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Western blotting of T24 cell lysates treated with chemical cross-linking

The radiosensitizing effect of erlotinib in HER2 knocked-down NTUB1 cells also emphasizes the synergism between EGFR and HER2 in determining radiosensitivity

Western blotting of T24 cell lysate precipitated with EGFR antibody

The representative pictures of annexin V-stained T24 and NTUB1 cells under fluorescence microscopy

Background: In the treatment of esophageal squamous cell carcinoma (ESCC), the optimal use of 18fluorodeoxyglucose positron emission tomography (PET) in measuring metabolic tumor response is undetermined. We launched a phase II trial to evaluate early metabolic response to one-cycle induction chemotherapy in patients with locally advanced ESCC. Methods: ESCC patients in stage classification T3N0, N1M0, or M1a (American Joint Committee on Cancer, 6th edition) received one-cycle chemotherapy comprising paclitaxel, cisplatin, and 24-h infusional 5-fluorouracil and leucovorin on days 1 and 8, followed by neoadjuvant chemoradiotherapy, 40 Gy, with paclitaxel/cisplatin and then esophagectomy. PET was performed at baseline and day 14 of chemotherapy. The primary endpoint was pathologic complete response (pCR). We hypothesized early metabolic responders with >35% reduction in maximum standardized uptake value (SUVmax), would have better pCR Results. Results: Sixty-six patients were enrolled. The median progression-free survival (PFS) and overall survival (OS) were 16 months (95% confidence interval [CI], 9–27) and 22 months (16–40), respectively. The early metabolic response rate was 55%; and the pCR rate was 34% in the esophagectomy population. The early metabolic response was not associated with pCR or survival. In an exploratory analysis, the postchemotherapy SUVmax was an independent prognostic factor for pCR, PFS, and OS. Conclusion: Our study failed to validate the predefined early metabolic response for pCR to neoadjuvant chemoradiotherapy in locally advanced ESCC patients. However, postchemotherapy SUVmax may be prognostic and predictive, and warrants further study. Keywords: Esophageal squamous cell carcinoma, Neoadjuvant chemoradiotherapy, Metabolic response, Positron emission tomography, Pathological complete response

One of the hallmarks of cancer is increased cell proliferation. Measurements of cell proliferation by estimation of DNA synthesis with several radiolabeled nucleosides have been tested to assess tumor growth. Deoxycytidine can be phosphorylated by deoxycytidine kinase (dCK) and is incorporated into DNA. This study evaluated a radiofluorinated deoxycytidine analog, 5-[18F]fluoro-2′-deoxycytidine ([18F]FdCyd), as a proliferation probe and compared it with 5-[18F]fluoro-2′-deoxyuridine ([18F]FdUrd), 3′-deoxy-3'-[18F]fluorothymidine ([18F]FLT), and [18F]fluorodeoxyglucose ([18F]FDG) in a tumor-bearing mouse model. [18F]FdCyd was synthesized from two precursors by direct electrophilic substitution. The serum stability and partition coefficient of [18F]FdCyd were evaluated in vitro. Positron emission topography (PET) imaging of Lewis lung carcinoma (LLC)-bearing mice with [18F]FdCyd, [18F]FdUrd, [18F]FLT, and [18F]FDG were evaluated. [18F]FdCyd was stable in mouse serum and normal saline for up to 4 h. With all radiotracers except [18F]FLT, PET can clearly delineate the tumor lesion. [18F]FdCyd and [18F]FdUrd showed high accumulation in the liver and kidney. The SUV and tumor-to-muscle (T/M) ratios derived from PET imaging of the radiotracers were [18F]FDG > [18F]FdCyd > [18F]FdUrd > [18F]FLT. Selective retention in tumors with a favorable tumor/muscle ratio makes [18F]FdCyd a protential candidate for further investigation as a proliferation imaging agent.

Introduction [ 18 F]Fluoromethylcholine ([ 18 F]FCH) is a potent tumors imaging agent. In order to fulfill the demand of pre-clinical and clinical studies, we have developed an automated high yield one-pot synthesis of this potent tumors imaging agent. Methods [ 18 F]FCH was synthesized using a modified TRACERlab Fx FN module. Briefly, dibromomethane (10% in CH 3 CN) was fluorinated with K[ 18 F]/K 2.2.2 in a glassy carbon reaction vessel at 120° C for about 5 min to generate [ 18 F]fluorobromomethane ([ 18 F]FBM). The resulting [ 18 F]FBM was then bubbling (He, 700 mL/min) through four Sep-Pak® Silica Plus Long cartridges to react with dimethylaminoethanol (10% DMAE in 0.3 mL DMSO) which was pre-loaded on Sep-Pak® C 18 Plus Short cartridge. The [ 18 F]FCH was purified by solid-phase extraction (SPE) using one Sep-Pak® C 18 Plus Short and one Sep-Pak® CM Plus Short in series. The quality of [ 18 F]FCH synthesized by this method was verified by HPLC and TLC as compared to authentic sample. Results Using this improved one-pot method, the RCY of [ 18 F]FCH was 18.8 ± 2.1% (EOB, n = 27) in a synthesis time of 49 ± 5 min from EOB. The radiochemical purity of [ 18 F]FCH was greater than 90% and the residual DMAE concentration in the final product was less than 10 ppm. Conclusions This optimized method could fulfill the demand of [ 18 F]FCH for both pre-clinical and clinical studies, especially for nearby study sites without a cyclotron.

PURPOSE This study determined the prognostic value of volumetric parameters derived from pretreatment F-FDG and F-DOPA PET/CT of neuroblastoma and their correlation with clinical and histopathologic features. PATIENTS AND METHODS A total of 25 children with neuroblastoma underwent pretreatment F-FDG and F-DOPA PET/CT within 4 weeks. The SUVmax of primary tumors on F-FDG and F-DOPA PET were recorded as SUVFDG and SUVDOPA, respectively. For volumetric parameters of primary tumors, 40% of SUVmax was used to generate volume of interest. If the 40% of SUVmax was below 2.5, an SUV threshold of 2.5 was used instead. Metabolic tumor volume (MTV), total lesion glycolysis (TLG), dopaminergic tumor volume (DTV), and total lesion F-DOPA activity (TLDA) were recorded as F-FDG and F-DOPA volumetric parameters. All indices were compared between groups distinguished by survival status and clinical features, including bone marrow involvement, lymph node metastasis, amplification of the MYCN oncogene, invasive features on anatomic images, and risk categories. The Kaplan-Meier method and log-rank test were used to compare the survival curves between groups. RESULTS The median follow-up period was 28.2 months. Nonsurvivors (20%) tended to have lower SUVDOPA, DTV, and TLDA (P ≤ 0.05), and higher SUVFDG, MTV, and TLG (all P < 0.05). Lower F-DOPA uptake is associated with bone marrow and lymph node metastases (all P < 0.05). Higher F-FDG uptake is associated with MYCN amplification (all P < 0.05) and anatomic invasive features of tumors such as vascular encasement or adjacent organ invasion (TLG, P = 0.05). Only volumetric indices (DTV, TLDA, MTV, and TLG) significantly differed among risk groups (all P < 0.05). CONCLUSIONS Pretherapeutic F-DOPA and F-FDG PET provided complementary information, and both can be served for risk stratification. Volumetric indices of F-DOPA and F-FDG PET correlate more highly with risk grouping.

