Acoustic attenuation by contrast agent microbubbles in superficial tissue markedly diminishes petechiae bioeffects in deep tissue

OBJECTIVE: To measure how ultrasound attenuation by contrast agent microbubbles (MBs) in superficial tissue affects petechiae creation in underlying deep tissue. MATERIALS AND METHODS: Studies using Sprague-Dawley rats were approved by the Animal Care and Use Committee. MBs were injected intravenously, and 12 ultrasound pulses (100 sinusoids of 1 MHz ultrasound per pulse) were applied through the skin overlying the hindlimb adductors at intervals of 10 or 60 seconds. In some groups, the skin was resected and immediately returned without re-establishing vascular connections. Muscle petechiae were counted. RESULTS: Applying ultrasound through unperfused skin after bolus and continuous intravenous MB injection yielded, respectively, 30-fold and 3.5-fold more petechiae than for perfused skin. Surprisingly, petechiae/mm2 decreased with a higher MB dosage [0.12 +/- 0.05 (1 x 10 MBs/g) vs. 0.04 +/- 0.02 (3 x 10 MBs/g)] when ultrasound was applied through perfused skin. In contrast, petechiae/mm2 was approximately proportional to MB dosage for unperfused skin [0.17 +/- 0.10(5) (1 x 10 MBs/g) vs. 0.42 + 0.14 (3 x 10(5) MBs/g)]. In comparison to MB-free controls, MB solutions in this concentration range reduced the peak-negative pressure of ultrasound by 65% to 85%. CONCLUSIONS: Acoustic attenuation by MBs in skin markedly reduces petechiae creation in deep muscle. Petechiae inhibition is dependent on [MB]2.1 and, therefore, dominates the otherwise proportional relationship between petechiae and [MB] in muscle. The drop of peak-negative pressure below a critical microvessel rupturing threshold is the probable mechanism for petechiae inhibition. These results indicate that high MB doses could, paradoxically, reduce the potential for petechiae creation and may have important bearing on the design of contrast ultrasound-based therapeutics.     

Influences of Microbubble Diameter and Ultrasonic Parameters on In Vitro Sonothrombolysis Efficacy

PurposeTo quantify the effects of microbubble (MB) size, elasticity, and pulsed ultrasonic parameters on in vitro sonothrombolysis (ultrasound [US]-mediated thrombolysis) efficacy.Materials and MethodsMonodispersive MBs with diameters of 1 μm or 3 μm were exposed to pulsed US (1 MHz or 3 MHz) to lyse rabbit blood clots. Sonothrombolysis efficacy (clot mass loss) was measured as functions of MB size and concentration, ultrasonic frequency and intensity, pulse duration (PD), pulse repeat frequency (PRF), and duty factor.ResultsSonothrombolysis at 1 MHz was more effective using 3-μm MBs and at 3 MHz using 1-μm MBs. Sonothrombolysis was more effective at 1 MHz when≥75% of MBs remained intact, especially for 3-μm MBs; improving sonothrombolysis by increasing PRF from 100 Hz to 400 Hz at 3 MHz was associated with increasing 3-μm MB survival. However, 60% of 1-μm MBs were destroyed during maximal sonothrombolysis at 3 MHz, indicating that considerable MB collapse may be required for sonothrombolysis under these conditions.ConclusionsThe ability to control MB size and elasticity permits using a wide range of US parameters (eg, frequency, intensity) to produce desired levels of sonothrombolysis. Comparable, maximal sonothrombolysis efficacy was achieved at 20-fold lower intensity with 3-μm MBs (0.1 W/cm²) than with 1-μm MBs (2.0 W/cm²), a potential safety issue for in vivo sonothrombolysis. US parameters that maximized MB survival yielded maximal sonothrombolysis efficacy except with 1-μm MBs at 3 MHz where most MBs were destroyed.     

Gene transfer with echo-enhanced contrast agents: comparison between Albunex, Optison, and Levovist in mice--initial results

