Cette recherche a été financée par la National Science Foundation Grant n ° 0651981 et le Alexander et Margaret Stewart confiance que le Cancer Center de l&#39;Université Johns Hopkins. Les auteurs tiennent à souligner la contribution du Dr Rhoda Alani à la CISR et l&#39;étude du patient ainsi que l&#39;aide et le soutien du Dr Kang Sewon et son ministère au cours de l&#39;étude du patient.

1. Installation <ol><li> Une salle d&#39;examen température contrôlée équipé d&#39;une caméra infrarouge et un ordinateur pour l&#39;acquisition d&#39;images infrarouges et de stockage ainsi que d&#39;une carte d&#39;acquisition de données connecté à un ordinateur sont présentés dans Fig.1a. </li><li> La température ambiante et la température de surface de la peau sont contrôlées par les thermocouples fixés à une carte d&#39;acquisition de données au cours de l&#39;examen d&#39;un patient et les données de mesure sont enregistrées sur l&#39;ordinateur. </li></ol> 2. Image Acquisition <ol><li> Depuis la lésion ne peut être détecté dans l&#39;image thermique sans l&#39;effet de refroidissement, un marqueur adhésif carré est utilisé pour localiser la lésion pigmentée d&#39;intérêt et ses environs (Fig. 1b). </li><li> Nous acquérons une image vive lumière de la lésion pigmentée et la fenêtre adhésive avec un appareil photo numérique (Canon PowerShot G11) (Fig. 1b). </li><li> Un dermatoscope connecté à un appareil photo numérique (DermLite Foto System) est utilisée pour capturer l&#39;image de la lumière polarisée. </li><li&gt; Nous acquérons une image infrarouge en régime permanent avec un midwave Merlin (3-5 um) caméra infrarouge montre la Figure 1a, c. </li><li> On applique un courant d&#39;air froid à la surface de la peau du patient contenant la lésion et une région de diamètre 50 mm environnant pendant toute la durée d&#39;une minute. </li><li> Après une minute, nous supprimons ce stress de refroidissement pour permettre à la peau de réchauffer à la température ambiante dans les 3-4 minutes (phase de récupération thermique) (Fig. 1c-d). </li><li> Pendant la phase de récupération thermique, des images infrarouges de la lésion pigmentée sont saisies toutes les 2 secondes (Fig. 1c-d). </li><li> Toutes les images IR (en plus de la lumière blanche et lumière polarisée images) prises au cours de l&#39;étude sont enregistrées et stockées à l&#39;aide du logiciel Labview. </li></ol> 3. Traitement d&#39;image <ol><li> Les images infrarouges sont analysées à l&#39;aide d&#39;un code dédié Matlab afin d&#39;obtenir des données précises distributions transitoires de la température sur la surface de la peau. À cette fin, nous intro dui re plusieurs étapes d&#39;étalonnage et d&#39;un système d&#39;analyse d&#39;images multimodal. </li><li> On commence par l&#39;application d&#39;un algorithme de détection de point de repère à l&#39;image de lumière afin de localiser les coins du marqueur adhésif. Ensuite, nous avons identifié les points correspondants dans l&#39;image de référence IR. </li><li> Afin de compenser involontaire du corps / la branche mouvement du patient, nous utilisons ces points de repères dans un modèle de mouvement quadratique pour aligner la séquence d&#39;images IR pendant la phase de récupération. </li><li> Nous utilisons le marcheur aléatoire, un algorithme de segmentation d&#39;image interactive où l&#39;utilisateur peut guider la segmentation spatiale par des points de semences de placer, pour créer une image de masque délimitant la lésion. </li><li> Une fois déterminée la forme de la lésion, nous identifions la région correspondante dans chacune des images infrarouges enregistrés. </li><li> On sélectionne des points aléatoires à l&#39;intérieur de la lésion et à distance de la lésion soit la lésion et le tissu sain, respectivement. </li><li> Nous compsont la réponse transitoire thermique de la peau saine et la réponse de la lésion. </li><li> Nous préparons un tableau montrant toutes les données: numérique, dermoscopie, code couleur des images IR de la lésion et ses environs enregistré dans des conditions ambiantes et 2 secondes après l&#39;excitation de refroidissement, et la réponse thermique transitoire de la lésion et le tissu sain. </li></ol> 4. Les résultats représentatifs: <img src="/files/ftp_upload/2679/2679fig1.jpg" alt="Figure 1" /> Figure 1. a) Les SIRH système infrarouge d&#39;imagerie dans la salle d&#39;essais cliniques, b) photographie de la surface supérieure du corps avec une grappe de lésions pigmentées et le cadre modèle appliqué pour l&#39;imagerie, c) référence de l&#39;image infrarouge de la région à température ambiante, d) la même zone après refroidissement et e) la section agrandie de la lésion mélanome et ses environs <img src="/files/ftp_upload/2679/2679fig2.jpg" alt="Figure 2" /><br/&gt; Figure 2. Salle d&#39;examen avec notre système d&#39;imagerie thermique. <img src="/files/ftp_upload/2679/2679fig3.jpg" alt="Figure 3" /> La figure 3. Refroidissement et la lésion du tissu cutané environnant par soufflage d&#39;un courant d&#39;air froid à partir d&#39;un tube à vortex.