Performing quantitative small-animal PET with an arterial input function has been considered technically challenging. Here, we introduce a catheterization procedure that keeps a rat physiologically stable for 1.5 mo. We demonstrated the feasibility of quantitative small-animal 18F-FDG PET in rats by performing it repeatedly to monitor the time course of variations in the cerebral metabolic rate of glucose (CMRglc). Methods: Aseptic surgery was performed on 2 rats. Each rat underwent catheterization of the right femoral artery and left femoral vein. The catheters were sealed with microinjection ports and then implanted subcutaneously. Over the next 3 wk, each rat underwent 18F-FDG quantitative small-animal PET 6 times. The CMRglc of each brain region was calculated using a 3-compartment model and an operational equation that included a k*4. Results: On 6 mornings, we completed 12 18F-FDG quantitative small-animal PET studies on 2 rats. The rats grew steadily before and after the 6 quantitative small-animal PET studies. The CMRglc of the conscious brain (e.g., right parietal region, 99.6 ± 10.2 μmol/100 g/min; n = 6) was comparable to that for 14C-deoxyglucose autoradiographic methods. Conclusion: Maintaining good blood patency in catheterized rats is not difficult. Longitudinal quantitative small-animal PET imaging with an arterial input function can be performed routinely.

Myocardial perfusion imaging (MPI) with SPECT is a well-established tool for the diagnosis of coronary artery disease (CAD). However, soft-tissue attenuation is a common artifact that limits the diagnostic accuracy of MPI. The aim of this study was to determine whether attenuation correction (AC) improved the diagnostic performance of MPI, using coronary angiography as a reference standard. Methods: MEDLINE and EMBASE were searched until March 2015 for studies evaluating AC MPI for the diagnosis of CAD. Methodologic quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies tool. For each study, the sensitivity, specificity, and diagnostic odds ratio, along with 95% confidence intervals (CIs), were calculated to determine the diagnostic accuracy of AC versus non–attenuation-corrected (NAC) MPI. A bivariate mixed-effects model was applied for pooling the data. Results: Of 201 articles, 17 studies (1,701 patients) were identified, including 5 studies that used CT AC, 12 studies that used radionuclide source AC (RAC), and 15 studies that reported NAC results. The pooled sensitivities across studies were 0.80 (95% CI, 0.64–0.91), 0.85 (95% CI, 0.81–0.88), 0.84 (95% CI, 0.79–0.88), and 0.80 (95% CI, 0.75–0.85) for CT AC, RAC, all AC, and NAC, respectively. The pooled specificities were 0.83 (95% CI, 0.71–0.91), 0.81 (95% CI, 0.73–0.86), 0.80 (95% CI, 0.74–0.85), and 0.68 (95% CI, 0.61–0.74). Both sensitivities and specificities resulted in a pooled diagnostic odds ratio of 20 (95% CI, 12–34), 24 (95% CI, 13–43), 22 (95% CI, 13–35), and 9 (7–11). Significant differences in specificity and diagnostic odds ratios were noted when AC (including CT AC, RAC, and all AC) was compared with NAC. Conclusion: The results from this study suggested that AC should be applied to MPI to improve the diagnosis of CAD, especially the specificity.

Identification of right ventricular (RV) abnormalities is important in patients with suspected coronary artery disease (CAD). RV activity can be better visualized on myocardial single-photon emission computerized tomography (SPECT) using a higher sensitivity cadmium-zinc-telluride (CZT) detector. The aim of this study was to investigate the clinical significance of RV/left ventricular (LV) uptake ratios during exercise thallium-201 SPECT using CZT detectors.A total of 102 patients underwent treadmill ECG-gated SPECT, coronary angiography, and echocardiography. SPECT myocardial perfusion was interpreted using a 17-segment model and a 0-4-point scale. RV/LV uptake ratios were calculated on the basis of maximum counts per pixel within the entire RV and LV walls. The relationships between RV/LV uptake ratio and gated SPECT, presence of CAD (≥50% stenosis in the left main or ≥70% in the main branches), demographics, and echocardiographic parameters were analyzed.Stress RV/LV ratios correlated positively with the presence of left main or multivessel disease, and tricuspid regurgitation maximum pressure gradient. After multivariate regression, stress/rest RV/LV ratios correlated positively with mitral flow deceleration time, age, female sex, and use of β-blockers.RV/LV uptake ratios on the basis of exercise myocardial perfusion SPECT imaging using CZT cameras are useful for the detection of severe CAD and could serve as an indicator of pulmonary hypertension and LV diastolic dysfunction.

[18F]T807 is a potent tau protein imaging agent. In order to fulfill the demand from preclinical and clinical studies, we developed an automated one-pot two-step synthesis of this potent tau imaging agent and studied its stability, and dosimetry in mice and monkeys. We also conducted a preliminary study of this imaging agent in humans. Using this one-pot two-step method, the radiochemical yield (RCY) of [18F]T807 was 20.5 ± 6.1% (n = 15) at the end of bombardment (EOB) in a synthesis time of 70±5 min. The chemical and radiochemical purities were >90% and the specific activities were 151 ± 52 GBq/μmol. The quality of [18F]T807 synthesized by this method met the U.S. Pharmacopoeia (USP) criteria. The stability test showed that the [18F]T807 injection was stable at room temperature for up to 4 h after the end of synthesis (EOS). The estimated effective dose of the [18F]T807 injection extrapolated from monkeys was 19 μSv/MBq (n = 2), while the estimated effective doses of the [18F]T807 injection extrapolated from fasted and non-fasted mice were 123 ± 27 (n = 3) and 94 ± 19 (n = 4) μSv/MBq, respectively. This one-pot two-step automated method produced the [18F]T807 injection with high reproducibility and high quality. PET imaging and radiation dosimetry evaluation in mice and Formosan rock monkeys suggested that the [18F]T807 injection synthesized by this method is suitable for use in human PET imaging studies. Thus, this method could fulfill the demand for the [18F]T807 injection in both preclinical and clinical studies of tauopathies, especially for nearby study sites without cyclotrons.

