Enhanced Extraction of Blood and Tissue Time-Activity Curves in Cardiac Mouse FDG PET Imaging by Means of Constrained Nonnegative Matrix Factorization.

We propose an enhanced method to accurately retrieve time-activity curves (TACs) of blood and tissue from dynamic 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) positron emission tomography (PET) cardiac images of mice. The method is noninvasive and consists of using a constrained nonnegative matrix factorization algorithm (CNMF) applied to the matrix (A) containing the intensity values of the voxels of the left ventricle (LV) PET image. CNMF factorizes A into nonnegative matrices H and W, respectively, representing the physiological factors (blood and tissue) and their associated weights, by minimizing an extended cost function. We verified our method on 32 C57BL/6 mice, 14 of them with acute myocardial infarction (AMI). With CNMF, we could break down the mouse LV into myocardial and blood pool images. Their corresponding TACs were used in kinetic modeling to readily determine the [18F]FDG influx constant (Ki) required to compute the myocardial metabolic rate of glucose. The calculated Ki values using CNMF for the heart of control mice were in good agreement with those published in the literature. Significant differences in Ki values for the heart of control and AMI mice were found using CNMF. The values of the elements of W agreed well with the LV structural changes induced by ligation of the left coronary artery. CNMF was compared with the recently published method based on robust unmixing of dynamic sequences using regions of interest (RUDUR). A clear improvement of signal separation was observed with CNMF compared to the RUDUR method.

Chemical synthesis of fluorinated and iodinated 17ß-HSD3 inhibitors and evaluation for imaging prostate cancer tumors and tissue biodistribution.

Prostate cancer is the most common cancer among men and the development of new therapeutic agents is needed for its treatment and/or diagnosis. 17ß-hydroxysteroid dehydrogenase type 3 (17ß-HSD3) is involved in the production of androgens, which stimulates the proliferation of prostate cancer cells. Piperazinomethyl-androsterone sulfonamide derivatives were developed as 17ß-HSD3 inhibitors and the concentration of a representative sulfonamide derivative (compound 1) was found to accumulate in prostate tumor tissues relatively to plasma in a mouse xenograft experiment. This finding gives us the opportunity to specifically target the prostate cancer tumors through the development of a radiolabelled version of compound 1 toward targeted molecular radiotherapy or radioimaging diagnosis. The chemical synthesis of fluorinated and iodinated analogs of compound 1 was achieved, leading to a series of compounds with similar levels of inhibition as the initial candidate. From 17ß-HSD3 inhibition activity, molecular modeling and mouse plasma-concentration studies, the most promising compound of this series was selected, its 18F-radiolabelled version (18F-3) synthesized, and imaging/biodistribution studies engaged. When injected in mice, however, 18F-3 uptake in the target tissues (LNCaP[17ß-HSD3] tumors and testicles) was not sufficient to allow their visualization by positron emission tomography. Plasma concentration values of compounds 3-8 administered orally, however, showed that the para-iodo compound 7 is the most metabolically stable and could therefore be an interesting alternative for radiolabelling and radiotreatment.

Profiles of Recruits Entering Army Basic Training in New Zealand.

