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Introduction Tris(4,6-dimethoxysalicylaldimine)- N , N ′-bis(3-aminopropyl)- N , N ′-ethylenediamine (BAPEN), a tris(salicylaldimine) derivative, is a heart positron emission tomography (PET) agent when labeled with 68 Ga. However, its labeling requires complicated and time-consuming procedures. In this study, the authors formulated a new BAPEN kit for convenient 68 Ga labeling. Methods BAPEN (0.25 mg) kits were prepared by dispensing its solution in 1 M sodium acetate buffer (pH 5.5) into sterile vials and lyophilization. The prepared kits were labeled with generator-eluted 68 Ga in 0.1 N HCl. Stability in human serum was tested. Expiration date was determined by accelerated testing according to US Food and Drug Administration guidelines. A Biodistribution study was performed in normal mice after injection via tail vein. Results The prepared kits achieved radiolabeling efficiencies in excess of 95% and showed a shelf-life of 98 days at 25°C and 64.3 months at 4°C. 68 Ga-BAPEN was found to be stable in human serum at 37°C for at least 1 h. Furthermore, a biodistribution study revealed high heart uptake (10.8% ID/g, 1 h). Conclusions The authors developed a BAPEN kit for convenient labeling with 68 Ga. The 68 Ga-BAPEN showed high stability and excellent biodistribution results in normal mice, which is required for myocardial PET imaging.

Background Iodine is an intrinsic element of thyroid hormone, which is essential for childhood growth and development. The Ideal Breast Milk (IBM) cohort study aims to evaluate the effects of maternal iodine status during pregnancy and lactation on maternal thyroid function, offspring growth and development, and offspring thyroid function. Methods The IBM cohort study recruited pregnant women from Seoul National University Hospital between June 2016 and August 2017, followed by enrollment of their offspring after delivery. For the maternal participants, iodine status is evaluated by urinary iodine concentration (UIC) and dietary records in the third trimester and at 3 to 4 weeks and 12 to 15 months postpartum. For the child participants, cord blood sampling and UIC measurements are performed at birth. At 3 to 4 weeks of age, UIC and breastmilk iodine concentrations are measured. At 12 to 15 months of age, growth and development are assessed and measurements of UIC, a thyroid function test, and ultrasonography are performed. Results A total of 198 pregnant women in their third trimester were recruited. Their mean age was 35.1±3.5 years, and 78 (39.4%) of them were pregnant with twins. Thirty-three (16.7%) of them had a previous history of thyroid disease. Conclusion Korea is an iodine-replete area. In particular, lactating women in Korea are commonly exposed to excess iodine due to the traditional practice of consuming brown seaweed soup postpartum. The study of the IBM cohort is expected to contribute to developing guidelines for optimal iodine nutrition in pregnant or lactating women.

We examined whether human sodium/iodide symporter (hNIS) radioiodine gene therapy can modulate the phenotype of cancer cells and enhance the killing activities of CTLs in a mouse tumor model. Various doses of I-131 (75, 300, 600, 1,200, and 2,400 μCi/5 mL) were incubated with hNIS-expressing colon cancer (CT26/hNIS) and parental cells (CT26), and numbers of MHC class I and Fas-expressing cells were determined by fluorescence-activated cell sorting (FACS). In addition, CT26/hNIS or CT26 tumor-bearing mice were treated with 1,200 μCi of I-131, and percentages of MHC class I and Fas-expressing tumor cells were determined by FACS. The levels of tumor-infiltrating CD8+IFNγ+ and CD11c+CD86+ cells and CTL killing activities were measured in CT26/hNIS tumor-bearing mice (treated with PBS or 1,200 μCi of I-131) by FACS and lactate dehydrogenase assay, respectively. MHC class I and Fas gene expressions were markedly upregulated in CT26/hNIS cells, but not in CT26 cells, in an I-131 dose-dependent manner. The level of MHC class I and Fas-expressing cancer cell were 4.5-fold and 2.1-fold higher in CT26/hNIS tumors than in CT26 tumors, respectively (P < 0.01). Interestingly, numbers of tumor-infiltrating CD8+IFNγ+ cells and CD11c+CD86+ cells were 5-fold and 2.5-fold higher in I-131–treated tumors than in PBS tumors, respectively (P < 0.001). Furthermore, CTL assays showed significantly more specific tumor cell lysis in I-131 tumors than in PBS tumors (P < 0.01). Our findings suggest that hNIS radioiodine gene therapy can generate tumor-associated immunity in tumor microenvironments and enhance the killing activities of CTLs. Mol Cancer Ther; 9(1); 126–33

To evaluate the role of p53 in radionuclide gene therapy, we investigated the cytotoxic effect of (131)I and (188)Re following cotransfection of the sodium iodide symporter (NIS) and wild-type p53 (wt-p53) genes into cancer cells.The NIS gene was transfected to human anaplastic thyroid carcinoma cells (ARO) expressing mutant p53 (mt-p53) using liposomes. The uptakes of (125)I and (188)Re were measured in the transfected (ARO-N) and wild-type cell lines (ARO). A recombinant adenovirus-5 vector containing a CMV promoter and wt-p53 cDNA, called Ad-p53, was established and transduced to ARO and ARO-N cells. After incubating cells with (131)I and (188)Re, the survival rate of each cell line was measured using a clonogenic assay. For radionuclide gene therapy in an animal model, Ad-p53 was injected directly into ARO and ARO-N tumours which were transplanted to nude mice. Two days later, (188)Re or saline was injected intraperitoneally into the mice, and the tumours were measured using a calliper for 4 weeks.In ARO-N cells, the uptakes of (125)I and (188)Re were 505.16+/-21.30 pmol/10(6) cells and 13,875.20+/-504.85 cpm/10(6) cells at 30 min, respectively. There was no difference between the survival rates of ARO cells and ARO-N cells after incubation with (131)I or (188)Re. When Ad-p53 was transduced to ARO-N cells, the survival rate of wt-p53-expressing ARO-N cells incubated with (131)I (18.5 MBq/5 ml) and (188)Re (18.5 MBq/5 ml) decreased to 48.8+/-18.4% and 32.6+/-23.5%, respectively. In the nude mice experiment, ARO and ARO-N tumours gradually grew up to six to eight times larger than the initial volume. ARO and ARO-N tumours transduced with Ad-p53 continued to grow. However, the ARO-N tumours treated with Ad-p53 and 185 MBq of (188)Re regressed to 20% of the initial volume.Growth of ARO-N tumour treated with (131)I or (188)Re was significantly inhibited by Ad-p53 transduction in vivo as well as in vitro. Transfection of the NIS gene into human anaplastic thyroid cancer induced the accumulation of beta-emitter radionuclides, and cotransfection with a wt-p53 gene enhanced the cytotoxic effect.

This study was performed to evaluate the clinical value of (18)F-fluorodeoxyglucose (FDG) positron-emission tomography (PET)/computed tomography (CT) for cancer screening in Korean asymptomatic people.Between February 2004 and December 2006, 1,587 asymptomatic individuals underwent FDG PET/CT as part of a cancer screening program with some other diagnostic tests at the healthcare center of our hospital. After excluding patients with a history of malignant tumor, 1,336 subjects were enrolled. All PET/CT images were visually analyzed. In subjects showing positive findings for PET/CT or other screening tests, further diagnostic tests and pathological confirmation were performed.Of the 1,336 subjects, malignant tumors were found in 16 participants (1.2%, thyroid cancer: 9, lung cancer: 2, stomach cancer: 2, and others: 4). There were 47 cases (3.6%) of positive PET/CT findings--11 cases were true positive (thyroid cancer: 8, lung cancer: 1, renal cancer: 1, and invasive thymoma), and 36 false positive, and five cases were false negative. The overall detection rate of PET/CT was 0.8%, and the sensitivity, specificity, positive-predictive value, and negative-predictive value of PET/CT were 68.8, 97.2, 23.4, and 99.6%, respectively.Fluorodeoxyglucose positron-emission tomography has the potential to detect various kinds of malignant tumors in cancer screening test, and the overall detection rate of PET/CT was 0.8%. FDG PET/CT can be a useful cancer screening modality with the selection of high-risk group and appropriate combination with other screening modalities.

The purpose of this study was to investigate the role of (18)F-fluorodeoxyglucose (FDG) PET in detecting high-grade meningioma and predicting the recurrence in patients with meningioma after surgical resection.Fifty-nine patients (27 men and 32 women) with intracranial meningioma who underwent preoperative FDG PET and subsequent surgical resection were enrolled. All patients underwent clinical follow-up for tumor recurrence with a mean duration of 34+/-20 months. The tumor to gray matter ratio (TGR) of FDG uptake was calculated and a receiver-operating characteristic (ROC) curve of the TGR was drawn to determine the cutoff value of the TGR for detection of high-grade meningioma. Further, univariate analysis with the log-rank test was performed to assess the predictive factors of meningioma recurrence.The TGR in high-grade meningioma (WHO grade II and III) was significantly higher than that in low-grade ones (WHO grade I) (p=0.002) and significantly correlated with the MIB-1 labeling index (r=0.338, p=0.009) and mitotic count of the tumor (r=0.284, p=0.03). The ROC analysis revealed that the TGR of 1.0 was the best cutoff value for detecting high-grade meningioma with a sensitivity of 43%, specificity of 95%, and accuracy of 81%. Of 59 patients, 5 (9%) had a recurrent event. In the log-rank test, the TGR, MIB-1 labeling index, presence of brain invasion, and WHO grade were significantly associated with tumor recurrence. The cumulative recurrence-free survival rate of patients with a TGR of 1.0 or less was significantly higher than that of patients with a TGR of more than 1.0 (p=0.0003)FDG uptake in meningioma was the significant predictive factor of tumor recurrence and significantly correlated with the proliferative potential of the tumor.

