Optical imaging of spontaneous breast tumors using protease sensing 'smart' optical probes

OBJECTIVE: The objective of this study was to determine if spontaneous breast cancer lesions can be detected by fluorescence reflectance imaging (FRI) and fluorescence mediated tomography (FMT) using protease-sensing optical probes. MATERIALS AND METHODS: Transgenic (FVB/N-TgN (WapHRAS)69Lin Y)) mice, which spontaneously develop breast cancer, were injected intravenously with a cathepsin-sensing fluorescent imaging probe. FRI and FMT were performed 24 hours after probe injection and region of interest (ROI) analysis was performed. Magnetic resonance images were acquired for anatomic coregistration with the FMT data. Moreover, correlative immunohistochemistry and fluorescence microscopy were performed. RESULTS: All tumor nodules were clearly delineated by FRI showing an average signal intensity of 380 +/- 106 AU. Similarly, tumors were clearly detected by FMT imaging. Immunohistochemistry confirmed cathepsin-B expression of primary tumors and fluorescence microscopy revealed a strong Cy 5.5 deposition in the tissue. CONCLUSIONS: FRI and FMT using "smart" protease sensing probes permits detection of experimental spontaneous breast cancers. Because the expression levels of various proteases correlate with patient outcome, this technique may not only help to detect, but also to differentiate breast cancers noninvasively.     

Steady-state blood volume measurements in experimental tumors with different angiogenic burdens a study in mice

PURPOSE: To experimentally validate the effectiveness of magnetic resonance (MR) imaging enhanced with long-circulating iron oxide for measurement of vascular volume fractions (VVFs) as indicators of angiogenesis in different experimental tumor models. MATERIALS AND METHODS: Tumors with differing degrees of angiogenesis-9L rodent gliosarcoma, DU4475 human mammary adenocarcinoma, HT1080 human fibrosarcoma, and EOMA hemangioendothelioma--were implanted in nude mice. Tumoral VVFs were measured at submillimeter voxel resolutions by using 1.5-T MR imaging. A technetium-labeled intravascular radiotracer was injected into a subset of the animals to validate the MR imaging measurements. Microvessel density and vascular endothelial growth factor (VEGF) also were measured. Statistical analysis was performed with analysis of variance. RESULTS: High-resolution multisection MR maps of tumor blood volume were obtained in all tumor models. Mean tumoral VVF differed significantly among the different tumors: 2.1% +/- 0.3 (standard error of mean) for 9L gliosarcoma, 3.1% +/- 0.4 for DU4475 mammary adenocarcinoma, 5.5% +/- 0.8 for HT1080 fibrosarcoma, and 6.6% +/- 0.9 for EOMA hemangioendothelioma (P <.01). There was a strong correlation between the MR imaging and radiotracer measurements. There was considerable intra- and intertumoral heterogeneity among the VVFs. MR imaging measurements were in accordance with conventional measurements of angiogenesis, such as microvessel density count and VEGF. CONCLUSION: Measurements of tumoral VVF at high-resolution MR imaging with long-circulating iron oxide are feasible and correlate with angiogenic burden in experimental tumor models.     

Antiangiogenic tumor treatment: early noninvasive monitoring with USPIO-enhanced MR imaging in mice

PURPOSE: To prospectively investigate steady-state blood volume measurements for early quantitative monitoring of antiangiogenic treatment with ultrasmall superparamagnetic iron oxide (USPIO)-enhanced magnetic resonance (MR) imaging. MATERIALS AND METHODS: The institutional animal care committee approved all experiments. HT-1080 fibrosarcoma-bearing nude mice were injected with a thrombogenic vascular targeting agent (VTA) (11 nude mice, 20 tumors) or saline (12 nude mice, 20 tumors). USPIO-enhanced (SH U 555C) MR imaging was performed after the VTA was administered. USPIO-induced changes in tissue R2* (DeltaR2*) were measured with a T2-weighted dual-echo echo-planar imaging sequence, and the vascular volume fraction (VVF) was calculated. Parametric DeltaR2* maps were analyzed with respect to tumor perfusion patterns. Correlative histologic analysis was performed for grading of tissue thrombosis, and tissue perfusion was quantified with fluorescent microbeads. Unpaired Student t test and Spearman nonparametric correlation coefficient were used for statistical analysis. RESULTS: The DeltaR2* values were significantly (P < .001) reduced shortly after treatment initiation (mean DeltaR2*, 0.017 msec(-1) +/- 0.0014 [standard error] in control animals vs 0.005 msec(-1) +/- 0.0007 in animals that received VTA), which was also reflected by a decrease in the VVF (2.47% +/- 0.18 vs 0.41% +/- 0.48, P < .001). Histologic analysis revealed various degrees of tumor thrombosis after VTA treatment that correlated inversely with the DeltaR2* values (r = -0.83). Moreover, tumor perfusion measurements corroborated the MR results, indicating a significant reduction in tissue perfusion after VTA treatment (mean tissue fluorescence, 570.4 arbitrary units [au] per gram +/- 27 vs 161.7 au/g +/- 17; P < .05). CONCLUSION: USPIO-enhanced MR imaging enables early monitoring of antiangiogenic treatment of tumors.     