A series of diphenylsulfide derivatives with various substitutions at the 4-position on phenyl ring A and different lengths of the 2-fluoroethoxy-substituted side-chain at the 4′-position on ring B were synthesized and evaluated as potential positron emission tomography (PET) imaging agents for serotonin transporters (SERT). These ligands exhibited high SERT binding affinities (Ki = 0.11–1.3 nM) and the 4-methyl-substituted (4-Me) compounds 7a and 8a displayed excellent selectivity for SERT versus norepinephrine transporters (NET) (392- and 700-fold, respectively). In the parallel artificial membrane permeability assay (PAMPA), these ligands demonstrated moderate to high brain penetration, and the 4-Me analogs showed higher BBB permeability than the corresponding 4-F analogs. The 2-fluoroethoxy-substituted ligands showed higher metabolic stability and lower lipophilicity than 4-F-ADAM. [18F]7a-c were readily prepared using an automatic synthesizer and exhibited significant uptake and slow washout in rat brains. At 120 min after iv injection, [18F]7a exhibited the highest uptake in the midbrain, whereas [18F]7b exhibited the highest uptake in the hypothalamus and midbrain. After treatment with citalopram, a SERT-selective ligand, the uptake of [18F]7a in the hypothalamus and striatum was significantly decreased. The potent and highly selective SERT binding and the selective and reversible accumulation in SERT-rich brain regions suggested that [18F]7a is a promising lead for the further development of novel [18F]-labeled PET imaging agents for SERT binding sites in the brain.

One of the hallmarks of cancer is increased cell proliferation. Measurements of cell proliferation by estimation of DNA synthesis with several radiolabeled nucleosides have been tested to assess tumor growth. Deoxycytidine can be phosphorylated by deoxycytidine kinase (dCK) and is incorporated into DNA. This study evaluated a radiofluorinated deoxycytidine analog, 5-[

Neuroblastoma (NB) is a heterogeneous childhood cancer that requires multiple imaging modalities for accurate staging and surveillances. This study aims to investigate the utility of positron emission tomography (PET) with 18F-fluorodeoxyglucose (FDG) and 18F-fluoro-dihydroxyphenylalanine (FDOPA) in determining the prognosis of NB. During 2007-2014, forty-two NB patients (male:female, 28:14; median age, 2.0 years) undergoing paired FDG and FDOPA PET scans at diagnosis were evaluated for the maximum standardized uptake value (SUV(max)) of FDG or FDOPA by the primary tumor. Patients with older age, advanced stages, or MYCN amplification showed higher FDG and lower FDOPA SUV(max) (all P < 0.02). Receiver operating characteristics analysis identified FDG SUV(max) ≥ 3.31 and FDOPA SUV(max) < 4.12 as an ultra-high-risk feature (PET-UHR) that distinguished the most unfavorable genomic types, i.e. segmental chromosomal alterations and/or MYCN amplification, at a sensitivity of 81.3% (54.4%-96.0%) and a specificity of 93.3% (68.1%-99.8%). Considering with age, stage, MYCN status, and anatomical image-defined risk factor, PET-UHR was an independent predictor of inferior event-free survival (multivariate hazard ratio, 4.9 [1.9-30.1]; P = 0.012). Meanwhile, the ratio between FDG and FDOPA SUV(max) (G:D) correlated positively with HK2 (Spearman's ρ = 0.86, P < 0.0001) and negatively with DDC (ρ = -0.58, P = 0.02) gene expression levels, which might suggest higher glycolytic activity and less catecholaminergic differentiation in NB tumors taking up higher FDG and lower FDOPA. In conclusion, the intensity of FDG and FDOPA uptake on diagnostic PET scans may predict the tumor behavior and complement the current risk stratification systems of NB.

Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare autosomal recessive disease that impairs synthesis of dopamine and serotonin. Children with AADC deficiency exhibit severe motor, behavioral, and autonomic dysfunctions. We previously generated an IVS6+4AT knock-in mouse model of AADC deficiency (Ddc(KI) mice) and showed that gene therapy at the neonatal stage can rescue this phenotype. In the present study, we extended this treatment to systemic therapy on young mice. After intraperitoneal injection of AADC viral vectors into 7-day-old Ddc(KI) mice, the treated mice exhibited improvements in weight gain, survival, motor function, autonomic function, and behavior. The yfAAV9/3-Syn-I-mAADC-treated mice showed greater neuronal transduction and higher brain dopamine levels than AAV9-CMV-hAADC-treated mice, whereas AAV9-CMV-hAADC-treated mice exhibited hyperactivity. Therefore, neurotransmitter-deficient animals can be rescued at a young age using systemic gene therapy, although a vector for preferential neuronal expression may be necessary to avoid hyperactivity caused by this treatment.

Background and Aim Sonic Hedgehog (SHH) is a regulator in tumorigenesis of hepatocellular carcinoma (HCC). This study aimed to determine whether radiation-induced SHH signaling occurs in HCC and whether SHH inhibitor acts as a radiosensitizer. Methods The in vitro effects of combining SHH ligand (recombinant human SHH) or inhibitor (cyclopamine) with irradiation were evaluated in the human HCC cell lines, Huh-7 and PLC/PRF/5, and murine cell line BNL. Cell survival and apoptosis were measured using a colony formation assay, annexin-V staining, and poly (ADP-ribose) polymerase activation. Western blotting and immunofluorescence staining were used to detect protein expression. The in vivo response to radiotherapy and/or cyclopamine was tested in BALB/c mice bearing an orthotopic allogeneic tumor. Results Treatment of HCC cells with irradiation and SHH ligand had a protective effect on clonogenic cell survival. Treatment with irradiation and cyclopamine was a more potent inhibitor of cell proliferation than either modality alone. The antiproliferative activity of cyclopamine was attributable to apoptosis induction. Radiation dose-dependently upregulated the expression of Gli-1 (a transcription factor induced by SHH), and this effect was observed mainly in the nucleus. When combined with cyclopamine, irradiation inhibited Gli-1 and increased DNA double-strand breakage. Radiotherapy increased SHH and Gli-1 expression in allogeneic tumor. When compared with radiotherapy alone, cyclopamine with radiotherapy reduced the mean tumor size of orthotopic tumors by 67% (P