Noninvasive imaging of specific mRNAs in living subjects promises numerous biological and medical applications. Common strategies use fluorescently or radioactively labelled antisense probes to detect target mRNAs through a hybridization mechanism, but have met with limited success in living animals. Here we present a novel molecular imaging approach based on the group I intron of Tetrahymena thermophila for imaging mRNA molecules in vivo. Engineered trans-splicing ribozyme reporters contain three domains, each of which is designed for targeting, splicing, and reporting. They can transduce the target mRNA into a reporter mRNA, leading to the production of reporter enzymes that can be noninvasively imaged in vivo. We have demonstrated this ribozyme-mediated RNA imaging method for imaging a mutant p53 mRNA both in single cells and noninvasively in living mice. After optimization, the ribozyme reporter increases contrast for the transiently expressed target by 180-fold, and by ten-fold for the stably expressed target. siRNA-mediated specific gene silencing of p53 expression has been successfully imaged in real time in vivo. This new ribozyme-based RNA reporter system should open up new avenues for in vivo RNA imaging and direct imaging of siRNA inhibition.

The maximal safe dose (MSD) on the basis of bone marrow irradiation levels allows the delivery of a large amount of I-131 to thyroid cancer tissue. The efficacy of MSD therapy in differentiated metastatic thyroid cancers that persisted after conventional fixed dose therapy is investigated.Forty-seven differentiated thyroid carcinoma patients with non-responsive residual disease despite repetitive fixed dose I-131 therapy were enrolled in this study. Their postoperative pathologies were 43 papillary carcinomas and 4 follicular carcinomas. The MSD was calculated with the Memorial Sloan-Kettering Cancer Center protocol using serial blood samples. The MSDs were administered at intervals of 6 months. Treatment responses were evaluated using I-131 whole-body scans and serum thyroglobulin measurements.The mean calculated MSD was 12.5 +/- 2.1 GBq (339.6 +/- 57.5 mCi). Of the 46 patients, 7 (14.9%) showed complete remission, 15 (31.9%) partial remission, 19 (40.4%) stable disease, and 6 (12.8%) disease progression. Of the patients who showed complete or partial remission, 15 (65%) showed response after the first MSD session and 6 (26%) showed response after the second session. Twenty-nine patients (62%) experienced transient cytopenia after therapy, but three did not recover to the baseline level.The maximal safe dose provides an effective means of treatment in patients who failed to respond adequately to conventional fixed dose therapy. I-131 MSD therapy can be considered in patients who fail fixed dose therapy.

Integrated positron emission tomography-computed tomography (PET-CT) with fluoro-2-deoxy-D-glucose (FDG) is a noninvasive technique that can depict different primary and metastatic tumor types on the basis of the increased glucose metabolism of malignant tissue. Some investigators believe that FDG PET is useful in detecting metastatic pelvic lymph nodes (LNs) especially in locally-advanced cervical cancer. The results of clinical studies, however, have been controversial, and the predictive value of PET-CT for the assessment of pelvic LNs in early-stage cervical cancer is unclear. This retrospective study examined the accuracy and feasibility of preoperative FDG PET-CT for primary pelvic LN staging in patients with operable cervical cancer. Between 2003 and 2007, the investigators identified through medical records 34 patients with invasive International Federation of Gynecology and Obstetrics (FIGO) stage IA2-IIB cervical cancer who underwent preoperative FDG PET-CT before type II or III radical hysterectomy and pelvic lymphadenectomy. The sensitivity, specificity, accuracy, and positive and negative predictive values for metastasis detection with PET-CT were calculated in relation to results of histological analysis following surgery. Seventeen of the 34 patients (50.0%) had pelvic LN metastases. The median number of harvested LNs was 29. Analysis of the accuracy of PET-CT scan in cervical cancer showed that overall region-specific sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of PET-CT were 36.4, 98.8, 85.7, 88.9, and 88.7%, respectively. The recurrence rates among patients with positive preoperative PET-CT scans were significantly higher than those with negative scans (3/8 versus 1/26, respectively, P = 0.03). These data show that preoperative integrated PET-CT has limited sensitivity, but high specificity in predicting pelvic LN metastasis in the staging of invasive cervical cancer, and may be useful for identifying patients at higher risk for cervical cancer recurrence.

We conducted a molecular imaging and gene therapy method in alpha-fetoprotein (AFP)-producing hepatocellular carcinoma (HCC) by tumor-specific expression of the human sodium/iodide symporter (hNIS) using an AFP promoter.The tumor-specific expression of hNIS gene by the AFP enhancer/promoter was constructed as pcDNA3-AFP/hNIS. The pcDNA3-AFP/hNIS was stably transfected to human HCC (Huh-7/AN) and rat glioma cells (C6/AN). Functional hNIS expression was confirmed by radioiodine uptake. The mRNA and protein-expression level of hNIS were measured. Biodistribution of 131I was evaluated, and scintigraphic images of 99mTc were obtained in xenografted mice. A clonogenic assay was performed by 131I. And, the in vivo therapeutic effect of 131I was evaluated in xenografted mice.In Huh-7/AN cells, iodine was highly accumulated and completely blocked by perchlorate. The protein and mRNA expression levels were correlated with iodine uptake. Radioiodine uptake in Huh-7/AN tumors was higher than those of control tumors and clearly visualized. The survival rate was significantly decreased in Huh-7/AN cells by 131I. Moreover, a growth of Huh-7/AN tumors was inhibited by 131I in mice.AFP-producing hepatoma can be targeted and treated with radionuclides and hNIS, using AFP enhancer/promoter. This targeted hNIS gene therapy and molecular imaging have the potential to be used in the management of AFP-producing HCC.

Molecular imaging includes proteomic, metabolic, cellular biologic process, and genetic imaging. In a narrow sense, molecular imaging means genetic imaging and can be called molecular-genetic imaging. Imaging reporter genes play a leading role in molecular-genetic imaging. There are 3 major methods of molecular-genetic imaging, based on optical, MRI, and nuclear medicine modalities. For each of these modalities, various reporter genes and probes have been developed, and these have resulted in successful transitions from bench to bedside applications. Each of these imaging modalities has its unique advantages and disadvantages. Fluorescent and bioluminescent optical imaging modalities are simple, less expensive, more convenient, and more user friendly than other imaging modalities. Another advantage, especially of bioluminescence imaging, is its ability to detect low levels of gene expression. MRI has the advantage of high spatial resolution, whereas nuclear medicine methods are highly sensitive and allow data from small-animal imaging studies to be translated to clinical practice. Moreover, multimodality imaging reporter genes will allow us to choose the imaging technologies that are most appropriate for the biologic problem at hand and facilitate the clinical application of reporter gene technologies. Reporter genes can be used to visualize the levels of expression of particular exogenous and endogenous genes and several intracellular biologic phenomena, including specific signal transduction pathways, nuclear receptor activities, and protein-protein interactions. This technique provides a straightforward means of monitoring tumor mass and can visualize the in vivo distributions of target cells, such as immune cells and stem cells. Molecular imaging has gradually evolved into an important tool for drug discovery and development, and transgenic mice with an imaging reporter gene can be useful during drug and stem cell therapy development. Moreover, instrumentation improvements, the identification of novel targets and genes, and imaging probe developments suggest that molecular-genetic imaging is likely to play an increasingly important role in the diagnosis and therapy of cancer.

