysine, glutamic acid, glutamine, etc.), nucleotides (adenosine triphosphate, guanosine triphosphate, uridine triphosphate, uridine monophosphate, etc.), membrane metabolites [glycerophosphocholine (GPC), phosphocoline (PC), phosphoethanolamine (PE), choline] were unambiguously assigned in both the involved and noninvolved ALN. The concentration of PC/GPC (p = 0.002) was significantly higher in the involved compared to noninvolved nodes. In addition, the concentration of glycolytic product Lac (p = 0.0001) was also found to be significantly higher in involved nodes. Increased concentration of membrane metabolites PC/GPC may be attributed to increased membrane synthesis in malignant cells and, therefore, suggests the presence of metastatic cells in lymph nodes. The higher concentration of Lac is indicative of the presence of malignant cells that derive energy via anaerobic glycolytic pathway. Present results demonstrate the potentials of in vitro proton NMR in detecting malignant cells in ALN and such studies may have an important bearing in determining the prognosis of breast cancer patients.     

Fluorescence diagnostics of metastatic lesion of regional lymph nodes upon surgical treatment of breast cancer

We have performed intraoperative fluorescence diagnostics of 60 patients for metastases of breast cancer to regional lymph nodes. All the patients were divided into two groups. The first group consisted of 50 patients, which were surgically treated at the first stage. The second group consisted of ten patients, which underwent combined treatment. At the first stage, they received from two to four courses of neoadjuvant chemotherapy, and, at the second stage, the surgical intervention was performed. The intraoperative fluorescence diagnostics was performed using the preparation alasens (precursor of protoporphyrin IX in the human organism). The occurrence of fluorescence of alasens-induced protoporphyrin IX was determined visually and using the local fluorescence spectroscopy method. Altogether, 498 lymph nodes were examined: 408 in the first group and 90 in the second one. For the first group, the sensitivity of the method was found to be 87.2%, and its specificity, 94.8%; in the second group, these parameters were determined to be 77 and 78%, respectively. The first experience of the application of the intraoperative fluorescence diagnostics of metastatic lesion of lymph nodes in patients with breast cancer has shown its high efficiency and application potential.     

Whole-body diffusion-weighted imaging vs. FDG-PET for the detection of non-small-cell lung cancer. How do they measure up?

Objective

To compare the diagnostic efficacy of whole-body diffusion-weighted imaging (WB-DWI) and [18F] fluoro-2-D-glucose PET/CT(FDG-PET/CT)for assessment of non–small cell lung cancer (NSCLC) patients.

Materials and Methods

A group of 56 patients (21 female, 35 male; 35–76 years) with NSCLC proved by pathologic examination or follow-up imaging findings was set as reference standards, and all patients underwent both WB-DWI at 1.5T (MAGNETOM Avanto) and PET/CT (Biograph 16). For WB-DWI, a free breathing diffusion-weighted single-shot spin-echo epi-sequence in five-stations (head-neck, thorax, abdomen, pelvis-thigh) was used. Each station-series contained 30 contiguous axial slices. Imaging parameters: FOV 360×360 mm, matrix size 128×80. B-values: 0 and 1000 s/mm² applied along x, y and z, 5 averages, acquisition time: 2.23 min/series, total: 11.55 min. The efficacy of WB-DWI and PET/CT were determined in a blinded reading by two radiologists and two nuclear medicine physicians using pathology and size change during follow up exams as the reference standard.

Results

Primary tumors (n=56 patients) were correctly detected in 56 (100%) patients by both PET/CT and WB-DWI. Ninety-six lymph nodes metastases were determined with pathologic and follow-up examinations. Sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value (NPV) being for lymph node metastases: 91%, 90%, 90%, 96%, 80% with WB-DWI and 98%, 97%, 97%, 99%, 93% with PET-CT, other metastases: 90%, 95%, 92%, 97%, 83% with WB-DWI and 98%, 100%, 98%, 100%, 95% with PET-CT). Differences in the accuracy of lymph node metastasis detection between PET/CT and WB-DWI (P=.031) were significant. The differences were not statistically significant for detection of other metastases.