PURPOSE: To determine if commercially available echo-enhanced microbubble contrast agents could be used to increase gene transfection efficiency by means of relatively low-intensity ultrasound-mediated microbubble destruction in skeletal muscles. MATERIALS AND METHODS: Three types of ultrasound microbubble contrast agents (0.01 mL of albumin [Albunex] and human albumin [Optison] and 10 mg/mL of SH U 508A [Levovist]) were each separately mixed with the reporter plasmid DNA (25 microg) encoding green fluorescent protein (GFP) prior to intramuscular injection into the quadriceps muscle of a mouse thigh bilaterally (seven mice per contrast agent). One of the muscle sites that was injected with plasmid DNA was irradiated with low-intensity therapeutic ultrasound (1 MHz) at an intensity of 2.0 W/cm2 for 2 minutes. Mice were sacrificed 7 days after ultrasound treatment for gene expression assay. The number of GFP-expressing muscle fibers was counted. Statistical significance was determined with a two-tailed Student t test. P <.05 was considered to indicate statistically significant difference. RESULTS: Muscle tissue exposed to ultrasound with air-filled Albunex or Levovist microbubbles revealed no difference in the number of GFP-expressing muscle fibers compared with the control non-ultrasound-exposed muscle. Albumin-coated octafluoropropane gas-filled Optison microbubbles showed a 10-fold increase in the number of GFP-expressing fibers (P <.05). CONCLUSION: Low-intensity ultrasound with echo-enhanced Optison induced efficient gene transfer unlike that with Albunex or Levovist.     

Pharmacodynamics of streptavidin-coated cyanoacrylate microbubbles designed for molecular ultrasound imaging

OBJECTIVES: To assess the pharmacodynamic behavior of cyanoacrylate, streptavidin-coated microbubbles (MBs) and to investigate their suitability for molecular ultrasound imaging. MATERIALS AND METHODS: Biodistribution of MBs was analyzed in tumor-bearing mice using gamma-counting, immunohistochemistry, flow cytometry, and ultrasound. Further, vascular endothelial growth factor receptor 2-antibody coupled MBs were used to image tumor neovasculature. RESULTS: After 1 minute >90% of MBs were cleared from the blood and pooled in the lungs, liver, and spleen. Subsequently, within 1 hour a decent reincrease of MB-concentration was observed in the blood. The remaining MBs were removed by liver and spleen macrophages. About 30% of the phagocytosed MBs were intact after 48 hours. Shell fragments were found in the kidneys only. No relevant MB-accumulation was observed in tumors. In contrast, vascular endothelial growth factor receptor 2-specific MBs accumulated significantly within the tumor vasculature (P < 0.05). CONCLUSIONS: The pharmacokinetic behavior of streptavidin-coated cyanoacrylate MBs has been studied. In this context, the low amount of MBs in tumors after >5 minutes is beneficial for specific targeting of angiogenesis.     

机械创伤使大鼠心肌对缺血/再灌注损伤的敏感性增加

目的 观察机械创伤大鼠对心肌缺血/再灌注损伤的敏感性.方法 用Noble -Collip创伤仪制备机械创伤模型,采用完全随机分组方法将Wistar大鼠分为假创伤组、创伤组、假创伤+假手术组、假创伤+缺血/再灌组和创伤+缺血/再灌组,于创伤后1周行缺血/再灌注.采用BL - 410生物信号记录分析系统记录大鼠左心室收缩压(left ventricular systolic pressure,LVSP)、左心室压力上升和下降最大速率(±dp/dtmax)等心功能数据;采用双抗体夹心ABC -ELISA法检测大鼠血清肌酸激酶同工酶MB( CK - MB)、心肌肌钙蛋白(cTnI)水平;缺血/再灌注后心脏通过伊文思蓝染色、2,3,5 -氯化三苯基四氮唑(TTC)复染后,用Image - Pro Plus 6.0图像分析软件进行心肌梗死面积测定.结果 创伤+缺血/再灌注组在体心功能明显低于假创伤+缺血/再灌注组(P＜0.01);创伤+缺血再灌注组血清CK - MB和cTnI水平显著高于假创伤+缺血/再灌注组[ CK - MB:(4 960±588) ng/ml:(2 925±426) ng/ml,P＜0.01;cTnI:( 18.10 ±3.06) ng/ml:(6.67±1.57) ng/ml,P＜0.01];创伤+缺血/再灌注组心肌梗死面积明显大于假创伤+缺血/再灌注组[(36.70±7.42)％:(22.27±4.54)％,P＜0.01].结论机械创伤使大鼠心肌对缺血/再灌注损伤的敏感性增加.     