18F alpha-methyl tyrosine PET studies in patients with brain tumors.

We have developed 18F-labeled alpha-methyl tyrosine (FMT) for PET imaging. The aim of this study was to evaluate the clinical application potential of FMT for patients with brain tumors. METHODS: Eleven healthy volunteers and 20 patients with brain tumors were injected with 185 MBq (5 mCi) FMT. In 3 healthy volunteers, whole-body imaging and urinary and plasma analysis were conducted for the assessment of the biodistribution of FMT. The normal range of cortical standardized uptake value (SUV) as a reference for comparing tumor SUV of FMT was estimated by using PET data obtained at 30 min postinjection in 8 healthy volunteers. Dynamic PET scans were conducted for 100 min in 4 healthy volunteers and for 30 min in 15 patients with brain tumors. The 10-min static images in another 4 volunteers and all patients were obtained at 30 min postinjection. In 13 patients, FMT uptake in the brain tumor was compared with 18F-fluorodeoxyglucose (FDG). Tumor-to-normal cortex count (T/N) ratio and tumor-to-white matter count (T/W) ratio and SUVs of brain tumors were determined on FMT and FDG PET images. RESULTS: Approximately 1480 MBq (40 mCi) FMT were produced in one radiosynthesis. Percentage injected dose (%ID) of FMT in the brain ranged from 2.8% to 4.9%, and approximately 50%ID of FMT was excreted in urine during 60 min postinjection, of which 86.6% was unmetabolized FMT. A faint physiological brain uptake with SUV of 1.61 +/- 0.32 (mean +/- SD, n = 8) was observed in healthy volunteers. Tumor SUV of FMT ranged from 1.2 to 8.2, with mean value of 2.83 +/- 1.57 (n = 23), which was significantly higher than that of the cortical area in healthy volunteers (P < 0.01). T/N and T/W ratios of FMT were significantly higher than those of FDG (2.53 +/- 1.31 versus 1.32 +/- 1.46, P < 0.001; 3.99 +/- 2.10 versus 1.39 +/- 0.65, P < 0.0001, respectively). CONCLUSION: FMT, like other radiolabeled amino acids, can provide high-contrast PET images of brain tumors.     

Schwannoma of the extremities: the role of PET in preoperative planning.

The aim of this study was to determine the relative utility of various preoperative diagnostic imaging modalities for the evaluation of benign schwannoma, including positron emission tomography (PET) utilising fluorine-18 fluoro-2-deoxy-D-glucose (FDG) and fluorine- 18 alpha-methyl tyrosine (FMT). computed tomography (CT), magnetic resonance imaging (MRI) and digital subtraction angiography (DSA). We retrospectively reviewed imaging findings in 22 patients with 25 histopathologically documented benign schwannomas of the extremities. Pre-operative imaging included: FDG-PET (n=22), FMT-PET (n=17), MRI (n=25), CT (n=16) and DSA (n=17). All 22 lesions examined by PET with FDG and/or FMT showed accumulation. The standardised uptake values (SUVs) for FDG-PET for the 22 examined tumours ranged from 0.33 to 3.7, and eight of them (36.4%) were assessed as malignant on the basis of their uptake. The SUVs for FMT ranged from 0.44 to 1.47, and 15 out of the 17 evaluated (88.2%) showed values indicating the tumour to be benign. CT demonstrated variable attenuation and contrast enhancement. MRI signal characteristics were relatively consistent: iso-signal or darker than skeletal muscle on T1-weighted and isosignal or brighter than subcutaneous fat on T2-weighted images. The venous tumour staining depicted on DSA was found to be significantly correlated with FDG accumulation. All tumours but one were treated by surgical enucleation. One tumour suspected to be malignant on the basis of imaging findings was treated with primary wide resection. Although CT, MRI and PET studies are all useful for the detection and localisation of schwannoma, our findings suggest that, among the imaging modalities studied, FMT-PET may be the most reliable technique for the differentiation of benign schwannoma from malignancy.     