Conclusions

WB-DWI is a feasible clinical technique for the assessment of NSCLC, lymph nodes and metastastic spread with high sensitivity and accuracy, but it was limited in the evaluation of neck lymph node metastases and small metastastic lung nodules.     

衰减全反射傅里叶变换红外光谱技术的临床应用研究进展

系统回顾了衰减全反射傅里叶变换红外光谱(attenuated total reflection Fourier transform infrared, ATR-FTIR)技术在临床医学领域中的应用研究进展。ATR-FTIR光谱技术具有实时、简单、无创扫描生物组织样品的优势,通过结合多模式识别统计学方法并对照临床及病理学诊断结果,可对生物组织样本进行判别分析。目前,对甲状腺、乳腺,以及肺组织等新鲜离体组织的光谱学检测中,ATR-FTIR技术可达到较高的判别敏感性、准确性与特异性;尤其在对甲状腺和乳腺疾病患者的前哨淋巴结活检的研究中,应用ATR-FTIR技术,可对良、恶性组织进行敏感性、准确性与特异性均较高的鉴别研究,对辅助临床诊断具有潜在的重要价值。此外,利用ATR-FTIR光谱技术对生物体液进行光谱分析并进一步诊断疾病的研究也在逐步开展。由于血清中所含成分的改变对于提示疾病状况具有重要价值,因而血清ATR-FTIR光谱学诊断方法成为研究热点,已有应用ATR-FTIR技术对脑胶质细胞瘤、心肌梗死、肾衰、阿尔茨海默病、卵巢癌与子宫内膜癌等多种疾病进行分析的研究报道。目前已可通过ATR-FTIR技术对卵巢癌进行疾病分期的判别诊断,该技术有望成为卵巢癌筛查的重要方法。由于血清ATR-FTIR光谱学技术具有检测快速、判别准确、性价比高等优势,有望成为今后生物医学发展的重要方向之一。     

Quantitative multivoxel proton chemical shift imaging of the breast

The study of focal pathology by single-voxel magnetic resonance spectroscopy (MRS) is hampered by the impossibility to study tissue heterogeneity or compare the metabolite signals in breast lesion directly to those in unaffected tissue. Multivoxel MRS studies, while potentially allowing for truly quantitative tissue characterization, have up to now also been far from quantitative with, for example, the signal-to-noise ratio of the choline (Cho) signal serving as measure of tumor activity. Shown in this study is that in a standard clinical setting with a regular 1.5-T magnetic resonance scanner, it is possible to perform quantitative multivoxel MRS. With the use of literature values for the T1 and T2 relaxation times of Cho and water in fibroglandular breast tissue and tumors, one can determine the concentrations of Cho in different tumor compartments and surrounding tissues in two brief multivoxel MRS measurements. This opens excellent perspectives to quantitative diagnostic and follow-up studies of focal pathology such as lesions suspected of breast cancer.     

Controlled mechanical ventilation to detect regional lymph node metastases in esophageal cancer using USPIO-enhanced MRI; comparison of image quality

BackgroundArtifacts caused by respiratory motion or ventilation-induced chest movements are a major problem for thoracic MRI, as they can obscure important anatomical structures such as lymph node metastases. We compared image quality of routine breathhold with intermittent apnea during controlled mechanical ventilation of patients under general anesthesia as the ideal situation without respiratory motion in the detection and characterization of regional lymph nodes in esophageal cancer.MethodsIn this prospective study, 10 patients treated for esophageal cancer underwent ultrasmall superparamagnetic iron oxide (USPIO) enhanced MRI scans. Before neoadjuvant therapy, MRI scans were acquired with a routine breathhold technique. After neoadjuvant therapy, patients were scanned under general anesthesia immediately prior to surgery with controlled mechanical ventilation. The image quality was compared using a Likert scale questionnaire based on visibility of anatomical structures and image artifacts.ResultsMRI with controlled mechanical ventilation and prolonged controlled apnea of 4 min was safe and feasible. All cardio-respiratory monitoring parameters remained stable during the apnea phases. Mediastinal and upper abdominal lymph nodes down to 2 mm in size could be visualized with all sequences. All image quality criteria, including visibility of thoracic structures and regional lymph nodes were scored higher using the controlled ventilation sequences compared to the routine breathhold phase.ConclusionUSPIO-enhanced MRI with controlled mechanical ventilation is superior to routine breathhold MRI in visualizing lymph nodes, which warrants new motion reduction techniques to use MRI for the detection of lymph node metastases in patients with esophageal cancer.     

