bei Marc Cain dabei sind Carolina Vera, Alexandra Neldel und Anna BrГјggemann. Anna Loos, Mirja DuMont, Judith Rakers, Alexandra Neldel, Franziska.bei Marc Cain dabei sind Carolina Vera, Alexandra Neldel und Anna BrГјggemann. Anna Loos, Mirja DuMont, Judith Rakers, Alexandra Neldel, Franziska. Dynamic small animal PET scans were performed for 1h p. The inner vessel diameter of the brachiocephalic trunk was measured and normalized by body weight. When combined with models of peripheral, tumor, blood something the hard way 2019 amusing wash-out bladder distributions, this kinetic imaging data can be transformed into a physiologically based pharmacokinetic model of agent distribution that provides an estimate of the pK values between the various compartments. Furthermore, the analysis of ER oscillation revealed continue reading ovariectomy does not fully disrupt the fluctuation observed in intact, cycling mice indicating the existence learn more here mechanisms other than circulating hormone responsible for ER transcriptional activity. In this communication we report visit web page efficient method for endosomal escape of paramagnetic Gd III chelates speaking, fear the walking dead staffel 2 bs.to apologise yields to a marked improvement of the efficiency in click the following article labeling. Plats: Högstadieskolor runtom i landet. Receptor affinity measurements were anna brГјggemann using radioligand assays and autoradiographic methods. It is therefore not surprising that today Verschmitztes lГ¤cheln has become one of the most important modalities for structural imaging.
Kirchberger, N. Kawakami, J. Dowden, F. Schmid, K. Dornmair, M. Hohenegger, A. Autoaggressive effector T cells in the course of experimental autoimmune encephalomyelitis visualized in the light of two-photon microscopy.
J Neuroimmunol. Instant effect of soluble antigen on effector T cells in peripheral immune organs during immunotherapy of autoimmune encephalomyelitis.
Live imaging of effector cell trafficking and autoantigen recognition within the unfolding autoimmune encephalomyelitis lesion. J Exp Med.
We applied this technique in order to visualize in living Lewis rat the fate of genetically labeled MBP specific effector T cells in the menigeal vessels during the initial phase of experimental autoimmune encephalomyelitis, a classical model of multiple sclerosis.
We observed that the incoming cells remained in close association with pial blood vessels, crawling on surfaces within the outline of the vessels.
This behavior was specific to the CNS: in peripheral organs, for example in peripheral nerves, muscle or subcutaneous tissue, MBP T cells mainly rolled along the inner surface of the vessels.
After diapedesis, the cells continued their scan on the abluminal vascular surface and the underlying leptomeningeal pial membrane. Here they established contact with local meningeal phagocytes and these interactions were crucial to induce the production of proinflammatory and to trigger tissue invasion.
For obvious reasons In is not an ideal nuclear imaging probe label. In addition we extended our clinical studies with a DTPA-modified peptide for higher specific activity labeling with In.
Methods: Internalisation, biodistribution and imaging studies were performed in the Rip1Tag2 mouse model and the corresponding cell line.
Kidney blocking was studied with poly-glutamic acid and Gelofusine. Due to the long residence time in the tumor intraoperative localisation with an endoscopic gamma probe was possible even 2 weeks i.
Conclusions: These very promising pharmacokinetic data show that the two new imaging probes are good candidates for clinical translation: Especially the 99mTc-labeled peptide may increase the availability and distribution of the method whereas the 68Ga derivative may even allow to image and localise very small lesions Acknowledgement: Oncosuisse grant No OSC and grants from the Novartis Foundation and Swiss National Science Foundation are gratefully achnowledged.
Normal organs showed much lower uptake resulting in a high lesion-to-background ratio. Kidney uptake was high and comparable to the tumor uptake.
Tumors with Some of the localisations such as an ectopic insulinoma could imaging life References: 1. Glucagonlike peptide 1-receptor scans to localize occult insulinomas.
N Engl J Med. Here we present the feasibility of using glycosylphosphatidylinositol anchored avidin av-GPI 1 as a novel reporter for multimodal in vivo imaging.
Expressed on the extracelluar side of cell membranes, av-GPI can be targeted with biotinylated imaging probes. Induced by tumor hypoxia to mediate the adaptation of cells to low oxygen tensions these transcription factors play an important role in cancer progression.
Typically, the expression of HIF in human cancer patients is associated with more aggressive tumor phenotypes and poor patient prognosis.
Imaging HIF activity in live tumors hence provides an important tool to study the mechanisms leading to its activation in cancer.
In additional experiments, biotinylated 67Ga-DOTA was shown to specifically label avidin expressing cells in vitro. Conclusions: Overall, we demonstrate the utility of av-GPI as a reporter for in vivo imaging of HIF transcriptional activity in an optical, fluorescence reflectance approach.
In vitro binding studies with 67Ga-DOTA showed a high specificity of the probe in targeting av-GPI in cells, which implies that this reporter can indeed be combined with different imaging modalities.
Results: Fluorescence stainings of av-GPI expressing cells demonstrated that this protein is specifically expressed on the extracellular side of cell membranes.
In vivo fluorescence imaging showed a specific uptake of a biotinylated dye alexabiocytin in the tumor from 60 minutes after contrast injection, whilst there was no accumulation of an unbiotinylated control probe alexacadaverine.
On ex vivo tissue sections alexa biocytin was found to co-localize with References: 1. Pinaud, F. Journal of the American Chemical Society 2.
Wanner, R. Blood 3. Raleigh, J. Although VAP-1 was identified more than 15 years ago, the leukocyte ligand has remained unknown until very recently.
Last year it was shown that Siglec sialic acid-binding immunoglobulinlike lectin expressed on a subpopulation of lymphocytes can bind to VAP-1 and serve as its substrate .
According to phage display screening and structural modeling also Siglec-9 expressed on granulocytes and monocytes interacts with VAP In this study, we investigated a Siglec-9 peptide as a potential imaging tool in positron emission tomography PET.