Evaluation of hemangioma by positron emission tomography: role in a multimodality approach

PURPOSE: The relative utility of various preoperative diagnostic imaging modalities for the evaluation of hemangioma of the extremities, including positron emission tomography (PET) (using 18F-fluoro-2-deoxy-D-glucose [FDG] and fluorine-18 alpha-methyltyrosine [FMT]), computed tomography (CT), magnetic resonance imaging (MRI), and digital subtraction angiography (DSA), was investigated. METHODS: Imaging findings in 16 patients with 16 histopathologically documented hemangiomas of the extremities were retrospectively reviewed. Preoperative imaging included: FDG-PET (n = 16), FMT-PET (n = 12), MRI (n =16), CT (n =11), and DSA (n =14). RESULTS: All 16 lesions examined by PET with FDG and/or FMT showed accumulation. The standardized uptake values (SUVs) for FDG-PET for the 16 examined tumors ranged from 0.7 to 1.67; for FMT-PET, they ranged from 0.14 to 1.00. The SUVs with both tracers indicated the benign nature of the tumor. Computed tomography demonstrated variable attenuation and phleboliths in two patients. The MRI signal characteristics were relatively consistent: heterogeneous signals were slightly higher than those of skeletal muscle on T1-weighted images and brighter than those of subcutaneous fat on T2-weighted images. The pooling and cotton-wool staining depicted in DSA was found to be significantly correlated with FDG accumulation, suggesting that localized blood retention-induced ischemia may accelerate anaerobic glycolysis, which leads to high FDG uptake. CONCLUSION: Although plain radiography, CT, MRI, and angiography may provide anatomic extent and be pathognomonic, FDG-PET and FMT-PET may be the most reliable among the studied imaging modalities for differentiating benign hemangiomas from other soft tissue tumors, especially malignant neoplasms.     

Stunned,infarcted, and normal myocardium in dogs: simultaneous differentiation by using gadolinium-enhanced cine MR imaging with magnetization transfer contrast

PURPOSE: To simultaneously differentiate stunned, infarcted, and normal myocardial regions by using gadolinium-enhanced cine magnetic resonance (MR) imaging with magnetization transfer contrast. MATERIALS AND METHODS: Twelve dogs were imaged on days 1 and 8 after transient 90-minute coronary artery occlusion. A magnetization transfer contrast with echo-train readout (MTET) MR sequence was performed before and 30 minutes after gadolinium contrast enhancement. Ex vivo analysis consisted of MR imaging, microsphere blood flow analysis, and triphenyltetrazolium chloride (TTC) staining. A paired two-tailed t test was used to compare wall thickening from day 1 to day 8. Linear regression and Bland-Altman analyses were used to compare infarct size depicted with MTET imaging with that seen on TTC-stained tissue. RESULTS: Severe wall motion abnormalities were detected in all dogs. At TTC analysis, seven dogs had evidence of myocardial infarction and five had evidence of stunned myocardium. The mean percentages of left ventricular wall thickening in infarcted, stunned, and remote myocardial regions were 2% +/- 4 (SD), 4% +/- 8, and 33% +/- 5, respectively. Wall thickening did not improve in the infarcted zones, but it improved to nearly normal levels in the stunned region 1 week after induced occlusion (mean, 40% +/- 8; P <.02). MTET images clearly depicted infarcted myocardium as brighter than both the normal and stunned myocardial regions but darker than the blood pool. In vivo MTET infarct volume correlated with ex vivo TTC analysis data (y = 1.01x + 0.00, R = 0.98, standard error of the estimate = 0.019). CONCLUSION: One day after myocardial ischemia, MTET during one MR imaging examination enabled simultaneous differentiation of infarcted, stunned, and normal myocardial regions on the basis of gadolinium enhancement and regional function.     

