Tumour expression of leptin is associated with chemotherapy resistance and therapy-independent prognosis in gastro-oesophageal adenocarcinomas.

BACKGROUND: Cytotoxic chemotherapy remains the main systemic therapy for gastro-oesophageal adenocarcinoma, but resistance to chemotherapy is common, resulting in ineffective and often toxic treatment for patients. Predictive biomarkers for chemotherapy response would increase the probability of successful therapy, but none are currently recommended for clinical use. We used global gene expression profiling of tumour biopsies to identify novel predictive biomarkers for cytotoxic chemotherapy. METHODS: Tumour biopsies from patients (n=14) with TNM stage IB-IV gastro-oesophageal adenocarcinomas receiving platinum-based combination chemotherapy were used as a discovery cohort and profiled with Affymetrix ST1.0 Exon Genechips. An independent cohort of patients (n=154) treated with surgery with or without neoadjuvant platinum combination chemotherapy and gastric adenocarcinoma cell lines (n=22) were used for qualification of gene expression profiling results by immunohistochemistry. A cisplatin-resistant gastric cancer cell line, AGS Cis5, and the oesophageal adenocarcinoma cell line, OE33, were used for in vitro validation investigations. RESULTS: We identified 520 genes with differential expression (Mann-Whitney U, P<0.020) between radiological responding and nonresponding patients. Gene enrichment analysis (DAVID v6.7) was used on this list of 520 genes to identify pathways associated with response and identified the adipocytokine signalling pathway, with higher leptin mRNA associated with lack of radiological response (P=0.011). Similarly, in the independent cohort (n=154), higher leptin protein expression by immunohistochemistry in the tumour cells was associated with lack of histopathological response (P=0.007). Higher leptin protein expression by immunohistochemistry was also associated with improved survival in the absence of neoadjuvant chemotherapy, and patients with low leptin protein-expressing tumours had improved survival when treated by neoadjuvant chemotherapy (P for interaction=0.038). In the gastric adenocarcinoma cell lines, higher leptin protein expression was associated with resistance to cisplatin (P=0.008), but not to oxaliplatin (P=0.988) or 5fluorouracil (P=0.636). The leptin receptor antagonist SHLA increased the sensitivity of AGS Cis5 and OE33 cell lines to cisplatin. CONCLUSIONS: In gastro-oesophageal adenocarcinomas, tumour leptin expression is associated with chemoresistance but a better therapy-independent prognosis. Tumour leptin expression determined by immunohistochemistry has potential utility as a predictive marker of resistance to cytotoxic chemotherapy, and a prognostic marker independent of therapy in gastro-oesophageal adenocarcinoma. Leptin antagonists have been developed for clinical use and leptin and its associated pathways may also provide much needed novel therapeutic targets for gastro-oesophageal adenocarcinoma.

Comparison of fluorodeoxyglucose uptake in symptomatic carotid artery and stable femoral artery plaques.

BACKGROUND: Fluorine-18-labelled fluoroxdeoxyglucose (FDG) positron emission tomography (PET) has been used to evaluate atherosclerotic plaque metabolic activity, and through its uptake by macrophages is believed to have the potential to identify vulnerable plaques. The aims were to compare FDG uptake in carotid plaques from patients who had sustained a recent thromboembolic cerebrovascular event with that in femoral artery plaques from patients with leg ischaemia, and to correlate FDG uptake with the proportion of M1 and M2 macrophages present. METHODS: Consecutive patients who had carotid endarterectomy for symptomatic, significant carotid stenosis and patients with severe leg ischaemia and significant stenosis of the common femoral artery underwent FDG-PET and histological plaque analysis. The voxel with the greatest activity in the region of interest was calculated using the Patlak method over 60 min. Plaques were dual-stained for CD68, and M1 and M2 macrophage subsets. RESULTS: There were 29 carotid and 25 femoral artery plaques for study. The maximum dynamic uptake was similar in carotid compared with femoral plaques: median (range) 9·7 (7·1-12·2) versus 10·0 (7·4-16·6) respectively (P = 0·281). CD68 macrophage counts were significantly increased in carotid compared with femoral plaques (39·5 (33·9-50·1) versus 11·5 (7·7-21·3) respectively; P < 0·001), as was the proportion of M1 proinflammatory macrophages. The degree of carotid stenosis correlated with the maximum dynamic FDG uptake (rs = 0·48, P = 0·008). CONCLUSION: FDG uptake was no greater in symptomatic carotid plaques than in the less inflammatory femoral plaques. In patients on statin therapy. FDG uptake occurred in areas of significant arterial stenosis, irrespective of the degree of inflammation.