磁共振体素内不相干性运动在肌骨系统肿瘤定性诊断中的作用

目的：评价磁共振体素内不相干性运动（IVIM）参数在肌骨系统肿瘤定性诊断中的价值。方法：本研究中的38例肌骨系统肿瘤的患者,均采用1．5T MR 扫描仪进行检查,在常规扫描后进行 IVIM 扫描。IVIM 使用的9个 b 值分别为0,20,40,60,80,100,250,500和750 s／mm2。分别测量病变区和作为对照组的正常肌肉组织 IVIM 参数 ADC、ADCslow 、ADCfast 及 PF 值。根据病理结果,将病变分为良性（12例）、交界性（14例）及恶性肿瘤（12例）,并对三组肿瘤之间以及肿瘤与对照组之间 ADC、ADCslow 、ADCfast 及 PF 值的差异进行统计学分析。结果：交界性肿瘤的 ADC 及 ADCslow 值分别为（1．02±0．15）×10－3 mm2／s 和（1．02±0．16）×10－3 mm2／s,明显低于对照组（1．38±0．20）×10－3 mm2／s 和（1．38±0．17）×10－3 mm2／s,差异具有统计学意义（P ＝0．001）。恶性肿瘤的 ADC 及 ADCslow 值分别为（0．90±0．13）×10－3 mm2／s和（0．88±0．14）×10－3 mm2／s,明显低于对照组（1．48±0．12）×10－3 mm2／s 和（1．48±0．09）×10－3 mm2／s,差异具有统计学意义（P ＜0．001）。而良性肿瘤的 ADC 及 ADCslow 与对照组之间差异无统计学意义（P ＝0．564,0．480）。ADCfast 以及 PF 值在肿瘤与对照组之间无统计学差异。交界性、恶性肿瘤的 ADC 和 ADCslow 值明显低于良性肿瘤,差异具有统计学意义（P 值均＜0．001）,但交界性肿瘤与恶性肿瘤之间的 ADC 和 ADCslow 值差异无统计学意义。良性、交界性和恶性肿瘤之间的 ADCfast 和 PF 值的差异没有统计学意义。结论：IVIM 参数 ADC 和 ADCslow 有助于区分良性肿瘤与交界性、恶性肿瘤,提高肌骨系统肿瘤的诊断准确性。     

256层CT评价心肌桥及其与冠状动脉粥样硬化的关系

目的:用256层CT冠脉成像技术评价心肌桥(myocardial bridge,MB)的发生率及其解剖特征与冠状动脉粥样硬化(athorosclerosis,AS)的关系。方法:收集行256层CT冠状动脉成像564例患者的资料,评价MB的有无、部位、长度、厚度、伴发的AS情况及MB解剖特点与AS关系。结果:564例检出MB 131例,159个,总发生率为23.2%(131/564),男性发生率28.8%(90/313)高于女性16.3%(41/251)(χ2=12.048 5,P=0.005)。第一对角支(D1)为最常累及的部位,其次为左前降支(LAD)。完全性MB 83个,不完全性MB 76个;完全性MB长度和厚度中位数分别为27.7mm(范围2.0～71.1mm)和2.4mm(范围1～6.9mm);其长度与厚度间无相关性(R=0.061 1,P=0.106 6)。不完全性MB的长度中位数为24.1mm(范围3.9～57.0mm)。桥血管及桥血管远段血管均未见AS改变。桥血管近段血管AS程度在LAD组高于D1组(χ2=7.576 6,P<0.05),完全性MB组高于不完全性MB组(χ2=7.484 9,P<0.05)。结论:256层CT冠脉成像技术能无创、清晰、可靠地显示活体内MB的存在及解剖特点,并能评估桥血管及其毗邻血管情况。     

Enhanced gene delivery into skeletal muscles with ultrasound and microbubble techniques

RATIONALE AND OBJECTIVES: This experiment was directed to explore the effects of ultrasound microbubbles on gene structure in vitro and green fluorescent protein (GFP) plasmid transfer into skeletal muscles in vivo. By establishing a rat ischemic hind limb model, the effects of ultrasound-mediated microbubble destruction on vascular endothelial growth factor (VEGF) gene transfection to skeletal muscles were also studied in vivo. MATERIALS AND METHODS: Ultrasound irradiation was applied on the mixture of microbubbles and GFP plasmid in vitro. Gel electrophoresis was used to detect the effects of ultrasound and microbubbles on GFP plasmid. For in vivo experiments, ultrasound irradiation was applied on the hind limb after directly injecting microbubbles into the hind limb of Wistar rats. Directly after treatment, the skeletal muscles were harvested to observe the microstructure. We also studied the transfer rate of GFP plasmid DNA into the skeletal muscles of rats by applying ultrasound and microbubble technique. Furthermore, a naked VEGF plasmid was applied to study the feasibility of angiogenesis by using rats ischemia models. RESULTS: Gel electrophoresis of plasmid DNA showed that there was no difference between the groups. By studying the hematoxylin and eosin stained pictures of the skeletal muscles, we found that ultrasound irradiation of skeletal muscle after injection of microbubbles could cause the exudation of the red blood cells, whereas it had no effects on the microstructure of muscle fibers. In vivo experiments showed that an ultrasound microbubble could enhance the transfer of plasmid DNA to the skeletal muscles. CONCLUSIONS: The ultrasound-mediated microbubble technique provides an effective noninvasive method for gene therapy.     