Background and purposes The aim of the study was to examine the patterns of lymph node metastases from esophageal squamous cell carcinoma (ESCC) and compare the laterality of lymphatic metastasis in cervical, supraclavicular, and paratracheal areas using F-FDG PET/CT. Patients and methods The data of 75 patients who underwent F-FDG PET/CT for staging of ESCC between January 2011 and March 2012 were reviewed. Fourteen groups of lymph nodes from the neck to abdomen were defined. Lateralization of the upper thoracic lymph nodes was defined in reference to the midline of the trachea. Frequencies of positive lymph nodes were used to determine the pattern of lymphatic spread and compare the lateralization of metastases in the cervical and upper thoracic regions. Results The right paratracheal region was the most frequent site of metastasis among all patients. Left paratracheal and paragastric nodes were more frequent in upper and lower third ESCC, respectively. Upward and downward lymphatic spread was equal in mid third ESCC. In all patients, there was a trend toward more frequent lymph node metastasis on the right side than the left side for the supraclavicular and paratracheal regions. Further stratified analysis with tumor location found that right paratracheal node metastasis was significantly associated with mid third ESCC (P = 0.03). Remote nodal metastasis was found in 10.5% of patients with upper third ESCC and 13% of patients with lower third ESCC, respectively. Remote nodal metastasis was associated with higher SUV of the primary tumor (P = 0.02) and worse survival (P = 0.03). Receiver operating characteristic analysis showed a cutoff SUV of 14.8 for predicting remote lymph node metastases. Conclusions PET/CT provides important information before radiotherapy planning. Mid and lower third ESCC tends to metastasize to the right paratracheal/supraclavicular lymph nodes. Remote nodal metastases on PET/CT correlated with higher primary tumor SUV and worse survival.

PET imaging with (18)F-FDOPA has been successfully applied in the diagnosis and surveillance of neuroblastoma (NB) by targeting the overexpression of aromatic L-amino acid decarboxylase. This study aims to assess the impact of residual (18)F-fluoride on (18)F-FDOPA PET in NB and to implement a method to maintain low (18)F-fluoride content in future studies.Automatic synthesis of (18)F-FDOPA was based on the electrophilic method using TRACERlab FXFE module. Radio-TLC was employed to determine residual (18)F-fluoride levels. We analyzed the impact of residual (18)F-fluoride on the images of 35 patients undergoing (18)F-FDOPA PET at initial diagnosis and/or follow-ups of NB.In 35 batches of (18)F-FDOPA products from 9/28/2010 to 07/27/2011, the mean residual (18)F-fluoride level was 4.4 % (range 0.2-19.1 %). Residual (18)F-fluoride level ≥4.0 % was associated with dense uptake in the growth plates, skull, and pelvis on PET scans, which may interfere with the interpretation of (18)F-FDOPA imaging in NB. By applying stringent restraints in (18)F-FDOPA production, including regular renewal of reagents, exclusive use of NH4OH, and timely replacement of HPLC column, the incidence of (18)F-FDOPA batches with residual (18)F-fluoride level ≥4.0 % was reduced from 33 to 4 % (P 0.0001) during 7/30/2011-4/29/2013.By monitoring residual (18)F-fluoride levels and keeping stringent restraint procedures, low (18)F-fluoride content was achieved in most batches of (18)F-FDOPA, which diminished false-positive skeletal uptake. An appropriate upper limit of (18)F-fluoride level is suggested to be included in the criteria of routine quality control of (18)F-FDOPA productions.

Blockade of EGFR has been proved useful in enhancing the effect of radiotherapy, but the advantages of new-generation EGFR tyrosine kinase inhibitors (TKI) in radiosensitization are not well known. We used two human bladder cancer cells with wild-type EGFR to study the synergism between irradiation and afatinib (an EGFR/HER2 dual kinase inhibitor) or erlotinib (an EGFR kinase inhibitor). Here, we showed that afatinib has better radiosensitizing effect than erlotinib in increasing cancer cell killing, the percentage of apoptotic cells, and DNA damage. Afatinib is also superior to erlotinib in combining radiation to decrease tumor size, inhibit glucose metabolism, and enhance apoptotic proteins in vivo. Finally, erlotinib suppressed cell growth and induced more DNA damage in bladder cancer cells transfected with HER2 shRNA, but not in control vector-treated cells. In conclusion, concomitant blockade of radiation-activated EGFR and HER2 signaling by a new-generation EGFR TKI better inhibits the growth of bladder cancer cells both in vitro and in vivo. The absence of radiosensitization by EGFR inhibition alone and the greater radiosensitizing effect of EGFR inhibitor in HER2 knocked down cells suggest the synergism between HER2 and EGFR in determining radiosensitivity. The regained radiosensitizing activity of erlotinib implies that with proper HER2 inhibition, EGFR tyrosine kinase is still a potential target to enhance radiotherapy effect in these seemingly unresponsive bladder cancer cells. Mol Cancer Ther; 14(3); 810–20. ©2015 AACR.

Background and Aim Instant oatmeal has been proposed as a good alternative to the standardized low-fat egg white test meal for gastric emptying studies. We aim to establish normal values of oatmeal-based gastric emptying scintigraphy and test its correlation with gastroparesis symptoms in the Chinese population. Methods This study prospectively enrolled 60 healthy volunteers, 30 functional dyspepsia and 30 diabetes patients with gastroparesis symptoms. All participants were evaluated using the Gastroparesis Cardinal Symptom Index. Each participant ingested instant oatmeal mixed with 1 mCi of 99mTc diethylenetriaminepentacetic acid, and serial imaging was immediately acquired for 3 h in the supine position using a left anterior oblique projection. Time-activity curves were generated and quantitative parameters were determined. Normal values were established from healthy volunteers and further applied in the symptomatic patients. Results All participants finished the test meal and tolerated the procedure well. All gastric emptying parameters were not significantly affected by age or gender. Values above the 95th percentile of T1/2, gastric retention at 1, 2, and 3 h (85 min, 65%, 28%, and 8%, respectively) were indicative of delayed gastric emptying. Values below the 5th percentile of gastric retention at 0.5 and 1 h (40% and 15%, respectively) were indicative of rapid gastric emptying. The lower gastric retention limit at 0.5 and 1 h were 40% and 15%, respectively. Four (13.3%) diabetes and four (13.3%) functional dyspepsia patients had delayed emptying while three diabetes patients (10%) had rapid emptying. Gastric emptying parameters correlated best with vomiting (r = 0.621) and nausea (r = 0.566) in diabetes patients. Conclusions We established normal values of oatmeal-based gastric emptying scintigraphy and observed good correlation with cardinal gastroparesis symptoms in the Chinese population.