INTRODUCTION: A high incidence of musculoskeletal injuries is sustained by army recruits during basic training. Describing recruits' personal, lifestyle, and physical performance characteristics at the entry to training can help identify existing intrinsic risk factors that may predispose some recruits to injury. Identifying modifiable and preventable intrinsic risk factors may contribute to lower recruit injury and associated burdens during the course of basic training. The aim of this study was to therefore describe the profile of New Zealand Army recruits upon entry to basic training using personal, lifestyle, and physical performance characteristics. METHODS: New Zealand Army male and female recruits from two intakes in the same year were invited to participate. Recruits' data on personal (sex, age, height, and weight), lifestyle (self-reported responses to the Military Pre-training Questionnaire comprising physical and injury history, diet, alcohol, and smoking status) and physical performance characteristics (2.4-km timed run, weight-bearing dorsiflexion lunge test, and the Y Balance TestTM for lower limb dynamic stability) were collected and analyzed. RESULTS: Participants included 248 New Zealand Army recruits: 228 males (91.9%), 20 females (8.1%), average age of 20.3 ± 2.8 years. Findings indicated 30.9% of recruits reported injury in the 12 months prior to training commencing, with 44.8% of those injuries in the lower limbs. Pre-entry alcohol consumption was higher than recommended and 20.1% of recruits identified as current smokers. Recruits who passed the 2.4-km timed run included 53.8% of males and 28.6% of females. Weight-bearing dorsiflexion lunge test performance was within a normal range (right = 10.3 ± 3.3 cm), however limb asymmetry (>1.5 cm) was present with 30.9% of recruits. For the Y Balance TestTM for dynamic lower limb stability, 70% of female recruits had high posterolateral reach asymmetry (8.1 ± 6.0 cm), while normalized composite reach scores were low (right) for male (92.2 ± 8.1%) and female recruits (89.0 ± 7.5%). CONCLUSIONS: New Zealand Army recruits entering basic training were predominantly active young males, reported few injuries in the previous year, had higher than recommended alcohol consumption and a minority were smokers. The majority of recruits had low aerobic fitness, average ankle dorsiflexion range, and low dynamic lower limb stability. While a number of adverse characteristics identified are potentially modifiable, more research is required to identify an association to musculoskeletal injury risk in New Zealand Army recruits. Describing the profile of recruits entering training, particularly recruits at risk of injury is one of the first steps in injury prevention.

Cross-Species Physiological Assessment of Brain Estrogen Receptor Expression Using 18F-FES and 18F-4FMFES PET Imaging.

PURPOSE: A retrospective analysis was performed of preclinical and clinical data acquired during the evaluation of the estrogen receptor (ER) PET tracer 4-fluoro-11ß-methoxy-16α-[18F]-fluoroestradiol (4FMFES) and its comparison with 16α-[18F]-fluoroestradiol (FES) in mice, rats, and humans with a focus on the brain uptake. PROCEDURES: Breast cancer tumor-bearing female BALB/c mice from a previous study and female Sprague-Dawley rats (control and ovariectomized) were imaged by 4FMFES or FES-PET imaging. Immediately after, low-dose CT was performed in the same bed position. Semi-quantitative analysis was conducted to extract %ID/g data. Small cohorts of mice and rats were imaged with 4FMFES in an ultra-high-resolution small animal PET scanner prototype (LabPET II). Rat brains were dissected and imaged separately with both PET and autoradiography. In parallel, 31 breast cancer patients were enrolled in a clinical phase II study to compare 4FMFES with FES for oncological assessment. Since the head was included in the field of view, brain uptake of discernable foci was measured and reported as SUVMax. RESULTS: Regardless of the species studied, 4FMFES and FES uptake were relatively uniform in most regions of the brain, except for bilateral foci at the base of the skull, at the midsection of the brain. Anatomical localization of the PET signal using CT image fusion indicates that the signal origins from the pituitary in all studied species. 4FMFES yielded lower pituitary uptake than FES in patients, but an inverse trend was observed in rodents. 4FMFES pituitary contrast was higher than FES in all assessed groups. High-resolution small animal imaging of the brain of rats and mice revealed a supplemental signal anterior to the pituitary, which is likely to be the medial preoptic area. Dissection data further confirmed those findings and revealed additional signals corresponding to the arcuate and ventromedial nuclei, along with the medial and cortical amygdala. CONCLUSION: 4FMFES allowed visualization of ER expression in the pituitary in humans and two different rodent species with better contrast than FES. Improvement in clinical spatial resolution might allow visualization and analysis of other ER-rich brain areas in humans. Further work is now possible to link 4FMFES pituitary uptake to cognitive functions.

Intratumoral 18F-FLT infusion in metabolic targeted radiotherapy.