Results: The Siglec-9 peptide binding to the enzymatic groove of VAP-1 could specifically detect inflammation in rat and tumor in mouse.
Competition experiments with excess of unlabeled Siglec-9 peptide revealed 3-fold lower tumor uptake in mice.
In a rat model, the Inflammation-to-muscle ratio of 68Ga-Siglec-9 peptide was 5. The VAP-1 specificity was further tested with competition assay in mice bearing melanoma xenografts by PET imaging and autoradiography.
In vivo imaging of inflammation was examined in a rat model. All in vivo studies were confirmed by ex vivo measurements.
Conclusions: Our results show that the Siglec-9 peptide detects VAP-1 in inflammation and tumor vasculature in animal models and it may also have potential in imaging of these diseases in patients.
We hypothesized that the combination of invisible near-infrared NIR fluorescent light and ex vivo injection could solve remaining problems in the field.
A total of 32 SLN mappings were performed in pigs, in vivo and ex vivo after oncologic resection, using an identical injection technique.
Guided by these results, SLN mappings were performed in ex vivo tissue specimens of 6 colorectal cancer patients undergoing resection Figure 1.
No false negatives were found. SLNs were identified within 5 minutes after injection of the fluorescent dye. Conclusions: The current study shows proof of principle that ex vivo NIR fluorescence-guided SLN mapping can provide high-sensitivity, rapid, and accurate identification of SLNs in colon and rectum.
This will permit optimized, non-FDA-approved NIR fluorescent lymphatic tracers to be translated immediately to the clinic.
Lucignani G. However, important parameters such as the distribution of the injected cells, cell survival, target organ localisation, cell proliferation and differentiation cannot be evaluated in vivo.
Here we propose multiple labelling protocols for in vivo visualisation by MRI, and Optical fluorescence imaging of murine neural stem cells mNSC originating from the subventricular zone of the adult murine brain in spinal cord injury animal models.
Methods: mNSC were isolated and cultured as described. Labelled cells were injected into the tail vein of a spinal cord injury murine model and followed by MRI for a month to visualize their distribution.
On the contrary, the percentage of iron-positive cells increased in proportion to the PS content in the medium. In both cases, labelled cells were able to give rise to floating neurospheres imaging life as observed by optical microscopy after further 5 days of culture, demonstrating their maintenance of the self-renewal capability.
Immune fluorescence analysis for Nestin confirmed these data. Scintigraphic techniques confirmed initial distribution to lung, spleen and liver while MRI showed iron signal due to stem cell localization into the lesion site at 21 and 28 days after injection.
Neural stem cells, infected with the viral vector PLW, were detected up to one week after i. Infected cells intramedullary injected as control were detected at the same timepoint as well.
Conclusions: These results showed that adult neural stem cells can be efficiently labelled with different molecules without significantly perturbing physiological stem cell features and self-renewal capability.
This labelling protocols can be applied for the in vivo visualisation by MRI, scintigraphy, and Luminescence imaging of the distribution of stem cells after their transplantation into murine model of disease.
Bottai D et al. Mol Med Politi LS et al. De Vocht N. However, a profound knowledge of cell implant migration, differentiation and survival will be necessary to understand the physiological mechanisms involved in regeneration of injured brain tissue.
For this, the development of multimodal cell imaging techniques, both in vivo and post-mortem, is currently of highest importance.
Methods: We optimised a multimodal labelling strategy for bone marrow-derived stromal cells BMSC , based on i genetic modification with both the eGFP and Luciferase reporter genes, and ii endocytic uptake of blue fluorescent magnetite-containing micron-sized particles MPIO.
Conclusions: We here demonstrate that combining three labelling strategies allows unambiguous identification of MSC following implantation in mouse brain.
Dr Gadella personally supervises a research team on spatiotemporal cell signaling. His team wants to understand how cells can achieve and maintain a local signal in order to drive morphogenesis, to define new cytoskeletal anchorage or vesicle-docking sites.
The focus is on signal flow across and in the plane of the membrane of living mammalian cells.
To this end genetic encoded fluorescent biosensors are employed and the in situ molecular interactions between signaling molecules including phospholipid-second messengers, receptors, G-proteins and downstream targets are analyzed.
By multiparameter imaging approaches several signaling events are visualized and quantified simultaneously in individual living cells with sub-second temporal and submicron spatial resolution.
The in situ cellular imaging research heavily depends on advanced bioimaging. To this end advanced automated microscopy approaches such as fluorescence resonance energy transfer FRET microscopy including bleaching-, spectral-, ratio- and fluorescence lifetime-imaging approaches , fast live cell microscopy including spinning disk, line-scanning and controlled light exposure confocal microscopy, and total internal reflection microscopy TIRF , and photochemical microscopy approaches such as photoactivation, uncaging, fluorescence recovery after photobleaching FRAP , fluorescence loss in photobleaching microscopy FLIP , fluorescence correlation spectroscopy FCS , cross-correlation FCCS , lifetime-correlation FCLS have been implemented developed and applied to quantifying cell signalling phenomena.
The Gadella lab has numerous inter national collaborations in the field of signal transduction Dr. Hoffman, Univ. Wymann, Univ. Basle; K.
Jalink, NKI Amsterdam and collaborations on probe development dr. Chudakov, Moscow and collaborations on advanced light microscopy prof.
Houtsmuller, Erasmus MC; dr. Gerritsen, Utrecht; dr. Dobrucki, Univ. Krakow; prof. Carmo Fonseca, Lisbon.
Recently he was appointed as national coordinator of the advanced light microscopy activities of the Netherlands for the new large EU infrastructure ESFRI programme EuroBioimaging that recently entered the startup phase.
In addition, the lifetime screening method also yielded several other CFP lifetime variants with identical spectra.
