Analysis of 13 C and 14 C labeling in pyruvate and lactate in tumor and blood of lymphoma-bearing mice injected with 13 C- and 14 C-labeled pyruvate.

Measurements of hyperpolarized 13 C label exchange between injected [1-13 C]pyruvate and the endogenous tumor lactate pool can give an apparent first-order rate constant for the exchange. The determination of the isotope flux, however, requires an estimate of the labeled pyruvate concentration in the tumor. This was achieved here by measurement of the tumor uptake of [1-14 C]pyruvate, which showed that <2% of the injected pyruvate reached the tumor site. Multiplication of this estimated labeled pyruvate concentration in the tumor with the apparent first-order rate constant for hyperpolarized 13 C label exchange gave an isotope flux that showed good agreement with a flux determined directly by the injection of non-polarized [3-13 C]pyruvate, rapid excision of the tumor after 30 s and measurement of 13 C-labeled lactate concentrations in tumor extracts. The distribution of labeled lactate between intra- and extracellular compartments and the blood pool was investigated by imaging, by measurement of the labeled lactate concentration in blood and tumor, and by examination of the effects of a gadolinium contrast agent and a lactate transport inhibitor on the intensity of the hyperpolarized [1-13 C]lactate signal. These measurements showed that there was significant export of labeled lactate from the tumor, but that labeled lactate in the blood pool produced by the injection of hyperpolarized [1-13 C]pyruvate showed only relatively low levels of polarization. This study shows that measurements of hyperpolarized 13 C label exchange between pyruvate and lactate in a murine tumor model can provide an estimate of the true isotope flux if the concentration of labeled pyruvate that reaches the tumor can be determined.

Prospective study evaluating the relative sensitivity of 18F-NaF PET/CT for detecting skeletal metastases from renal cell carcinoma in comparison to multidetector CT and 99mTc-MDP bone scintigraphy, using an adaptive trial design.

BACKGROUND: The detection of occult bone metastases is a key factor in determining the management of patients with renal cell carcinoma (RCC), especially when curative surgery is considered. This prospective study assessed the sensitivity of (18)F-labelled sodium fluoride in conjunction with positron emission tomography/computed tomography ((18)F-NaF PET/CT) for detecting RCC bone metastases, compared with conventional imaging by bone scintigraphy or CT. PATIENTS AND METHODS: An adaptive two-stage trial design was utilized, which was stopped after the first stage due to statistical efficacy. Ten patients with stage IV RCC and bone metastases were imaged with (18)F-NaF PET/CT and (99m)Tc-labelled methylene diphosphonate ((99m)Tc-MDP) bone scintigraphy including pelvic single photon emission computed tomography (SPECT). Images were reported independently by experienced radiologists and nuclear medicine physicians using a 5-point scoring system. RESULTS: Seventy-seven lesions were diagnosed as malignant: 100% were identified by (18)F-NaF PET/CT, 46% by CT and 29% by bone scintigraphy/SPECT. Standard-of-care imaging with CT and bone scintigraphy identified 65% of the metastases reported by (18)F-NaF PET/CT. On an individual patient basis, (18)F-NaF PET/CT detected more RCC metastases than (99m)Tc-MDP bone scintigraphy/SPECT or CT alone (P = 0.007). The metabolic volumes, mean and maximum standardized uptake values (SUV mean and SUV max) of the malignant lesions were significantly greater than those of the benign lesions (P < 0.001). CONCLUSIONS: (18)F-NaF PET/CT is significantly more sensitive at detecting RCC skeletal metastases than conventional bone scintigraphy or CT. The detection of occult bone metastases could greatly alter patient management, particularly in the context when standard-of-care imaging is negative for skeletal metastases.

Detecting treatment response in a model of human breast adenocarcinoma using hyperpolarised [1-13C]pyruvate and [1,4-13C2]fumarate.

BACKGROUND: The recent introduction of a dynamic nuclear polarisation technique has permitted noninvasive imaging of tumour cell metabolism in vivo following intravenous administration of (13)C-labelled cell substrates. METHODS: Changes in hyperpolarised [1-(13)C]pyruvate and [1,4-(13)C(2)]fumarate metabolism were evaluated in both MDA-MB-231 cells and in implanted MDA-MB-231 tumours following doxorubicin treatment. RESULTS: Treatment of MDA-MB-231 cells resulted in the induction of apoptosis, which was accompanied by a decrease in hyperpolarised (13)C label flux between [1-(13)C]pyruvate and lactate, which was correlated with a decrease in the cellular NAD(H) coenzyme pool. There was also an increase in the rate of fumarate conversion to malate, which accompanied the onset of cellular necrosis. In vivo, the decrease in (13)C label exchange between pyruvate and lactate and the increased flux between fumarate and malate, following drug treatment, were shown to occur in the absence of any detectable change in tumour size. CONCLUSION: We show here that the early responses of a human breast adenocarcinoma tumour model to drug treatment can be followed by administration of both hyperpolarised [1-(13)C]pyruvate and [1,4-(13)C(2)]fumarate. These techniques could be used, therefore, in the clinic to detect the early responses of breast tumours to treatment.