Patient Preparation and Radiation Protection Guidance for Adult Patients Undergoing Radioiodine Treatment for Thyroid Cancer in the UK.

Radioactive iodine is a highly effective treatment for thyroid cancer and has now been used in clinical practice for more than 80 years. In general, the treatment is well tolerated. However, it can be logistically quite complex for patients due to the need to reduce iodine intake and achieve high levels of thyroid-stimulating hormone prior to treatment. Radiation protection precautions must also be taken to protect others from unnecessary radiation exposure following treatment. It has been well documented by thyroid cancer patient support groups that there is significant variation in practice across the UK. It is clear that some patients are being asked to observe unnecessarily burdensome restrictions that make it more difficult for them to tolerate the treatment. At the instigation of these support groups, a multidisciplinary group was assembled to examine the evidence and generate guidance on best practice for the preparation of patients for this treatment and the management of subsequent radiation protection precautions, with a focus on personalising the advice given to individual patients. The guidance includes advice about managing particularly challenging situations, for example treating patients who require haemodialysis. We have also worked together to produce a patient information leaflet covering these issues. We hope that the guidance document and patient information leaflet will assist centres in improving our patients' experience of receiving radioactive iodine. The patient information sheet is available as Supplementary Material to this article.

Determination of normal regional left ventricular function from cine-MR images using a semi-automated edge detection method.

A semi-automated edge detection method for the delineation of the endo- and epicardial borders of the left ventricle from cine MR images has been developed. The feasibility of this was demonstrated by processing end diastolic and end systolic ECG-gated images of four short axis images in 10 healthy subjects. The first derivative method combined with a 2D weighted polynomial fitting procedure was used to determine the endo- and epicardial borders, which then allowed determination of the wall motion, wall thickening, and ejection fraction, of the left ventricle. The results show that the end-systolic radial wall motion varies from (32+/-8)% to (76+/-12)%, and wall thickening from (0.60+/-0.46) cm to (1.26+/-0.50) cm. An average ejection fraction of (69+/-6)% was found which agrees well with literature values. The method described, for the delineation of the borders, reduces considerably the long and tedious operator time inherent in manual measurement and greatly increases the reproducibility of the measurements.

Application of the principle of optical phase-contrast microscopy to velocity phase-encoded MRI of blood flow in the aorta.

A new method of presenting magnetic resonance phase information is described and an example of its application given in the context of velocity phase-encoded MRI of blood flow in the aorta. The method takes as its starting point Zernike's technique of phase contrast microscopy. It exploits the parallel between the transform plane in Fourier optics and kappa-space in MRI. In the example described two datasets are acquired, one with and the other without velocity encoding, as in conventional phase-encoded velocity imaging. A dataset is formed which is corrected for unwanted phase variations caused by static field inhomogeneity. The method then effectively combines phase and magnitude information into a single image. The technique is complementary to existing methods of displaying phase information.

Automatic detection of coronary artery disease in myocardial perfusion SPECT using image registration and voxel to voxel statistical comparisons.

The purpose of this study was to compare the performance of automatic detection of coronary artery disease (CAD) with that of expert observers. A male and female normal image template was constructed from normal stress technetium-99m single photon emission computed tomography (SPECT) studies. Mean and standard deviation images for each sex were created by registering normal studies to a standard shape and position. The test group consisted of 104 patients who had been routinely referred for SPECT and angiography. The gold standard for CAD was defined by angiography. The test group studies were registered to the respective templates and the Z-score was calculated for each voxel. Voxels with a Z-score greater than 5 indicated the presence of CAD. The performance of this method and that of three observers were compared by continuous receiver operating characteristic (CROC) analysis. The overall sensitivity and specificity for automatic detection were 73% and 92%, respectively. The area (Az) under the CROC curve (+/-1 SE) for automatic detection of CAD was 0.88+/-0.06. There was no statistically significant difference between the performances of the three observers in terms of Az and that of automatic detection (P> or =0.25, univariate Z-score test). The use of this automated statistical mapping approach shows a performance comparable with experienced observers, but avoids inter-observer and intra-observer variability.

Comparison of methods for quantification of transient ischaemic dilation in myocardial perfusion SPET.