干扰血管内皮生长因子表达联合γ射线对食管癌细胞移植瘤的影响

目的 研究反义cDNA干扰血管内皮生长因子(VEGF)表达联合γ射线对裸鼠移植瘤的影响,并观察各组裸鼠肿瘤病理组织学和细胞生物学特征的变化,为VEGF基因治疗联合放射治疗食管癌提供理论依据。方法 将32只Balb/c/nu裸鼠随机分为4组:对照组、单纯照射组、反义组和反义+照射组,使用已转染和未转染反义VEGFcDNA TE-1食管癌细胞株,分别建立裸鼠爪垫移植瘤模型, 瘤体直径0.8~1.0 cm,⁶⁰Co γ射线18 Gy单次照射单纯照射组与反义+照射组裸鼠移植瘤,对比观察各组移植瘤生长情况,检测移植瘤组织中VEGF mRNA和蛋白的表达,并分析肿瘤组织中细胞凋亡情况。结果 与未转染两组比较,转染两组瘤体生长速度慢,成瘤潜伏期延长(t=13.898, P<0.01)。反义组肿瘤体积为(1207.50±97.07)mm³,反义+照射组为(1057.5±91.50)mm³,两者差异无统计学意义(t=1.124,P>0.05),而此2组与对照组(5442.50±185.08)mm³ 和单纯照射组(2922.50±152.773)mm³ 体积间差异有统计学意义(t=9.475~21.238,P<0.01)。反义组与反义+照射组VEGF mRNA和蛋白表达较对照组和单纯照射组差异有统计学意义(F=387.394、13.519, P<0.01)。结论 抗VEGF治疗可有效地抑制裸鼠移植瘤的生长,但单纯抑制VEGF表达对增加裸鼠移植瘤放射治疗增敏效果有限,需进一步研究。     

Visualization of Hepatic Uptake Transporter Function in Healthy Subjects by Using Gadoxetic Acid-enhanced MR Imaging

Purpose: To determine if genetic polymorphisms of liver-specific human organic anion transporting polypeptide (OATP) 1B1 and OATP1B3 influence cellular uptake of gadoxetic acid in vitro and if functionally relevant polymorphisms are confounders for liver enhancement by gadoxetic acid in healthy subjects. Materials and Methods: This study received ethics approval, and all subjects provided written informed consent. Cellular uptake of gadoxetic acid by OATP1B1 and OATP1B3 and their frequent genetic variants was measured by using stable transfected embryonic kidney HEK293 cells. Liver signal intensity at gadoxetic acid-enhanced MR imaging and pharmacokinetics of gadoxetic acid were evaluated in 36 healthy carriers of SLCO1B1/1B3 wild-type alleles (n = 10), SLCO1B1*1b/*1b (n = 8), SLCO1B1*15/*15 (n = 7), SLCO1B1*5/*15 (n = 1), SLCO1B1*1a/*5 (n = 6), and SLCO1B3*4/*4 (n = 4) by using T1-weighted MR imaging and liquid chromatography tandem mass spectrometry. Results: Transport activity for gadoxetic acid was increased in cells transfected with SLCO1B1c.388A>G (12.8 pmol/[mg·min]6 3.53, P = .001) but decreased in cells with SLCO1B1c.388A>G/521T>C (3.11 pmol/[mg·min] ± 0.918, P = .004) compared with cells with nonvariant transporter (6.32 pmol/[mg·min] ± 2.73). Compared with activity of cells transfected with the nonvariant SLCO1B3 (7.43 pmol/[mg·min] ± 2.43), SLCO1B3c.699G>A was a gain-of-function variant (15.1 pmol/[mg·min] ± 5.52, P = .002), whereas SLCO1B3c.334T>G (0.364 pmol/[mg·min] ± 0.125, P = .0001) and SLCO1B3c.1564G>T (0.295 pmol/[mg·min] ± 0.247, P = .0001) were variants with lower function. Liver enhancement with gadoxetic acid was reduced in subjects with OATP1B1*1a/*5 compared with wild-type subjects and those with OATP1B1*1b/*1b (area under enhancement curve, 3-480 minutes in arbitrary units [au]; 20.7 au ± 6.85 vs 36.5 au ± 8.08 [P = .006] vs 34.6 au ± 8.92 [P = .026]). The OATP1B3*4 polymorphism was not of functional relevance. No pharmacokinetic characteristics of gadoxetic acid were influenced by genetic polymorphisms of OATP1B1 and OATP1B3. Conclusion: Liver-specific OATP1B1 and OATP1B3 are uptake carriers for gadoxetic acid in subjects. Genetic polymorphisms of OATP1B1 are signal confounders in gadoxetic acid-enhanced liver MR imaging. ? RSNA, 2012 Supplemental material: http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.12112061/-/DC1.     