An introduction to functional and molecular imaging with MRI.

Magnetic resonance imaging (MRI) has been applied to many aspects of functional and molecular imaging. Many of the parameters used to produce image contrast in MRI are influenced by the local chemical environment around the atoms being imaged; these parameters can be exploited to probe the molecular content of tissues and this has been shown to have many applications in radiology. Diffusion-weighted imaging is a well-established method for measuring small changes in the molecular movement of water that occurs following the onset of ischaemia and in the presence of tumours. Exogenous contrast agents containing gadolinium or iron oxide have been used to image tissue vascularity, cell migration, and specific biological processes, such as cell death. MR spectroscopy is a technique for measuring the concentrations of tissue metabolites and this has been used to probe metabolic pathways in cancer, in cardiac tissue, and in the brain. Several groups are developing positron-emission tomography (PET)-MRI systems that combine the spatial resolution of MRI with the metabolic sensitivity of PET. However, the application of MRI to functional and molecular imaging is limited by its intrinsic low sensitivity. A number of techniques have been developed to overcome this which utilize a phenomenon termed hyperpolarization; these have been used to image tissue pH, cellular necrosis, and to image the lungs. Although most of these applications have been developed in animal models, they are increasingly being translated into human imaging and some are used routinely in many radiology departments.

The Prostate Health Index adds predictive value to multi-parametric MRI in detecting significant prostate cancers in a repeat biopsy population.

Both multi-parametric MRI (mpMRI) and the Prostate Health Index (PHI) have shown promise in predicting a positive biopsy in men with suspected prostate cancer. Here we investigated the value of combining both tests in men requiring a repeat biopsy. PHI scores were measured in men undergoing re-biopsy with an mpMRI image-guided transperineal approach (n = 279, 94 with negative mpMRIs). The PHI was assessed for ability to add value to mpMRI in predicting all or only significant cancers (Gleason ≥7). In this study adding PHI to mpMRI improved overall and significant cancer prediction (AUC 0.71 and 0.75) compared to mpMRI + PSA alone (AUC 0.64 and 0.69 respectively). At a threshold of ≥35, PHI + mpMRI demonstrated a NPV of 0.97 for excluding significant tumours. In mpMRI negative men, the PHI again improved prediction of significant cancers; AUC 0.76 vs 0.63 (mpMRI + PSA). Using a PHI≥35, only 1/21 significant cancers was missed and 31/73 (42%) men potentially spared a re-biopsy (NPV of 0.97, sensitivity 0.95). Decision curve analysis demonstrated clinically relevant utility of the PHI across threshold probabilities of 5-30%. In summary, the PHI adds predictive performance to image-guided detection of clinically significant cancers and has particular value in determining re-biopsy need in men with a negative mpMRI.

A randomised controlled trial comparing two methods of teaching medical students trauma and orthopaedics: traditional lectures versus the "donut round".

OBJECTIVE: To assess whether a new form of teaching, the 'donut round', is as good at imparting factual knowledge as interactive lectures in both the short-term and the long-term. DESIGN: Randomised controlled trial. SETTING: University of Oxford Medical School. PARTICIPANTS: 106 fifth year clinical medical students taught half of their A&E/trauma course by donut round and half by lecture. MAIN OUTCOME MEASURES: The results of multiple choice questions (MCQs) divided according to how the material was taught. Three MCQ papers were set: one at the end of a four-week course, one approximately 10 weeks later and a final exam approximately 17 months after the first. RESULTS: At the first MCQ, the average result for questions taught by donut round was 41.0 (out of 50) and for those taught by conventional lecture was 40.1. At 10 weeks these averages fell to 36.3 and 37.3 and at 17 months they were 38.7 and 38.1, respectively. None of these pairs were significantly different. Ratios were calculated for each candidate by dividing their donut round score by their lecture score. The average ratios for the first, second and third MCQ papers were: 1.029, 1.007 and 1.027, respectively, and were not significantly different. The individual ratios of all candidates in all three MCQs were plotted against their equivalent total mark. The calculated linear regression showed a statistically significant advantage of donut rounds over lectures in those candidates who scored a total mark less than 89 (n=260, p=0.02). CONCLUSIONS: Donut rounds are at least as good as lectures in imparting factual knowledge and may provide a selective advantage to weaker students.

Comparing the accuracy of initial head CT reporting by radiologists, radiology trainees, neuroradiographers and emergency doctors.

