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.

68Ga-DOTATATE Prepared from Cyclotron-Produced 68Ga: An Integrated Solution from Cyclotron Vault to Safety Assessment and Diagnostic Efficacy in Neuroendocrine Cancer Patients.

Cyclotron production of 68Ga is a promising approach to supply 68Ga radiopharmaceuticals. To validate this capability, an integrated solution for a robust synthesis of 68Ga-DOTATATE prepared from cyclotron-produced 68Ga was achieved. A retrospective comparison analysis was performed on patients who underwent PET/CT imaging after injection of DOTATATE labeled with 68Ga produced by a cyclotron or eluted from a generator to demonstrate the clinical safety and diagnostic efficacy of the radiopharmaceutical as a routine standard-of-care diagnostic tool in the clinic. Methods: An enriched pressed 68Zn target was irradiated by a cyclotron with a proton beam set at 12.7 MeV for 100 min. The fully automated process uses an in-vault dissolution system in which a liquid distribution system transfers the dissolved target to a dedicated hot cell for the purification of 68GaCl3 and radiolabeling of DOTATATE using a cassette-based automated module. Quality control tests were performed on the resulting tracer solution. The internal radiation dose for 68Ga-DOTATATE was based on extrapolation from rat biodistribution experiments. A retrospective comparison analysis was performed on patients who underwent PET/CT imaging after injection of DOTATATE labeled with cyclotron- or generator-produced 68Ga. Results: The synthesis of 68Ga-DOTATATE (20.7 ± 1.3 GBq) with high apparent molar activity (518 ± 32 GBq/µmol at the end of synthesis) was completed in 65 min, and the radiopharmaceutical met the requirements specified in the European Pharmacopoeia monograph on 68Ga-chloride (accelerator-produced) solution for radiolabeling. 68Ga-DOTATATE was stable for at least 5 h after formulation. The dosimetry calculated with OLINDA for cyclotron- and generator-produced 68Ga-DOTATATE was roughly equivalent. The SUVmean or SUVmax of tumoral lesions with cyclotron-produced 68Ga-DOTATATE was equivalent to that with generator-produced 68Ga. Among physiologic uptake levels, a significant difference was found in kidneys, spleen, and stomach wall, with lower values in cyclotron-produced 68Ga-DOTATATE in all cases. Conclusion: Integrated cyclotron production achieves reliable high yields of clinical-grade 68Ga-DOTATATE. The clinical safety and imaging efficacy of cyclotron-produced 68Ga-DOTATATE in humans provide supporting evidence for its use in routine clinical practice.

Nasal respiratory support and tachypnea and oral feeding in full-term newborn lambs.

Newborn infants with respiratory difficulties frequently require nasal respiratory support such as nasal continuous positive airway pressure (nCPAP) or high-flow nasal cannulae (HFNC). Oral feeding of these infants under nasal respiratory support remains controversial out of fear of aspiration and cardiorespiratory events. The main objective of this study was to evaluate the safety of oral feeding under different types of nasal respiratory support in newborn lambs without or with tachypnea. Eight lambs aged 4-5 days were instrumented to record sucking, swallowing, respiration, ECG, oxygen saturation, and arterial blood gases. Each lamb was given two bottles of 30 mL of milk with a pause of 30 s under videofluoroscopy in four conditions [no respiratory support, nCPAP 6 cmH2O, HFNC 7 L/min, HFNCCPAP (= HFNC 7 L/min + CPAP 6 cmH2O)] administered in random order. The study was conducted in random order over 2 days, with or without standardized tachypnea induced by thoracic compression with a blood pressure cuff. Generalized linear mixed models were used to compare the four nasal respiratory supports in terms of safety (cardiorespiratory events and aspiration), sucking-swallowing-breathing coordination, and efficacy of oral feeding. Results reveal that no nasal respiratory support impaired the safety of oral feeding. Most of the few laryngeal penetrations we observed occurred with HFNCCPAP. Nasal CPAP modified sucking-swallowing-breathing coordination, whereas the efficiency of oral feeding decreased under HFNCCPAP. Results were similar with or without tachypnea. In conclusion, oral feeding under nasal respiratory support is generally safe in a term lamb, even with tachypnea.NEW & NOTEWORTHY The practice of orally feeding newborns suffering from respiratory problems while on nCPAP or HFNC remains controversial for fear of triggering cardiorespiratory events or aspiration pneumonia, or aggravating chronic lung disease. The present results show that bottle-feeding is generally safe in full-term lambs under nasal respiratory support, both without and with tachypnea.