Controlled delivery of Gd-containing liposomes to lymph nodes: surface modification may enhance MRI contrast properties

Surface modification of liposomes containing Gd-diethylenetriaminepentaacetyl-phosphatidylethanolamine (Gd-DTPA-PE) as an amphiphilic paramagnetic label influences their contrast properties as MRI agents. Liposomes with the mean diameter ca.220 nm and 10% mol content of Gd-DTPA-PE were modified with dextran (molecular weight 6 kDa) or polyethyleneglycol (PEG, molecular weight 5 kDa). The preparations obtained were used for MR visualization of axillary and subscapular lymph nodes after SC injection into rabbit's paw. Dextran-modified liposomes were found to have limited or no effect on lymph node/muscle MR signal intensity ratio during the first 2 h after the administration when compared to nonmodified plain liposomes, whereas the liposomes modified with PEG demonstrated a 3-3.5-fold enhancement of lymph node signal. However, the partial biodistribution studies with the ¹¹¹In-labeled liposomes revealed that PEG-modified vesicles accumulated in the axillary lymph node at half the amount comparing with nonmodified and dextran-modified liposomes. The phenomenon found might be explained by noticeably increased relaxivity of PEG-modified Gd-liposomes.     

脑科学研究中的质子磁共振波谱方法

核磁共振（NMR）波谱作为一种操作简单、高效且重复性良好的技术手段，在脑科学领域的应用日益广泛，尤其是离体核磁共振氢谱（¹H NMR）和活体核磁共振氢谱（¹H MRS）.¹H NMR和¹H MRS在实验设计、样品预处理、数据处理等方面各有优劣、各擅胜场.本综述主要从适用范围、样品预处理、数据处理分析等方面，对二者在方法学层面的研究现状进行总结和讨论，以期为脑科学领域的研究者进行大脑代谢相关研究时提供一定的参考和帮助.     

FTIR结合主成分分析法对直肠癌转移淋巴结的研究

应用傅里叶变换红外光谱(FTIR)联合主成分分析法(PCA),分析直肠癌转移淋巴结的谱学特征,并对直肠癌转移淋巴结和非转移淋巴结进行线性判别分析。80例直肠癌转移淋巴结和80例未转移淋巴结进行FTIR光谱分析,计算峰强并进行主成分分析,得出在波数4 000～1 700 cm-1范围主成分1(Principal components 1,PC1)是3 260 cm-1,PC2为1 740 cm-1。波数1 700～1 000 cm-1范围,PC1为1 640 cm-1,PC2为1 080 cm-1,将良、恶性淋巴结光谱3 260,1 740,1 640,1 080 cm-1相对峰强比(I/I_{1 460})和波数1 080和1 300 cm-1进行t检验,良、恶性结果差异有统计学意义(p<0.05),表明癌转移淋巴结中蛋白含量、蛋白的形成、氨基酸增多;脂肪含量明显减少与癌组织中无氧酵解脂肪含量减少有关。将相对峰强比(I_{1 080}/I_{1 460}, I_{1 640}/I_{1 460}, I_{3 260}/I_{1 460}, I_{1 740}/I_{1 460}, n=160)进行PCA聚类分析,结果显示可以将良恶性淋巴结鉴别,良性淋巴结聚类在第一和四象限,恶性淋巴结聚类在二和三象限。将相对峰强比、1 080和1 300 cm-1进行线性判别分析(LDA),将25例淋巴结作为验证集进行分析,得出PCA/LDA模型的敏感度是87.5%,特异度是88.5%。结果表明傅里叶变换红外光谱分析技术可成为术中原位、在体和快速诊断直肠癌淋巴结转移的一种简便方法。     

