Supplementary Figure S3: Treatment related toxicity.

Supplementary Figure S1-S3 legends

Supplementary Figure S2: Liver function tests changes after hepatic intraarterial therapy. Liver chemistry (alanine aminotransferase [ALT]

Supplementary Figure S1: Quantitative volumetric image analysis.

Purpose Alternating magnetic field (AMF) tissue interaction models are generally not validated. Our aim was to develop and validate a coupled electromagnetic and thermal model for estimating temperatures in large organs during magnetic nanoparticle hyperthermia (MNH). Materials and methods Coupled finite element electromagnetic and thermal model validation was performed by comparing the results to experimental data obtained from temperatures measured in homogeneous agar gel phantoms exposed to an AMF at fixed frequency (155 +/- 10 kHz). The validated model was applied to a three-dimensional (3D) rabbit liver built from computed tomography (CT) images to investigate the contribution of nanoparticle heating and nonspecific eddy current heating as a function of AMF amplitude. Results Computed temperatures from the model were in excellent agreement with temperatures calculated using the analytical method (error < 1%) and temperatures measured in phantoms (maximum absolute error <2% at each probe location). The 3D rabbit liver model for a fixed concentration of 5 mg Fe/cm(3) of tumor revealed a maximum temperature similar to 44 degrees C in tumor and similar to 40 degrees C in liver at AMF amplitude of similar to 12 kA/m (peak). Conclusion A validated coupled electromagnetic and thermal model was developed to estimate temperatures due to eddy current heating in homogeneous tissue phantoms. The validated model was successfully used to analyze temperature distribution in complex rabbit liver tumor geometry during MNH. In future, model validation should be extended to heterogeneous tissue phantoms, and include heat sink effects from major blood vessels.

Neuroendocrine tumors (NET) are a group of neoplasms with neuroendocrine differentiation affecting a wide range of organs. Functional NETs present with symptoms due to the particular hormone produced. Functional NETs are usually small at diagnosis and therefore can be challenging to diagnose. In contrast, non-functioning NETs are generally larger and present with mass effect. Imaging plays an indispensable role in diagnosis, staging and management of patients with NETs. The optimal modality and technique for imaging of NETs depend on the location of primary and metastatic lesions. Regardless of the imaging modality, dynamic contrast-enhanced imaging is essential for evaluation of NETs. In general, CT scan is typically the primary imaging modality for evaluating NETs. MRI is used as a complementary modality, being superior to other modalities to assess liver metastasis. Nuclear medicine imaging is also widely used in NET assessment.

Aim In liver CT perfusion, the dual-input maximum slope (DI-MS) method is commonly used to estimate perfusion to aid diagnosis of tumors. The DI-MS method relies on a model that assumes the splenic time-to-peak (TTP) separates arterial and portal venous perfusion, and occurs prior to venous perfusion. In this preclinical study, we examined how the timeliness of splenic TTP affects DI-MS perfusion calculations of liver tumors. Materials and Methods We analyzed imaging data obtained from 11 New Zealand White rabbits bearing a single implanted VX2 tumor in liver. A liver 320-slice CT perfusion protocol (5,400 images per study) was used to generate images. Times for arterial and portal slopes were recorded, and hepatic arterial perfusion (HAP), portal perfusion (HPP) and perfusion index (HPI) for liver and tumor were separately calculated using manual and automated methods. T-test comparisons and Bland-Altman plot analyses were performed. Results Mean tumor TTP occurred at 9.79 s (SD=3.41) and splenic TTP at 9.75 s (SD=4.47, p=0.98). In 3/11 (27.27%) cases, tumor SP occurred prior to spleen (mean difference=1.33 s, SD=1.15 s). In these cases, mean automated HPP values were 43.8% (SD=52.48) higher compared to manually computed ones. There were statistically significant differences between automated and manual methods for normal liver and tumor HPI and HPP (p<0.01 and p<0.0001, respectively), but not HAP values (p=0.125 and p=0.78, respectively). There was also a statistically significant variation between methods for tumor HPP and HPI (p=0.001, respectively). Conclusion In 320-slice CT perfusion of liver in this preclinical model, we observed that tumor TTP occurred prior to splenic TTP in 27.27% of tumors in liver. This temporal relationship affects tumor perfusion calculations and should be identified to address potential deviations of model assumptions.