表观扩散系数值鉴别良恶性骨肿瘤及肿瘤样病变的价值

目的 探讨MR DWI的ADC值鉴别良、恶性骨肿瘤及肿瘤样病变的价值.方法 对18例良性骨肿瘤及肿瘤样病变和26例恶性骨肿瘤行DWI.采用单激发EPI序列,3个扩散敏感梯度,b值分别为0.500、1000 s/nun2.在ADC图上测量每个病变的最低、最高和整体ADC值.结果 良性骨肿瘤及肿瘤样病变的最低ADC值[(1.28±0.49)×10-3mm2/s]高于恶性骨肿瘤[(0.92±0.35)×10-3mm2/s,t=2.839,P<0.01],整体ADC值[(1.62±0.51)×10-3mm2/s]也高于恶性骨肿瘤[(1.21±0.36)×10-3mm2mm/s,t=3.092,P<0.01],但两者都有很大重叠.良性骨肿瘤及肿瘤样病变的最高ADC值[(2.02±0.55)×10-3mm2/s]与恶性骨肿瘤的最高ADC值[(1.71±0.65)×10-3mm2/s]差异无统计学意义(t:1.669,P＞0.05).去掉以囊腔为主的骨囊肿及动脉瘤样骨囊肿病例,则良性骨肿瘤及肿瘤样病变的最低、最高和整体ADC值分别为(1.11±0.31)×10-3mm2/s、(1.88±0.49)×10-3mm2/s和(1.45±0.35)×10-3mm2/s,与恶性骨肿瘤比较差异无统计学意义(t值分别为1.728、0.964、2.012,P值均>0.05).结论 ADC值不能鉴别良、恶性骨肿瘤及肿瘤样病变.     

In vivo “hot spot” MR imaging of neural stem cells using fluorinated nanoparticles

To optimize ¹⁹F MR tracking of stem cells, we compared cellular internalization of cationic and anionic perfluoro‐15‐crown‐5‐ether (PFCE) nanoparticles using cell culture plates with different surface coatings. The viability and proliferation of anionic and cationic PFCE‐labeled neural stem cells (NSCs) did not differ from unlabeled cells. Cationic PFCE nanoparticles (¹⁹F T1/T2 = 580/536 ms at 9.4 Tesla) were superior to anionic particles for intracellular fluorination. Best results were obtained with modified polystyrene culture dishes coated with both carboxylic and amino groups rather than conventional carboxyl‐coated dishes. After injecting PFCE‐labeled NSCs into the striatum of mouse brain, cells were readily identified in vivo by ¹⁹F MRI without changes in signal or viability over a 2‐week period after grafting. These results demonstrate that neural stem cells can be efficiently fluorinated with cationic PFCE nanoparticles without using transfection agents and visualized in vivo over prolonged periods with an MR sensitivity of approximately 140 pmol of PFCE/cell. Magn Reson Med 60:1506–1511, 2008. © 2008 Wiley‐Liss, Inc.     

Myocardial bridging as evaluated by 16 row MDCT

Purpose

The purpose of this study is to find out the prevalence, appearance and clinical symptoms of myocardial bridging (MB) by MDCT coronary angiography (CTA).

Materials and methods

A total of 280 (50 females) consecutive patients followed with coronary artery disease or postoperative stent and bypass control, underwent CTA performed by 16-MDCT scanner between January 2006 and April 2006. Short axis multiplanar reformatted images were evaluated. MBs were classified as complete and incomplete bridges with respect to continuity of the myocardium over the tunneled segment of left anterior descending artery (LAD) in interventricular groove and the cut-off value is 1.3 mm. Patients diagnosed with MB on CTA who had prior catheter angiography studies were re-evaluated for the presence of MB.

Results

One hundred and twenty MBs [98 (81.6%) on LAD, 2 (1.6%) on diagonal branch, 11 (9.1%) on obtuse marginal, 4 (3.3%) on right coronary artery, 5 (4.1%) on ramus intermedius artery] were detected in 108 (38.5%) patients. Eighty-five (70.8%) of bridged segments in 79 (28.2%) patients were complete and the rest [35 (29.2%) in 34 (12.1%) patients] were incomplete. In 12 patients two MBs (either on different arteries or on the same artery) were detected. The length of bridged segments in patients with complete and incomplete MBs varied between 4–50.9 mm (mean 18 mm) and 4–37.3 mm (mean 13.6 mm), respectively, and the depth of myocardium over the artery ranged between 1–6.4 mm (mean 2.3 mm) and 1–1.2 mm (mean 1 mm), respectively. Thirty (27.7%) out of 108 patients, in whom MB was detected on CTA, were found to have correlative catheter angiography studies, retrospectively and MB was detected only in 4 (13.3%) out of 30 patients.