Assessment of bone marrow angiogenesis in patients with acute myeloid leukemia by using contrast-enhanced MR imaging with clinically approved iron oxides: initial experience

PURPOSE: To prospectively assess bone marrow (BM) angiogenesis in patients with acute myeloid leukemia (AML) by using iron oxide-enhanced magnetic resonance (MR) imaging. MATERIALS AND METHODS: The study was institutional ethics committee approved. Informed signed consent was obtained from each study participant. The requirement for informed consent for use of data from a reference database was waived. Eleven patients (seven women, four men; mean age, 53 years+/-4.40 [standard deviation]) with an initial diagnosis of AML were enrolled in the study and underwent T2*-weighted two-echo echo-planar MR imaging of the pelvis before and after intravenous injection of a clinically approved iron oxide blood-pool contrast agent. Six healthy control subjects (one woman, five men; mean age, 35 years+/-2.31) were examined with the same MR protocol. The iron oxide-induced change in R2* relaxation rate (DeltaR2*) was calculated, and the vascular volume fraction (VVF) of the BM was derived by dividing the DeltaR2* of the BM by the DeltaR2* of the muscle. Parametric DeltaR2* maps were calculated to visualize vessel distribution. Patients underwent BM biopsy for correlative determination of microvessel density (MVD) and vascular endothelial growth factor (VEGF). Differences in DeltaR2*, VVF, VEGF, and MVD were compared by using the Wilcoxon rank sum test. RESULTS: DeltaR2* maps showed prominent areas of highly vascularized BM in the patients with AML, whereas the control subjects had moderately vascularized BM with homogeneous vessel distribution. Quantitative analysis revealed VVF values to be significantly higher in patients with AML than in control subjects: The mean VVF in the pelvis was 9.18%+/-1.54 for patients versus 3.91%+/-0.61 for control subjects (P=.010). In accordance with MR results, MVD (P=.009) and VEGF expression (P=.017) were significantly elevated in the AML group compared with values in the control group. CONCLUSION: Iron oxide-enhanced MR imaging enables assessment of BM angiogenesis in patients with AML.     

裸鼠胃癌移植瘤的近红外荧光成像

目的：通过对非特异性探针Cy5.5在裸鼠体内分布及显像研究,探讨近红外荧光成像对胃癌的早期诊断及动态监测价值。方法：用MGC-803细胞株建立胃癌动物模型,进行早期、实时活体及离体显像实验。结果：近红外荧光成像可检测早期胃癌移植瘤的平均大小为2.807mm×3.045mm,与游标卡尺测得的肿瘤大小呈直线相关,r=0.924,P〈0.05。Cy5.5主要分布在肿瘤组织,主要代谢器官为肾脏;注入探针30min后,裸鼠肿瘤部位成像清晰,荧光寿命、荧光强度均高于对照部位（P〈0.05）。60min后,肿瘤区的荧光强度始高于血液（P〈0.05）。90min时达峰值。肿瘤部位的平均荧光寿命为（3.1376±0.9894）ns,明显高于对侧部位（P〈0.05）。结论：近红外荧光成像可用于胃癌的早期诊断及瘤体的动态监测。     

Optical imaging of matrix metalloproteinase-2 activity in tumors: feasibility study in a mouse model.

PURPOSE: To develop an optical imaging method to determine the expression level of tumoral matrix metalloproteinase-2 (MMP-2) in vivo. MATERIALS AND METHODS: An optical contrast agent was developed that was highly activatable by means of MMP-2-induced conversion. Signal characteristics of the probe were quantified ex vivo with a recombinant enzyme. Animal tumor models were established with MMP-2-positive (human fibrosarcoma cell line, n = 4) and MMP-2-negative (well-differentiated mammary adenocarcinoma, n = 4) tumor cell lines. Both tumors were implanted into nude mice and were optically imaged after intravenous administration of the MMP-2-sensitive probe. RESULTS: The MMP-2-sensitive probe was activated by MMP-2 in vitro, producing up to an 850% increase in near-infrared fluorescent signal intensity. This activation could be blocked by MMP-2 inhibitors. MMP-2-positive tumors were easily identified as high-signal-intensity regions as early as 1 hour after intravenous injection of the MMP-2 probe, while contralateral MMP-2-negative tumors showed little to no signal intensity. A nonspecific control probe showed little to no activation in MMP-2-positive tumors. CONCLUSION: It is feasible to image MMP-2 enzyme activity in vivo by using near-infrared optical imaging technology and "smart" matrix metalloproteinase-sensitive probes.     