Colorectal cancer (CRC) is one of the most common cancers in the world. The most important determinant of survival and prognosis is the stage and presence of metastasis. The liver is the most common location for CRC metastasis. The only curative treatment for CRC liver metastasis (CRLM) is resection; however, many patients are ineligible for surgical resection of CRLM. Locoregional treatments such as ablation and intra-arterial therapy are also available for patients with CRLM. Assessment of response after chemotherapy is challenging due to anatomical and functional changes. Antiangiogenic agents such as bevacizumab that are used in the treatment of CRLM may show atypical patterns of response on imaging. It is vital to distinguish patterns of response in addition to toxicities to various treatments. Imaging plays a critical role in evaluating the characteristics of CRLM and the approach to treatment. CT is the modality of choice in the diagnosis and management of CRLM. MRI is best used for indeterminate lesions and to assess response to intra-arterial therapy. PET-CT is often utilized to detect extrahepatic metastasis. State-of-the-art imaging is critical to characterize patterns of response to various treatments. We herein review the imaging characteristics of CRLM with an emphasis on imaging changes following the most common CRLM treatments.

The goal of this study was to investigate the role of Lipiodol as a tumor-specific imaging biomarker to determine therapeutic efficacy of cTACE and investigate its inter-dependency with tumor perfusion using radiological-pathological correlation in an animal model of liver cancer. Methods: A total of N=36 rabbits were implanted in the left lobe of the liver with VX2 tumors, treated with cTACE using doxorubicin suspended in Lipiodol, and randomly sacrificed at 24 h, 7 days, or 20 days post-TACE. Unenhanced and contrast-enhanced CT scans including a perfusion protocol were obtained before cTACE and immediately before sacrifice. Tumor vascularity and Lipiodol deposition within tumors and hepatic tissue (non-target deposits) were quantified using 3D quantitative assessment tools and measurements of arterial flow, portal flow, and perfusion index (PI). After sacrifice histologic staining, including hematoxylin and eosin (H&E), CD31, and Oil Red O (ORO) were performed on tumor and liver samples to evaluate necrosis, microvascular density (MVD), and Lipiodol retention over time. Transmission electron microscopy (TEM) was performed to assess Lipiodol deposition and clearance over time. Results: All cTACE procedures were carried out successfully except for one, which was excluded from further analysis. Twenty-four hours post-TACE, tumor PI (p=0.04) was significantly decreased, which was maintained at 7 days (p=0.003), but not at 20 days (p=0.4). A strong correlation (R2 = 0.894) was found between the volume of enhancing tumor tissue at baseline and Lipiodol-positive tumor volume post-TACE. Both ORO and TEM showed deposition of Lipiodol across all imaging time points within the VX2 tumors. However, gradual and ultimately near-complete Lipiodol washout was observed over time in the non-tumoral liver. MVD decreased between 24 h and 7 days post-TACE, and then increased 20 days post-TACE (both p<0.01). Conclusions: Our data provide radiology-pathology evidence for the function of Lipiodol as a theranostic, tumor-specific drug delivery agent because it is both imageable and tumor-seeking, whereby it is preferentially taken up and retained by tumor cells. Those tumor-specific functions also enable Lipiodol to act as an imaging biomarker for the therapeutic efficacy of cTACE. Together with volumetric quantification of tumor vascularization on CT, Lipiodol could be used as a predictor of a patient's response to cTACE and contribute to the therapeutic management of patients with liver cancer.