Neurofibrillary tangles are associated with cognitive dysfunction, and hippocampal atrophy with increased CSF tau markers. However, the plasma tau levels of Alzheimer's disease (AD) have not been well studied. We investigated plasma tau by using an immunomagnetic reduction assay in 20 patients with mild cognitive impairment (MCI) due to AD, 10 early AD dementia, and 30 healthy eld- ers (HE). All received a 3D-brain MRI scan and a set of cognitive function test. We explored their rela- tionships with both brain structure and cognitive functions. Images were analyzed to determine the brain volumes and gray matter densities. Patients with MCI or early AD had significantly increased plasma tau levels compared with HE. Plasma tau levels were negatively associated with the perform- ance of logical memory, visual reproduction, and verbal fluency; also negatively associated with vol- ume of total gray matter, hippocampus, amygdala; and gray matter densities of various regions.

Given the promising control of bladder cancer achieved by combined chemotherapy/radiotherapy with selective transurethral resection, obstacles remain to the treatment of unresectable bladder cancer. The aim of this study was to determine whether targeting epidermal growth factor receptor (EGFR)/human epidermal growth factor receptor 2 (HER2) can radiosensitise a murine bladder tumour (MBT-2) cell line. Cell survival, expression of signal proteins and cell cycle changes in MBT-2 cells treated in vitro and in vivo with afatinib, an irreversible EGFR/HER2 inhibitor, plus radiotherapy were investigated by colony formation assay, Western blot assay and flow cytometry, respectively. Ectopic xenografts were established by subcutaneous injection of MBT-2 cells in C3H/HeN mice. Mice were randomised into 4 groups to receive afatinib (10 mg/kg/day on day 1–7) and/or radiotherapy (15 Gy on day 4). Positron emission tomography (PET) on day 8 was used to evaluate the early treatment response. Afatinib (200–1000 nM) increased cell killing by radiation (0–10 Gy). Pre-treatment of irradiated cells with afatinib inhibited radiation-activated HER2 and EGFR phosphorylation. As compared to either treatment alone, the combination increased the level of the cleavage form of poly (ADP-ribose) polymerase, the expression of phospho-γH2AX and the percentage of cells in subG1 phase (indicating enhanced induction of apoptosis), and decreased tumour metabolism and inhibited tumour growth by 64%. Afatinib has therapeutic value as a radiosensitiser of murine bladder cancer cells. The synergism between afatinib and radiation likely enhances DNA damage, leading to increased cell apoptosis.

[[Using this improved one-pot method, the RCY of [This optimized method could fulfill the demand of [

Several strategies to improve the efficacy of radiation therapy against hepatocellular carcinoma (HCC) have been investigated. One approach is to develop radiosensitizing compounds. Because histone deacetylase 4 (HDAC4) is highly expressed in liver cancer and known to regulate oncogenesis through chromatin structure remodeling and controlling protein access to DNA, we postulated that HDAC4 inhibition might enhance radiation's effect on HCC cells. HCC cell lines (Huh7 and PLC5) and an ectopic xenograft were pretreated with HDAC inhibitor or short hairpin RNA to knock down expression of HDAC4 and then irradiated (2.5-10.0 Gy). We evaluated cell survival by a clonogenic assay; apoptosis by Annexin V immunofluorescence; γH2AX, Rad51, and HDAC4 by immunofluorescence staining; HDAC4, Rad51, and ubiquitin-conjugating enzyme 9 (Ubc9) in HCC cell nuclei by cell fractionation and confocal microscopy; physical interaction between HDAC4/Rad51/Ubc9 by immunoprecipitation; and the downstream targets of HDAC4 knockdown by immunoblotting. Both HDAC4 knockdown and HDAC inhibitor enhanced radiation-induced cell death and reduced homologous recombination repair of DNA double-strand breaks and protein kinase B activation, leading to increased apoptosis. HDAC4 knockdown with or without an HDAC inhibitor significantly delayed tumor growth in a radiation-treated xenograft model. Radiation stimulated nuclear translocation of Rad51 in an HDAC4-dependent manner and the binding of Ubc9 directly to HDAC4, which led to Ubc9 acetylation. Moreover, these effects were accompanied by HDAC4/Ubc9/Rad51 complex dissociation through inhibiting nuclear translocation. Conclusion: HDAC4 signaling blockade enhances radiation-induced lethality in HCC cells and xenografts. These findings raise the possibility that HDAC4/Ubc9/Rad51 complex in DNA repair may be a target for radiosensitization of HCC. (Hepatology 2018;67:586-599).

Background/Purpose: LMBD1 protein, a type IV-B plasma membrane protein possessing nine putative trans-membrane domains, was previously demonstrated at cellular level to play a critical part in the signaling cascade of insulin receptor through its involvement in regulating clathrin-mediated endocytosis. However, at physiological level, the significance of LMBD1 protein in cardiac development remains unclear. Methods: To understand the role of Lmbrd1 gene involved in the cardiac function, heterozygous knockout mice were used as an animal model system. The pathological outcomes were analyzed by micro-positron emission tomography, ECG acquisition, cardiac ultrasound, and immunohistochemistry. Results: By studying the heterozygous knockout of Lmbrd1 (Lmbrd1+/−), we discovered that lack of Lmbrd1 not only resulted in the increase of cardiac-glucose uptake, pathological consequences were also observed. Here, we have distinguished that Lmbrd1+/− is sufficient in causing cardiac diseases through a pathway independent of the recessive vitamin B12 cblF cobalamin transport defect. Lmbrd1+/− mice exhibited an increase in myocardial glucose uptake and insulin receptor signaling that is insensitive to the administration of additional insulin. Pathological symptoms such as cardiac hypertrophy, ventricular tissue fibrosis, along with the increase of heart rate and cardiac muscle contractility were observed. As Lmbrd1+/− mice aged, the decrease in ejection fraction and fraction shortening showed signs of ventricular function deterioration. Conclusion: The results suggested that Lmbrd1 gene not only plays a significant role in mediating the energy homeostasis in cardiac tissue, it may also be a key factor in the regulation of cardiac function in mice. Keywords: Cardiac hypertrophy, Disease animal model, Gene knockout, Mouse Lmbrd1 gene, Human LMBRD1 gene, LMBD1 protein