We apply the different probes for the study of GPCR-triggered signaling in single mammalian cells. In the other direction, the spectroscopical axis in microscopes can also be used to obtain novel GFP variants with optimized properties.
Here we report a screening method that, in addition to fluorescence intensity, quantifies the excited state lifetime of a fluorescent protein, providing a direct measure for the quantum yield of the fluorescent protein.
The novel approach was used to screen a library of cyan fluorescent protein CFP variants yielding the brightest cyan fluorescent protein variant described thus far which we dubbed mTurquoise.
However, there are currently no transgenic animal models that would image specifically one or another neurological disease.
Typically, a transgenic reporter animal highlights the activity of a gene promoter that is specifically active in one or another cell type during a certain physiological process, upon response to injury, along the course of neurodegeneration or during a regenerative response.
For example, currently available models allow the elucidation of several aspects of neurodegeneration: 1 Inflammatory events can be visualized using transgenic animals with the TLR2 promoter controlling the expression of reporter genes; herein, these reporters are up-regulated in response to the onset imaging life of an inflammatory response hand in hand with endogenous TLR2 and microglial activation Lalancette-Hebert, Gowing et al.
This presentation will provide a summary and an overview on the current state of transgenic models in the field of neurological diseases.
In order to specifically target certain neuronal cell populations we have recently developed conditional lentiviral and AAV viral vectors based on Cremediated recombination.
Application of these viral vectors in specific cre-transgenic mouse strains allows to specifically label endogenous neural stem cells in the subventricular zone or dopaminergic cells in the substantia nigra.
This approach holds promise to non-invasively follow up endogenous neurogenesis and neurodegeneration over time. We are using lentiviral vectors and adeno-associated viral AAV vectors for stable overexpression and suppression of gene expression in rodent brain.
Stereotactic injection of these viral vectors into different brain regions of mouse and rat brain induces overexpression or inhibition of disease-related genes, which can be used for target validation, functional genomics and generation of disease models.
We have invested over the last years into reporter gene technology and non-invasive imaging in rodent brain.
Increases in microvessel density MVD have been reported to occur in the peri-infarct zone of stroke patients, which was correlated with increased survival time .
However, noise in the MR images can significantly distort the results of the method. We aim to apply the method to monitor angiogenesis after stroke in rats and to validate this by immunohistochemistry.
Areas with increased Q could be detected close to the infarct in a few of the rats as early as 7 days and in most of the rats at 14 days post MCAO.
Conclusions: Our results show that scanner noise can severely limit the capabilities of SSCE MRI but a set of optimal scan parameters can minimize this problem.
The method has been shown to monitor potential changes of the microvasculature longitudinally and non-invasively in the ischemic rat brain.
It will in the future allow to study neo-angiogenesis in the course of stem cell-based therapies after cerebral lesions. Arrow indicates an area with high Q value.
Corot and P. Robert is gratefully acknowledged. Krupinski et al, Stroke ; 2. Jensen et al. Wu et al. IA administration of AMNP-labeled cells was performed at the time of reperfusion in all tMCAO animals and in 3 shams animals 4 million in 1-ml except for one sham: 1 million in 1-ml.
Animals were sacrificed after completion of the MRI exams and brains were prepared for immunohistological analysis with Prussian blue for iron detection and Ox for macrophage detection.
Results: IA administration in tMCAO group lead to heterogeneous results: 3 rats died following injection, 2 had cells detectable only in the temporalis muscle and 7 had cells detectable in the ipsilateral parenchyma but with heterogeneous patterns: 3 had a widespread persistent hypointense signal distribution, while 4 had only few local spots of signal loss on follow-up scans.
Immunohistological analysis is in progress to ascertain iron-labeled macrophage localization. Conclusions: Our results were consistent with that of previous studies2, 4 showing that IA delivery route efficiently brought a large number of cells to the brain soon after transplantation, but severely increased mortality.
More importantly, lesions observed in the sham group undemonstrated to date to our knowledge suggested that this high mortality rate resulted from cell embolisation in cerebral vessels leading to formation or worsening of the ischemic lesion.
This result points out a serious limitation for the translation of this cell delivery route into the clinics. Surg Neurol.
Li L, et al. J Cereb Blood Flow Metab Brain Res. Walczak P, et al. It is well established that cerebral ischemia results in a complex inflammatory cascade that mainly involves cells from the mononuclear phagocyte system1, although the beneficial or deleterious effect of this activation following stroke is still a matter of controversy.
Furthermore, therapeutic benefits gained from cell-based therapy depend on migration and localization of grafted cells within the target tissue that is closely related to the cell delivery route2 and therapeutic time window3 chosen for the therapy.
The aim of this study was to assess the feasibility of in vivo early intra-arterial IA bone marrow-derived macrophage BMDM administration for acute focal ischemic stroke treatment, using multiparametric magnetic resonance imaging MRI.
Despite numerous clinical trials, tissue plasminogen activator tPA induced thrombolysis remains the only treatment of the acute phase.
However there is growing body of evidences that parenchymal tPA has deleterious effects including increased risk of intra cranial hemorrhages and neurotoxicity.
Accordingly, increasing data of the literature suggest that BBB specific permeability to tPA could be critical for brain outcome following thrombolysis  .
Conclusions: Non invasive imaging of inactivated paramagnetic tPA could allow clinicians to include stroke patients for thrombolysis with objective brain imaging data independently of the time post-stroke onset.
Figure 1. Ex-vivo NIRF brain imaging of fluorescent tPA and albumin administered at 1 or 4 hours after permanent middle cerebral artery occlusion in mice.
Results: Here in a model of permanent middle cerebral artery occlusion in mice, we show that intravenous tPA aggravates ischemic lesion size and BBB permeability if administrated at 4 hours but not at 1 hour after ischemia.
In parallel, although BBB remains impermeable to albumin, we provide evidences both by microscopic epifluorescence and near infrared fluorescence NIRF brain imaging of a passage of tPA across the BBB only when administrated at 4 hours post ischemia.