We compared the F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) findings of brain metastasis between patients with non-small-cell lung cancer (NSCLC) and small cell lung cancer (SCLC). A whole-body FDG and a brain PET were performed in 48 patients (31 men, 17 women; 57 ± 9 years, 42 NSCLC, 6 SCLC), who had brain metastasis on magnetic resonance (MR). All primary lung lesions were detected by FDG-PET and confirmed pathologically. We analyzed the PET findings, lesion sizes, and the pathological result of primary lung cancer. Of the 48 patients, 31 (64.6%) showed hypermetabolic lesions on FDG-PET of the brain image, and 14 (29.2%) showed hypometabolic lesions. Three patients (6.3%) had both hypermetabolic and hypometabolic lesions. On the lesion-based analysis, 74 lesions (67.3%) showed hypermetabolism on FDG-PET, and 36 lesions (32.7%) showed hypometabolism. All primary lung lesions were hypermetabolic on FDG-PET. When the FDG findings of metastatic brain lesions were analyzed with the pathological types of primary lung cancer, NSCLC was more frequently associated with hypermetabolic metastatic brain lesions than SCLC (80% and 26.7%, respectively, P < 0.01). On comparing the sizes of metastatic lesions between SCLC (1.3 ± 1.2 cm) and NSCLC (1.8 ± 1.2 cm), lesions of

Three-dimensional fibrous scaffolds provide an environment that enhances transplanted stem cell survival in vivo and facilitates imaging their localization, viability, and growth in vivo. To assess transplanted stem cell viability on biocompatible polymer scaffolds in vivo, we developed in vivo imaging systems for evaluation of implanted viable neural stem cells (NSC) and mesenchymal stem cells (MSC) on scaffolds using luciferase or sodium/iodide symporter (NIS) genes. Firefly luciferase stably expressing-C6 cell was established (C6-Fluc). The human neural stem cell, F3, was infected with adenoviral vector carrying luciferase gene (F3-Fluc) and MSC expressing NIS controlled by ubiquitin C promoter using lentiviral vector was established by treating blasticidine for 2 weeks (MSC-NIS). Chitosan and poly l-lactic acid (PLLA) scaffolds were used for in vivo image. In vivo expression of luciferase and human NIS was examined by bioluminescence image or 99mTc-pertechnetate gamma camera image, respectively. The cell/scaffold complex was implanted into subcutaneous or abdominal area of BALB/C nude mouse. For quantitative evaluation of cell viability, regions of interest were drawn on 99mTc-pertechnetate scintigraphy by manual. The gradual increase of luciferase activity was observed in C6-Fluc seeded with chitosan according to the increase in the number of cells. C6-Fluc/chitosan complex subcutaneously implanted into nude mice showed longitudinal bioluminescence image until 34 days. Luciferase image of abdominal-injected C6-Fluc/PLLA complex was saturated in only 14 days, showing great cell growth due to abundant nutrients. F3 cells showed well-incorporated pattern with fibrous chitosan scaffold using scanning electron microscopy. F3 infected with Ad-Fluc showed >100-fold higher luciferase activity than luciferase activity in F3. Cell-number-dependent increase of luciferase activity was shown in F3-Fluc seeded on chitosan. F3-Fluc incorporation into chitosan after abdominal injection was clearly visible on bioluminescence image up to 11 days. Radionuclide imaging showed higher uptake by MSC-NIS on PLLA scaffolds than by MSC-NIS not seeded on a scaffold. Quantitative data showed significantly better survival of MSC-NIS on PLLA scaffolds than without scaffold at 72 h post-implantation, which concurred with histologic findings. These results suggest that NSC-Fluc and MSC-NIS cells incorporated within polymer scaffolds can be monitored on a long-term basis by serial in vivo imaging. We believe that a biocompatible scaffold-based imaging system could be used to assess stem cell viabilities in a non-invasive way to aid the development of regenerative therapeutics.

To establish stable and long-term gene expression in vitro and in vivo, we developed a lentiviral vector system carrying sodium iodide symporter (hNIS) gene under UbC promoter, and transfected this into a colon cancer cell line. The in vitro and in vivo kinetics of radioiodine and [99mTc]-pertechnetate were then investigated, and the therapeutic effect of 1-131 was evaluated in this system. The hNIS gene was transferred into CT26 cells using lentivirus containing UbC promoter. In vitro iodide uptake and efflux were measured in CT26-hNIS cells at various time points. In addition, scintigraphic images were acquired at 30 min after injecting [99mTc]-pertechnetate i.p. into Balb/C mice for 27 days after CT26-hNIS induction. Biodistribution studies were performed at 10 and 30 min and at 1.5, 6 and 24 h after [99mTc]-pertechnetate injections, and the therapeutic effects of radioiodine were investigated by measuring tumor size using a caliper or by quantifying tumor radioactivity levels in scintigraphic images. The iodide uptakes of CT26-hNIS tumors were 10-fold greater than those of CT26 tumors. In addition, iodide uptake was completely blocked by 100 microM potassium perchlorate. The accumulation of [99mTc]-pertechnetate in hNIS expressing tumor cells was found to be positively related to tumor growth. In biodistribution studies, the %ID/g values of CT26-hNIS were 84.0 +/- 4.5 at 1.5 h and 40.8 +/- 3.9 at 24 h and these were approximately 60 times greater than those of CT26 at these time points. Tumor growth in mice treated with 131I was retarded until 46 days post-tumor challenge. The devised lentiviral vector system carrying hNIS controlled by UbC promoter was found to be suitable for the long-term monitoring and radionuclide therapy of cancer in living organism.

Firefly luciferase (Fluc) has been widely used as a reporter gene. The aim of this study was to investigate immune response to luciferase protein after an intradermal injection of pcDNA3.1-Fluc in immunocompetent BALB/c mice. We observed bioluminescence at injection sites from one to seven days post-injection when pcDNA3.1-Fluc was intradermally injected into ear-pinnae. To observe induced immune response, the percentages of CD8+IFNgamma+ cells in the draining lymphoid cells of immunocompetent BALB/c mice immunized by pcDNA3.1-Fluc were measured. And the tumor growths of CT26/Fluc in pcDNA3.1-Fluc group were monitored by observing bioluminescent signals and measuring tumor mass, and these were compared with those of the pcDNA3.1 group in immunocompetent BALB/c mice and immunodeficient Nu/Nu mice. In the immunocompetent BALB/c mice, percentages of CD8+IFNgamma+ cells in the pcDNA3.1-Fluc group were higher than those in the pcDNA3.1 group. Ten days after tumor inoculation, tumor growth inhibition was found in the pcDNA3.1-Fluc group, but not in the pcDNA3.1 group in the immunocompetent BALB/c mice. No significant difference in tumor growth inhibition was observed when CT26/Fluc was injected into immunodeficient Nu/Nu mice. In terms of cytokine profiles of draining lymphoid cells of immunized mice, IFNgamma protein levels in the pcDNA3.1-Fluc group were higher than in pcDNA3.1 group animals among the immunocompetent BALB/c mice. In conclusion, Fluc induced a Th1 immune response to Fluc protein delivered by injecting pcDNA3.1-Fluc into immunocompetent BALB/c mice. We suggest that immune response to the Fluc gene is cautionary in preclinical or clinical trials involving the Fluc gene, and that the immunologic potential of firefly luciferase as a naked DNA may be useful in cancer immunotherapy.

FDG uptake mediated by glucose transporter type 1 (Glut-1) and tumor proliferative activity assessed by Ki-67 expression provide prognostic information in patients with non-small-cell lung cancer (NSCLC). Here, we compared the prognostic significances of FDG uptake, and of Glut-1 and Ki-67 expressions in patients with NSCLC.NSCLC patients (n=53, F:M=16:37, age 61.9+/-12.1 years) who underwent curative resection after FDG-PET were enrolled. Thirty-one patients had stage I, 15 stage II, and 7 stage III disease. Patients were treated by surgery only (n=12), surgery plus adjuvant oral chemotherapy (n=32), or surgery plus adjuvant intravenous chemo- or radio-therapy (n=9). Maximum standardized FDG uptake values (maxSUV), and the Glut-1 and Ki-67 expressions of resected tumors were analyzed for correlations and relations with tumor recurrence. The median follow-up duration was 15 months.Thirteen (24.5%) of the 53 patients experienced recurrence during a median follow-up of 8 months and significant correlations were found between maxSUV, Glut-1, and Ki-67 expressions (r=0.48-0.79, p0.001). Univariate analysis revealed that disease-free survival (DFS) was significantly correlated with maxSUV (7 versusor =7, p=0.001), % Ki-67 expression (25% versusor =25%, p=0.047), tumor size (3 cm versusor =3 cm, p=0.027), and tumor cell differentiation (well/moderate versus poor, p=0.011). However, multivariate Cox proportional analysis identified maxSUV as the only determinant of DFS (p=0.005). Patients with a maxSUV ofor =7 (n=14) had a significantly lower 1-year DFS rate (57.1%) than those with a maxSUV of7 (n=39, 89.7%).FDG uptake is more valuable than Glut-1 or Ki-67 expression in terms of predicting prognosis in patients with resected NSCLC.