OBJECTIVES: Demand for out-of-hours cranial CT imaging is increasing and some departments have considered addressing this shortfall by allowing non-radiologists to provisionally report imaging studies. The aim of this work was to assess whether it is appropriate for non-radiologists to report head CTs by comparing the misreporting rates of those who regularly report head CTs with two groups of non-radiologists who do not usually report them: neuroradiographers and emergency doctors. METHODS: 62 candidates were asked to report 30 head CTs, two-thirds of which were abnormal, and the results were compared by non-parametric statistical analysis. RESULTS: There was no evidence of a difference in the score between neuroradiographers, neuroradiologists and general consultant radiologists. Neuroradiographers scored significantly higher than senior radiology trainees, and the emergency doctors scored least well. CONCLUSION: The results of this preliminary study show that appropriately trained neuroradiographers are competent at reporting the range of abnormalities assessed with this test and that their misreporting rates are similar to those who already independently report these studies.

Postmortem fetal organ volumetry using magnetic resonance imaging and comparison to organ weights at conventional autopsy.

OBJECTIVES: Following perinatal death, organ weights at autopsy may provide evidence of growth restriction and pulmonary hypoplasia. Whilst postmortem magnetic resonance imaging (MRI) may provide comparable information to autopsy about structural abnormalities, its ability to provide reproducible data about organ size has yet to be determined. We examined the feasibility of using postmortem MRI to provide estimates of organ size and weight. METHODS: Twenty-five fetuses of gestational age from 16 to 40 weeks underwent postmortem MRI prior to autopsy. Fetal lung, brain and liver volume estimations were performed by two observers using the stereology technique on postmortem MRI slices. Fetal lung, brain and liver weights were recorded at autopsy. Organ volume estimates and autopsy organ weights were compared using regression analysis, and estimates of fetal organ densities made. Interobserver variability was assessed using a Bland-Altman plot. Receiver-operating characteristics curve (ROC) analysis compared MRI brain : liver volume ratios to autopsy brain : liver weight ratios. RESULTS: A linear relationship between organ volume estimates and organ weight was observed. Estimated densities for the fetal brain, liver and lung were 1.08 g/cm(3), 1.15 g/cm(3) and 1.15 g/cm(3), respectively. Interobserver 5th and 95th percentile limits of agreement for fetal brain, liver and lung were - 5.4% to + 7.9%, - 11.8% to + 8.3% and - 14.3% to + 8.7%, respectively. For MRI organ volumes to detect a brain weight : liver weight ratio > or = 4, ROC analysis demonstrated an area under the curve of 0.61, with an optimal cut-off of 4.1. CONCLUSION: Postmortem MRI organ volumetry can be used to estimate weights of major fetal organs. This may increase the information obtained from a minimally-invasive perinatal autopsy, particularly in the context of pulmonary hypoplasia and intrauterine growth restriction, where differential organ growth plays a major part in assessment of the underlying pathology.

Osmotic 'detubulation' in frog muscle arises from a reversible vacuolation process.

Isolated Rana temporaria sartorius muscle fibres were subject to introduction and subsequent withdrawal of 400 mM extracellular glycerol, exposures to high divalent ion concentrations and then cooling. Tubular detachment was then assessed through changes in the action potential afterdepolarization. (1) The rapid (5-10 min) rather than slow cooling step (30 min) produced a gradual (30 min) development of detubulation arrested by the subsequent replacement of glycerol and reversed by addition of 350 mM sucrose. Such osmotic agents influenced neither resting potentials of intact or detubulated fibres nor action potentials in intact fibres. (2) Full tubular detachment was achieved by 40 min. Laser epifluorescence microscopy demonstrated an accompanying tubular vacuolation through its trapping of a Rhodamine dye. (3) Subsequent re-additions (at 10-80 min) of glycerol restored the afterdepolarization in 30% of detubulated fibres and correspondingly reduced vacuolation. Sustained (> 60 min) exposures to 350 mM sucrose, applied between 30-60 min, both reversed tubular isolation in 70% of detubulated fibres and abolished tubular vacuolation. Finally, results from transient (10-30 min) sucrose exposures resembled the consequences of sustained applications of glycerol, suggesting that detubulation and its reversal result from an osmotic mechanism. (4) Nevertheless, irreversible changes developed after 70-80 min in 70% of detubulated fibres, a process hastened by slow cooling steps in the initial osmotic stress. The present study thus correlates morphological and electrophysiological consequences of applying osmotic shock to skeletal muscle for the first time. It additionally differentiates reversible and irreversible components of detubulation. Finally, it suggests that detubulation results from the similarly reversible vacuolation observed under comparable osmotic conditions, and that such vacuolation can eventually lead to irreversible detubulation.