Benchenane et al. Roussel et al. Therefore, the enzyme should be considered as a candidate target for therapeutic pharmacological intervention.
Methods: High resolution light- and electronmicroscopic immunocytochemistry, in situ hybridization, in situ zymography; use of transgenic animals; antibody microarrays Acknowledgement: The work was supported by Polish-Norwegian Research Grant Results: Our study revealed MMP-9 as a novel synaptic enzyme, and a key pathogenic factor in two distinct animal models of TLE: kainateevoked-epilepsy and pentylenetetrazole PTZ kindling-induced epilepsy.
Moreover, confocal- and immunoelectron-microscopic analyses demonstrated that MMP-9 associated with hippocampal dendritic spines bearing asymmetric excitatory synapses.
In addition, both MMP-9 protein levels as well as its enzymatic activity became strongly increased upon seizures. Furthermore, MMPdeficiency diminished seizure-evoked pruning of dendritic spines, and it decreased aberrant synapse formation following mossy-fibers sprouting.
We then asked whether MMP-9 could play an important role also in human intractable epilepsy. Accordingly, we have studied the expression and localization of MMP-9 in samples of human epileptic brain tissue, obtained upon surgical treatment of childhood intractable epilepsy.
By antibody microarrays, we found MMP-9, but none of the other seven MMPs studied, to be consistently upregulated in epileptic lesions, as compared to autopsy brain tissue.
By immunohistochemistry of epileptic tissue, increased MMP-9 localized specifically to affected neurons. J Cell Biol , Konopacki et al.
Recently, MMP-9, a matrix metalloproteinase, has been implicated in synaptic plasticity, long-term potentiation and learning and memory formation.
We asked, whether MMP-9 might play a pathogenic role in epileptogenesis. The traditional risk assessment relies on clinical, biological and conventional imaging tools.
However, they fall short in predicting near future events particularly in patients with unstable carotid artery disease or coronary artery disease.
The plaque instability is dictated in part by plaque morphology, which in turn is influenced by pathophysiologic mechanisms at the cellular and molecular level.
In current clinical practice, anatomic imaging modalities such as intravascular ultrasound, highresolution magnetic resonance imaging can identify several morphologic features supporting the unstable plaque, but give little or no information regarding molecular and cellular mechanisms.
Methods: We aimed to review 1 the role of relevant biological, factors and advances in atherosclerosis biology, that illuminate key biological aspects of atherosclerosis, including macrophage activity, protease activity, lipoprotein presence, apoptosis, and angiogenesis imaging life 2 imaging agent chemistry chemical biology screens nanotechnology, 3 recent advances in the field of molecular imaging that have led to the development of novel paramagnetic and superparamagnetic targeted contrast agents that bind exclusively to cells such as macrophages, or molecules such as albumin, fibrin, angiogenesis markers Conclusions: A multimodal assessment of plaque instability involving the combination of systemic markers, high resolution imaging and molecular imaging targeting inflammatory and thrombotic components and the potential of new drugs targeting plaque stabilization may lead to a new stratification of the atherothrombotic risk and to a better prevention of atherothrombotic stroke or myocardial infarction.
Conclusions: PSOP could be useful sensors for the in vivo identification of vulnerable plaques in the future. Results: Optical imaging with activatable fluorescent smart probes has become the modality of choice for experimental in vivo detection of protease activity.
Recently we introduced a novel highrelaxivity nanosensors that are suitable for in vivo imaging of protease activity by MRI.
Upon specific protease cleavage, the nanoparticles rapidly switch from a stable low-relaxivity stealth state to become adhesive, aggregating high-relaxivity particles.
Up to date the identification of vulnerable atherosclerotic plaques prone to rupture has not been solved for the clinics.
The pathological processes leading to destabilization of such plaques are highly complex and involve inflammatory action and pathological tissue remodeling of the extracellular matrix performed by proteases.
Therefor the imaging of specific protease activity by high resolution MR imaging could be an important tool to find dangerous plaques that need urgent treatment.
Karczmarczyk U. Such a high cardiovascular mortality is associated with significantly accelerated atherosclerosis in ESRD mainly due to intense inflammatory state.
Therefore serum concentrations of inflammatory agents that promote atherosclerosis are often used to evaluate the cardiovascular risk. In all cases ultrasound examination of the carotid arteries was performed to obtain information about localization and morphology of atherosclerosis plaques and intima-media thickness IMT measurement.
Furthermore, levels of selected proinflammatory factors, atherogenic markers and calcium-phosphate balance parameters were measured.
Conclusions: Scintigraphy with the use of labeled IL-2 can be a tool for inflamed atherosclerotic vulnerable plaque visualization within common carotid arteries in ESRD patients.
Quantitative results of the carotid arteries scintigraphy with labeled IL-2 correlate with serum concentration of selected cardiovascular risk markers Acknowledgement: This work is supported by the Polish Committee for Scientific Research KBN within Research Project 2 P05B 28 References: 1.
Annovazzi A et al; 99mTc-interleukin-2 scintgraphy for the in vivo imaging of vulnerable atheroslerotic plaque.
Radiogallium has been used for decades for in vivo imaging of inflammation. Purpose of this study was to explore the uptake of 68Ga-chloride in atherosclerotic plaques in mice.
Control mice were fed with regular chow. The biodistribution of the tracer was evaluated, and aortic cryosections were further analysed by digital autoradiography.
Subsequently, the autoradiographs were combined with histological and immunohistological analysis of sections .
Results: According to the autoradiography analysis, the radioactivity uptake in atherosclerotic plaques was higher compared to healthy vessel wall ratio 1.
Some 68Ga-radioactivity was also detected in calcified regions of plaques. Autoradiography signal co-localized with macrophages as demonstrated by Mac-3 immunohistochemistry.