BACKGROUND: The goal of targeted radiotherapy (TRT) is to administer radionuclides to tumor cells, while limiting radiation exposure to normal tissues. 3'-Deoxy-3'-[18F]-fluorothymidine (18F-FLT) is able to target tumor cells and emits a positron with energy appropriate for local (~ 1 mm range) radiotherapy. In the present work, we investigated the potential of TRT with a local administration of 18F-FLT alone or in combination with 5-fluorouracil (5FU), which acts as a chemotherapeutic agent and radiosensitizer. Treatment efficiency of 18F-FLT combined or not with 5FU was evaluated by intratumoral (i.t.) infusion into subcutaneous HCT116 colorectal tumors implanted in nu/nu mice. The tumor uptake and kinetics of 18F-FLT were determined and compared to 2-deoxy-2-[18F]-fluoro-D-glucose (18F-FDG) by dynamic positron emission tomography (PET) imaging following i.t. injection. The therapeutic responses of 18F-FLT alone and with 5FU were evaluated and compared with 18F-FDG and external beam radiotherapy (EBRT). The level of prostaglandin E2 (PGE2) biosynthesis was measured by liquid chromatography/tandem mass spectrometry (LC/MS/MS) in order to determine the level of inflammation to healthy tissues surrounding the tumor, after i.t. injection of 18F-FLT, and compared to EBRT. RESULTS: We found that i.t. administration of 18F-FLT offers (1) the highest tumor-to-muscle uptake ratio not only in the injected tumor, but also in distant tumors, suggesting potential for concurrent metastases treatment and (2) a sixfold gain in radiotherapeutic efficacy in the primary tumor relative to EBRT, which can be further enhanced with concurrent i.t. administration of the radiosensitizer 5FU. While EBRT stimulated PGE2 production in peritumoral tissues, no significant increase of PGE2 was measured in this area following i.t. administration of 18F-FLT. CONCLUSION: Considering the biochemical stability of 18F-FLT and the physical properties of localized 18F, this study shows that TRT via intratumoral infusion of 18F-FLT and 5FU could provide a new effective treatment option for solid tumors. Using this approach in a colorectal tumor model, the tumor and its metastases could be efficiently irradiated locally with much lower doses absorbed by healthy tissues than with i.t. administration of 18F-FDG or conventional EBRT.

The loss of P2X7 receptor expression leads to increase intestinal glucose transit and hepatic steatosis.

In intestinal epithelial cells (IEC), it was reported that the activation of the P2X7 receptor leads to the internalization of the glucose transporter GLUT2, which is accompanied by a reduction of IEC capacity to transport glucose. In this study, we used P2rx7 -/- mice to decipher P2X7 functions in intestinal glucose transport and to evaluate the impacts on metabolism. Immunohistochemistry analyses revealed the presence of GLUT2 at the apical domain of P2rx7 -/- jejunum enterocytes. Positron emission tomography and biodistribution studies demonstrated that glucose was more efficiently delivered to the circulation of knockout animals. These findings correlated with increase blood glucose, insulin, triglycerides and cholesterol levels. In fact, P2rx7 -/- mice had increased serum triglyceride and cholesterol levels and displayed glucose intolerance and resistance to insulin. Finally, P2rx7 -/- mice developed a hepatic steatosis characterized by a reduction of Acaca, Acacb, Fasn and Acox1 mRNA expression, as well as for ACC and FAS protein expression. Our study suggests that P2X7 could play a central role in metabolic diseases.

A Novel Positron Emission Tomography (PET) Approach to Monitor Cardiac Metabolic Pathway Remodeling in Response to Sunitinib Malate.