The purpose of this study was to compare six methods of measuring the left ventricular (LV) transient ischaemic dilation (TID) ratio during stress-rest myocardial perfusion single-photon emission tomography (SPET). The TID ratio was defined as the mean LV short-axis area at stress divided by the mean LV area of similar slices at rest. The centre of the LV wall was defined as either the maximum, mean or median of the radial short-axis count profiles. The area within the endocardial wall was also calculated for each definition of the LV wall centre. We identified 50 consecutive patients undergoing dipyridamole technetium-99m-tetrofosmin SPET imaging and angiography. Continuous receiver operating characteristic (CROC) analysis showed no significant difference between the six methods in terms of identifying severe coronary artery disease (P >0.47). Algorithms using the mean or the median value in the profile were significantly more robust than those using the maximum (P <0.0005). TID measured by all the algorithms is an indicator of severe coronary disease (P < 0.05). The algorithms compared provide a repeatable, quantitative and specific measure of the TID ratio.

An assessment of perfusion deficits in decompression illness using 99Tcm HMPAO SPET and statistical parametric mapping.

AIM: The aim of this study was to use statistical parametric mapping (SPM) to investigate differences in the regional cerebral blood flow (rCBF) of decompression illness (DCI) and non-diving control subjects. METHODS: Twenty DCI subjects were imaged using 99Tcm hexamethylproplylene amine oxime (HMPAO) single photon emission tomography (SPET) within days of their incident of DCI, and on a second occasion between 2 and 27 months after their DCI event. SPM was used to make a comparison between decompression illness studies and a second group of non-diving control studies for both early and late imaging of DCI. A paired comparison of early and late imaging was also performed testing for correlation between time of imaging and changes in the rCBF pattern. RESULTS: The comparison between control subjects and early imaging of decompression illness showed a large, significant (P = 0.002), irregularly shaped, multi-focal area of reduced uptake in the left occipito-parieto-temporal region of the brain. Comparisons between control and repeat decompression illness imaging studies showed two areas of reduced uptake. The first was similar to that found in the initial comparison although it was less significant (P = 0.035). The second region was similar but contra-lateral to the first region with significance P < 0.001. An attempt to correlate any changes in rCBF with the time that elapsed between each set of imaging found no areas of significant correlation. CONCLUSION: 99Tcm HMPAO SPET images of decompression illness display areas of reduced perfusion, which persist in the majority of cases.

A variable threshold edge-detector for improved quantitation of gated tomographic imaging of the left ventricular blood pool.

The accurate measurement of left ventricular volume from tomographic MUGA studies is difficult due to the limited resolving power of the gamma camera, which causes errors in the detection of the correct ventricular boundaries. Therefore, the use of fixed threshold or second-derivative edge-detectors results in overestimates at small volumes. A variable threshold edge-detection technique was developed to overcome this. Computer-simulated short-axis slices through the heart over a range of left ventricular dimensions were convolved by the Point Spread Response Function of the system to model the acquired image. The maximum pixel value and the threshold value required to detect the true ventricular edge from each simulation were then combined into a look-up table for the calculation of the required threshold value. As the dimension of the ventricle decreased, the threshold value chosen to detect the ventricular edge increased. Left ventricular volumes and ejection fraction measurements were calculated for seven patients using cine-MRI as the gold-standard technique for validation of the proposed method. The single photon emission tomographic studies were analysed using both the standard second-derivative edge-detection software and the proposed variable threshold technique. The variable threshold technique was shown to increase significantly the accuracy of ventricular volume measurements and ejection fraction calculations. The average error in the measurement of volumes was reduced from 41.4 +/- 45.1% to 18.5 +/- 14.6% and the accuracy of ejection fraction measurement was increased from 29.7 +/- 4.6% to 11.3 +/- 6.9%.

Automatic classification of DMSA scans using an artificial neural network.

DMSA imaging is carried out in nuclear medicine to assess the level of functional renal tissue in patients. This study investigated the use of an artificial neural network to perform diagnostic classification of these scans. Using the radiological report as the gold standard, the network was trained to classify DMSA scans as positive or negative for defects using a representative sample of 257 previously reported images. The trained network was then independently tested using a further 193 scans and achieved a binary classification accuracy of 95.9%. The performance of the network was compared with three qualified expert observers who were asked to grade each scan in the 193 image testing set on a six point defect scale, from 'definitely normal' to 'definitely abnormal'. A receiver operating characteristic analysis comparison between a consensus operator, generated from the scores of the three expert observers, and the network revealed a statistically significant increase (α < 0.05) in performance between the network and operators. A further result from this work was that when suitably optimized, a negative predictive value of 100% for renal defects was achieved by the network, while still managing to identify 93% of the negative cases in the dataset. These results are encouraging for application of such a network as a screening tool or quality assurance assistant in clinical practice.