Support Vector Machine on Fluorescence Landscapes for Breast Cancer Diagnostics

Excitation-emission matrices (EEM) and total synchronous fluorescence spectra (SFS) of normal and malignant breast tissue specimens are measured in UV-VIS spectral region to serve as data inputs in development of Support Vector Machine (SVM) based breast cancer diagnostics tool. Various input data combinations are tested for classification accuracy using SVM prediction against histopathology findings to discover the best combination regarding diagnostics sensitivity and specificity. It is shown that with EEM data SVM provided 67?% sensitivity and 62?% specificity diagnostics. With SFS data SVM provided 100?% sensitivity and specificity for a several input data combinations. Among these combinations those that require minimal data inputs are identified.     

Diagnostic value of breast MRI for predicting metastatic axillary lymph nodes in breast cancer patients: diffusion-weighted MRI and conventional MRI

Purposes

To evaluate the diagnostic value of diffusion-weighted MRI (DWI) and combination of conventional MRI and DWI to predict metastatic axillary lymph nodes in breast cancer.

Materials and methods

Two hundred fifty-two breast cancer patients with 253 axillae were included. The morphological parameters on axial T2-weighted images without fat saturation and apparent diffusion coefficient (ADC) values were retrospectively analyzed. An independent t-test/chi-square test and receiver operating characteristics (ROC) curve analysis were used.

Results

On conventional MRI, short and long axis length, maximal cortical thickness, relative T2 value, loss of fatty hilum (p < 0.001 for each), and eccentric cortical thickening (p < 0.003) were statistically significantly different between the metastatic and nonmetastatic groups. The short axis to long axis ratio was not a statistically significant parameter. The ADC value was significantly different between the 2 groups, with an AUC that was higher than that of conventional MR parameters (AUC, 0.815; threshold, ≤ 0.986 × 10–3 mm²/sec; sensitivity, 75.8%; specificity, 83.9%). Using the adopted thresholds for each parameter, a total number of findings suggesting malignancy of 4 or higher was determined as the threshold, with high specificity (90.1%).

Conclusion

Using conventional MRI and DWI, we can evaluate the axilla in breast cancer with high specificity.     

Use of USPIO-induced magnetic susceptibility artifacts to identity sentinel lymph nodes and lymphatic drainage patterns. I. dependence of artifact size with subcutaneous Combidex® dose in rats

Subcutaneously administered Combidex^® contrast agent produced characteristic magnetic susceptibility artifacts in gradient-echo (GE) images of rat brachial and axillary lymph nodes. These artifacts were useful in the rapid location and identification of normal sentinel lymph nodes. A linear dose response was observed with maximum artifact size in transverse images and was used noninvasively to study lymphatic drainage patterns.     

Equivalent cross relaxation rate image for decreasing a false negative case of sentinel lymph node biopsy

In the breast carcinamas, sentinel lymph node biopsy (SLNB) attracts attention as technique to be settled by axillary lymph node metastasis, but existence of a false negative case is a problem. Equivalent cross relaxation rate image (ECRI) is the measurement method that we can evaluate a change of organization structure quantitatively by magnetic resonance imaging. We executed axillary ECRI as a purpose with decreasing a false negative case of SLNB. ECRI related to a macroscopic image, and it was possible to evaluate yes or no of axillary lymph node metastasis. ECRI is the useful method to evaluate adaptability of SLNB.     