In both mice strains, the highest level of radioactivity was found in the urine and blood. Conclusions: We observed a moderate but significantly higher 68Ga-chloride uptake in the aortic plaques of atherosclerotic mice.
While the uptake of 68Ga-chloride was promising in this animal model, the slow blood clearance may limit the usability of 68Ga-chloride in clinical imaging of atherosclerotic plaques.
Iodine based blood pool CT contrast agent and a fluorescent probe for active cathepsins were used. CT and FMT datasets were fused using automated marker detection and registration.
Group 1 consisted of 5 mice with an age of more than 40 weeks fed with normal chow. For statistical testing the Kruskal Wallis test was used.
Results: All mice of group 1 had calcified plaques at the aortic arch and the brachiocephalic trunk. One calcified plaque was found at the bifurcation of the left common carotid artery.
No calcified plaques were found in the other two groups. The inner vessel diameter of the brachiocephalic trunk was measured and normalized by body weight.
In all groups high activity was also found in axillary lymph nodes and some activity was found in lymph nodes along the jugular veins.
Inflammation and remodelling of the vessel wall in the large arteries is reflected by a decrease in relative vessel diameter, which holds true for young mice fed with cholesterol diet and old mice fed with normal chow.
In contrast, cathepsin activity as a measure of macrophage infiltration is capable of indicating the inflammatory stage at early and imaging life References: 1.
Conclusions: Circulation times of the iodinated emulsions as CT contrast agents can be further extended when co-injecting liposomes.
The liposomes are likely taken up by the RES system, thereby slowing down the clearance of the iodinated emulsion in a dose dependant manner.
As liposomes are well tolerated with little toxicity at high doses, the above strategy can be exploited to formulate well tolerated long circulating iodinated contrast agents without increasing the administered iodine dose.
Results: Fig. Previously, we prepared iodinated emulsion-based CT contrast agents that remain in the blood pool for several hours .
The primary blood clearance occurs via uptake in the reticuloendothelial system RES , which is known to be dose dependant.
Here, we tested if circulation times of these iodinated emulsions can possibly be extended by a co-injection with liposomes to slow down RES uptake of the contrast agents.
Interest in the application of microfluidics to PET radiosynthesis has come about primarily because miniaturised reaction systems have the potential to address several key challenges in PET radiochemistry, including increasing the speed, reducing the scale and improving the processing of radiolabelling procedures.
Miller, J. Miller, A. Gee, Curr. On six of the nine experimental days a baseline measurement was followed by a displacement measurement in which fenfluramine 1.
On three of the nine experimental days a baseline measurement was followed by a pretreatment measurement in which fenfluramine 5.
For determination of the fenfluramine effect the binding potential BPND was calculated for different time frames with the cerebellum as reference region.
Results: Administration of fenfluramine had no evident effect on radioactivity in the cerebellum. After administration of fenfluramine 1.
In a classical displacement paradigm after bolus administration of radioligand fenfluramine caused a dose-dependent reduction in specific binding ratios.
The aim of this study was to confirm our previous findings by using a bolus plus continuous infusion approach in monkey.
We will present these results and discuss the similarities and differences regarding the specificity and the sensitivity of the brain uptake.
The most well-known tracer used for this molecular imaging is the [11C]-PIB. Numerous studies with PIB have demonstrated that the properties of this tracer permit the visualization and the quantification of amyloid deposits, but its [11C] labelling limits considerably its potential use due to its short half-life.
A strong effort have been performed by different groups in order to develop a tracer labelled with [18F] for amyloid plaques. Several 18F amyloid binding compounds belonging to different families have been described.
The conjugates were labeled with 64Cu using 0. Receptor affinity measurements were performed using radioligand assays and autoradiographic methods.
The radiolabeled conjugates were evaluated in vitro and in vivo in tumor-bearing nude mice, using the HEK-sst2 cell line. All the radioconjugates showed substantially high and receptor mediated uptake by HEK-sst2 cells.
Biodistribution studies with nude mice showed high uptake of the radioconjugates in HEK-sst2 xenografts at 1h p. The radioactivity in tumor persisted at 4h p.
The study infers that triazacyclononane and cross-bridged cyclam based chelating systems might be ideal for 64Cu labeling of biomolecules intended to be used in targeted PET imaging and radionuclide therapy but PHENDAM may be a new 64Cu chelating system of interest for protein labeling.
A rapid washout in all regions was observed, indicating a high non-specific binding. Halldin C et al; Synapse. Gd-L1A has been delivered to tumor areas in these mice models and signal enhancement monitored over time and compared to that found in non-tumor bearing tissue.
Gd-DO3A has been used as a control. A significant signal enhancement has been found in the tumor area of mice treated with Gd-L1A, with wash-out kinetics of the contrast agent consistent with its internalisation by tumor cells.
Conclusions: These results show that Gd-L1A is suitable to image EPTs, that are in turn sensitive and responsive to the extracellular redox.
Int J Radiat Biol. Severe hypoxia and acidosis has been related to aggressive cell phenotype, poor clinical outcome and low likelihood of patient survival.
Theories assuming that very little renewing potential is identified within the adult CNS have been contravened, new promising sources of myelinogenic cells for transplantation purposes have been characterized, and new cell-replacement strategies have been proposed and established.
A better understanding of the dynamics of endogenous remyelination has been achieved, and insights concerning the process of remyelination driven by site-specific myelin-forming cell transplantation have been discovered.
Somatic stem cells might represent therefore an alternative and promising area of investigation with some potential in its essence.
However, most of the results with stem cells as therapeutic weapons for MS have consistently challenged the sole and limited view that stem cells therapeutically work exclusively throughout cell replacement.
Indeed, the transplantation of somatic non-hematopoietic stem cells promotes substantial CNS repair via a number of bystander mechanisms, mainly exerted by undifferentiated stem cells releasing in vivo a milieu of tissue-trophic and immune modulatory molecules, whose release is likely to be temporally and spatially orchestrated by specific micro environmental cues.