Sunitinib is a tyrosine kinase inhibitor approved for the treatment of multiple solid tumors. However, cardiotoxicity is of increasing concern, with a need to develop rational mechanism driven approaches for the early detection of cardiac dysfunction. We sought to interrogate changes in cardiac energy substrate usage during sunitinib treatment, hypothesising that these changes could represent a strategy for the early detection of cardiotoxicity. Balb/CJ mice or Sprague-Dawley rats were treated orally for 4 weeks with 40 or 20 mg/kg/day sunitinib. Cardiac positron emission tomography (PET) was implemented to investigate alterations in myocardial glucose and oxidative metabolism. Following treatment, blood pressure increased, and left ventricular ejection fraction decreased. Cardiac [18F]-fluorodeoxyglucose (FDG)-PET revealed increased glucose uptake after 48 hours. [11C]Acetate-PET showed decreased myocardial perfusion following treatment. Electron microscopy revealed significant lipid accumulation in the myocardium. Proteomic analyses indicated that oxidative metabolism, fatty acid ß-oxidation and mitochondrial dysfunction were among the top myocardial signalling pathways perturbed. Sunitinib treatment results in an increased reliance on glycolysis, increased myocardial lipid deposition and perturbed mitochondrial function, indicative of a fundamental energy crisis resulting in compromised myocardial energy metabolism and function. Our findings suggest that a cardiac PET strategy may represent a rational approach to non-invasively monitor metabolic pathway remodeling following sunitinib treatment.

Transcriptional Changes Associated with Long-Term Left Ventricle Volume Overload in Rats: Impact on Enzymes Related to Myocardial Energy Metabolism.

Patients with left ventricle (LV) volume overload (VO) remain in a compensated state for many years although severe dilation is present. The myocardial capacity to fulfill its energetic demand may delay decompensation. We performed a gene expression profile, a model of chronic VO in rat LV with severe aortic valve regurgitation (AR) for 9 months, and focused on the study of genes associated with myocardial energetics. Methods. LV gene expression profile was performed in rats after 9 months of AR and compared to sham-operated controls. LV glucose and fatty acid (FA) uptake was also evaluated in vivo by positron emission tomography in 8-week AR rats treated or not with fenofibrate, an activator of FA oxidation (FAO). Results. Many LV genes associated with mitochondrial function and metabolism were downregulated in AR rats. FA ß-oxidation capacity was significantly impaired as early as two weeks after AR. Treatment with fenofibrate, a PPARα agonist, normalized both FA and glucose uptake while reducing LV dilation caused by AR. Conclusion. Myocardial energy substrate preference is affected early in the evolution of LV-VO cardiomyopathy. Maintaining a relatively normal FA utilization in the myocardium could translate into less glucose uptake and possibly lesser LV remodeling.

[(18)F]-fluorodeoxyglucose positron emission tomography of the cat brain: A feasibility study to investigate osteoarthritis-associated pain.

The objective of this pilot study was to investigate central nervous system (CNS) changes related to osteoarthritis (OA)-associated chronic pain in cats using [(18)F]-fluorodeoxyglucose ((18)FDG) positron emission tomography (PET) imaging. The brains of five normal, healthy (non-OA) cats and seven cats with pain associated with naturally occurring OA were imaged using (18)FDG-PET during a standardized mild anesthesia protocol. The PET images were co-registered over a magnetic resonance image of a cat brain segmented into several regions of interest. Brain metabolism was assessed in these regions using standardized uptake values. The brain metabolism in the secondary somatosensory cortex, thalamus and periaqueductal gray matter was increased significantly (P ≤ 0.005) in OA cats compared with non-OA cats. This study indicates that (18)FDG-PET brain imaging in cats is feasible to investigate CNS changes related to chronic pain. The results also suggest that OA is associated with sustained nociceptive inputs and increased activity of the descending modulatory pathways.

Endurance training or beta-blockade can partially block the energy metabolism remodeling taking place in experimental chronic left ventricle volume overload.