For noninvasive monitoring of cellular status by dual reporters, a dual membrane protein reporter system was developed and its in vivo applicability was examined. Human sodium iodide symporter (hNIS) and mutant dopamine D(2) receptor (D(2)R) transgenes were chosen considering their complementarity.pIRES-hNIS/D(2)R containing NIS and D(2)R linked with an internal ribosomal entry site (IRES) was constructed and transfected into human hepatoma SK-Hep1 and rat glioma C6 cells. The cell lines stably expressing hNIS and D(2)R (named SK-ND and C6-ND) were produced, which was confirmed by messenger RNA expression of reporter genes. The functional activities of hNIS and D(2)R were measured by (125)I uptake assay and (3)H-spiperone receptor-binding assays. A biodistribution study was performed on SK-ND tumor-bearing mice using (99m)Tc-pertechnetate and (3)H-spiperone. In vivo hNIS expression was examined using (99m)Tc-pertechnetate gamma-camera imaging and, D(2)R expression was examined using a (3)H-spiperone autoradiographic study.(125)I uptake of SK-ND and C6-ND cell lines showed a maximum 97-fold and 43-fold increase, respectively, which were completely inhibited by KClO(4). Specific (3)H-spiperone binding to SK-ND and C6-ND cell homogenates was observed, which were completely inhibited by (+)-butaclamol. Among the dual reporter gene-expressing cell lines, the activities of both reporters were inversely correlated with each other. Competition assay of hNIS-expressing cells by D(2)R vector transfection and D(2)R-expressing cells by hNIS vector transfection showed a dose-dependent decrease of hNIS and D(2)R activities, respectively. In the biodistribution study, (99m)Tc-pertechnetate accumulated 10-fold and (3)H-spiperone accumulated 4-fold more in SK-ND tumors than that in parental SK tumors. In vivo imaging of (99m)Tc-pertechnetate persisted until 5 wk after the cell graft in SK-ND tumors. Autoradiographic study of brain tissues from these mice also revealed an accumulation of (3)H-spiperone in SK-ND tumors.We developed a dual membrane-bound positron and gamma-imaging reporter system of hNIS and D(2)R. We observed its reporting capability in vitro and in vivo and elucidated that these 2 membrane protein reporters competed with each other in their expression. Although we expect that hNIS and D(2)R transgenes can complement each other as a dual reporter system, we suggest that one needs to validate the ratio of expression of the 2 membrane protein reporter transgenes for cellular status tracking.

Human sodium iodide symporter (hNIS) is a transmembrane protein that actively transports iodide ions into thyroid cells. hNIS is over-expressed in some cases of the thyroid cancers compared with the surrounding normal tissues and has been considered to be an attractive target for immunotherapy. The aim of this study is to determine the feasibility of utilizing the hNIS antigenic protein in enhanced-antigen-associated immunotherapy using image analysis with a gamma counter. To accomplish this, minimalistic immunogenically defined gene expression (MIDGE), either plain or coupled to a nuclear localization signal (NLS) peptide, was used as a vector system. Vaccination with MIDGE/hNIS, MIDGE/hNIS-NLS and pcDNA3.1/hNIS produced a significant increase in the number of hNIS-associated IFN-gamma-secreting CD8(+) T cells, with MIDGE/hNIS having the strongest effect. In addition, immunization with the hNIS encoding vectors induced antigen-mediated antitumor activity against NIS-expressing CT26 tumors in vivo, with the highest tumor free rate (100%) and lowest tumor growth being observed up to 40 days after the CT26/NIS tumor challenge with MIDGE/hNIS than those resulting from other immunization groups. Tumor progression could be followed noninvasively and repetitively by monitoring levels of hNIS gene expression in the tumors using scintigraphic image analysis. Overall, hNIS has a potential use as an antigen for immunization approaches, and vaccination with MIDGE/hNIS vectors is an effective means of generating hNIS-associated immune responses in mice.

The purpose of this study was to determine the ovarian findings on integrated positron emission tomography/computed tomography scans during follow-up in cervical cancer patients with ovarian transposition.We retrospectively reviewed the clinical data and integrated 18F-fluorodeoxyglucose positron emission tomography/computed tomography of women with ovarian transposition during radical hysterectomy for cervical cancer between December 2003 and March 2006.Eighty-four premenopausal women had ovarian transposition performed during the study period. Twelve positron emission tomography/computed tomography scans from 11 patients were registered for the current study and three women were diagnosed with metastasis: two in lung and one in pelvis. Two patients complained of menopausal symptoms during follow-up, and the hormonal tests were consistent with it. In the current study, an integrated positron emission tomography/computed tomography scan detected one patient with right lower abdominal mass with increased 18F-fluorodeoxyglucose uptake, which was identified as a transposed right ovary. Clinical information of ovarian transposition was helpful in interpretation of the lesion. In the patient, transposed ovary was associated with increased fluorodeoxyglucose uptake, with standard uptake values ranging from 3.7 to 5.5. Other positron emission tomography/computed tomography scans did not show abnormal uptake of 18F-fluorodeoxyglucose.Transposed ovary in premenopausal women may appear on integrated positron emission tomography/computed tomography scan as a mass with increased 18F-fluorodeoxyglucose uptake, which may be associated with preserved ovarian function. Clinical information regarding transposition should be noted in order not to interpret these as recurrent or metastatic lesions.

To prospectively compare the diagnostic accuracy of ultrasmall superparamagnetic iron oxide (USPIO)-enhanced magnetic resonance (MR) imaging and integrated positron emission tomography-computed tomography (PET/CT) for the depiction of lymph node metastasis in an animal model, with histologic findings as the reference standard.This experiment was approved by the local animal care committee. VX2 carcinoma was implanted into the thighs of 11 rabbits 4 weeks before the imaging study. T2- and T2*-weighted MR examinations were performed 24 hours after USPIO administration, followed by integrated PET/CT. USPIO-enhanced MR imaging and PET/CT analysis for the evaluation of the presence of metastasis in iliac lymph nodes were performed independently by two radiologists and two nuclear medicine physicians, respectively, without histopathologic knowledge. Results were evaluated by using receiver operating characteristic (ROC) analysis, and sensitivities and specificities were compared by using a Z test.Metastases were histopathologically confirmed in 22 of 62 iliac lymph nodes. USPIO-enhanced MR imaging showed a significantly greater area under the ROC curve than did PET/CT (0.984 vs 0.852; P=.023). The respective sensitivity and specificity for the detection of lymph node metastasis were 91% (20 of 22) and 95% (38 of 40) for USPIO-enhanced MR imaging and 64% (14 of 22) and 98% (39 of 40) for PET/CT. In terms of sensitivity, a significant difference was found between USPIO-enhanced MR imaging and PET/CT, particularly for nodal metastasis of less than 5 mm (86% [six of seven] vs 0% [zero of seven]; P=.031), whereas the specificity of the two imaging modalities was similar (P=.226).USPIO-enhanced MR imaging results in higher diagnostic accuracy for depicting lymph node metastasis than does PET/CT.

(18)F-FDG PET is an effective method of predicting recurrence of hepatocellular carcinoma (HCC) after liver transplantation. We compared recently introduced metabolic and volumetric (18)F-FDG PET/CT indices with the current clinicopathologic predictors for ability to predict recurrence.In total, 110 HCC patients who underwent (18)F-FDG PET and liver transplantation were enrolled. On PET, SUVs and tumor-to-background ratios (TBRs) were measured as metabolic activity indices. Various metabolic tumor volumes and uptake-volume products (UVP) were also measured as volumetric indices. The ability of these indices and other clinicopathologic factors to predict recurrence was compared.All metabolic and volumetric indices were significant for recurrence prediction on receiver-operating-characteristic curve analyses (P0.001). On univariate survival analyses, all PET indices-as well as tumor size, tumor number, the Milan criteria, tumor grade, vascular invasion, and T-stage-were significant factors. However, on multivariate analyses, tumor size, tumor grade, maximum TBR, and UVP calculated by inferior vena cava activity were significant factors (P = 0.004, 0.014, 0.009, and 0.021, respectively). When the Milan criteria and PET factors were included in the multivariate analysis, the Milan criteria (P = 0.029), maximum TBR (P0.001), and UVP (P = 0.016) were significant.Volumetric and metabolic activity indices of (18)F-FDG PET are effective predictors of posttransplantation HCC recurrence. In addition to clinicopathologic factors, these indices need to be considered in the selection of candidates for liver transplantation.

Dopaminergic degeneration is a hallmark of Parkinson's disease (PD), which causes various symptoms affected by corticostriatal circuits. So far, the relationship between cortical changes and dopamine loss in the striatum is unclear. Here, we evaluate the gray matter (GM) changes in accordance with striatal dopaminergic degeneration in PD using hybrid PET/MR. Sixteen patients with idiopathic PD underwent (18) F-FP-CIT PET/MR. To measure dopaminergic degeneration in PD, binding ratio (BR) of dopamine transporter in striatum was evaluated by (18) F-FP-CIT. Voxel-based morphometry (VBM) was used to evaluate GM density. We obtained voxelwise correlation maps of GM density according to the striatal BR. Voxel-by-voxel correlation between BR maps and GM density maps was done to evaluate region-specific correlation of striatal dopaminergic degeneration. There was a trend of positive correlation between striatal BR and GM density in the cerebellum, parahippocampal gyri, and frontal cortex. A trend of negative correlation between striatal BR and GM density in the medial occipital cortex was found. Voxel-by-voxel correlation revealed that the positive correlation was mainly dependent on anterior striatal BR, while posterior striatal BR mostly showed negative correlation with GM density in occipital and temporal cortices. Decreased GM density related to anterior striatal dopaminergic degeneration might demonstrate degeneration of dopaminergic nonmotor circuits. Furthermore, the negative correlation could be related to the motor circuits of posterior striatum. Our integrated PET/MR study suggests that the widespread structural progressive changes in PD could denote the cortical functional correlates of the degeneration of striatal dopaminergic circuits. Hum Brain Mapp 37:1710-1721, 2016. © 2016 Wiley Periodicals, Inc.