Conclusion

MDCT coronary angiography is a non-invasive, efficient method in the diagnosis of MB avoiding the procedural risks that catheter angiography carries. MDCT coronary angiography allows direct visualization of the bridge itself and may thus give the opportunity to differentiate between complete and incomplete myocardial bridges.     

In-vivo identification of tumor multidrug resistance with 3H-colchicine [corrected]

Multidrug resistance (MDR) is a major obstacle in the clinical treatment of cancer with natural-product anticancer agents. Identification of MDR in vivo could be important in the design of chemotherapeutic regimens. As a first step in developing radiolabeled drugs to detect MDR, we measured the in vivo distribution of radiolabel from [ring C, methoxy-3H]-colchicine ([3H]-CHC) in immunosuppressed mice bearing xenografts of colchicine-resistant and sensitive tumor cell lines. Experiments were done at trace (1 microgram/kg) and LD50 (4 mg/kg) dose levels. Activity concentration/injected dose was more than twice as great in sensitive as in resistant tumors (p less than 0.01) at 60 min following retroorbital injection of [3H]-CHC. There was no significant difference in activity distribution between trace- and high-dose injections for any of the tissues sampled. Chromatographic analysis of plasma and tumor extracts demonstrated extensive extravascular metabolic degradation of [3H]-CHC. The ratio of [3H]-CHC concentration of injected dose between sensitive and resistant tumors was 3:1 (p less than 0.05), due primarily to protein-bound [3H]-CHC. This preliminary study demonstrates that it is possible to distinguish multidrug resistant from sensitive tumors in vivo on the basis of radiolabel uptake from an injected MDR drug. Colchicine, labeled with 11C at the [ring C]-methoxy group, may be useful as a radiopharmaceutical for quantitative identification of MDR in human tumors using PET.     

Radiolabeled alpha(v)beta3 integrin antagonists: a new class of tracers for tumor targeting.

The alpha(v)beta3 integrins play an important role during tumor metastasis and tumor-induced angiogenesis. Targeting of this receptor may provide information about the receptor status of the tumor and enable specific therapeutic planning. Cyclo(-Arg-Gly-Asp-D-Phe-Val-) has been shown to be a selective alpha(v)beta3 integrin antagonist with high affinity. In this study we describe the synthesis and biological evaluation of [125I]-3-iodo-D-Tyr4-cyclo(-Arg-Gly-Asp-D-Tyr-Val-) ([125I]P2), [125I]-3-iodo-Tyr5-cyclo(-Arg-Gly-Asp-D-Phe-Tyr-) ([125I]P4) and the negative control peptide [1251]-3-iodo-D-Tyr4-cyclo(-Arg-D-Ala-Asp-Tyr-Val-) ([125I]P6). METHODS: Peptides were assembled on a solid support using fluorenylmethoxycarbonyl amino acid coupling protocols. Radioiodination was performed using the iodogen method. The in vitro binding assays were performed using isolated, immobilized alphaIIbeta3 and alpha(v)beta3 integrins. Expression of the alphaVbeta3 receptor on the different tumors was validated by immunohistochemical methods using alpha(v) and alpha(v)beta3 specific antibodies. For biodistribution studies, nude mice with melanoma M21 or mammary carcinoma MaCaF and BALB/c mice with osteosarcoma were used. RESULTS: The in vitro binding assays demonstrate that the introduction of tyrosine and subsequent iodination have no influence on the high affinity and selectivity for alpha(v)beta3. Immunohistochemical staining clearly indicates the presence of the alpha(v)beta3 integrins on the tumor tissue of the melanoma and the osteosarcoma. Pretreatment and displacement studies show specific binding of [125I]P2 on melanoma M21-bearing nude mice and osteosarcoma-bearing BALB/c mice but less specific binding on mammary carcinomas. [125I]P2 exhibits fast elimination kinetics. The accumulation in the tumor 10 min postinjection is 2.07 +/- 0.32 %ID/g for the melanoma M21 and 3.50 +/- 0.49 %ID/g for the osteosarcoma and decreases to 1.30 +/- 0.13 %ID/g and 2.03 +/- 0.49 %ID/g 60 min postinjection, respectively. [125I]P4 shows even faster elimination kinetics, resulting in a tumor accumulation of 0.40 +/- 0.10 %ID/g 60 min postinjection for the osteosarcoma-bearing BALB/c mice. Both peptides reveal predominately hepatobiliary excretion. For [1251]P2, this also is confirmed by autoradiography. The negative control peptide [125I]P6 shows no specific activity accumulation. CONCLUSION: [125I]P2 exhibits high affinity and selectivity for the alpha(v)beta3 integrin in vitro and in vivo and, thus, represents the first radiolabeled alpha(v)beta3 antagonist for the investigation of angiogenesis and metastasis in vivo.     