BACKGROUND: Patients with chronic aortic valve regurgitation (AR) causing left ventricular (LV) volume overload can remain asymptomatic for many years despite having a severely dilated heart. The sudden development of heart failure is not well understood but alterations of myocardial energy metabolism may be contributive. We studied the evolution of LV energy metabolism in experimental AR. METHODS: LV glucose utilization was evaluated in vivo by positron emission tomography (microPET) scanning of 6-month AR rats. Sham-operated or AR rats (n = 10-30 animals/group) were evaluated 3, 6 or 9 months post-surgery. We also tested treatment intervention in order to evaluate their impact on metabolism. AR rats (20 animals) were trained on a treadmill 5 times a week for 9 months and another group of rats received a beta-blockade treatment (carvedilol) for 6 months. RESULTS: MicroPET revealed an abnormal increase in glucose consumption in the LV free wall of AR rats at 6 months. On the other hand, fatty acid beta-oxidation was significantly reduced compared to sham control rats 6 months post AR induction. A significant decrease in citrate synthase and complex 1 activity suggested that mitochondrial oxidative phosphorylation was also affected maybe as soon as 3 months post-AR.Moderate intensity endurance training starting 2 weeks post-AR was able to partially normalize the activity of various myocardial enzymes implicated in energy metabolism. The same was true for the AR rats treated with carvedilol (30 mg/kg/d). Responses to these interventions were different at the level of gene expression. We measured mRNA levels of a number of genes implicated in the transport of energy substrates and we observed that training did not reverse the general down-regulation of these genes in AR rats whereas carvedilol normalized the expression of most of them. CONCLUSION: This study shows that myocardial energy metabolism remodeling taking place in the dilated left ventricle submitted to severe volume overload from AR can be partially avoided by exercise or beta-blockade in rats.

[(11)C]-Acetoacetate PET imaging: a potential early marker for cardiac heart failure.

UNLABELLED: The ketone body acetoacetate could be used as an alternate nutrient for the heart, and it also has the potential to improve cardiac function in an ischemic-reperfusion model or reduce the mitochondrial production of oxidative stress involved in cardiotoxicity. In this study, [(11)C]-acetoacetate was investigated as an early marker of intracellular damage in heart failure. METHODS: A rat cardiotoxicity heart failure model was induced by doxorubicin, Dox(+). [(14)C]-Acetoacetate, a non-positron (ß-) emitting radiotracer, was used to characterize the arterial blood input function and myocardial mitochondrial uptake. Afterward, [(11)C]-acetoacetate (ß+) myocardial PET images were obtained for kinetic analysis and heart function assessment in control Dox(-) (n=15) and treated Dox(+) (n=6) rats. The uptake rate (K1) and myocardial clearance rate (k2or kmono) were extracted. RESULTS: [(14)C]-Acetoacetate in the blood was increased in Dox(+), from 2 min post-injection until the last withdrawal point when the heart was harvested, as well as the uptake in the heart and myocardial mitochondria (unpaired t-test, p <0.05). PET kinetic analysis of [(11)C]-acetoacetate showed that rate constants K1, k2 and kmono were decreased in Dox(+) (p <0.05) combined with a reduction of 24% of the left ventricular ejection fraction (p <0.001). CONCLUSION: Radioactive acetoacetate ex vivo analysis [(14)C], and in vivo kinetic [(11)C] studies provided evidence that [(11)C]-acetoacetate can assess heart failure Dox(+). Contrary to myocardial flow reserve (rest-stress protocol), [(11)C]-acetoacetate can be used to assess reduced kinetic rate constants without requirement of hyperemic stress response. The proposed [(11)C]-acetoacetate cardiac radiotracer in the investigation of heart disease is novel and paves the way to a potential role for [(11)C]-acetoacetate in cardiac pathophysiology.

A randomized controlled trial to assess the effect of self-paced walking on task-specific anxiety in cardiac rehabilitation patients.