Improved visualization of vessels and hepatic tumors by micro-computed tomography (CT) using iodinated liposomes

OBJECTIVE: The goal of this study was to determine whether iodinated liposomes are a suitable tracer for mice microvessel and liver imaging by preclinical computed tomography (CT). MATERIALS AND METHODS: Iodinated liposomes were evaluated for vessel and liver imaging. A first group of nude mice was imaged by micro-CT after i.v. injection of liposomes at 1 or 2 gI/kg body weight (b.w.) for intervals up to 24 hours. A second group of mice bearing liver micrometastases was imaged after injection of liposomes at 2 gI/kg b.w. for intervals up to 24 hours. RESULTS: Vascular enhancements of 120 +/- 8 and 322 +/- 20 Hounsfield unit (HU) were obtained after injection of liposomes at 1 or 2 gI/kg b.w., respectively. This enhancement decreased with a blood half-life of 135 +/- 10 and 86 +/- 9 minutes, respectively. Liver enhancement of 157 +/- 5 and 235 +/- 23 HU were obtained after injection of iodinated liposomes at 1 and 2 gI/kg b.w., respectively. Liver micrometastases (250 microm) were detectable after injection of iodinated liposomes at 2 gI/kg b.w. CONCLUSIONS: Iodinated liposomes are a suitable contrast agent for vessels and liver imaging by micro-CT allowing clear vascular enhancement and detection of small liver metastases.     

Prediction of antiangiogenic treatment efficacy by iron oxide enhanced parametric magnetic resonance imaging

RATIONALE AND OBJECTIVES: Tools for monitoring modern target-specific antiangiogenic and antivascular therapies are highly desirable because treatment strategies are time consuming, expensive, and yet sometimes ineffective. Therefore, the aim of this experimental study was to evaluate the predictive value of steady-state ultrasmall particles of iron oxide (USPIO; SH U 555 C)-enhanced magnetic resonance imaging (MRI) for early assessment of antivascular tumor-treatment effectiveness. METHODS: Mice were inoculated with an HT-1080 fibrosarcoma xenograft and subjected to target-specific antivascular therapy using a selective thrombogenic vascular-targeting agent (truncated tissue factor fused to RGD peptide) or saline as control. Four to 8 hours after treatment, the USPIO-induced change in the transverse relaxation rate DeltaR2* was measured by MRI, and the vascular volume fraction (VVF) was calculated by calibrating DeltaR2* of the tumor by DeltaR2* of muscle tissue. Treatment response was defined by histologic grading of vascular thrombosis and tumor necrosis. RESULTS: After thrombogenic treatment, half of the HT-1080 xenograft-bearing animals showed only minor (=nonresponder) whereas the other half showed extensive tumor thrombosis (=responders). For responders, a significant decrease of DeltaR2* and VVF was observed compared with the control group (DeltaR2*: controls: 16 +/- 1 s-1 vs. responder: 4 +/- 2 s-1; P < 0.001) whereas DeltaR2* and VVF remained nearly unchanged for nonresponders (DeltaR2*: nonresponder 14 +/- 2 s-1). VVF and DeltaR2* values correlated inversely with the histologic grading of vascular thrombosis and tumor necrosis (VVF: r = -0.8; DeltaR2*: r = -0.71; P < 0.01). CONCLUSION: USPIO-enhanced MRI allows a noninvasive, early assessment of treatment efficacy of thrombogenic vascular-targeting agents.     

Differentiation of alveolitis and pulmonary fibrosis with a macromolecular MR imaging contrast agent.

The ability of macromolecular contrast agent (polylysine-[gadopentetate dimeglumine]) to allow differentiation of pulmonary fibrosis and alveolitis at magnetic resonance imaging was investigated. Lung damage was induced by means of left bronchial instillation of 200 micrograms of cadmium chloride. Rats were imaged 3 hours (early alveolitic stage, n = 5), 24 hours (late alveolitic stage, n = 5), and 8 days (fibrotic stage, n = 5) later. Rats imaged 3 hours after cadmium chloride instillation demonstrated a gradually increasing contrast enhancement over 45 minutes (from 314% +/- 110 to 476% +/- 69 over baseline [P less than .01]), indicating a leak of paramagnetic macromolecules from the intravascular into the extravascular spaces. Conversely, lung enhancement remained virtually constant after injection of contrast material in contralateral control lungs and in damaged lungs imaged 24 hours and 8 days after cadmium chloride instillation. Furthermore, the enhancement in the fibrotic lung was lower (170% +/- 50) than that in the alveolitic and control lungs (320% +/- 65 and 298% +/- 61, respectively), indicating a decrease in plasma volume in the fibrotic lung. A macromolecular contrast agent can facilitate the differentiation between the exudative and fibrotic phases of interstitial lung disease.     