Neuroimaging has been used to examine developmental changes of the brain. While PET studies revealed maturation-related changes, maturation of metabolic connectivity of the brain is not yet understood. Here, we show that rat brain metabolism is reconfigured to achieve long-distance connections with higher energy efficiency during maturation. Metabolism increased in anterior cerebrum and decreased in thalamus and cerebellum during maturation. When functional covariance patterns of PET images were examined, metabolic networks including default mode network (DMN) were extracted. Connectivity increased between the anterior and posterior parts of DMN and sensory-motor cortices during maturation. Energy efficiency, a ratio of connectivity strength to metabolism of a region, increased in medial prefrontal and retrosplenial cortices. Our data revealed that metabolic networks mature to increase metabolic connections and establish its efficiency between large-scale spatial components from childhood to early adulthood. Neurodevelopmental diseases might be understood by abnormal reconfiguration of metabolic connectivity and efficiency. DOI: http://dx.doi.org/10.7554/eLife.11571.001, eLife digest The brain consumes a great deal of a sugar called glucose, which is delivered to the brain through blood vessels. Active regions of the brain need more glucose, and so the brain has a metabolic network that controls when and where glucose is metabolized. Yet precisely how this metabolic network changes during brain development is not yet understood. Choi et al. have now monitored the patterns of glucose metabolism in the brains of awake rats as they matured from 'childhood' to early adulthood. The experiments involved injecting the rats with radioactive glucose, and then using a technique called positron emission tomography (commonly known as 'PET scan') to monitor the metabolism of these radioactive sugar molecules in the animals’ brains. Choi et al. showed that the patterns of glucose consumption in the brain shift drastically as the rats mature. Importantly, the findings showed that these shifts in glucose metabolism seem to support the activity of long distance connections that develop as the brain matures. The findings also showed that the increased long distance connections were energy efficient. The results suggest that these metabolic changes are likely a way of maintaining high-energy efficiency that is crucial for the brain to perform normally. Finally, in addition to revealing the changes involved in normal brain development, these findings may have implications in neurological and psychiatric disorders in which the brain fails to achieve efficient metabolic networks as it matures. DOI: http://dx.doi.org/10.7554/eLife.11571.002

Purpose Human sodium/iodide symporter (hNIS) protein is a membrane glycoprotein that transports iodide ions into thyroid cells. The function of this membrane protein is closely regulated by post-translational glycosylation. In this study, we measured glycosylation-mediated changes in subcellular location of hNIS and its function of iodine uptake. Methods HeLa cells were stably transfected with hNIS/tdTomato fusion gene in order to monitor the expression of hNIS. Cellular localization of hNIS was visualized by confocal microscopy of the red fluorescence of tdTomato. The expression of hNIS was evaluated by RT-PCR and immunoblot analysis. Functional activity of hNIS was estimated by radioiodine uptake. Cyclic AMP (cAMP) and tunicamycin were used to stimulate and inhibit glycosylation, respectively. In vivo images were obtained using a Maestro fluorescence imaging system. Results cAMP-mediated Glycosylation of NIS resulted in increased expression of hNIS, stimulating membrane translocation, and enhanced radioiodine uptake. In contrast, inhibition of glycosylation by treatment with tunicamycin dramatically reduced membrane translocation of intracellular hNIS, resulting in reduced radioiodine uptake. In addition, our hNIS/tdTomato fusion reporter successfully visualized cAMP-induced hNIS expression in xenografted tumors from mouse model. Conclusions These findings clearly reveal that the membrane localization of hNIS and its function of iodine uptake are glycosylation-dependent, as our results highlight enhancement of NIS expression and glycosylation with subsequent membrane localization after cAMP treatment. Therefore, enhancing functional NIS by the increasing level of glycosylation may be suggested as a promising therapeutic strategy for cancer patients who show refractory response to conventional radioiodine treatment.

In-pentetreotide has been used for neuroendocrine tumors expressing somatostatin receptors. Recently, (68)Ga-DOTATOC PET has been used with the advantage of high image quality. In this study, we compared quantitative indices between (111)In-pentetreotide SPECT/CT and (68)Ga-DOTATOC PET/CT.Thirteen patients diagnosed with neuroendocrine tumors were prospectively recruited. Patients underwent (111)In-pentetreotide scans with SPECT/CT and (68)Ga-DOTATOC PET/CT before treatment. The number and location of lesions were analyzed on both imaging techniques to compare lesion detectability. Additionally, the maximal uptake count of each lesion and mean uptake count of the lungs were measured on both imagings, and target-to-normal lung ratios (TNR) were calculated as quantitative indices.Among 13 patients, 10 exhibited lesions with increased uptake on (111)In-pentetreotide SPECT/CT and/or (68)Ga-DOTATOC PET/CT. Scans with SPECT/CT detected 19 lesions, all of which were also detected on PET/CT. Moreover, 16 additional lesions were detected on PET/CT (6 in the liver, 9 in the pancreas and 1 in the spleen). PET/CT exhibited a significantly higher sensitivity than SPECT/CT (100 % vs. 54 %, P 0.001). TNR was significantly higher on PET/CT than on SPECT/CT (99.9 ± 84.3 vs. 71.1 ± 114.9, P 0.001) in spite of a significant correlation (r = 0.692, P = 0.01).Ga-DOTATOC PET/CT has a higher diagnostic sensitivity than (111)In-pentetreotide scans with SPECT/CT. The TNR on PET/CT is higher than that of SPECT/CT, which also suggests the higher sensitivity of PET/CT. (111)In-pentetreotide SPECT/CT should be used carefully if it is used instead of (68)Ga-DOTATOC PET/CT.

Context: Previous studies on the extent to which radioactive iodine (RAI) therapy for thyroid cancer increases the risk of subsequently developing breast cancer have given conflicting results. Objective: This study aimed to evaluate the effect of RAI treatment on breast cancer development and recurrence among female patients with primary thyroid cancer. Design: This was a retrospective cohort study. The risk of subsequent breast cancer associated with RAI and its dose in hazard ratios (HRs) with 95% confidential intervals (CIs) were calculated using time-dependent Cox proportional hazard models. Patients: A total of 6150 patients with thyroid cancer enrolled between 1973 and 2009 were followed until December 2012. Of these, 3631 (59.0%) received RAI therapy. During the follow-up period, 99 primary breast cancers were diagnosed. Main Outcome Measure: Risk of breast cancer development according to RAI therapy and RAI dose during treatment for primary thyroid cancer. Results: RAI therapy did not significantly increase the incidence of subsequent breast cancer among female patients (hazard ratio [HR], 0.49; 95% confidence interval [CI], 0.22–1.06) when a 2-year latency period was accounted for. High-dose RAI (≥120 mCi) was associated with a reduced incidence of subsequent breast cancer (HR, 0.17; 95% CI, 0.05–0.62) in the cohort with a 2-year latency period. Conclusions: The long-term follow-up results of this study suggest that RAI treatment for patients with thyroid cancer may not increase the risk or recurrence of breast cancer.

Aim: We tried to develop a multimodal iron oxide nanoparticles (IO NP) imaging probe by an encapsulation method using specific amphiphiles for 68Ga-labeling and lymph node-targeting. Materials & methods: Nanoparticles (NPs) were encapsulated with a solution containing polysorbate 60 and the amphiphiles. The prepared NPs were labeled with 68Ga and tested in vitro and in vivo. Results: Prepared 1,4,7-triazacyclononane-1,4,7-triacetic acid-IO-Mannose (NOTA-IO-Man) showed a narrow size distribution, and no significant aggregation or degradation under harsh conditions. The relaxivity coefficient of 68Ga-NOTA-IO-Man was higher than that of ferumoxide. The accumulation of 68Ga-NOTA-IO-Man in the lymph node after injection into rat's footpad was confirmed by both positron emission tomography and MRI. Conclusion: We successfully developed PET/MRI dual-modality imaging probe targeting lymph nodes by using the facile encapsulation method.