// Hsiu-Hao Chang 1 , Yen-Lin Liu 1, 2 , Meng-Yao Lu 1 , Shiann-Tarng Jou 1 , Yung-Li Yang 1, 3 Dong-Tsamn Lin 1, 3 , Kai-Hsin Lin 1 , Kai-Yuan Tzen 4 , Ruoh-Fang Yen 4 , Ching-Chu Lu 4 , Chia-Ju Liu 4 , Steven Shinn-Forng Peng 5 , Yung-Ming Jeng 6 , Shiu-Feng Huang 7 , Hsinyu Lee 8, 9 , Hsueh-Fen Juan 8, 10 , Min-Chuan Huang 11 , Yung-Feng Liao 12 , Ya-Ling Lee 13, 14 , Wen-Ming Hsu 15 1 Department of Pediatrics, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan 2 Department of Pediatrics, Taipei Medical University Hospital, Taipei, Taiwan 3 Department of Laboratory Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan 4 Department of Nuclear Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan 5 Department of Medical Imaging, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan 6 Department of Pathology, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan 7 Division of Molecular and Genomic Medicine, National Health Research Institutes, Miaoli County, Taiwan 8 Department of Life Science, National Taiwan University, Taipei, Taiwan 9 Institutes of Zoology, National Taiwan University, Taipei, Taiwan 10 Institutes of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan 11 Graduate Institute of Anatomy and Cell Biology, National Taiwan University College of Medicine, Taipei, Taiwan 12 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan 13 Department of Nursing, National Taiwan University College of Medicine, Taipei, Taiwan 14 Department of Nursing, Central Taiwan University of Science and Technology, Taichung, Taiwan 15 Department of Surgery, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan Correspondence to: Ya-Ling Lee, email: yallee@ntu.edu.tw Wen-Ming Hsu, email: billwmhsu@gmail.com Keywords: chemotherapy, high-risk, multidisciplinary team care, neuroblastoma, outcomes Received: June 30, 2016 Accepted: December 01, 2016 Published: December 10, 2016 ABSTRACT We assessed the impact of a multidisciplinary team care program on treatment outcomes in neuroblastoma patients. Newly diagnosed neuroblastoma patients received treatment under the Taiwan Pediatric Oncology Group (TPOG) N2002 protocol at the National Taiwan University Hospital beginning in 2002. A multidisciplinary team care approach that included nurse-led case management for patients treated under this protocol began in January 2010. Fifty-eight neuroblastoma patients, including 29 treated between 2002 and 2009 (Group 1) and 29 treated between 2010 and 2014 (Group 2), were enrolled in the study. The 5-year overall survival (OS) and event-free survival (EFS) rates for all 58 patients were 59% and 54.7%, respectively. Group 2 patients, who were treated after implementation of the multidisciplinary team care program, had better 3-year EFS ( P = 0.046), but not OS ( P = 0.16), rates than Group 1 patients. In a multivariate analysis, implementation of the multidisciplinary team approach was the only significant independent prognostic factor for neuroblastoma patients. In further subgroup analyses, the multidisciplinary team approach improved EFS, but not OS, in patients with stage 4 disease, those in the high-risk group, and those with non-MYCN amplified tumors. These data indicate a multidisciplinary team care approach improved survival outcomes in high-risk neuroblastoma patients. However, further investigation will be required to evaluate the long-term effects of this approach over longer follow-up periods.

18F-Fluorodeoxyglucose (FDG) positron emission tomography (PET)/CT has the potential to track vascular inflammation and monitor therapeutic response. The purpose of this study was to determine the association between arterial inflammation, calcification and serological biomarkers in subjects with atherosclerosis, and to assess their therapeutic response to 12-week atorvastatin treatment. Forty-three statin-naive subjects with atherosclerosis received atorvastatin (40 mg/day) for 12 weeks and underwent 18F-FDG PET/CT, coronary calcification and abdominal adipose tissue volume measurements. A panel of serological biomarkers was analysed. Arterial inflammation was measured at seven arterial segments and normalized to venous FDG activity to produce target to background ratios (TBR). Thirty-four subjects without cardiovascular disease who repeated PET 1–4 years apart for routine health check-ups were retrospectively evaluated for comparison. The baseline mean TBR values in atherosclerotic patients were positively correlated with age (R = 0.36), body mass index (R = 0.54), abdominal visceral adipose tissue volume (R = 0.65), coronary calcification score (R = 0.40), levels of low-density lipoprotein cholesterol (R = 0.54), matrix metalloproteinase (MMP)-9 (R = 0.46) and fatty acid binding protein 4 (FABP4) (R = 0.67, all p

Cardiac allograft vasculopathy (CAV) is the major determinant of long-term survival after heart transplantation. We aimed to evaluate the efficacy of PET as a noninvasive way to assess the early stages of CAV.Twenty-seven consecutive patients (20 men and 7 women; mean age +/- SD, 46 +/- 12 y) who had normal results on coronary angiography and normal left ventricular systolic function (ejection fractionor= 60%) were enrolled at 2.5 +/- 2.1 y after transplantation. Myocardial blood flow (MBF) was assessed using dynamic (13)N-ammonia PET at rest and during adenosine-induced hyperemia, and myocardial perfusion reserve (MPR) was calculated as the ratio of hyperemic MBF to resting MBF. Regional (13)N-ammonia PET was assessed using a 5-point scoring system. The intravascular ultrasound (IVUS) measurements for the extent of intimal hyperplasia, including plaque volume index (calculated as [total plaque volume/total vessel volume] x 100%) and maximum area of stenosis, were compared with MPR by linear regression analysis.In 27 angiographically normal cardiac transplant recipients, MBF at rest and during adenosine stress and MPR of the left anterior descending artery distribution correlated strongly with the other 2 coronary artery distribution territories (ror= 0.97, P0.0001). Summed stress score and summed difference score showed a moderate inverse correlation with MPR (r = -0.41 and -0.49, respectively; P0.05) but not with IVUS measurements. MPR correlated inversely with plaque volume index (r = -0.40, P0.05) but not with maximal luminal stenosis as assessed by IVUS. In addition, MPR and IVUS measurements gradually inversely changed after heart transplantation (all P0.05).This study confirms that CAV is a progressive process, diffusely involving the epicardial and microvascular coronary system. Plaque burden as determined by IVUS agrees well with MPR as assessed by PET in recipients with normal coronary angiography results. This finding suggests that dynamic (13)N-ammonia PET is clinically feasible for the early detection of CAV and can be used as a reliable marker of disease progression.