Usefulness of gated myocardial perfusion SPECT imaging soon after exercise to identify postexercise stunning in patients with single-vessel coronary artery disease

BACKGROUND: This study determines the value of gated single photon emission computed tomography (SPECT) imaging soon after exercise to identify patients with single-vessel disease and exercise-induced prolonged myocardial dysfunction (ie, postischemic stunning). METHODS AND RESULTS: We examined 19 normal individuals and 52 patients with single-vessel disease by use of 2-day technetium 99m tetrofosmin exercise/rest gated SPECT imaging. Sequential imaging was started 10, 30, and 50 minutes after exercise. The ejection fraction (EF) values were calculated with the Cedars-Sinai program. The participants were classified as follows: group A (normal individuals, n = 19), group B (individuals with coronary stenosis without Q-wave infarction, n = 18), group C (individuals with Q-wave infarction without myocardial ischemia, n = 15), and group D (individuals with Q-wave infarction and ischemia, n = 19). The post-stress EF values at 10 minutes (69.8% +/- 9.6% and 59.8% +/- 11.8%, respectively) were higher in groups A and C than those at 30 minutes (67.6% +/- 10.2% and 57.2% +/- 11.3%, respectively) ( P < .05) but were lower in group B (61.7% +/- 9.2%) than both the 30- and 50-minute values (64.2% +/- 9.5% and 64.6% +/- 9.4%, respectively; P < .05). The EF value did not significantly change in group D. CONCLUSIONS: Tc-99m gated SPECT imaging soon after exercise is superior to conventional late imaging to discriminate patients with single-vessel disease and postexercise stunning.     

Development and validation of a novel technique for murine first-pass radionuclide angiography with a fast multiwire camera and tantalum 178

BACKGROUND: The use of genetically altered mice as a model system to study cardiovascular disease has created a need for accurate and quantitative assessment of murine ventricular function. To address this very challenging problem, we have developed a technique of murine first-pass radionuclide angiography using pinhole imaging and the short-lived isotope tantalum 178 (Ta-178) with a high-speed multiwire proportional camera (MPC). METHODS AND RESULTS. An MPC was fitted with a pinhole lens of 2-mm-diameter aperture positioned 15 cm from the camera face. The short-lived isotope Ta-178 (half-life 9.3 minutes) was generated from the tungsten 178 (W-178) (half-life 21.7 days)/Ta-178 generator and concentrated on site to an injection volume of 15 to 20 microL. Mice were imaged in the supine position with the chest wall 3 mm from the camera pinhole aperture, and images were acquired at 160 frames per second after a rapid bolus injection of Ta-178. In the absence of a true gold standard, the technology was validated with measurements in control mice and mice with surgically ligated left anterior descending arteries (LADs). In addition, the effects of pharmacologic intervention with verapamil and with dobutamine were observed. Finally, peak aortic velocity measurements obtained with this technology were compared with those obtained with echocardiographic Doppler ultrasonography, the only available quantitative comparator. There was a significant decrease in the mean left ventricular ejection fraction (LVEF) between normal mice (62% +/- 4.6% [mean +/- SEM], n = 12) and mice with experimentally induced myocardial infarction produced by surgical LAD ligation (22% +/- 2.0%, n = 41; P <.01). The LVEF decreased from 51% +/- 5.8% to 37% +/- 3.5% in a group of normal mice receiving verapamil (P <.05, n = 8) and increased from 34% +/- 2.2% to 43% +/- 2.3% in a group of LAD-ligated mice receiving dobutamine (P <.01, n = 48). Peak camera sensitivity during first pass was 25,000 cps/mCi injected. Intraobserver and interobserver variability of LVEF was studied, yielding r = 0.9639 and 0.9529 and SE of the estimate 2.6% and 3.1%, respectively. Reproducibility in serial studies was excellent (r = 0.92, SE of the estimate 5.18). CONCLUSIONS: This study demonstrates the development and use of a promising new method that uses the short-lived radioisotope Ta-178 and MPC for noninvasive quantification of murine ventricular function, that produces accurate and highly reproducible results, and that can be applied in multiple serial studies.     

Gamma camera dual imaging with a somatostatin receptor and thymidine kinase after gene transfer with a bicistronic adenovirus in mice