Performance evaluation of the mouse version of the LabPET II PET scanner.

The LabPET II is a new positron emission tomography technology platform designed to achieve submillimetric spatial resolution imaging using fully pixelated avalanche photodiodes-based detectors and highly integrated parallel front-end processing electronics. The detector was designed as a generic building block to develop devices for preclinical imaging of small to mid-sized animals and for clinical imaging of the human brain. The aim of this work is to assess the physical characteristics and imaging performance of the mouse version of LabPET II scanner following the NEMA NU4-2008 standard and using high resolution phantoms and in vivo imaging applications. A reconstructed spatial resolution of 0.78 mm (0.5 µ l) is measured close to the center of the radial field of view. With an energy window of 350 650 keV, the system absolute sensitivity is 1.2% and its maximum noise equivalent count rate reaches 61.1 kcps at 117 MBq. Submillimetric spatial resolution is achieved in a hot spot phantom and tiny bone structures were resolved with unprecedented contrast in the mouse. These results provide convincing evidence of the capabilities of the LabPET II technology for biomolecular imaging in preclinical research.

Automated radiosynthesis of 68Ga for large-scale routine production using 68Zn pressed target.

Gallium-68 (68Ga) has attracted increasing interest in recent years due to the expanding clinical applications of 68Ga-based radiopharmaceuticals (Rahbar et al., 2017). 68Ga is mainly produced via 68Ge/68Ga generators that are limited in yield by the 68Ge activity (typically up to 1.85 GBq at calibration time). With the increased-demand of 68Ga in nuclear medicine for positron emission tomography (PET) imaging, there is a need for more efficient and robust production methods to obtain larger amounts of [68Ga]GaCl3 with high radionuclidic and radiochemical purity and apparent molar activity (AMA) for facilitating the distribution of 68Ga-based radiopharmaceuticals. The objectives of this study were to develop a fast and efficient process for the preparation of 68Zn-based solid targets and to optimize the critical parameters for the automated radiosynthesis of [68Ga]GaCl3 for large-scale routine production.

Publisher Correction: Dichotomic effects of clinically used drugs on tumor growth, bone remodeling and pain management.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Dichotomic effects of clinically used drugs on tumor growth, bone remodeling and pain management.

Improvements in the survival of breast cancer patients have led to the emergence of bone health and pain management as key aspects of patient's quality of life. Here, we used a female rat MRMT-1 model of breast cancer-induced bone pain to compare the effects of three drugs used clinically morphine, nabilone and zoledronate on tumor progression, bone remodeling and pain relief. We found that chronic morphine reduced the mechanical hypersensitivity induced by the proliferation of the luminal B aggressive breast cancer cells in the tumor-bearing femur and prevented spinal neuronal and astrocyte activation. Using MTT cell viability assay and MRI coupled to 18FDG PET imaging followed by ex vivo 3D µCT, we further demonstrated that morphine did not directly exert tumor growth promoting or inhibiting effects on MRMT-1 cancer cells but induced detrimental effects on bone healing by disturbing the balance between bone formation and breakdown. In sharp contrast, both the FDA-approved bisphosphonate zoledronate and the synthetic cannabinoid nabilone prescribed as antiemetics to patients receiving chemotherapy were effective in limiting the osteolytic bone destruction, thus preserving the bone architecture. The protective effect of nabilone on bone metabolism was further accompanied by a direct inhibition of tumor growth. As opposed to zoledronate, nabilone was however not able to manage bone tumor-induced pain and reactive gliosis. Altogether, our results revealed that morphine, nabilone and zoledronate exert disparate effects on tumor growth, bone metabolism and pain control. These findings also support the use of nabilone as an adjuvant therapy for bone metastases.