In this view, the therapeutic plasticity of stem cell can be viewed imaging life as the capacity of stem cells to adapt their fate and function s to specific environmental needs occurring as a result of different pathological conditions.
Conclusions: While further studies are certainly required to assess the overall safety, efficacy and in vivo therapeutic plasticity of NPCs, the great challenge for any future human application of NPC-based protocols in MS will be to develop more reliable and reproducible approaches optimizing both tissue trophic as well as immune regulatory capacities of stem cells for functional and anatomical rescuing of myelin architecture in MS patients.
Detailed review of the most recent data on the highly peculiar immune regulatory stem cell signature will be here provided. Aigner L. Like in the in vitro situation, the presence of MSCs in the lesioned spinal cord promoted oligodendroglial and inhibited astroglial differentiation of endogenous progenitors.
In animal models, systemic or local transplantation of MSCs provided functional improvement. The therapeutic effects of MSCs are derived from their immuno-modulatory activity and from the fact that MSCs secrete a number of neuroprotective cytokines.
Moreover, there is evidence that MSCs might be able to influence the pool of endogenous stem or progenitor cells.
Along this line, we have recently demonstrated that MSCs secrete activities that promote oligodendroglial fate and differentiation of neural progenitors Rivera et al.
Dresch C. Current knowledge supports a T cell mediated autoimmune pathogenesis targeting myelin components or myelin-producing cells.
Immunization of susceptible animals with myelin antigens or transfer of myelin antigen-reactive T cells induces experimental autoimmune encephalomyelitis EAE , an inflammatory disorder of the CNS which closely resembles MS.
Because the ethiology of MS is not yet completely understood, there is no curative treatment available at present. After reinfusion, the modified HSC will give rise to all cells of the immune system including antigen expressing dendritic cells.
Results: We demonstrated the effectiveness of this strategy for inducing myelin oligodendrocyte glycoprotein MOG -specific tolerance in an EAE model in mice.
Conclusions: We confirmed the potential of lentivirus vectors that transcriptionally target transgene imaging life expression to dendritic cells for antigen-specific tolerance induction in autoimmune diseases.
We will further investigate the mechanisms of vector mediated tolerance induction, in particular the involvement of regulatory T cells and cytokines.
The strategy presented here is particularly promising for clinical applications. Moreover, these tools will also prove useful for studying disease mechanisms and for addressing fundamental questions in immunity and tolerance.
Acknowledgement: Swiss-MS Society. BMCs were transplanted into a congenic mouse CD The level of engraftment was measured 2 months later by measuring CD LucF expression was evaluated in vivo 6 hours post heating by BLI.
The remaining mice did not exhibit any light emission. A week later lightemitting mice did not produce light anymore.
Conclusions: The bioluminescent chimera mice express the lucF reporter under transcriptional control of a thermosensitive promoter in hematopoietic cells.
This model allows for studying gene delivery to tumor using circulating blood cells. MRgHIFU heating reveals both expected inside and around the tumor and unexpected contralateral leg activation patterns further to be explored.
CMT cells implanted into engrafted mice formed tumors ranging from 5 and 10 mm in one month. In the present study, we investigate an in vivo strategy to deliver a transgene around a tumor and to control the expression of the transgene.
This strategy includes bone marrow engraftment of genetically engineered cells into a wild type mouse to create a chimera with nucleated circulating blood cells CBC expressing the firefly luciferase lucF reporter gene under the transcriptional control of a thermosensitive promoter.
Later on, a subcutaneous tumor is induced and as part of the physiological inflammatory process, CBC accumulate into and around the tumor.
Clerici M. The aim of this study is the development and the evaluation of a tumour-specific DC vaccine, tested on a transgenic murine model of breast cancer MMTV-v-Ha-Ras.
Methods: Total bone marrow cells were extracted from wt mice. Tumour lysates from breast cancer lesions of transgenic mice were used to load immature DCs iDCs , and maturation was monitored by flow cytometry.
Stimulatory activity of Ag-loaded DCs and migratory ability were evaluated in vitro. MNP labelled and Ag loaded DCs were then injected into the footpad of a transgenic tumour bearing mice.
Vitality was not significantly affected by both labelling strategies. MRI evidenced the presence of an hypointense signal in both axillary and popliteal LN, in relation to the injection site, 4h after cell injection.
Signal was visible up to 24 h after DC injection. Ex vivo analysis of different organs showed a statistically significant higher signal within the LN omolateral to the injection site in comparison with the controlateral one.
Conclusions: Labelling protocols do not perturb DC physiology and functionality. Dynamic in vivo MRI and SPET imaging monitoring of DCs distribution will shed light on the fundamental parameters responsible for anti-neoplasic efficacy, while the use of clinically approved MNPs and tracers will speed up the transfer from pre-clinical studies to clinical trials.
Nature Biotech, ,23 11 2. Baumjohann D et al. Immunobiology ; 3. Lucignani G et al. Baekelandt V. BLI scans were performed at 16, 58 and days after surgery.
Time-activity curves TAC and parametric binding potential maps were determined. The animals were sacrificed and perfused and double immunohistochemical staining against CB 2 and eGFP of the brain slices was performed.
Results: The observed CB 2 binding potential increased over time and reached a maximum in right striatum between 18 and 58 days after vector injection in the brain.
The time-activity curves persistently expressed an increased uptake in right striatum compared to control left striatum and cerebellum.
In contrast, only eGFP expression was seen in the contralateral hemisphere. However, for the brain no successful PET reporter systems are available with low endogenous background gene expression and good blood-brain-barrier BBB penetration of the PET probe.
The aim of this study was to develop a new PET reporter gene system which can be applied to the brain. Initial experiments showed unfavourable in vitro characteristics and low labelling yields.