PURPOSE: To compare two systems for assessing gene transfer to cancer cells and xenograft tumors with noninvasive gamma camera imaging. MATERIALS AND METHODS: A replication-incompetent adenovirus encoding the human type 2 somatostatin receptor (hSSTr2) and the herpes simplex virus thymidine kinase (TK) enzyme (Ad-hSSTr2-TK) was constructed. A-427 human lung cancer cells were infected in vitro and mixed with uninfected cells at different ratios. A-427 tumors in nude mice (n = 23) were injected with 1 x 10(6) to 5 x 10(8) plaque-forming units (pfu) of Ad-hSSTr2-TK. The expressed hSSTr2 and TK proteins were imaged owing to internally bound, or trapped, technetium 99m ((99m)Tc)-labeled hSSTr2-binding peptide (P2045) and radioiodinated 2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl-5-iodouracil (FIAU), respectively. Iodine 125 ((125)I)-labeled FIAU was used in vitro and iodine 131 ((131)I)-labeled FIAU, in vivo. The (99m)Tc-labeled P2045 and (125)I- or (131)I-labeled FIAU were imaged simultaneously with different window settings with an Anger gamma camera. Treatment effects were tested with analysis of variance. RESULTS: Infected cells in culture trapped (125)I-labeled FIAU and (99m)Tc-labeled P2045; uptake correlated with the percentage of Ad-hSSTr2-TK-positive cells. For 100% of infected cells, 24% +/- 0.4 (mean +/- SD) of the added (99m)Tc-labeled P2045 was trapped, which is significantly lower (P <.05) than the 40% +/- 2 of (125)I-labeled FIAU that was trapped. For the highest Ad-hSSTr2-TK tumor dose (5 x 10(8) pfu), the uptake of (99m)Tc-labeled P2045 was 11.1% +/- 2.9 of injected dose per gram of tumor (thereafter, dose per gram), significantly higher (P <.05) than the uptake of (131)I-labeled FIAU at 1.6% +/- 0.4 dose per gram. (99m)Tc-labeled P2045 imaging consistently depicted hSSTr2 gene transfer in tumors at all adenovirus doses. Tumor uptake of (99m)Tc-labeled P2045 positively correlated with Ad-hSSTr2-TK dose; (131)I-labeled FIAU tumor uptake did not correlate with vector dose. CONCLUSION: The hSSTr2 and TK proteins were simultaneously imaged following dual gene transfer with an adenovirus vector.     

Mitral or aortic regurgitation: quantification of regurgitant volumes with cine MR imaging

A new, rapid magnetic resonance (MR) imaging method, cine MR imaging, was used to determine the regurgitant fraction (RF) in patients with left-sided regurgitant lesions. Right and left ventricular stroke volumes were determined with cine MR imaging and a modified Simpson formula in ten healthy volunteers and 23 patients known to have either predominant mitral (n = 17) or aortic (n = 6) regurgitation. RFs evaluated at cine MR imaging were compared in healthy persons and patients with mild, moderate, or severe regurgitation demonstrated at angiography (n = 10) and Doppler echocardiography (n = 13). Cine MR imaging depicted regurgitant blood flow in all 29 regurgitant lesions in 23 patients as areas of low signal intensity within the regurgitant chamber. The RF was 4% +/- 7% in healthy subjects and 12% +/- 12% in those with mild, 35% +/- 14% in those with moderate, and 63% +/- 5% in those with severe regurgitation. The RFs determined by two observers were similar.     

Fluorescent bacterial magnetic nanoparticles as bimodal contrast agents

OBJECTIVES: The purpose of this study was to assess whether fluorochrome-coupled bacterial magnetic nanoparticles can be used as bimodal contrast agent for both magnetic resonance imaging (MRI) and near-infrared fluorescence optical (NIRF) imaging of cultured macrophages. MATERIALS AND METHODS: Bacterial magnetic nanoparticles (magnetosomes, particle diameter: 42 nm) were harvested from Magnetospirillum gryphiswaldense and characterized by using MRI. After covalent coupling to the fluorescent dye DY-676 (lambdaabs./lambdaem.= 676 nm/701 nm, Dyomics, Jena, Germany), the fluorescent magnetosomes were analyzed by fluorescence-activated cell sorting. Subsequently, murine macrophages J774 were incubated with the bimodal contrast agent (3 hours) and examined by a whole-body near infrared small animal imaging system as well as by using a 1.5 T clinical MR system. Moreover, labeled cells were characterized using confocal laser scanning microscopy (CLSM) and ultrathin section transmission electron microscopy. RESULTS: Characterization of the nanoparticles by MRI revealed R1 and R2 relaxivities of 3.2 mMs and 526 mMs, respectively. Fluorochrome-coupled magnetosomes exhibited increased fluorescence intensities at wavelengths >670 nm. Macrophages that were incubated with the contrast agent showed a significant fluorescence emission in the near infrared range as imaged with a whole body NIR imaging system, FACS analysis and CLSM. Moreover, CLSM data showed the greatest fluorescence intensities within intracellular compartments and colocalized with the magnetosomes. With MRI, both T1 and T2 relaxation times were substantially shortened at concentrations greater than 600 cells/microL. DISCUSSION AND CONCLUSION: Macrophages could be labeled with fluorescent magnetosomes, and they were successfully imaged using both a 1.5 T MR scanner as well as with NIRF optical methods. The use of this bimodal contrast agent for diagnostic purposes may benefit from the excellent spatial resolution of the MRI and the high sensitivity of the fluorescence imaging.     