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.

[(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.

A Longitudinal Low Dose µCT Analysis of Bone Healing in Mice: A Pilot Study.

Low dose microcomputed tomography (µCT) is a recently matured technique that enables the study of longitudinal bone healing and the testing of experimental treatments for bone repair. This imaging technique has been used for studying craniofacial repair in mice but not in an orthopedic context. This is mainly due to the size of the defects (approximately 1.0 mm) in long bone, which heal rapidly and may thus negatively impact the assessment of the effectiveness of experimental treatments. We developed a longitudinal low dose µCT scan analysis method combined with a new image segmentation and extraction software using Hounsfield unit (HU) scores to quantitatively monitor bone healing in small femoral cortical defects in live mice. We were able to reproducibly quantify bone healing longitudinally over time with three observers. We used high speed intramedullary reaming to prolong healing in order to circumvent the rapid healing typical of small defects. Bone healing prolongation combined with µCT imaging to study small bone defects in live mice thus shows potential as a promising tool for future preclinical research on bone healing.

Imaging performance of LabPET APD-based digital PET scanners for pre-clinical research.

The LabPET is an avalanche photodiode (APD) based digital PET scanner with quasi-individual detector read-out and highly parallel electronic architecture for high-performance in vivo molecular imaging of small animals. The scanner is based on LYSO and LGSO scintillation crystals (2×2×12/14 mm3), assembled side-by-side in phoswich pairs read out by an APD. High spatial resolution is achieved through the individual and independent read-out of an individual APD detector for recording impinging annihilation photons. The LabPET exists in three versions, LabPET4 (3.75 cm axial length), LabPET8 (7.5 cm axial length) and LabPET12 (11.4 cm axial length). This paper focuses on the systematic characterization of the three LabPET versions using two different energy window settings to implement a high-efficiency mode (250­650 keV) and a high-resolution mode (350­650 keV) in the most suitable operating conditions. Prior to measurements, a global timing alignment of the scanners and optimization of the APD operating bias have been carried out. Characteristics such as spatial resolution, absolute sensitivity, count rate performance and image quality have been thoroughly investigated following the NEMA NU 4-2008 protocol. Phantom and small animal images were acquired to assess the scanners' suitability for the most demanding imaging tasks in preclinical biomedical research. The three systems achieve the same radial FBP spatial resolution at 5 mm from the field-of-view center: 1.65/3.40 mm (FWHM/FWTM) for an energy threshold of 250 keV and 1.51/2.97 mm for an energy threshold of 350 keV. The absolute sensitivity for an energy window of 250­650 keV is 1.4%/2.6%/4.3% for LabPET4/8/12, respectively. The best count rate performance peaking at 362 kcps is achieved by the LabPET12 with an energy window of 250­650 keV and a mouse phantom (2.5 cm diameter) at an activity of 2.4 MBq ml−1. With the same phantom, the scatter fraction for all scanners is about 17% for an energy threshold of 250 keV and 10% for an energy threshold of 350 keV. The results obtained with two energy window settings confirm the relevance of high-efficiency and high-resolution operating modes to take full advantage of the imaging capabilities of the LabPET scanners for molecular imaging applications.

Mammary cancer bone metastasis follow-up using multimodal small-animal MR and PET imaging.