Ex vivo proton MR spectroscopy (1H-MRS) for evaluation of human gastric carcinoma

The present study was performed to determine the characteristics of the biochemical metabolites related to gastric cancer using ex vivo (1)H magnetic resonance spectroscopy (MRS), and to assess the clinical usefulness. A total of 35 gastric specimens resected during surgery for gastric cancer were used to compare MR spectra. A 1.5-T (64-MHz) clinical MR imager equipped with facilities for spectroscopy was used to obtain MR spectra from 33 gastric specimens. High-resolution (1)H nuclear magnetic resonance (NMR) spectra of the remains of two specimens were also examined with a 9.4-T (400-MHz) NMR spectrometer. Localized spectroscopic measurements were performed in two layers of gastric tissue, the proper muscle layer and the composite mucosa/submucosa layer. T(2) FSE and 3D SPGR images were used to determine the voxel size and the location for MRS data collection. MR spectra were obtained using the single-voxel PRESS technique with parameters of TR/TE = 2000/30 ms, NA = 256, and voxel size = 3 x 3 x 3 mm(3) (27 microL). Cancerous and noncancerous gastric tissues in the voxel were determined by histopathological analysis. On 9.4-T ex vivo NMR spectroscopy, the following metabolite peaks were found: lipids at 0.9 ppm (CH(3)) and 1.3 ppm (CH(2)); alanine (beta-CH(3)) at 1.58 ppm; N-Acetyl neuraminic acid (NANA: sialic acid) at 2.03 ppm; and glutathione at 2.25 ppm in normal gastric tissue layers. In the 1.5-T MR system, broad and featureless spectral peaks of the various metabolites in normal human gastric tissue were observed at 0.9 ppm, 1.3 ppm, 2.0 ppm, and 2.2 ppm regardless of gastric tissue layer. In specimens (Borrmann type III) with tubular adenocarcinoma, resonance peaks were observed at 1.26 ppm, 1.36 ppm (doublet of lactate), and 3.22 ppm (choline). Cancer lesions showed decreased levels of lipid peaks, showing the significant lactate doublet peaks, and increased intensity of the choline peak as compared with noncancerous gastric tissue. We found that decreased levels of lipids and increases in lactate and choline peaks in gastric tissue were markers for malignancy in gastric lesions. Information provided by ex vivo (1)H MRS, together with the development of in vivo (1)H MRS with high field strength and high resolution, may be very useful for the diagnosis of gastric cancer in clinical situation.     

N-Acetyl peak in MR spectra of intracranial metastatic mucinous adenocarcinomas

Absence of N-acetylaspartate (NAA) is one important diagnostic criterion of MR spectroscopy (MRS) that may suggest that an intracranial mass lesion is a metastasis. We report two cases of histopathology-confirmed intracranial metastatic mucinous adenocarcinoma, which predominantly showed a large metabolite peak at 2.0 ppm, mimicking an NAA peak of normal brain tissue. This finding could be of help in the interpretation of MRS in cases of intracranial enhancing mass lesions, metastases or gliomas.     

Stationary interphases with extended alkyl chains: A comparative study on chain order by solid-state NMR spectroscopy

Stationary interphases with long n-alkyl chains (n = 18, 22, 30, 34) have been examined by solid-state NMR spectroscopy. The determination of the silane functionality and the degree of cross-linking of silane ligands on the silica surface was performed by ²⁹Si CP/MAS NMR spectroscopy. High-speed ¹H MAS and ¹³C CP/MAS NMR spectroscopy were utilized to assess alkyl chain order and mobility of the different bonded phases. For this purpose, ¹H NMR line widths and ¹³C chemical shifts have been evaluated. It is shown that stationary phase order and rigidity increase with alkyl chain length. In addition, the temperature-dependent trans/gauche conformational change occurs at higher temperatures for a polymeric C₃₄ phase compared with a C₃₀ sorbent. This behaviour is discussed in the context of previously reported Chromatographic (HPLC) shape selectivity differences.     