PURPOSE: Cardiac rehabilitation can reduce overall anxiety. However, task-specific anxiety is yet to be investigated in the cardiac patient. This study investigates the effect of an outdoor walking intervention (WI) in alleviating the high degree of task-specific anxiety in cardiac patients. METHODS: Participants (N = 22, mean age ± SD = 62.0 ± 10.8 years, 9 women), who had experienced a cardiac event and exhibited a moderate to high level of anxiety for outdoor walking (anxiety score ≥ 7 on a modified version of the Hospital Anxiety and Depression Scale), were initially assessed during an incremental shuttle walk test and a self-paced 1-mile walk. Heart rate and the ratings of perceived exertion were monitored during both tests. Participants also completed an exercise self-efficacy (ESE) questionnaire. Following this, participants were randomized to either a 3 sessions per week, 4 weeks, self-paced WI on a predetermined variable topographic course, or to a control group (CG; 30-minute stationary cycling, 3 sessions per week for 4 weeks). Identical assessments (Incremental Shuttle Walk Test, Self-Paced 1-Mile Walk, Hospital Anxiety and Depression Scale, ESE) were used postintervention. The 2 groups were compared for anxiety, ESE, and fitness by analysis of variance. RESULT: The WI group exhibited a significantly greater decrease in task-specific anxiety (51%; P < .01), increased self-efficacy (6.6%; P < .001) and improved fitness (P < .05) in comparison with CG. CONCLUSION: This study demonstrated that task familiarization reduced the task-specific anxiety associated with outdoor walking in cardiac patients and, as such, may help in changing exercise behavior patterns in patients undergoing cardiac rehabilitation.

Angiotensin II-converting enzyme inhibition improves survival, ventricular remodeling, and myocardial energetics in experimental aortic regurgitation.

BACKGROUND: Aortic valve regurgitation (AR) is a volume-overload disease causing severe eccentric left ventricular (LV) hypertrophy and eventually heart failure. There is currently no approved drug to treat patients with AR. Many vasodilators including angiotensin-converting enzyme inhibitors have been evaluated in clinical trials, but although some results were promising, others were inconclusive. Overall, no drug has yet been able to improve clinical outcome in AR and the controversy remains. We have previously shown in an animal model that captopril (Cpt) reduced LV hypertrophy and protected LV systolic function, but we had not evaluated the clinical outcome. This protocol was designed to evaluate the effects of a long-term Cpt treatment on survival in the same animal model of severe aortic valve regurgitation. METHODS AND RESULTS: Forty Wistar rats with AR were treated or untreated with Cpt (1 g/L in drinking water) for a period of 7 months to evaluate survival, myocardial remodeling, and function by echocardiography as well as myocardial metabolism by µ positron emission tomography scan. Survival was significantly improved in Cpt-treated animals with a survival benefit visible as soon as after 4 months of treatment. Cpt reduced LV dilatation and LV hypertrophy. It also significantly improved the myocardial metabolic profile by restoring the level of fatty acids metabolic enzymes and use. CONCLUSIONS: In a controlled animal model of pure severe aortic valve regurgitation, Cpt treatment reduced LV remodeling and LV hypertrophy and improved myocardial metabolic profile and survival. These results support the need to reevaluate the role of angiotensin-converting enzyme inhibitors in humans with AR in a large, carefully designed prospective clinical trial.

A taxometric analysis of problem gambling data from a South African national urban sample.

We investigate the question whether problem gambling (PG) in a recent South African sample, as measured by the Problem Gambling Severity Index (PGSI), is dimensional or categorical. We use two taxometric procedures, Mean Above Minus Below A Cut (MAMBAC) and Maxim Covariance (MAXCOV), to investigate the taxonic structure of PG as constructed by the PGSI. Data are from the 2010 South African National Urban Prevalence Study of Gambling Behavior. A representative sample of the urban adult population in South Africa (N = 3,000). Responses are to the 9 item PGSI. MAMBAC provided positive but modest evidence that PG as measured by the PGSI was taxonic. MAXCOV pointed more strongly to the same conclusion. These analyses also provide evidence that a PGSI cutoff score of 10 rather than the standard 8 may be called for. PG as constructed by the PGSI may best be thought of as categorical, but further studies with more theory based measurements are needed to determine whether this holds in a wider range of samples and for other screens. A higher cutoff score may be called for on the PGSI when it is used for research purposes to avoid false positives.