EANM/ESC guidelines for radionuclide imaging of cardiac function.

Radionuclide imaging of cardiac function represents a number of well-validated techniques for accurate determination of right (RV) and left ventricular (LV) ejection fraction (EF) and LV volumes. These first European guidelines give recommendations for how and when to use first-pass and equilibrium radionuclide ventriculography, gated myocardial perfusion scintigraphy, gated PET, and studies with non-imaging devices for the evaluation of cardiac function. The items covered are presented in 11 sections: clinical indications, radiopharmaceuticals and dosimetry, study acquisition, RV EF, LV EF, LV volumes, LV regional function, LV diastolic function, reports and image display and reference values from the literature of RVEF, LVEF and LV volumes. If specific recommendations given cannot be based on evidence from original, scientific studies, referral is given to "prevailing or general consensus". The guidelines are designed to assist in the practice of referral to, performance, interpretation and reporting of nuclear cardiology studies for the evaluation of cardiac performance.

EANM/ESC procedural guidelines for myocardial perfusion imaging in nuclear cardiology.

The European procedural guidelines for radionuclide imaging of myocardial perfusion and viability are presented in 13 sections covering patient information, radiopharmaceuticals, injected activities and dosimetry, stress tests, imaging protocols and acquisition, quality control and reconstruction methods, gated studies and attenuation-scatter compensation, data analysis, reports and image display, and positron emission tomography. If the specific recommendations given could not be based on evidence from original, scientific studies, we tried to express this state-of-art. The guidelines are designed to assist in the practice of performing, interpreting and reporting myocardial perfusion SPET. The guidelines do not discuss clinical indications, benefits or drawbacks of radionuclide myocardial imaging compared to non-nuclear techniques, nor do they cover cost benefit or cost effectiveness.

Comparison of four magnetization preparation schemes to improve blood-wall contrast in cine short-axis cardiac imaging.

Improvements in short-axis blood-myocardium contrast in the heart with the use of four magnetization preparation schemes applied before the imaging sequence are demonstrated. Gradient-echo cine cardiac images are acquired and compared at 0.95 T incorporating T2, T1rho, magnetization transfer, and double inversion (black blood) preparations in a series of volunteer studies over the first 550 ms of the cardiac cycle. T2 and T1rho preparations exhibit improvements of 100% and above in image contrast. Magnetization transfer preparation exhibits improvements of 50% in image contrast, whereas an initial improvement (50%) followed by a large loss in contrast is observed using the black blood preparation. Improvements in contrast are dependent on tissue relaxation parameters and therefore are suitable for studies involving patients exhibiting poor in-flow enhancement of blood caused by poor heart function.

Dobutamine magnetic resonance imaging as a predictor of myocardial functional recovery after revascularisation.

OBJECTIVE: To assess the use of dobutamine magnetic resonance imaging (MRI) as a preoperative predictor of myocardial functional recovery after revascularisation, comparing wall motion and radial wall thickening analyses by observer and semi-automated edge detection. PATIENTS: 25 men with multivessel coronary disease and resting wall motion abnormalities were studied with preoperative rest and stress MRI. MAIN OUTCOME MEASURES: Observer analysis for radial wall thickening was compared with a normal range, while wall motion analysis used a standard four point scale. Semi-automated analysis was performed using an edge detection algorithm. Segments displaying either improved or worsened thickening or motion with dobutamine were considered viable. Postoperative rest images were performed 3-6 months after coronary artery bypass grafting (CABG) for comparison. RESULTS: For observer analysis the values for sensitivity and specificity were 50% and 72% for wall motion, with respective values of 50% and 68% for thickening. With semi-automated edge detection the figures for motion were 60% and 73%, with corresponding values of 79% and 58% for thickening. Combining thickening and motion for the semi-automated method to describe any change in segmental function yielded a sensitivity of 71% and specificity of 70%. CONCLUSIONS: Dobutamine MRI is a reasonably good predictor of myocardial functional recovery after CABG. The use of semi-automated edge detection analysis improved results.