Exploiting our experience in the development of small peptides, i. We have synthesized approximately monomeric and dimeric peptides and evaluated their binding characteristics on Jurkat cells and transfected CMS5 cells stably expressing the hCXCR4 receptor.
We could demonstrate that N-methylated peptides showed an improved IC50 when compared to the nonmethylated analogues.
A variety of linkers were investigated allowing for conjugation of chelators without affecting the affinity of the entire molecule.
A variety of dimers were synthesized, the linkers and linker length optimized and the generated constructs assessed.
C4-C16 dicarbonic acid spacers showed a continuously increasing affinity in the series C4 6. The first of a series of promising 68Ga-labeled peptides is renally excreted leading to high tumor-to-background contrast and high contrasting small animal PET-imaging approx.
Compared to the other CXCR4-probes described, the small cyclic peptides developed offer significant advantages and will allow for broadening state-ofthe-art high contrast peptide receptor imaging to another important class of GPCRs.
In addition, CXCL12 induces recruitment of progenitor cells, which allows for tumor angiogenesis. A high CXCR4 density has also been reported for cancer stem cells.
Consequently, CXCR4 directed therapies are currently under development aiming to inhibit tumor growth and metastasis by blocking CXCL12 binding to CXCR4 using antibodies, large and small peptides as well as small molecules.
However under certain conditions the effects of US for drug delivery last for hours after the exposure. The present work is the first in vitro attempt to confirm and quantitatively assess the temporal window for the US-mediated intracellular drug delivery by live-cell imaging.
Methods: Cell-impermeable optical chromophores with fluorescence intensity increasing fold upon intercalation with nucleic acids served as smart agents for reporting cellular uptake.
Opticell chambers with a monolayer of C6 cells were subjected to ultrasound in the presence of microbubbles followed by varying delays between 0 and 24 hours before addition of Sytox Green optical contrast agent.
Micro- and macroscopic fluorescence imaging was used for qualitative and quantitative analysis. Conclusions: Optical imaging showed that temporal window of increased membrane permeability is much longer than previously suggested.
In the absence of US only occasional cells with the compromised membrane show the uptake of cell-impermeable Sytox green A.
However the effects of the US on membrane permeability can last up to 24h B , with the fluorescence intensity increasing as the time between US application and fluorophore administration shortens C-F.
Results: Strong enhancement of the fluorescence signal upon binding to nucleic acids allowed efficient visualization of the local effect of US on internalization of cell-impermeable intercalating dyes.
Only cells exposed to ultrasound showed the effect. Ultrasound Med Biol. IntegriSenseTM NIR signal was measured in vivo 24 and 48 hours after probe injection and its localization in inflamed aortas was further evaluated by ex-vivo imaging and histology.
Biodistribution studies in mice with xenograft tumors showed a specific uptake of IntegriSenseTM by tumor cells and supported using IntegriSenseTM to monitor tumor growth in animal models.
In vivo measurements with FMT in mice atherosclerosis models detected a longitudinal increase in IntegriSenseTM fluorescence in the thorax area that was three to five fold higher when compared with control animals.
This NIR signal originated from atherosclerotic lesions in the aortic arch, carotid, and subclavian arteries as confirmed by ex vivo imaging of the dissected vessels and histopathology.
For instance, IntegriSenseTM can be used to monitor tumor growth as well as to detect inflammation in atherosclerosis plaques.
The ability of this biomarker to report on several important cellular and physiological functions makes it a tool of choice to evaluate preclinically the therapeutic potential of novel anticancer and atherosclerosis drugs.
In contrast optical molecular imaging allows the noninvasive monitoring and quantification of fluorescent probes in vivo with less expensive equipment that can be installed at multiple research sites.
In order to take full advantage of the rich biology of integrins, a partnership has been established between Merck Research Laboratories and VisEn Medical to develop a novel near-infrared NIR fluorescent integrin probe, IntegriSenseTM.
The pharmacological and biological characterization of IntegriSenseTM, a peptidomimetic antagonist-based molecule with improved specificity and binding affinity will be presented together with research applications in the oncology and atherosclerosis research fields.
The imaging process in animal models requires a reporter construct that leads to production of luciferase enzyme. The most commonly used reporter for this purpose is a construct that can express firefly luciferase 1.
We will also demonstrate that this substrate can be used to non-invasively observe the apoptotic affects of chemotherapy several days before they are able to be detected by traditional methods 5.
Methods: In this study, liver apoptosis was induced in luciferase reporter mice by a single i. No photon emission was observed in organs not affected by the apoptotic treatment.
A complete analysis of pro-apoptotic effects induced by the treatment was carried out by ex vivo imaging acquisition of photon emission in several dissected imaging life tissues.
This investigation confirmed that light emission observed in vivo was produced selectively by liver and adipose tissues.
The present application carried out in reporter mice genetically engineered to develop specific cancers will open the way to novel, more predictive approaches for the study of anti-cancer treatments that will enable researchers to measure drug efficacy in space and time, providing relevant information to be rapidly translated to human therapy.
Drug Discov. Biserni A. CE had a major effect in the genital and hepatic areas shown by a significant increase of ER activity.
As expected the effect of SERMs was significant in the skeletal limbs where ER activity started to be significantly increased only after day In the body areas affected by treatments, we observed time-dependent changes: i.
This observation prompted us to perform a more in-depth analysis of ER activity by measuring photon emission daily. This second analysis demonstrated that ER activity oscillates in time with a period and amplitude that is tissue specific and is differentially affected by the hormonal treatments.
Furthermore, the analysis of ER oscillation revealed that ovariectomy does not fully disrupt the fluctuation observed in intact, cycling mice indicating the existence of mechanisms other than circulating hormone responsible for ER transcriptional activity.
The treatments evaluated partially restored the physiological oscillatory behavior of the receptor.
Methods: The study was carried out in the ERE-Luc reporter mouse model where the luciferase reporter gene is expressed under the transcriptional control of an ER responsive promoter.