A spatiotemporal land-use regression model of winter fine particulate levels in residential neighbourhoods.

Residential wood burning can be a significant wintertime source of ambient fine particles in urban and suburban areas. We developed a statistical model to predict minute (min) levels of particles with median diameter of <1 µm (PM1) from mobile monitoring on evenings of winter weekends at different residential locations in Quebec, Canada, considering wood burning emissions. The 6 s PM1 levels were concurrently measured on 10 preselected routes travelled 3 to 24 times during the winters of 2008-2009 and 2009-2010 by vehicles equipped with a GRIMM or a dataRAM sampler and a Global Positioning System device. Route-specific and global land-use regression (LUR) models were developed using the following spatial and temporal covariates to predict 1-min-averaged PM1 levels: chimney density from property assessment data at sampling locations, PM2.5 "regional background" levels of particles with median diameter of <2.5 µm (PM2.5) and temperature and wind speed at hour of sampling, elevation at sampling locations and day of the week. In the various routes travelled, between 49% and 94% of the variability in PM1 levels was explained by the selected covariates. The effect of chimney density was not negligible in "cottage areas." The R(2) for the global model including all routes was 0.40. This LUR is the first to predict PM1 levels in both space and time with consideration of the effects of wood burning emissions. We show that the influence of chimney density, a proxy for wood burning emissions, varies by regions and that a global model cannot be used to predict PM in regions that were not measured. Future work should consider using both survey data on wood burning intensity and information from numerical air quality forecast models, in LUR models, to improve the generalisation of the prediction of fine particulate levels.

Positron emission tomography detection of human endothelial cell and fibroblast monolayers: effect of pretreament and cell density on 18FDG uptake.

BACKGROUND: The non-destructive assessment and characterization of tridimensional (3D) cell and tissue constructs in bioreactors represents a challenge in tissue engineering. Medical imaging modalities, which can provide information on the structure and function of internal organs and tissues in living organisms, have the potential of allowing repetitive monitoring of these 3D cultures in vitro. Positron emission tomography (PET) is the most sensitive non-invasive imaging modality, capable of measuring picomolar amounts of radiolabeled molecules. However, since PET imaging protocols have been designed almost exclusively for in vivo investigations, suitable methods must be devised to enable imaging of cells or tissue substitutes. As a prior step to imaging 3D cultures, cell radiotracer uptake conditions must first be optimized. METHODS: In this study, human umbilical vein endothelial cells (HUVEC) and human fibroblasts were cultured at different densities and PET was used to non-destructively monitor their glycolytic activity by measuring 18F-fluorodeoxyglucose (18FDG) uptake. Various cell preconditioning protocols were investigated by adjusting the following parameters to optimize 18FDG uptake: glucose starvation, insulin stimulation, glucose concentration, 18FDG incubation time, cell density and radiotracer efflux prevention. RESULTS: The conditions yielding optimal 18FDG uptake, and hence best detection sensitivity by PET, were as follows: 2-hour cell preconditioning by glucose deprivation with 1-hour insulin stimulation, followed by 1-hour 18FDG incubation and 15-minute stabilization in standard culture medium, prior to rinsing and PET scanning. CONCLUSIONS: A step-wise dependence of 18FDG uptake on glucose concentration was found, but a linear correlation between PET signal and cell density was observed. Detection thresholds of 36 ± 7 and 21 ± 4 cells were estimated for endothelial cells and fibroblasts, respectively.

[11C]Acetate rest-stress protocol to assess myocardial perfusion and oxygen consumption reserve in a model of congestive heart failure in rats.