UNLABELLED: Despite tremendous progress in the management of breast cancer, the survival rate of this disease is still correlated with the development of metastases-most notably, those of the bone. Diagnosis of bone metastasis requires a combination of multiple imaging modalities. MR imaging remains the best modality for soft-tissue visualization, allowing for the distinction between benign and malignant lesions in many cases. On the other hand, PET imaging is frequently more specific at detecting bone metastasis by measuring the accumulation of radiotracers, such as (18)F-sodium fluoride ((18)F-NaF) and (18)F-FDG. Thus, the main purpose of this study was to longitudinally monitor bone tumor progression using PET/MR image coregistration to improve noninvasive imaging-assisted diagnoses. METHODS: After surgical implantation of mammary MRMT-1 cells in a rat femur, we performed minimally invasive imaging procedures at different time points throughout tumor development. The procedure consisted of sequential coregistered MR and PET image acquisition, using gadolinium-diethylenetriaminepentaacetic acid (DTPA) as a contrast agent for MR imaging and (18)F-FDG, (11)C-methionine, and (18)F-NaF as molecular tracers for PET imaging. The animals were then euthanized, and complementary radiologic (micro-CT scans) and histologic analyses were performed. RESULTS: In this preclinical study, we demonstrated that coregistered MR and PET images provide helpful information in a rat mammary-derived bone cancer model. First, MR imaging provided a high-definition anatomic resolution that made the localization of bone resorption and tumor extension detectable between days 9 and 18 after the injection of cancer cells in the medullary channel of the femur. Indeed, the calculation of mean standardized uptake value (SUVmean) and maximal SUV (SUVmax) in bone and soft-tissue regions, as defined from the gadolinium-DTPA contrast-enhanced MR images, showed (18)F-NaF uptake modifications and increased (18)F-FDG or (11)C-methionine uptake in the bone and surrounding soft tissues. (18)F-FDG and (11)C-methionine were compared in terms of the magnitude of change in their uptake and variability. We observed that (11)C-methionine SUVmean variations in the tumor were more important than those of (18)F-FDG. We also found fewer interindividual variations using SUVmean as a quantitative parameter than SUVmax. CONCLUSION: This preclinical evaluation demonstrated that a PET/MR image coregistration protocol provided a powerful tool to evaluate bone tumor progression in a rat model of bone metastasis and that this protocol could be translated to improve the clinical outcome for metastatic breast cancer management.

PET-based geometrical calibration of a pinhole SPECT add-on for an animal PET scanner.

We developed SPECT imaging capability on an animal PET scanner to provide a cost effective option for animal SPECT imaging. The SPECT add-on sub-system was enabled by mechanically integrating a multiple-pinhole collimator in the PET detector ring. This study introduces a method to calibrate the geometrical parameters of the SPECT add-on using the PET imaging capability of the scanner. The proposed PET imaging-based calibration method consists of two steps: (1) paint the pinhole apertures of the collimator with a positron emitting radioactive solution; and (2) image the collimator inside the scanner in PET mode. The geometrical parameters of the multi-pinhole SPECT add-on can then be derived directly from a set of PET images by simple linear calculation and used in defining the SPECT system. The method was compared to our implementation of a SPECT calibration approach with model-based fitting of SPECT projection data. The procedure for carrying out the PET imaging-based calibration method is simpler and faster than that of our implementation of the SPECT model-based calibration method. Since it does not require model fitting, the uniqueness of the calibration result is warranted. Better quality SPECT images were reconstructed using the PET-derived calibration parameters rather than our implementation of the SPECT model-based calibration parameters. We conclude that the proposed PET imaging-based calibration method provides a highly effective means for enabling SPECT imaging on a PET scanner.

A dual tracer PET-MRI protocol for the quantitative measure of regional brain energy substrates uptake in the rat.