Objective To evaluate the value of 123I-2β-carbomethoxy-3β-(4-iodophenyl)-N-(3-fluoropropyl) nortropane (123I-FP-CIT) dopamine transporter single photon emission computed tomography (DAT-SPECT) to change management strategies of patients suspected of parkinsonism. Method This was an institutional review board–approved, retrospective study. DAT-SPECT scans ordered by movement disorder specialist and neurologists from 2011–2014 were reviewed. Clinical data and radiological reports of 173 patients suspected of parkinsonism were reviewed. The DAT-SPECT scan results were correlated with clinical assessment and treatment changes. Results A total of 173 patients (104 male and 69 female subjects; age, 64.4 ± 12.6 years) suspected of parkinsonism were included. Median duration of symptoms was 36 months (range, 1–480 months). Scans were most often requested when there was diagnostic uncertainty in clinical features (59.6%, 103/173) or to differentiate one other disease from parkinsonism such as Parkinson disease (PD) versus essential tremor (23.7%, 41/173), PD versus drug-induced parkinsonism (8.7%, 15/173), or PD versus psychogenic (6.4%, 11/173) or vascular (1.7%, 3/173) disorders. Patients were classified, according to the DAT-SPECT scanning results, as those with abnormal DAT-SPECT findings (59%, 102/173) and those with normal DAT-SPECT findings (41%, 71/173). In patients with normal DAT-SPECT findings, follow-up management data were available in 76.1% (54/71). The management changed in 39.4% (28/54) after DAT scan with starting a new appropriate medications or supportive therapy in 4.2% (3/28), withholding inappropriate dopaminergic treatment in 11.3% (8/28), or continuing observation in 23.9% (17/28). In patients with abnormal DAT-SPECT findings, follow-up management data were available in 78.4% (80/102). There was change in management of 37.3% (38/80), a new PD treatment was started in 89.5% (34/38). The dose of medication was adjusted in 5.3% (2/38), although the original treatment was not changed. Parkinson disease treatment was stopped in 2.6% (1/38) and discontinued in 2.6% (1/38) based on clinical decision of neurologists despite abnormal DAT-SPECT findings. Conclusions DAT-SPECT findings impacted treatment decisions in 44.7% of patients suspected of Parkinsonism.

Effects of high frequency spark plasma discharge as a time efficiency method in order to softening the natural hard water has been investigated experimentally. A very hard water sample with 331 ± 19 mg/l of CaCO3 hardness was used. The current and voltage of each spike was about 9.6 A and 3.5 kV respectively at 16 kHz frequency with 35 μs pulse width. Hard water was treated for 2, 4, 6, and 8 min. The concentration of CaCO3, Ca2+ ions, Mg2+ ions and pH as well as water conductivity was controlled before and after treatment. The concentration of CaCO3 dropped by 70%, after 8 min treatment. During the treatment, the pH had a fluctuation about 1.5 and finally remained in neutral state. Also the elemental composition, crystalline structure and morphology of the precipitates were identified. Molecular dynamics simulation revealed that the ozone and hydroxyl play important roles in the softening of the hard water.

Abstract Purpose: To evaluate safety and characterize anticancer efficacy of hepatic hypoxia-activated intra-arterial therapy (HAIAT) with evofosfamide in a rabbit model. Experimental Design: VX2-tumor-bearing rabbits were assigned to 4 intra-arterial therapy (IAT) groups (n = 7/group): (i) saline (control); (ii) evofosfamide (Evo); (iii) doxorubicin–lipiodol emulsion followed by embolization with 100–300 μm beads (conventional, cTACE); or (iv) cTACE and evofosfamide (cTACE + Evo). Blood samples were collected pre-IAT and 1, 2, 7, and 14 days post-IAT. A semiquantitative scoring system assessed hepatocellular damage. Tumor volumes were segmented on multidetector CT (baseline, 7/14 days post-IAT). Pathologic tumor necrosis was quantified using manual segmentation on whole-slide images. Hypoxic fraction (HF) and compartment (HC) were determined by pimonidazole staining. Tumor DNA damage, apoptosis, cell proliferation, endogenous hypoxia, and metabolism were quantified (γ-H2AX, Annexin V, caspase-3, Ki-67, HIF1α, VEGF, GAPDH, MCT4, and LDH). Results: cTACE + Evo showed a similar profile of liver enzymes elevation and pathologic scores compared with cTACE. Neither hematologic nor renal toxicity were observed. Animals treated with cTACE + Evo demonstrated smaller tumor volumes, lower tumor growth rates, and higher necrotic fractions compared with cTACE. cTACE + Evo resulted in a marked reduction in the HF and HC. Correlation was observed between decreases in HF or HC and tumor necrosis. cTACE + Evo promoted antitumor effects as evidenced by increased expression of γ-H2AX, apoptotic biomarkers, and decreased cell proliferation. Increased HIF1α/VEGF expression and tumor glycolysis supported HAIAT. Conclusions: HAIAT achieved a promising step towards the locoregional targeting of tumor hypoxia. The favorable toxicity profile and enhanced anticancer effects of evofosfamide in combination with cTACE pave the way towards clinical trials in patients with liver cancer. Clin Cancer Res; 23(2); 536–48. ©2016 AACR.