Technetium-99m-MRP20, a potential brain perfusion agent: in vivo biodistribution and SPECT studies in non-primate animals

MRP20 (N-(2(1H pyrolylmethyl]N'-(4-pentene-3-one-2] ethane-1,2-diamine) complexes with technetium-99m, yielding a neutral, lipophilic species. This compound has been characterized as [TcO(MRP20)]. Biologic investigation of [99mTc][TcO(MRP20)] in female rats showed 2.35% ID in the brain 30 min p.i. with no significant wash-out over 3 hr. A single-photon emission computed tomography (SPECT) study in a dog demonstrated rapid tracer uptake in the brain, reaching a maximum within 1 min, with 2.24% i.d. 15 min p.i., decreasing to 1.7% after 4 hr. The complex undergoes hydrolysis in vitro forming a cationic species. This is possibly the trapping mechanism in the brain in vivo. The main excretory route of [99mTc][TcO(MRP20)] is via the hepatobiliary tract. There is evidence of some "in vivo" cell labeling and soft-tissue uptake.     

磁共振弹性成像对肝脏局灶性良恶性肿瘤鉴别价值的初步研究

目的：探讨磁共振弹性成像(MRE)在肝脏良恶性肿瘤的诊断价值。方法对36例肝脏肿瘤患者(共39个病灶,其中肝细胞癌20个,血管瘤7个,胆管细胞癌5个,转移瘤3个,平滑肌脂肪瘤2个,癌肉瘤1个,巨淋巴增生1个)以及9例健康志愿者行 MRE。通过 FUNCTOOL 后处理获得肿瘤层面弹性图。测量、比较恶性肿瘤、良性肿瘤、恶性肿瘤周围组织、良性肿瘤周围组织和健康志愿者肝组织平均弹性值。结果恶性肿瘤平均弹性值[(7.39±1.70)kPa]明显高于良性肿瘤[(4.11±0.37)kPa,P <0.001]、恶性肿瘤周围组织[(3.50±0.73)kPa,P <0.001]、良性肿瘤周围组织[(2.61±0.45)kPa,P <0.001]及志愿者正常肝组织平均弹性值[(2.38±0.24)kPa,P <0.001]。恶性肿瘤周围组织平均弹性值[(3.50±0.73)kPa]明显高于良性肿瘤周围组织平均弹性值[(2.61±0.45)kPa, P <0.001]。良性肿瘤周围组织[(2.61±0.45)kPa]稍高于志愿者正常肝组织的平均弹性值[(2.38±0.24)kPa],两者无显著差异(P >0.05)。当取截断值为5.08 kPa 时,可鉴别区分良恶性肝肿瘤和正常肝实质。结论MRE 技术可用于肝脏局灶肿瘤性病变的诊断,有助于对肝脏良恶性肿瘤的鉴别。     

Preferential labeling of glial and meningial brain tumors with [2-(14)C]acetate.