We present a method for comparing the uptake of the brain's two key energy substrates: glucose and ketones (acetoacetate [AcAc] in this case) in the rat. The developed method is a small-animal positron emission tomography (PET) protocol, in which (11)C-AcAc and (18)F-fluorodeoxyglucose ((18)F-FDG) are injected sequentially in each animal. This dual tracer PET acquisition is possible because of the short half-life of (11)C (20.4 min). The rats also undergo a magnetic resonance imaging (MRI) acquisition seven days before the PET protocol. Prior to image analysis, PET and MRI images are coregistered to allow the measurement of regional cerebral uptake (cortex, hippocampus, striatum, and cerebellum). A quantitative measure of (11)C-AcAc and (18)F-FDG brain uptake (cerebral metabolic rate; µmol/100 g/min) is determined by kinetic modeling using the image-derived input function (IDIF) method. Our new dual tracer PET protocol is robust and flexible; the two tracers used can be replaced by different radiotracers to evaluate other processes in the brain. Moreover, our protocol is applicable to the study of brain fuel supply in multiple conditions such as normal aging and neurodegenerative pathologies such as Alzheimer's and Parkinson's diseases.

The ketogenic diet increases brain glucose and ketone uptake in aged rats: a dual tracer PET and volumetric MRI study.

Despite decades of study, it is still unclear whether regional brain glucose uptake is lower in the cognitively healthy elderly. Whether regional brain uptake of ketones (ß-hydroxybutyrate and acetoacetate [AcAc]), the main alternative brain fuel to glucose, changes with age is unknown. We used a sequential, dual tracer positron emission tomography (PET) protocol to quantify brain (18)F-fluorodeoxyglucose ((18)F-FDG) and (11)C-AcAc uptake in two studies with healthy, male Sprague-Dawley rats: (i) Aged (21 months; 21M) versus young (4 months; 4M) rats, and (ii) The effect of a 14 day high-fat ketogenic diet (KD) on brain (18)F-FDG and (11)C-AcAc uptake in 24 month old rats (24M). Similar whole brain volumes assessed by magnetic resonance imaging, were observed in aged 21M versus 4M rats, but the lateral ventricles were 30% larger in the 21M rats (p=0.001). Whole brain cerebral metabolic rates of AcAc (CMR(AcAc)) and glucose (CMR(glc)) did not differ between 21M and 4M rats, but were 28% and 44% higher, respectively, in 24M-KD compared to 24M rats. The region-to-whole brain ratio of CMR(glc) was 37-41% lower in the cortex and 40-45% lower in the cerebellum compared to CMR(AcAc) in 4M and 21M rats. We conclude that a quantitative measure of uptake of the brain's two principal exogenous fuels was generally similar in healthy aged and young rats, that the % of distribution across brain regions differed between ketones and glucose, and that brain uptake of both fuels was stimulated by mild, experimental ketonemia.

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.

Behavioral, medical imaging and histopathological features of a new rat model of bone cancer pain.

Pre-clinical bone cancer pain models mimicking the human condition are required to respond to clinical realities. Breast or prostate cancer patients coping with bone metastases experience intractable pain, which affects their quality of life. Advanced monitoring is thus required to clarify bone cancer pain mechanisms and refine treatments. In our model of rat femoral mammary carcinoma MRMT-1 cell implantation, pain onset and tumor growth were monitored for 21 days. The surgical procedure performed without arthrotomy allowed recording of incidental pain in free-moving rats. Along with the gradual development of mechanical allodynia and hyperalgesia, behavioral signs of ambulatory pain were detected at day 14 by using a dynamic weight-bearing apparatus. Osteopenia was revealed from day 14 concomitantly with disorganization of the trabecular architecture (µCT). Bone metastases were visualized as early as day 8 by MRI (T(1)-Gd-DTPA) before pain detection. PET (Na(18)F) co-registration revealed intra-osseous activity, as determined by anatomical superimposition over MRI in accordance with osteoclastic hyperactivity (TRAP staining). Pain and bone destruction were aggravated with time. Bone remodeling was accompanied by c-Fos (spinal) and ATF3 (DRG) neuronal activation, sustained by astrocyte (GFAP) and microglia (Iba1) reactivity in lumbar spinal cord. Our animal model demonstrates the importance of simultaneously recording pain and tumor progression and will allow us to better characterize therapeutic strategies in the future.