To validate the accuracy of wide-array CT to measure differences in regional liver and tumor perfusion in a rabbit model of liver cancer, during first-pass, contrast-enhanced wide-array CT. 7 NZW male liver VX2 bearing rabbits rabbits were anesthetized and IA injected with gold microspheres (15μm, 2 x 106 spheres, BioPal, Worcester, MA) in 30 sec, into the left ventricle under fluoroscopic guidance. while withdrawing the reference sample at 1 ml/min for 2 min, from the femoral artery. Rabbits were scanned in a wide-array 320-detector row volumetric CT scanner (Acquilion One, Toshiba, Japan) 2 hours before IA injection. Scanning parameters were: FOV = 22 cm, kV = 80, mA = 100, slice thickness = 0.5 mm, scan delay = 6 sec, intermittent scanning for 74 sec (arterial phase every 2 sec, portal venous phase every 3 sec). Rabbits were injected with 1.5 ml/kg of isoosmolar CM at 1 ml/sec, followed by a saline flush. CT data were analyzed using the Toshiba CT body perfusion software and calculated tumor and hepatic perfusion values of arterial flow (AF) portal flow (PF) and perfusion index (PI). All rabbits were euthanized after microsphere injection. Samples were weighed and subsequently analyzed together with the respective reference samples using neutron activation quantification. Regional blood flow for each tissue sample was calculated according to: RBF = Fref_(Asmp/Wsmp)/Aref, where Fref is reference flow ( = withdrawal rate), Asmp is activity sample, Wsmp is weight of sample and Aref is activity of reference. Pearson's correlation was used for correlating stable microsphere and CT perfusion flow values. Mean RBF for tumor was 1.15 (SD = 0.74), left liver 0.48 (0.35) and right liver 0.5 (SD = 0.45) ml/min/g. Mean tumor AF was 0.87 (SD = 0.28), left liver AF 0.56 (SD = 0.24), right liver AF 0.55 (SD = 0.21) ml/min/g. CT perfusion AF values were weakly (r = 0.6), but statistically significantly correlated (p = 0.04) to RBF values measured with stable microspheres. Wide-array CT perfusion AF values provide semiquantitative measurements of tumor regional blood flow in the rabbit model of liver cancer.