UNLABELLED: This study describes an [(11)C]acetate rest-stress method to obtain an indirect estimate of myocardial blood flow (MBF) and myocardial oxygen consumption (MVO(2)) in rats. Doxorubicin cardiotoxicity was used to test the usefulness of this approach for the assessment of congestive heart failure. METHODS: [(11)C]Acetate rest-stress studies have been used in clinical research to assess the capacity of the coronary arteries to respond to stress. In this article, we used this approach to assess the cardiotoxicity of doxorubicin in a rat model. The method was first validated in a group of healthy rats and then used to follow the effect of doxorubicin chemotherapy on cardiac function. The effect of doxorubicin on myocardial perfusion and oxygen consumption reserve was measured at rest and under dobutamine stimulation. RESULTS: Validation of the protocol showed a good correlation between the MBF and MVO(2) (r(2)=.68). The doxorubicin-treated group showed a significant (P=.04) decrease in cardiovascular perfusion reserve at 1.3±0.2 compared with the control animals at 1.6±0.2. Similar results were obtained for the MVO(2) reserve (treated 1.8±0.4 vs. controls 2.3±0.3; P=.02). CONCLUSIONS: We describe an [(11)C]acetate PET rest-stress protocol for the assessment of congestive heart failure in rats and its application to the follow-up of cardiotoxicity under doxorubicin chemotherapy. This is a rapid and reliable approach to the measurement of cardiac perfusion and oxygen consumption reserve that could be applied to the development of new strategies to reduce the cardiotoxicity of anthracycline.

Image-derived input function in dynamic human PET/CT: methodology and validation with 11C-acetate and 18F-fluorothioheptadecanoic acid in muscle and 18F-fluorodeoxyglucose in brain.

PURPOSE: Despite current advances in PET/CT systems, blood sampling still remains the standard method to obtain the radiotracer input function for tracer kinetic modelling. The purpose of this study was to validate the use of image-derived input functions (IDIF) of the carotid and femoral arteries to measure the arterial input function (AIF) in PET imaging. The data were obtained from two different research studies, one using (18)F-FDG for brain imaging and the other using (11)C-acetate and (18)F-fluoro-6-thioheptadecanoic acid ((18)F-FTHA) in femoral muscles. METHODS: The method was validated with two phantom systems. First, a static phantom consisting of syringes of different diameters containing radioactivity was used to determine the recovery coefficient (RC) and spill-in factors. Second, a dynamic phantom built to model bolus injection and clearance of tracers was used to establish the correlation between blood sampling, AIF and IDIF. The RC was then applied to the femoral artery data from PET imaging studies with (11)C-acetate and (18)F-FTHA and to carotid artery data from brain imaging with (18)F-FDG. These IDIF data were then compared to actual AIFs from patients. RESULTS: With (11)C-acetate, the perfusion index in the femoral muscle was 0.34+/-0.18 min(-1) when estimated from the actual time-activity blood curve, 0.29+/-0.15 min(-1) when estimated from the corrected IDIF, and 0.66+/-0.41 min(-1) when the IDIF data were not corrected for RC. A one-way repeated measures (ANOVA) and Tukey's test showed a statistically significant difference for the IDIF not corrected for RC (p<0.0001). With (18)F-FTHA there was a strong correlation between Patlak slopes, the plasma to tissue transfer rate calculated using the true plasma radioactivity content and the corrected IDIF for the femoral muscles (vastus lateralis r=0.86, p=0.027; biceps femoris r=0.90, p=0.017). On the other hand, there was no correlation between the values derived using the AIF and those derived using the uncorrected IDIF. Finally, in the brain imaging study with (18)F-FDG, the cerebral metabolic rate of glucose (CMRglc) measured using the uncorrected IDIF was consistently overestimated. The CMRglc obtained using blood sampling was 13.1+/-3.9 mg/100 g per minute and 14.0+/-5.7 mg/100 g per minute using the corrected IDIF (r ( 2 )=0.90). CONCLUSION: Correctly obtained, carotid and femoral artery IDIFs can be used as a substitute for AIFs to perform tracer kinetic modelling in skeletal femoral muscles and brain analyses.