Purpose The aim of this study was to investigate the clinical performance of whole-body (18)F-FDG PET/Dixon-volume-interpolated breath-hold examination (Dixon-VIBE), T1-weighted, and T2-weighted MRI protocol in patients with colorectal cancer. Patients and methods A total of 59 patients with colorectal cancer were enrolled in this study. Each patient had one of the following clinical conditions: initial stage before therapy, stage after neoadjuvant therapy, suspicious colorectal liver metastases, and colorectal liver metastases after chemotherapy. Fourteen patients had primary colorectal cancer, whereas 38 patients had a total of 132 hepatic lesions, 53 lesions existed before chemotherapy, and 79 lesions appeared after chemotherapy. The primary stage and metastases images were obtained using our PET/Dixon-VIBE/T1/T2 MRI protocol and were analyzed by 2 nuclear medicine physicians. Diagnostic accuracy was compared with contrast-enhanced MRI images, which were based on surgical pathology results. Results The sensitivity of our imaging protocol for primary colorectal cancer was 100% (14/14). T and N stage both showed 92.9% (13/14) accuracy. Of all 132 hepatic lesions, 115 metastatic lesions were analyzed, and 17 benign lesions were excluded (6 were during pretreatment cases, and 11 were during posttreatment cases). In pretreatment metastatic lesions (n = 47), the sensitivities of our protocol and dedicated MRI were 95.7% (45/47) and 100% (47/47), respectively. In posttreatment lesions (n = 68), sensitivities of our protocol and dedicated MRI were 75% (51/68) and 91.2% (62/68), respectively. Conclusions Whole-body PET/Dixon-VIBE/T1/T2 MRI protocol is clinically useful for TNM staging and chemonaive hepatic metastasis in colorectal cancer.

C-11 methionine (MET) PET is commonly used for diagnosing high-grade glioma (HGG). Recently, volumetric analysis has been widely applied to oncologic PET imaging. In this study, we investigated the prognostic value of metabolic tumor volume (MTV) on MET PET in HGG. A total of 30 patients with anaplastic astrocytoma (n = 12) and glioblastoma multiforme (n = 18) who underwent MET PET before treatment (surgery followed by chemo-radiotherapy) were retrospectively enrolled. Maximal tumor-to-normal brain ratio (TNRmax, maximum tumor activity divided by mean of normal tissue) and MTV (volume of tumor tissue that shows uptake >1.3-fold of mean uptake in normal tissue) were measured on MET PET. Adult patients were classified into two subgroups according to Radiation Therapy Oncology Group Recursive Partitioning Analysis (RTOG RPA) classification. Prognostic values of TNRmax, MTV and clinicopathologic factors were evaluated with regard to progression-free survival (PFS). Median PFS of all patients was 7.9 months (range 1.0–53.8 months). In univariate analysis, MTV (cutoff 35 cm3) was a significant prognostic factor for PFS (P = 0.01), whereas TNRmax (cutoff 3.3) and RTOG RPA class were not (P = 0.80 and 0.61, respectively). Treatment of surgical resection exhibited a borderline significance (P = 0.06). In multivariate analysis, MTV was the only independent prognostic factor for PFS (P = 0.03). MTV on MET PET is a significant and independent prognostic factor for PFS in HGG patients, whereas TNRmax is not. Thus, performing volumetric analysis of MET PET is recommended in HGG for better prognostication.

The function of membrane-localized sodium iodide symporter (NIS) determines the efficacy of radioiodine therapy in thyroid cancer. Here, we describe a dual mode reporter fused with human NIS (hNIS) and a red fluorescent protein named tandem dimeric Tomato (tdTomato) for the in vitro and in vivo imaging of hNIS protein expression, localization, and iodide uptake function. Human cervical epithelial adenocarcinoma cell line (HeLa)-hNIS/tdTomato cells were established by transducing a fusion gene expressing hNIS/tdTomato under the control of a cytomegalovirus promoter. Fluorescence imaging, confocal microscopy, and an 125I uptake assay were performed to validate the integrity of the fusion protein. Actinomycin D and cycloheximide were used to block newly synthesized hNIS proteins. In vivo images were acquired using a gamma camera and a Maestro fluorescence imaging device. The fluorescence intensity of membrane-localized hNIS and 125I uptake both were increased after heat shock. Scintigraphy and fluorescence imaging indicated specific accumulation of the hNIS/tdTomato fusion protein in xenografted tumors, supporting the utility of this system for in vivo monitoring of hNIS expression and activity. We developed a novel hNIS/tdTomato dual mode reporter that enables visualization of the expression, localization, and iodine uptake function of hNIS in vitro and in vivo.

The aim of this study was to investigate relationships between the immunohistochemical results and radioiodine scan and (18)F-FDG PET findings in papillary thyroid cancer (PTC) patients with recurrent cervical nodal metastases.A total of 46 PTC patients who had undergone a radioiodine scan and/or (18)F-FDG PET/CT and a subsequent operation on recurrent cervical lymph nodes were enrolled. Twenty-seven patients underwent (18)F-FDG PET/CT, 8 underwent radioiodine scans, and 11 underwent both scans. In all surgical specimens, the immunoexpressions of thyroglobulin (Tg), sodium-iodide symporter (NIS), glucose transporter 1 (Glut-1), and somatostatin receptor 1 and 2A (SSTR1 and SSTR2A) were assessed, and associations between these expressions and radioiodine scan and (18)F-FDG PET findings were evaluated.Of the 38 patients who underwent (18)F-FDG PET/CT, all patients with weak Tg expression had positive (18)F-FDG uptake, while only 45 % of the patients with moderate or strong Tg expression showed positive uptake (p = 0.01). The proportion of patients with positive (18)F-FDG uptake increased as the degree of Glut-1 expression with luminal accentuation increased. Of the 19 patients who underwent a radioiodine scan, the proportion with positive radioiodine uptake was greater among patients with strong NIS and SSTR2A expression than among patients expressing these markers at weak levels (p = 0.04 for all). All three patients with weak Tg expression were negative for radioiodine uptake.The (18)F-FDG uptakes of recurrent cervical nodes are related to strong Glut-1 expression with luminal accentuation and weak Tg expression, whereas radioiodine uptake is related to the strong expressions of NIS and SSTR2A.

Nowadays, the number of primary studies on fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) has been increasing rapidly. Thus, we updated meta-analysis to evaluate the test performance of FDG PET/CT for nodal staging in non-small cell lung cancer (NSCLC) including the most recent studies.We performed a systematic search of MEDLINE and EMBASE for English publications using keywords "positron emission tomography", "lung cancer", and "lymph node". All searches were limited to human studies. Inclusion criteria were studies of the initial nodal staging of NSCLC with PET/CT. The reasons for exclusion are as follows: (1) studies with PET, (2) previous therapy before PET/CT, (3) nodal staging not confirmed by histology, and (4) reviews, abstracts, and editorial materials. 786 articles were identified through database searching.28 studies including 3,255 patients and 11,887 lymph nodes (LN) were eligible for this study. The pooled sensitivity was 0.62 (95% CI 0.54-0.70), widely ranging from 0.13 to 0.98. The specificity ranged between 0.72 and 0.98 with an overall estimated specificity of 0.92 (0.88-0.95) for node-based data. The pooled sensitivity, specificity, positive and negative likelihood ratio were 0.67 (0.54-0.79), 0.87 (0.82-0.91), 5.20 (3.59-7.54), and 0.37 (0.25-0.55) for patient-based data. Studies from tuberculosis (Tb) endemic countries showed lower sensitivity (0.56 vs 0.68, p = 0.03) for node-based data and lower specificity (0.83 vs 0.89, p0.01) for patient-based ones.PET/CT has a high specificity, but low sensitivity for detecting LN metastasis in patients with NSCLC. Tb might be one of the main reasons for lower sensitivity of PET/CT in several countries. The primary clinicians of lung cancer should be aware of the possibility of hidden metastatic LNs in bilateral FDG uptake of mediastinal and hilar LNs, especially in the Tb endemic countries.

To generate a more efficient in vivo reporter and therapeutic gene, we optimized the coding sequence of the human sodium/iodide symporter (NIS) gene by replacing NIS DNA codons from wild type to new codons having the highest usage in human gene translation. The Codon Adaptation Index (CAI), representing the number of codons effective for human expression, was much improved (0.79 for hNIS, 0.97 for opt-hNIS). Both wild-type (hNIS) and optimized human NIS (opt-hNIS) were cloned into pcDNA3.1 and pMSCV vectors for transfection. Various cancer cell lines such as thyroid (TPC-1, FRO, B-CPAP), breast (MDA-MB-231), liver (Hep3B), cervical (HeLa), and glioma (U87MG) were transfected with pcDNA3.1/hNIS or pcDNA3.1/opt-hNIS. I-125 uptake by opt-hNIS-expressing cells was 1.6 similar to 2.1 times higher than uptake by wild-type hNIS-expressing cells. Stable cell lines were also established by retroviral transduction using pMSCV/hNIS or pMSCV/opt-hNIS, revealing higher NIS protein levels and I-125 uptake in opt-hNIS-expressing cells than in hNIS-expressing cells. Moreover, scintigraphic images from cell plates and mouse xenografts showed stronger signals from opt-hNIS-expressing cells than hNIS-expressing cells, and radioactivity uptake by opt-hNIS-expressing tumors was 2.3-fold greater than that by hNIS-expressing tumors. To test the efficacy of radioiodine therapy, mouse xenograft models were established with cancer cells expressing hNIS or opt-hNIS. I-131 treatment reduced tumor sizes of hNIS-and opt-hNIS-expressing tumors to 0.57-and 0.27-fold, respectively, compared to their sizes before therapy, suggesting an improved therapeutic effect of opt-hNIS. In summary, this study shows that codon optimization strongly increases hNIS protein levels and radioiodine uptake, thus supporting opt-hNIS as a more sensitive reporter and efficient therapeutic gene.