Purpose We aimed to evaluate the added value of performing fourth and subsequent follow-up 18F-FDG-PET/CT scans to clinical assessment and impact on the patient’s management in patients with melanoma. Methods This was a retrospective study of 232 biopsy-proven melanoma patients who underwent 18F-FDG-PET/CT scans. Of these, 71 patients had 4 or more follow-up 18F-FDG-PET/CT scans after completion of primary treatment, with a total of 246 fourth or subsequent follow-up PET/CT scans. The added value of each follow-up PET/CT scan to the patient’s clinical assessment and treatment management was established. Kaplan-Meier plots with a Mantel-Cox log-rank test were used to establish the patient’s overall survival. Results Of the 246 fourth and subsequent follow-up PET/CT scans, 61% (150/246) were negative for malignancy, and 39.0% (96/246) were positive for recurrence/metastases. FDG-PET/CT was helpful in identifying malignancy in 6.5% of the scans performed without prior clinical suspicion, which ruled out malignancy in 28.5% of the scans obtained with prior clinical signs suggestive of recurrence or for secondary therapy assessment. The PET/CT scan resulted in change of the patient’s management in approximately 16.7% (41/246) of the scans. Change in management was significantly greater in patients whose scans were done with prior clinical signs suggestive of malignancy, or for therapy assessment than without prior clinical suspicion (29.3% vs 4.1%; P < 0.0001). Statistically significant difference was seen in the overall survival between patients with at least 1 positive and all negative fourth and subsequent follow-up PET/CT scans at patient level (P = 0.001). Conclusions The fourth and subsequent 18F-FDG-PET/CT scans obtained after completion of primary treatment added value to clinical assessment in patients with melanoma. Patients with clinical signs suggestive of recurrence or metastases or being monitored for treatment response are more likely to benefit from the fourth or subsequent FDG PET/CT than those without prior clinical suspicion.

Purpose/objective: The aim of this study was to develop and investigate the properties of a magnetic iron oxide nanoparticle-ethiodised oil formulation for image-guided thermal therapy of liver cancer.Materials and methods: The formulation comprises bionised nano-ferrite (BNF) nanoparticles suspended in ethiodised oil, emulsified with polysorbate 20 (BNF-lip). Nanoparticle size was measured via photon correlation spectroscopy and transmission electron microscopy. In vivo thermal therapy capability was tested in two groups of male Foxn1(nu) mice bearing subcutaneous HepG2 xenograft tumours. Group I (n=12) was used to screen conditions for group II (n=48). In group II, mice received one of BNF-lip (n=18), BNF alone (n=16), or PBS (n=14), followed by alternating magnetic field (AMF) hyperthermia, with either varied duration (15 or 20min) or amplitude (0, 16, 20, or 24kA/m). Image-guided fluoroscopic intra-arterial injection of BNF-lip was tested in New Zealand white rabbits (n=10), bearing liver VX2 tumours. The animals were subsequently imaged with CT and 3 T MRI, up to 7 days post-injection. The tumours were histopathologically evaluated for distribution of BNF-lip.Results: The BNF showed larger aggregate diameters when suspended in BNF-lip, compared to clear solution. The BNF-lip formulation produced maximum tumour temperatures with AMF >20kA/m and showed positive X-ray visibility and substantial shortening of T1 and T2 relaxation time, with sustained intratumoural retention up to 7 days post-injection. On pathology, intratumoural BNF-lip distribution correlated well with CT imaging of intratumoural BNF-lip distribution.Conclusion: The BNF-lip formulation has favourable thermal and dual imaging capabilities for image-guided thermal therapy of liver cancer, suggesting further exploration for clinical applications.

This study aimed to evaluate the effects of micron sized non-thermal atmospheric pressure plasma inside the animal body on breast cancer tumor. The μ-plasma jet consists of micron sized hollow tube in which pure helium gas is ionized by high voltage (4 kV) and high frequency (6 kHz). The efficiency of the plasma treatment in killing cancer cells was first investigated by cell viability measurements of treated 4T1 cells using flow cytometry and cell cycle analysis. For exploration of the in vivo effects of the plasma treatment, the BALB/c mice inoculated by 4T1 cell lines were exposed subcutaneously to plasma for 3 minutes. In addition, H&E staining, TUNEL and Western blotting assays were performed in order to observed the effects of the non-thermal plasma on the tumor cells. The results showed that the efficiency of the plasma in suppression of the tumor growth is comparable to that of a typical chemotherapy drug. Moreover, the results indicated that the plasma induces apoptosis in the tumor tissue and increases the ratio of the apoptotic to anti-apoptotic protein expression. We believe that these findings presented herein may extend our knowledge of the mechanisms by which the plasma exerts its promising anti-cancer effects.