Acetate is preferentially transported into and metabolized by astrocytes, rather than synaptosomes or neurons, and labeled acetate is used as a glial reporter molecule to assess glial metabolism and glial-neuronal interactions. Because monocarboxylic acid transporter specificity might confer a phenotype to help localize, detect, and characterize brain tumors of glial origin, use of [2-(14)C]acetate and [(14)C]deoxyglucose (a glucose analog metabolized by all brain cells) was compared in rat and human brain tumors. METHODS: Cultured C6 glioma or U-373 glioblastoma/astrocytoma tumor cells were injected into the caudate nucleus of anesthetized CDF Fisher rats; 2--3 wk later, an intravenous pulse of [2-(14)C]acetate or [(14)C]deoxyglucose was given, and timed blood samples were drawn during the 5- or 45-min experiment, respectively. Local (14)C levels in the brain were assayed by quantitative autoradiography, and acetate uptake or glucose use was calculated. Uptake and metabolism of the [(14)C]acetate was also assayed in C6 glioma and human surgical tumor samples in vitro. RESULTS: [(14)C]Acetate uptake into rat brain C6 tumors was 9.9 +/- 2.1 mL/100 g/min, compared with 3.9 +/- 1.0 mL/100 g/min in contralateral tissue (n = 6; P < 0.001), and was much higher than that into other brain structures (e.g., 5:1 for white matter and 2:1 for cortical gray matter). Glucose use in C6 tumors was 111 +/- 34 micromol/100 g/min, versus 81 +/- 5 micromol/100 g/min in contralateral tissue (n = 6; P = 0.08); no left-right differences in glucose use or acetate uptake were seen in other brain structures. The tumor-to-contralateral-tissue ratio for acetate (2.3 +/- 0.3) exceeded that for deoxyglucose (1.4 +/- 0.5) (P < 0.05), indicating that acetate is a sensitive C6 glioma marker. [(14)C]Acetate uptake also demarcated a few 3-wk-old C6 tumors that had unlabeled necrotic cores. U-373 tumors were smaller than C6 tumors in rat brain and were detected equally well with [(14)C]acetate and [(14)C]deoxyglucose. In vitro uptake of [(14)C]acetate into human glioblastoma or meningioma tumors was higher than uptake into pituitary adenoma. Rat C6 and human tumors with high uptake metabolized acetate to acidic compounds and amino acids. CONCLUSION: Tumor imaging with radiolabeled acetate can help to localize and classify brain tumors. Transporter and metabolic substrate specificity are traits that can be exploited further for in vivo imaging of brain glial tumors.     

肝脏肿瘤的活体氢质子MR波谱和病理分析

目的 研究活体肝脏氢质子MRS(1H-MRS)评价肝脏局灶性病变的价值.方法 对53例肝肿瘤患者(54个直径＞4 cm的肿瘤)和19名正常志愿者进行1H-MRS检查,分别测量满足诊断需要的正常对照组(17名)和经病理证实的良性肝脏肿瘤组(8例)和肝细胞癌(HCC)组(25例)胆碱I唪/甘油三酯亚甲基峰(Cho/Lip)比值,对结果进行方差分析和两样本间Dunnett-t检验,并通过受试者工作特征(ROC)曲线评价1H-MRS诊断HCC的敏感性和特异性.结果 正常肝脏、良性肿瘤及HCC组的Cho/Lip比值分别为0.07±0.04、0.11±0.06和0.55±0.17(F=6.58,P＜0.05),HCC组高于正常对照组和良性肝脏肿瘤组(t值分别为2.99和2.32,P值均＜0.05),正常肝脏组和良性肿瘤组Cho/Lip比值差异无统计学意义(t=1.53,P＞0.05).ROC曲线下面积为0.77,1H-MRS对诊断HCC有较高诊断价值,当Cho峰与Lip峰比值界值点为0.1时,诊断HCC的敏感性和特异性分别为80.0%和62.5%.结论肝脏局灶性病变1H-MRS分析是可行的,对于鉴别IHCC和良性肝脏肿瘤有一定的临床参考价值.     

The synthesis, magnetic purification and evaluation of 99mTc-labeled microbubbles

Introduction

Ultrasound (US) contrast agents based on microbubbles (MBs) are being investigated as platforms for drug and gene delivery. A methodology for determining the distribution and fate of modified MBs quantitatively in vivo can be achieved by tagging MBs directly with ^99mTc. This creates the opportunity to employ dual-modality imaging using both US and small animal SPECT along with quantitative ex vivo tissue counting to evaluate novel MB constructs.

Methods

A ^99mTc-labeled biotin derivative (^99mTcL1) was prepared and incubated with streptavidin-coated MBs. The ^99mTc-labeled bubbles were isolated using a streptavidin-coated magnetic-bead purification strategy that did not disrupt the MBs. A small animal scintigraphic/CT imaging study as well as a quantitative biodistribution study was completed using ^99mTcL1 and ^99mTc-labeled bubbles in healthy C57Bl-6 mice.

Results

The imaging and biodistribution data showed rapid accumulation and retention of ^99mTc-MBs in the liver (68.2±6.6 %ID/g at 4 min; 93.3±3.2 %ID/g at 60 min) and spleen (214.2±19.7 %ID/g at 4 min; 213.4±19.7 %ID/g at 60 min). In contrast, ^99mTcL1 accumulated in multiple organs including the small intestine (22.5±3.6 %ID/g at 4 min; 83.4±5.9 %ID/g at 60 min) and bladder (184.0±88.1 %ID/g at 4 min; 24.2±17.7 %ID/g at 60 min).

Conclusion

A convenient means to radiolabel and purify MBs was developed and the distribution of the labeled products determined. The result is a platform which can be used to assess the pharmacokinetics and fate of novel MB constructs both regionally using US and throughout the entire subject in a quantitative manner by employing small animal SPECT and tissue counting.