Characterization of polyalkylvinyl ether phases by solid-state and suspended-state nuclear magnetic resonance investigations

Pure organic polyalkylvinyl ether phases were synthesized by suspension polymerization using different ratios and compositions of n-butylvinyl ether (C₄VE) and n-octadecylvinyl ether (C₁₈VE) with triethylene glycol divinyl ether or divinylbenzene as crosslinkers, respectively. These phases were investigated by means of solid-state ¹³C cross-polarization magic angle spinning nuclear magnetic resonance (NMR) spectroscopy and ¹H high-resolution magic angle spinning (HR MAS) NMR spectroscopy in suspended-state. A comparison of these two methods showed the substantial advantages of ¹H HR MAS NMR measurements. Structure elucidation was achieved using a 2D H,H-COSY NMR experiment performed under MAS conditions enabling full peak assignment of the ¹H NMR spectra of these phases. The dynamic behavior of the polyalkylvinyl ether phases was determined by employing temperature-dependent measurements of spin–lattice relaxation times (T₁) as well as accumulation of a 2D wide line separation NMR spectrum.     

塞曼调制磁旋转光谱线型的研究

在研究塞曼调制磁旋转光谱线型的过程中，发现了入射光场的偏振特性地谱线型的影响，从而解释了实验中出现的各种线型，其中包括一些难以解释的除二次微分线型外的其它线型，并且在实验中，得到了理论分析一致的结果，为塞曼调制旋转光谱，激光光谱技术，光谱测量精度的进一步改善提供了参考依据。     

Raman spectroscopic analysis differentiates between breast cancer cell lines

Breast cancer incident rates are increasing in women worldwide with the highest incidence rates reported in developing countries. Major breast cancer screening approaches like mammography, ultrasound, clinical breast examination (CBE) and magnetic resonance imaging (MRI) are currently used but have their own limitations. Optical spectroscopy has attained great attention from biomedical researchers in recent years due to its non‐invasive and non‐destructive detection approach. Chemometrics is one of the powerful tools used in spectroscopic research to enhance its sensitivity. Raman spectroscopy, a vibrational spectroscopic approach, has been used to explore the chemical fingerprints of different biological tissues including normal and malignant types. This approach was used to characterize and differentiate two breast cancer and one normal breast cell lines (MDA‐MB‐436, MCF‐7 and MCF‐10A) using dispersive Raman spectroscopy. Raman spectra of the cell lines have revealed that basic differences in the concentration of biochemical compounds such as lipids, nucleic acids and protein Raman peaks were found to differ in intensity, and principal component analysis (PCA) was able to identify variations that lead to accurate and reliable separation of the three cell lines. Linear discriminant analysis (LDA) model of three cell lines was predicted with 100% sensitivity and 91% specificity. We have shown that a combination of Raman spectroscopy and chemometrics are capable of differentiation between breast cancer cell lines. These variations may be useful in identifying new spectral markers to differentiate different subtypes of breast cancer although this needs confirmation in a larger panel of cell lines as well as clinical material. Copyright © 2015 John Wiley & Sons, Ltd.     

Dynamic sequential 3D gadolinium-enhanced MRI of the whole breast

Dynamic contrast-enhanced 2D MR imaging of the breast has shown high sensitivity and specificity for the detection and characterization of breast lesions. We investigated the ability of a dynamic fast 3D MR imaging technique that repeatedly scans the whole breast in 44-s intervals without an interscan delay time to obtain similar sensitivity and specificity as 2D imaging. Fifty-six patients scheduled for breast biopsy were entered into the study, and 83 lesions detected by 3D dynamic scanning were biopsied. Dynamic 3D contrast-enhanced breast imaging with subtraction detected and correctly classified all 23 cancers, and 44 of the 60 benign lesions yielding a sensitivity of 100%, a specificity of 73%, and a 100% predictive negative value. The enhancement profiles of metastatic lymph nodes were similar to those of primary cancer. This technique allowed detection of multifocal and multicentric lesions and did not require a priori knowledge of lesion location. These results indicate that dynamic contrast-enhanced 3D MRI of the whole breast is a useful and economically feasible method for staging breast cancer, providing a comprehensive noninvasive method for total evaluation of the breast and axilla in patients considering breast conservation surgery or lumpectomy.