Our study demonstrated that high resolution animal positron emission tomography (PET) can non-invasively assess the change in glucose metabolism of the central auditory pathway including the inferior colliculus and auditory cortex in the rat.The traditional in vitro approach with immunohistochemical staining or autoradiography to assess chronological changes or topographic arrangement of central auditory pathway required sacrificing a large number of animals. Inter-individual difference is also a major concern. Therefore, development of an in vivo animal model using PET imaging would be a rational method to overcome these shortcomings.Small animal PET scan using (18)F-fluorodeoxyglucose (FDG) as a functional marker was performed in rats. Each animal was serially scanned before and after unilateral cochlear ablation, with and without acoustic stimulation. The images were analyzed by the region of interest (ROI) method. Ratios of radioactivity at the inferior colliculus and auditory cortex and a referenced cerebral cortex between bilateral hemispheres were measured.These scans demonstrated several brain structures including the inferior colliculus (IC) and cortex (B). Moreover, unilateral cochlear ablation decreased the radioactivity at contralateral IC and auditory cortex (C) areas. Differences may reach 33% in IC and 27% in C, and average radioactivity ratios were 1.24+/-0.08 and 1.18+/-0.07 in selected adjacent sections, respectively.

We screened for mutations in the PARKIN, DJ-1, and PINK1 genes in a Taiwanese cohort (68 probands; 58 sporadic and 10 familial) with early-onset parkinsonism (EOP, onset

The GE FASTlab radiosynthesis module is routinely used for the production of [(18)F]FDG, utilizing the commercially available phosphate cassettes. Because of the observation of a white precipitate in the product vial before the product expiry time, we re-examined the quality of the produced [(18)F]FDG solution.Phosphate buffered [(18)F]FDG solution was synthesized on the FASTlab and analyzed at both National Taiwan University Hospital (NTUH) of Taiwan and Royal Brisbane and Women's Hospital (RBWH) of Australia. In addition to the standard product quality control (QC), the concentration of aluminum (Al(3+)) as probable cause of the precipitations in the [(18)F]FDG solution was analyzed by inductively coupled plasma mass spectrometry (ICP-MS at RBWH) and inductively coupled plasma optical emission spectrometry (ICP-OES at NTUH), and using three semi-quantitative methods at NTUH, Advantec(®) Alumi Check Test Strip, Quantofix(®) Aluminum Test Strip and MColortest™ Aluminum Test kit.The precipitates were observed in the [(18)F]FDG solution within 24 (NTUH) and 6 (RBWH) hours after the end of synthesis in 38-100 % of the batches, dependent on the batch of the FASTlab cassettes. Addition of metal-free HCl(aq) to aliquots of [(18)F]FDG containing precipitate, followed by ICP-MS analysis revealed Al(3+) concentrations of 70-80 ppm. Al(3+) concentrations of 10-12 ppm were detected in [(18)F]FDG batches that did not show any precipitation. In contrast, less than 5 ppm of the residual Al(3+) was detected by semi-quantitative methods in all batches.The US (USP), British (BP), European (EP) and Japanese (JP) pharmacopeias demand that [(18)F]FDG for injection should be clear and particulate free within the given shelf-life/expiration time. To avoid Al-phosphate precipitation within the product expiry time, FASTlab citrate cassettes, rather than phosphate cassettes, should be used for [(18)F]FDG production. Although testing for Al(3+) is not listed in the [(18)F]FDG monographs of the USP, BP and EP, residual Al(3+) levels should be considered in the interests of patient safety.

Objectives: We aimed to determine whether epicardial and intramyocardial arteries were involved concordantly in early cardiac allograft vasculopathy. Methods: Thirty consecutive recipients who had received cardiac transplantation more than 1 year before and had a normal coronary angiogram were enrolled for intravascular ultrasound (IVUS), fractional flow reserve, coronary flow reserve and dipyridamole thallium-201 single photon emission computed tomography. Graft failure including cardiac death and retransplantation served as the primary outcome. Results: Plaque volume index and maximum area stenosis calculated from IVUS measurements correlated with coronary flow reserve deterioration in a subgroup of patients with normal fractional flow reserve (n = 13; r = –0.80, p = 0.001 and r = –0.91, p Conclusion: In patients with physiologically normal epicardial coronary arteries, impairment of microvascular integrity correlates with the plaque burden measured by IVUS, suggesting the concordant involvement of both epicardial and resistant vessels in early cardiac allograft vasculopathy. Evident epicardial coronary narrowing on IVUS may predict graft failure in spite of normal coronary angiograms.

Objectives PET/CT with F-fluorodeoxyglucose can be used to image cellular metabolism and has been used for evaluating fever of unknown origin in adults. However, there are limited studies about the role of F-fluorodeoxyglucose PET/CT in evaluation of fever of unknown origin in critically ill children, especially those presenting with complicated underlying diseases under treatment. Here, we report our preliminary experience using F-fluorodeoxyglucose PET/CT in this specific group of patients. Design Retrospective observational study. Setting PICUs of a university hospital. Patients Nineteen critically ill children (mean age, 5.7 yr old) with complicated underlying diseases requiring intensive care support underwent F-fluorodeoxyglucose PET/CT to evaluate fever of unknown origin. The median hospitalized stay was 34 days (range, 15-235 d) and fever of at least 7 days (mean, 21.6 d; range, 7-52 d). The PET scan was advocated after all routine microbiology, and conventional imaging showed negative or inconclusive results. Interventions None. Measurements and main results The F-fluorodeoxyglucose PET/CT findings (blinded to the final clinical diagnosis) were compared with final histopathology, culture, serology results, or follow-up imaging. A final diagnosis was made in 16 patients (84.2%). F-fluorodeoxyglucose PET/CT accurately localized the source of fever in 14 patients, confers to a sensitivity of 87.5% (14 of 16; 95% CI, 0.604-0.978). A false-positive scan in a patient led to subsequent unnecessary investigations. Two false-negative F-fluorodeoxyglucose PET/CT images were later attributed to relapse of underlying disease in the bone marrow and renal abscesses, respectively. In the other two patients where F-fluorodeoxyglucose PET/CT also showed negative findings, fever subsided shortly thereafter without treatment. Conclusions Our preliminary experience suggests that F-fluorodeoxyglucose PET/CT may be clinically beneficial in evaluating fever of unknown origin in children with complicated underlying diseases mandating intensive support in ICUs if usual investigative methods are unsuccessful. Further large prospective studies are needed to validate these findings.