Background: There is increasing interest in cardiovascular imaging modalities in the detection of subclinical and clinically-manifested coronary artery disease (CAD) to improve cardiovascular outcome in these patients.Methods: SPECT/CT and PET/CT can be applied for the assessment of myocardial perfusion and myocardial blood flow (MBF) quantification in CAD detection and characterization, while CT is predominantly used to identify coronary plaque burden and epicardial narrowing. In addition, PET/CT plays an increasing role in the detection of the "vulnerable" plaque in the epicardial artery.Results: Imaging of myocardial perfusion with SPECT, SPECT/CT and PET/CT is a mainstay in clinical practice for the identification of flow-limiting epicardial lesions and risk stratification of patients with suspected or known CAD. In this direction, the concurrent ability of PET/CT to determine regional myocardial blood flow (MBF) in ml/g/min at rest and during pharmacologically- induced hyperemic flows allows the calculation of the myocardial flow reserve (MFR) that may unravel reductions in coronary vasodilator capacity, as functional precursor of the CAD process, monitor its response to preventive medical intervention, yield important prognostic information in subclinical-and clinically-manifested CAD, and contributes to identify the flow-limiting effect of single lesions in multivessel CAD. Adding noncontrast computed-tomography (CT) measurements of coronary artery calcifications has further improved the reclassification of cardiovascular risk in asymptomatic individuals with intermediate probability of the presence of CAD. With contrast CT, the non-invasive visualization of coronary vessels, CAD-related plaque burden and stenosis has become feasible. Yet, a definite identification of the "vulnerable plaque" is still a matter of ongoing research. PET/CT in conjunction with various positron-emitting radiotracer yields promise in the detection of the "vulnerable plaque," that, however, needs further clinical evaluation in CAD patients.Conclusion: Multimodality imaging in cardiovascular disease is likely to further advances and refine the identification and characterization of cardiovascular pathology in the near future.

PURPOSE:Embolotherapy using microshperes is currently performed with soluble contrast to aid in visualization. However, administered payload visibility dimishes soon after delivery due to soluble contrast washout, leaving the radiolucent bead's location unknown. The objective of our study was to characterize inherently radiopaque beads (RO Beads) in terms of physicomechanical properties, deliverability and imaging visibility in a rabbit VX2 liver tumor model.MATERIALS AND METHODS:RO Beads, which are based on LC Bead® platform, were compared to LC Bead. Bead size (light microscopy), equilibrium water content (EWC), density, X-ray attenuation and iodine distribution (micro-CT), suspension (settling times), deliverability and in vitro penetration were investigated. Fifteen rabbits were embolized with either LC Bead or RO Beads + soluble contrast (iodixanol-320), or RO Beads+dextrose. Appearance was evaluated with fluoroscopy, X-ray single shot, cone-beam CT (CBCT).RESULTS:Both bead types had a similar size distribution. RO Beads had lower EWC (60-72%) and higher density (1.21-1.36 g/cc) with a homogeneous iodine distribution within the bead's interior. RO Beads suspension time was shorter than LC Bead, with durable suspension (>5 min) in 100% iodixanol. RO Beads ≤300 µm were deliverable through a 2.3-Fr microcatheter. Both bead types showed similar penetration. Soluble contrast could identify target and non-target embolization on fluoroscopy during administration. However, the imaging appearance vanished quickly for LC Bead as contrast washed-out. RO Beads+contrast significantly increased visibility on X-ray single shot compared to LC Bead+contrast in target and non-target arteries (P=0.0043). Similarly, RO beads demonstrated better visibility on CBCT in target arteries (P=0.0238) with a trend in non-target arteries (P=0.0519). RO Beads+dextrose were not sufficiently visible to monitor embolization using fluoroscopy.CONCLUSION:RO Beads provide better conspicuity to determine target and non-target embolization compared to LC Bead which may improve intra-procedural monitoring and post-procedural evaluation of transarterial embolization.