To evaluate the accuracy of integrated positron emission tomography (PET) and computed tomography (CT) for depiction of suspected recurrent ovarian carcinoma after treatment, with use of clinical or histological findings as the reference standard.Seventy-seven women (median age, 51 years) with ovarian carcinoma treated with primary cytoreductive surgery followed by platinum-based combination chemotherapy were included, and [18F]fluorodeoxyglucose (FDG) PET/CT was performed for suspected recurrence. In all patients, imaging findings were compared with results of histological examination after surgical exploration or clinical follow-up to determine the diagnostic accuracy of PET/CT in the evaluation of disease status. Fisher's exact test was used to measure the ability of PET/CT to predict recurrent lesions.Forty-five (58.4%) of the 77 patients had documented recurrence during surgical exploration or clinical follow-up, while 32 (41.6%) had no evidence of recurrent tumour. Of the 45 patients with recurrent disease, 27 (60%) were confirmed to have recurrence by surgical biopsy. A correlation was found between PET/CT and histological or clinical analyses (kappa = 0.894). The overall sensitivity, specificity, accuracy, positive predictive value and negative predictive value of PET/CT were 93.3%, 96.9%, 94.8%, 97.7% and 91.2%, respectively. PET/CT modified the diagnostic or treatment plan in 19 (24.7%) patients, by leading to the use of previously unplanned therapeutic procedures in 11 (57.9%) patients and the avoidance of previously planned diagnostic procedures in eight (42.1%) patients.Integrated FDG PET/CT is a sensitive post-therapy surveillance modality for the detection of recurrent ovarian cancer; it aids decisions on treatment plans and may ultimately have a favourable impact on prognosis.

FDG uptake in NSCLC is related to glucose transporter type 1 (Glut-1) expression. Here, we investigated the direct causal relationship between FDG uptake and Glut-1 expression to determine the role of Glut-1 in FDG uptake by malignant and benign lymph nodes (LNs).Fifty-five curative lung resections in 53 NSCLC patients (male:female=36:17, age=62.0+/-11.8 years) were included. Maximum standardized uptake values (maxSUVs) of LNs in preoperative whole body FDG-PET and Glut-1 immunostaining results were compared.Of 316 pathologically confirmed LNs, 12.3% (39/316) were malignant, and in malignant LNs, FDG positive LNs were no different from FDG negative LNs in terms of size (15.0+/-6.7 mm vs 10.0+/-6.1mm, p0.05), or in terms of the proportion of LNs occupied by tumor (60.0+/-28.8% vs 39.2+/-38.4%, p0.05), but had greater percentages of Glut-1 positive cells in tumors (74.1+/-31.8% vs 22.7+/-18.7%, p0.01), and Glut-1 staining intensities (3.4+/-0.9 vs 1.8+/-1.3, p0.01). FDG negative malignant LNs featured cytoplasmic Glut-1 expression and adenocarcinoma. Glut-1 staining intensities were found to be significantly correlated with the maxSUVs of malignant LNs (rho=0.516, p0.05), but the percentages of Glut-1 positive cells in tumors were not (r=0.2072, p0.05). Analysis of FDG positive benign LNs showed that maxSUV was not correlated with degree of follicular hyperplasia, or Glut-1 expression (p0.05).Intense Glut-1 immunoreactivity was found to be proportionally related to the degree of FDG uptake by malignant LNs in NSCLC. However, the finding that Glut-1 expression in lymphoid hyperplasia showed no correlation with FDG uptake in benign LNs requires further investigation.

Background Oestrogen receptors are ligand-dependent transcription factors whose activity is modulated either by oestrogens or by an alternative signalling pathway. Oestrogen receptors interact via a specific DNA-binding domain, the oestrogen responsive element (ERE), in the promoter region of sensitive genes. This binding leads to an initiation of gene expression and hormonal effects. Objective To determine the transcriptional activity of the oestrogen receptor, we developed a molecular imaging system using sodium iodide symporter (NIS) as a reporter gene. Methods The NIS reporter gene was placed under the control of an artificial ERE derived from pERE-TA-SEAP and named as pERE-NIS. pERE-NIS was transferred to MCF-7, human breast cancer cells, which highly expressed oestrogen receptor-alpha with lipofectamine. Stably expressing cells were generated by selection with G418 for 14 days. After treatment of 17beta-oestradiol and tamoxifen with serial doses, the (125)I uptake was measured for the determination of NIS expression. The inhibition of NIS activity was performed with 50 micromol x l(-1) potassium perchlorate. Results The MCF7/pERE-NIS treated with 17beta-oestradiol accumulated (125)I up to 70-80% higher than did non-treated cells. NIS expression was increased according to increasing doses of 17beta-oestradiol. MCF7/pERE-NIS treated with tamoxifen also accumulated (125)I up to 50% higher than did non-treated cells. Potassium perchlorate completely inhibited (125)I uptake. When MDA-MB231 cells, the oestrogen receptor-negative breast cancer cells, were transfected with pERE-NIS, (125)I uptake of MDA-MB-231/pERE-NIS did not increase. Conclusion This pERE-NIS reporter system is sufficiently sensitive for monitoring transcriptional activity of the oestrogen receptor. Therefore, cis-enhancer reporter systems with ERE will be applicable to the development of a novel selective oestrogen receptor modulator with low toxicity and high efficacy.

The plasma membrane glycoprotein sodium/iodide symporter (NIS) is crucial for thyroid hormone biosynthesis and mediates the iodide uptake of thyrocytes. It has been shown that retinoic acid (RA) alters NIS gene expression in thyroid carcinoma lines and stimulates their iodide uptake. Here, we generated an ARO human thyroidal cancer cell line that expresses the NIS gene (ARO-NIS) and found that its baseline 125I uptake was threefold higher than that of its parental ARO cells. However, a 1-microM all-trans retinoic acid (tRA) treatment significantly increased this 125I uptake up to approximately approximately 6.5-fold on Day 3. tRA also elevated NIS mRNA expression in ARO-NIS cells, with peaks of expression being observed on Day 3. To investigate the underlying genomic mechanisms involved in these tRA-induced phenotypic changes, we subjected tRA-treated and untreated ARO-NIS cells to cDNA microarray analysis. Of 1152, genes spotted onto the microarray membrane, 18 were up-regulated (z ratio>2.0) and 33 were down-regulated (z ratio

The administration of naked DNA into animals is increasing as a research tool to develop DNA vaccine. To monitor the distribution and duration of gene expression of a DNA vaccine in living organisms, we used the naked DNA encoding firefly luciferase (F luc ) as an imaging reporter gene, and evaluated in vivo bioluminescent images in a murine model. We observed bioluminescence at the injection site and at inguinal lymph node from 10 h to 24 h post-injection when DNA vaccine encoding F luc (pcDNA3.1-F luc ) was injected into the bilateral posterior flanks in mice. F luc gene expressions at injection sites and unilateral posterior flank inguinal lymph node were also confirmed by RT-PCR. However, when pcDNA3.1-F luc was injected into the mid-dorsum bioluminescent signals were observed at the injection site for up to 14 days post-injection, but no bioluminescent signals were detected in inguinal lymph nodes. Concurrent mRNA expressions of F luc gene at injection sites but not at inguinal lymph nodes were confirmed by RT-PCR. These findings suggest that optical imaging using F luc could be useful for monitoring the location, intensity and duration of gene expression of naked DNA vaccines in living animals non-invasively and repetitively.

Thymic hyperplasia is frequently observed in Graves' disease. However, detectable massive enlargement of the thymus is rare, and the mechanism of its formation has remained elusive. This case showed dynamic changes in thymic hyperplasia on serial computed tomography images consistent with changes in serum thyrotropin receptor (TSH-R) antibodies and thyroid hormone levels. Furthermore, the patient's thymic tissues underwent immunohistochemical staining for TSH-R, which demonstrated the presence of thymic TSH-R. The correlation between serum TSH-R antibody levels and thymic hyperplasia sizes and the presence of TSH-R in her thymus suggest that TSH-R antibodies could have a pathogenic role in thymic hyperplasia.