Purpose Neurocognitive performance among patients with carotid artery stenosis or occlusion may deteriorate because of chronic cerebral hypoperfusion. Carotid artery stenting (CAS) has been reported to improve cerebral perfusion and neurocognitive function. The purpose of the study was to evaluate cerebral metabolism using F-fluorodeoxyglucose (FDG) positron emission tomography (PET) after CAS. Methods Nineteen consecutive patients (15 men, 69 ± 13 years) with carotid artery stenosis or occlusion and cerebral ischemia detected on brain perfusion computed tomography (CT) or magnetic resonance imaging (MRI) were enrolled. Four patients had bilateral lesions, and 15 subjects had previous ischemic stroke. Neurocognitive function (NCF) assessments and brain FDG PET scans were performed before and 19 ± 7 (12-31) months after the procedure. Results The procedural success rate of CAS was 70%. Two patients were excluded from the study because of procedural complications. No new cerebral ischemic events or neurologic deaths occurred during follow-up of 44 ± 11 (15-54) months. Significant improvements were observed in the Mini-Mental State Examination (before, 26.06 ± 3.32 versus after, 28.13 ± 2.8; P = 0.0016), the verbal fluency test (26.81 ± 7.82 versus 30.75 ± 9.58; P = 0.0378), and marginal upgrading in the Alzheimer Disease Assessment Scale-Cognitive Subtest (7.19 ± 7.59 versus 5.63 ± 5.90; P = 0.0523). Six of 9 patients who underwent successful CAS showed improvement of cerebral glucose metabolism. Of the 4 patients with recanalization failure, 2 exhibited decline in ipsilateral glucose metabolism. Cerebral FDG metabolism improved in patients with successful CAS (P = 0.038), although there was a weak correlation between interval change of NCF tests and brain FDG metabolism. Conclusions Successful CAS may improve long-term cerebral glucose metabolism and neurocognitive function in patients with chronic severe carotid stenosis or occlusion.

Purpose Myocardial perfusion imaging (MPI) using gated single-photon emission tomography (gSPECT) may underestimate the severity of coronary artery disease (CAD). This study aimed to evaluate the significance of combined parameters derived from gSPECT, as well as treadmill stress test parameters, in the detection of severe CAD. Methods A total of 211 consecutive patients referred for exercise MPI between June 2011 and June 2013 (who received invasive coronary angiography within six months after MPI) were retrospectively reviewed. Exercise MPI was performed with Bruce protocol and 201Tl injected at peak exercise. Gated SPECT was performed using a cadmium-zinc-telluride camera and processed by QPS/QGS software. Perfusion defect abnormalities such as sum stress score (SSS); sum difference score, algorithm-derived total perfusion deficits, transient ischemic dilatation ratios of end-diastolic volumes and end-systolic volumes, post-stress changes in ejection fraction, and lung/heart ratio (LHR) were calculated. Treadmill parameters, including ST depression (STD) at the 1st and 3rd minutes of recovery stage (1’STD and 3’STD), maximal STD corrected by heart rate increment (ST/HR), heart rate decline in 1st and 3rd minutes of recovery stage, recovery heart rate ratio (HR ratio), systolic and mean blood pressure ratios (SBP ratio and MAP ratio) during recovery phase were recorded. Diagnostic performances of these parameters were analyzed with receiver operating characteristic (ROC) analysis and logistic regression for detection of left main (≥ 50%) or 3-vessel disease (all ≥ 70% luminal stenosis) on invasive angiography. Results Among various MPI and treadmill parameters used for detection of severe CAD, SSS and ST/HR had the highest AUC (0.78, 0.73, p = NS) and best cut-off values (SSS > 6, ST/HR > 17.39 10-2mV/bpm), respectively. By univariate logistic regression, all parameters except 1’HRR, 3’HRR, SBP and MAP ratios increased the odds ratio of severe CAD. Only increased L/H ratio, 3’STD, and HR ratio remained significant after multivariate regression. The predicted values of combined MPI and treadmill parameters (LHR, 3’STD, and HR ratio) gave the best ROC (AUC: 0.91) than any individual parameter or parameter combination. Conclusions Of all treadmill and gSPECT parameters, the combination of MPI and treadmill parameters can offer better diagnostic performance for severe CAD.

Aim 9 9 m Tc-HL91 is a new hypoxia agent and can identify acutely ischaemic viable myocardium in a canine model using a standard gamma camera. The purpose of the study was to determine whether this tracer could be used todetect regional ischaemia in chronic ischaemic myocardium in a swine model. Method Using a porcine model of chronic myocardial ischaemia, five mini-pigs with proximal left anterior descending artery (LAD) stenoses were studied. Injection of 462.5 MBq (12.5mCi) 9 9 m Tc-HL91 was followed by imaging over 2 h. Coronary angiography and dipyridamole stress-re-injection 2 0 1 Tl single photon emission computed tomography (SPECT) were performed within 1 day of each other. Results None of the five pigs demonstrated positive hot 9 9 m Tc-HL91 uptake throughout the 2h imaging, whereas four of the five animals showed significant myocardial ischaemia on 2 0 1 Tl SPECT. The region of interest analysis of LAD/left circumflex artery count ratios at 1 and 2 h demonstrated similar 9 9 m Tc-HL91 uptake and retention in chronic ischaemic as well as non-ischaemic myocardium. Conclusion 9 9 m Tc-HL91 is inferior to 2 0 1 Tl in scintigraphic detection of chronic myocardial ischaemia.

a Department of Nuclear Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan b Department of Oncology, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan c Department of Nuclear Medicine and Cardiology Division of Cardiovascular Medical Center, Far Eastern Memorial Hospital, New Taipei City, Taiwan d National Yang-Ming University School of Medicine, Taipei, Taiwan