In this model, MARs Matrix Attachment Regions insulator sequences flanking the transgene guarantee a ubiquitous and tightly regulated expression of the biosensor making it suitable for pharmacological studies.
To measure ER state of activity, mice were subjected to in vivo imaging. Photon emission was measured by the use of a segmentation algorithm enabling the automatic quantification of photon emission in different body regions of mouse images.
At the end of the study a further analysis was carried out on tissue extracts by luciferase enzymatic assay. Conclusions: Our study: Provides evidence for a novel, long paced, oscillatory activity of ERs that is independent of ovarian function.
This oscillatory ER activity may be relevant for the whole body homeostatic control and may be of relevance for the identification of safer and more efficacious therapies.
Points to the importance of whole animal imaging for drug development and for the study spatio-temporally drug activity on targets.
Novel protocols for a wider application of BLI to the study of molecular events in vivo should be developed. At the whole organism level, circadian rhythms and pulsatility of hormone release may be responsible for maintaining a state of activity of IR that is critical for hormone action.
With hormone therapy, the reinstatement of the receptor oscillatory activity may be a key element for achieving the desired beneficial effect.
To date, the lack of appropriate models to study the response to pharmacological treatments in time has been a limiting factor in the study of spatiotemporal activity of drugs.
The availability of the ERE-Luc reporter mouse model where the application of BLI-based imaging methodologies allows one to study estrogen receptor activity in living animals, opens the way to a more in-depth analysis of the effect of estrogen therapy ET.
The aim of this study was to set up a protocol for the study of ET and SERM Selective Estrogen Receptor Modulator therapies to verify their ability to efficaciously restore the effects of the natural endocrine state in reproductive and non reproductive organs.
After fellowships in heart failure, transplantation, and cardiovascular imaging, he joined the Massachusetts General cardiology faculty.
At Hahnemann, he was the Thomas J. Narula is involved in clinical and basic research in the fields of heart failure and atherosclerosis, with major emphasis on development of novel noninvasive imaging techniques.
He has made vital contributions to the imaging of apoptotic cell death in heart muscle, and to the imaging of atherosclerotic plaques that are vulnerable to rupture.
His research is not limited by any one imaging modality; and uses integrated imaging approaches for better identification of cardiovascular pathology.
His research is funded, in part, by grants from the National Institutes of Health. Narula has authored more than research publications or presentations and edited 25 books and journal supplements.
He serves on various committees of the American Heart Association. After crossing the intimal border, infiltrating monocytes express receptors for chemoattractants such as MCP-1 or adhesion molecules , and subsequently develop scavenger receptors in the subintimal layers for ingestion of oxidized LDL.
Although multiple cellular processes can be targeted for the identification of plaque inflammation only a few molecular imaging strategies have been successfully employed for clinical imaging.
The inflammatory cells in plaques have high metabolic activity. In contrast to granulocytes, which carry a supply of glycogen to provide energy for their phagocytic activities, the macrophages in an atheroma require exogenous glucose for their metabolism.
Incidental uptake of a glucose analog, [18F]-FDG, commonly used to characterize malignant tumors, has been demonstrated in the aorta, carotid arteries and, rarely, the coronary vessels of patients with tumors.
Carotid endarterectomy specimens have demonstrated that uptake correlates with macrophage content, and is reduced in patients treated with statins.
A prospective study has demonstrated that FDG uptake can be seen in the coronary arteries if background FDG uptake in the myocardium is adequately suppressed.
Following coronary interventions, intense FDG uptake has been seen at the sites of stent implantations in patients with acute coronary events; no uptake was observeat the stent sites in patients with stable disease.
Another clinical approach with molecular imaging has utilized radiolabeled annexin A5 to target apoptotic macrophages in the atherosclerotic lesions.
A large proportion of macrophages in the attenuated fibrous caps of vulnerable and ruptured plaques undergo apoptosis.
The annexin uptake in the carotid vessels has been predominantly seen in symptomatic carotid disease. Annexin imaging of coronary vessels has not been attempted.
Various other molecules targeting evolution of receptors such as MCP-1 and scavenger receptors, as well as those targeting the products of inflammation such as metalloproteinases are being successfully employed for the imaging of atherosclerosis in experimental models.
Otherwise asymptomatic individuals with subclinical atherosclerosis almost always have a classic risk-factor profile and it is essential that they are identified before the occurrence of an acute coronary event.
The ability to recognize such individuals requires the development of noninvasive strategies that can localize unstable atherosclerotic lesions.
Plaques that are vulnerable to rupture demonstrate distinct histological characteristics, including large plaque and necrotic core volumes covered by attenuated fibrous caps and extensive remodeling of the vessel at the lesion site.
The morphologic features can be characterized by CT angiography and magnetic resonance imaging. This is mainly thanks to its high temporal and spatial resolution, enabling accurate determination of cardiac functional parameters.
Recently, it has become possible to use MR contrast agents for the evaluation of the mouse myocardial perfusion status. Molecular MR imaging techniques are being developed, allowing contrast agents to specifically target disease markers, providing additional information about the infarction.
The use of contrast agents in the mouse myocardium requires innovative mouse cardiac MRI techniques to quantify contrast agent kinetics and concentration in the myocardium.
Results: In this presentation I will present recent advances in the contrast-enhanced MR imaging of the mouse heart. First, I will discuss new MR technology, which enables the assessment of mouse myocardial perfusion.
This methodology involves intravenous injection of an MRI contrast agent, after which the first pass of the contrast agent through the heart is monitored in a time-series of images.
The combination of the high heart rate bpm , small heart size mm left ventricle LV diameter and fast systemic blood circulation time s require a very fast imaging protocol while preserving adequate spatial resolution to visualize the regional myocardial inflow of the contrast agent.
Secondly, I will discuss recent advances in the T1 quantification of the mouse myocardium, which enables quantification of contrast agent concentration.
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