Evaluation of the Extension of Cerebral Gliomas by Scintigraphy

BACKGROUND: Single photon emission computed tomography (SPECT) with 201Tl and 123I-alpha-methyl tyrosine (123I-IMT) are routine methods for the evaluation of brain tumors. 123I-IMT transport across the blood brain barrier is mediated by an amino acid carrier, 201Tl accumulation is analogous to cerebral potassium uptake. PATIENTS AND METHODS: To determine the differences in glioma extension as shown by the 2 methods, 17 patients with malignant gliomas were included in this comparative imaging study: astrocytoma III: n = 6, ependymoma III: n = 1, oligodendroglioma III: n = 1, glioblastoma IV: n = 9. The tomographic image sets were matched anatomically and the slices showing maximal tumor extension were identified in both image sets respectively. Tumor spread was compared visually and the tumor extension was quantified. RESULTS: In gliomas WHO III tumor extension was delineated significantly larger by 123I-IMT-SPECT than by 201Tl-SPECT (mean +/- SD: 816 +/- 281 pixels vs 600 +/- 220 pixels, n = 8, p < 0.05). The size of glioblastomas was shown in a comparable manner by the 2 methods (977 +/- 571 vs 1.051 +/- 588, n = 9, ns, p = 0.57), but there were considerable regional differences between the area of 201Tl uptake and amino acid retention. In the whole group a weak but significant negative correlation between intensity of 201Tl uptake on the one hand and a ratio of the area as depicted by 123I-IMT vs area as depicted by 201Tl on the other hand, was found (n = 17, r = 0.49, p < 0.05). Thus the differences in the delineation of areas became smaller with increasing 201Tl uptake. CONCLUSIONS: These preliminary data indicate that the extension of gliomas is depicted differently by the 2 methods. 123I-IMT-SPECT shows a larger tumor extension especially in gliomas WHO III. Since 201Tl uptake has previously been shown to correlate with disruption of the blood brain barrier, 123I-IMT-SPECT may delineate tumor parts without endothelial leakage. This additional information may be helpful in planning surgical or radiation therapy. The advantages of 123I-IMT in this respect decrease with increasing 201Tl uptake and with increasing malignancy.     

Catheter-based in vivo imaging of enzyme activity and gene expression: feasibility study in mice

PURPOSE: To construct and evaluate an interventional catheter-based imaging system for intravital monitoring of molecularly sensitive near-infrared fluorescent probes and optical marker genes. MATERIALS AND METHODS: An imaging device that was based on a miniaturized fiberoptic sensor (MIFS) was built in which images created with a 2.7-F fiberoptic catheter were relayed through a dichroic mirror, through a bandpass filter, and on two independent cameras. This system permitted simultaneous recording of white-light and fluorescent images. Spatial resolution, spectral transmissions, and sensitivity were determined in vitro. In vivo testing was performed in nude mice bearing intraperitoneal tumors that express green fluorescent protein and in a mouse model of ovarian carcinoma with enzyme-activatable near-infrared probes sensitive to tumoral protease activity. Signal intensity on images of tumors and that on images of normal tissue were measured and compared with t test. RESULTS: The catheter, which was advanced through an 18-gauge sheath, showed resolution of 7 line pairs per millimeter and detection limit for fluorochrome Cy5.5 of 1-10 pmol. Detection of endogenous green fluorescent protein gene expression was feasible in tumor nodules smaller than 1 mm in diameter (mean tumor signal intensity, 153.26 +/- 26.45 [SD], compared with that of adjacent nontumoral tissue of 36.73 +/- 11.69; P <.008). Similarly, activation of the near-infrared probe by tumoral proteases could be detected in peritoneal tumor seeds of ovarian cancer model with mean tumor signal intensity of 246.33 +/- 7.77 compared with that of adjacent nontumoral tissue of 41.56 +/- 18.64 (P <.001). Mean contrast-to-noise ratio in the near-infrared channel exceeded white-light contrast-to-noise ratio by a factor of 6.7 (P <.02). CONCLUSION: With this system, in vivo MIFS imaging of gene expression, enzyme activity, and potentially other molecular events is feasible, through direct interventional access to several organs and body cavities and potentially through transvascular approaches.