Several studies have shown that behavioral hyperactivity induced by psychomotor stimulants is prevented by ginseng saponins. In an attempt to investigate whether the effect of ginseng saponins is through their inhibitory action on the enhanced dopaminergic transmission by psychomotor stimulants, we examined the effects of ginseng total saponin (GTS) presynaptically on nicotine-induced dopamine (DA) release in the striatum of freely moving rats using in vivo microdialysis technique and postsynaptically on the in vitro and in vivo binding of [3H]raclopride to DA D2 receptors. Also, we examined the effects of GTS on nicotine-induced locomotor hyperactivity and on nicotine-induced Fos protein expression in the nucleus accumbens and striatum. Systemic pretreatment with GTS (100 and 400 mg/kg, intraperitoneally (i.p.)) resulted in a dose-dependent inhibition of locomotor hyperactivity induced by nicotine. GTS decreased nicotine-induced DA release in the striatum in a dose-dependent manner. However, GTS had no effects on resting levels of locomotor activity and extracellular DA in the striatum. GTS inhibited the in vitro binding of [3H]raclopride to rat striatal membranes with an IC50 of 5.14+/-1.09 microM. High doses of GTS (400 and 800 mg/kg, i.p.) resulted in decreases in the in vivo binding of [3H]raclopride in the striatum. GTS decreased nicotine-induced Fos protein expression in the nucleus accumbens and striatum, reflecting the inhibition by GTS of nicotine-induced enhancement of dopaminergic transmission. The results of the present study suggest that GTS acts not only on dopaminergic neurons directly or indirectly to prevent nicotine-induced DA release but also postsynaptically by binding to DA D2 receptors. This may explain the blocking effect of GTS on behavioral activation induced by nicotine and conceivably by other psychostimulants. Our data raise the possibility that GTS, by attenuating nicotine-induced enhancement of dopaminergic transmission, may prove to be a useful therapeutic agent for nicotine addiction and warrant further investigation on its effect on nicotine's rewarding property.

Algorithm-based parametric imaging of myocardial blood flow (MBF), as measured by H2(15)O PET, has been the goal of many research efforts. A method for generating parametric images of regional MBF by factor and cluster analysis on H2(15)O dynamic myocardial PET was validated by its comparison with gold-standard MBF values determined invasively using radiolabelled microspheres. Right and left ventricular blood pool activities and their factor images were obtained by the application of factor analysis to dynamic frames. By subtraction of the factor images multiplied by their corresponding values on the factors from the original dynamic images for each frame, pure tissue dynamic images were obtained, from which arterial blood activities were excluded. Cluster analysis that averaged pixels having time-activity curves with the same shape was applied to pure tissue images to generate parametric MBF images. The usefulness of this method for quantifying regional MBF was evaluated using canine experiment data. H2(15)O PET scans and microsphere studies were performed on seven dogs at rest and after pharmacological stress. The image qualities and the contrast of parametric images obtained using the proposed method were significantly improved over either the tissue factor images or the parametric images obtained using a conventional method. Regional MBFs obtained using the proposed method correlated well with those obtained by the region of interest method (r = 0.94) and by the microsphere technique (r = 0.90). A non-invasive method is presented for generating parametric images of MBF from H2(15)O PET, using factor and cluster analysis.

To determine the correlation between serum CEA level and the metabolic volume by FDG PET in postoperative patients with recurrent colorectal cancer, FDG PET was performed in 29 consecutive patients with recurrent or metastatic colorectal cancer whose CEA levels were higher than 5 ng/ml. A whole body emission scan was performed 60 minutes after injecting 370-555 MBq of F-18 FDG. "PET volume" and "PET metabolic volume" of tumors were measured on FDG PET images. Based on an isocontour plot of tumor mass at 2.5 SUV (standardized uptake value), the metabolically active tumor "PET volume" was calculated. "PET metabolic volume" was obtained by multiplying the "PET volume" by the mean SUV of the tumor. All recurrent or metastatic lesions were single or multiple lesions of measurable size (axial diameter > 1 cm, minimum "PET volume" 3.5 cm3), and were verified by operation or by other imaging modalities (CT or MRI). There was a linear associations between "PET volume" and serum CEA level. Further regression analysis by least squares showed a highly significant model with an equation of volume = 41.2 + 0.471 x CEA (r2 = 0.629). However, no such association was found between "PET metabolic volume" and serum CEA level according to the residual normality test. In conclusion, "PET volume" measured by FDG PET and serum CEA level in colorectal cancer are significantly correlated. Tumor volume determined by FDG PET can be used as an effective marker of tumor burden in postoperative patients with colorectal carcinoma.

Ischaemic myocardial dysfunction shows different time courses of functional recovery according to the pathophysiological characteristics of the dysfunction. In this study, we investigated the time course of functional recovery according to the preoperative reversibility of perfusion impairment on myocardial single-photon emission computed tomography (SPECT) after revascularisation surgery.Forty-eight patients (42 men and 6 women; mean age 59+/-9 years) who underwent revascularisation surgery were included in the study. 201Tl rest/dipyridamole stress (99m)Tc-sestamibi gated SPECT was performed 10+/-8 days before (preoperative), 105+/-13 days after (early follow-up) and 497+/-66 days after (late follow-up) surgery. Using a 20-segment model, segmental perfusion and thickening were quantified with automatic software. As an indicator of the reversibility of perfusion impairment, a reversibility score (RevS) was defined as a measure of rest minus stress perfusion values. Segmental dysfunction and functional recovery were defined from quantified thickening values. Function-recovered segments were divided into early recovery and late recovery groups, and preoperative perfusion status was compared in these groups. Function-recovered segments were also re-classified into high-RevS and low-RevS groups according to the preoperative RevS, and the time courses of functional recovery were investigated in each group.A total of 502 segments were included in the analysis and 263 were finally classified as function-recovered segments. Of these, 172 were in the early recovery and 91 in the late recovery group. In terms of preoperative perfusion status, RevS was 8.9+/-10.8 in the early recovery group and 5.4+/-11.0 in the late recovery group (P=0.01). When all 502 segments were classified by RevS, no difference in the proportion of final function recovery was observed between the high-RevS and the low-RevS group (54% vs 51%). However, the proportion of early recovery was higher in the high-RevS group (73%) than in the low-RevS group (57%) (P=0.01).Ischaemic dysfunctional myocardium with reversible perfusion impairment tends to recover function earlier after revascularisation surgery than myocardium with a persistent decrease in perfusion.

The purpose of this study was to compare the prognostic value of 11C-methionine (MET) and 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) in glioma patients.The study population comprised 47 patients with gliomas (19 glioblastoma, 28 others). Pretreatment magnetic resonance imaging, MET PET and FDG PET were performed within a time interval of 2 weeks in all patients. The uptake ratio and standard uptake values were calculated. Univariate and multivariate analyses were done to determine significant prognostic factors. Ki-67 index was measured by immunohistochemical staining, and compared with FDG and MET uptake in glioma.Among the several clinicopathological prognostic factors, tumour pathology (glioblastoma or not), age (or =60 or60 years), Karnofsky performance status (KPS) (or =70 or70) and MET PET (higher uptake or not compared with normal cortex) were found to be significant predictors by univariate analysis. In multivariate analysis, tumour pathology, KPS and MET PET were identified as significant independent predictors. The Ki-67 proliferation index was significantly correlated with MET uptake (r=0.64), but not with FDG uptake.Compared with FDG PET in glioma, MET PET was an independent significant prognostic factor and MET uptake was correlated with cellular proliferation. MET PET may be a useful biological prognostic marker in glioma patients.

Retinoic acids are natural derivatives of vitamin A, and play important roles in modulating tumor cell growth by regulating differentiation, thus suggesting the potential use of these derivatives in cancer therapy and prevention. To visualize the intranuclear responses of functional retinoic acid receptors, we have developed a dual-imaging reporter gene system based on the use of sodium/iodide symporter (NIS) and luciferase in cancer cell lines. NIS and luciferase genes were linked with an internal ribosome entry site, and placed under the control of an artificial cis -acting retinoic acid responsive element (pRARE/NL). After retinoic acid treatment, I-125 uptake by pRARE/NL transfected cells was found to have increased by up to about five times that of nontreated cells. The bioluminescence intensity of pRARE/NL transfected cells showed dose-dependency. In vivo luciferase images showed higher intensity in retinoic acid treated SK-RARE/NL tumors, and scintigraphic images of SK-RARE/NL tumors showed increased Tc-99m uptake after retinoic acid treatment. The NIS/luciferase imaging reporter system was sufficiently sensitive to allow the visualization of intranuclear retinoic acid receptor activity. This cis -enhancer imaging reporter system may be useful in vitro and in vivo for the evaluation of retinoic acid responses in such areas as cellular differentiation and chemoprevention.

Iodide uptake occurs across the membrane of thyroid cells via an active transporter, sodium/iodide symporter (NIS). Decreased NIS expression levels account for the reduced iodide uptake in thyroid carcinomas. We found that thyroid cancer patients with positive immunostaining for NIS responded to I-131 therapy better than did the patients with negative immunostaining. Thus, NIS gene can be used for radionuclide gene therapy. Targeted expression of functional NIS in cancer cells would enable these cells to concentrate iodide from plasma and would, therefore, offer the possibility of radioiodine therapy. We and others have shown that gene transfer of NIS into a variety of cell types confers increased radioiodine uptake up to several hundred-fold that of controls. Imaging reporter gene is useful in noninvasively determining the location, duration and magnitude of transgene expression in living animal. We could monitor not only exogenous gene expression, but also endogenous gene expression such as p53 and nuclear receptor activity such as retinoic acid receptor using cis-NIS reporter imaging gene system. This reporter gene imaging can also be used for monitoring cancer cells, stem cells and immune cells. In conclusion, the NIS has the potential to expand molecular nuclear medicine in the near future.