<ol>
	<li>Elder, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+progression,+early+diagnosis+and+prognosis+of+melanoma.">Tumor progression, early diagnosis and prognosis of melanoma.</a> Acta Oncol. 38, 535-547 (1999).</li><li>Wartman, D., Weinstock, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+we+overemphasizing+sun+avoidance+in+protection+from+melanoma.">Are we overemphasizing sun avoidance in protection from melanoma.</a> Cancer Epidemiol Biomarkers Prev. 17, 469-470 (2008).</li><li>Pirtini Cetingul, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Using high resolution infrared imaging to detect melanoma and dysplastic nevi [dissertation]. , (2010).</li><li>Jones, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+reappraisal+of+the+use+of+infrared+thermal+image+analysis+in+medicine.">A reappraisal of the use of infrared thermal image analysis in medicine.</a> IEEE Trans. Med. Imaging. 17, 1019-1027 (1998).</li><li>Anbar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+thermal+imaging+today-shifting+from+phenomenological+thermography+to+pathophysiologically+based+thermal+imaging.">Clinical thermal imaging today-shifting from phenomenological thermography to pathophysiologically based thermal imaging.</a> IEEE Eng. Med. Biol. Mag. 17, 25-33 (1998).</li><li>Anbar, M., Gratt, B. M., Hong, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermology+and+facial+telethermography.+Part+I:+history+and+technical+review.">Thermology and facial telethermography. Part I: history and technical review.</a> Dentomaxillofacial Radiology. 27, 61-67 (1998).</li><li>Jones, B. F., Plassmann, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digital+infrared+thermal+imaging+of+human+skin.">Digital infrared thermal imaging of human skin.</a> IEEE Eng. Med. Bio. 21, 41-48 (2002).</li><li>Qi, H., Diakides, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Infrared imaging in Medicine. , (2007).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+skin+lesions+from+the+transient+thermal+response+using+infrared+imaging+technique.">Identification of skin lesions from the transient thermal response using infrared imaging technique.</a> IEEE 5th Int. Symp. on Biomedical Imaging: From Nano to Macro 1-4. , 1219-1222 (2008).</li><li>Cetingul, P. i. r. t. i. n. i., M, ., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+the+thermal+signature+of+a+melanoma+lesion.">Quantification of the thermal signature of a melanoma lesion.</a> Int. Journal of Thermal Science. 50, 421-431 (2011).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+heat+transfer+model+of+skin+tissue+for+the+detection+of+lesions:+sensitivity+analysis.">A heat transfer model of skin tissue for the detection of lesions: sensitivity analysis.</a> Physics in Medicine and Biology. 55, 5933-5951 (2010).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+evaluation+of+skin+lesions+using+transient+thermal+imaging.">Quantitative evaluation of skin lesions using transient thermal imaging.</a> Proc. Int. Heat Transfer Conf. , (2010).</li></ol>

Les résultats suggèrent que, en appliquant le stress de refroidissement, nous avons amélioré les différences de température entre la lésion et le tissu sain environnant. En outre, en raison de petits mouvements du patient pendant l&#39;imagerie thermique, nous avons dû appliquer le suivi de mouvement correctement superposer des images pour mesurer les différences de température entre l&#39;état de référence et la distribution de la température lors de la récupération thermique. Sans le suivi de mouvement, nous n&#39;aurions pas été en mesure de détecter et de mesurer la différence de température entre la lésion maligne et le tissu sain. Ces résultats et la nécessité pour le mouvement précis de suivi expliquent les chercheurs difficultés rencontrées dans le passé lorsque l&#39;on tente de diagnostiquer le mélanome en utilisant l&#39;imagerie infrarouge basée sur les informations d&#39;état stationnaire seule et clairement démontrer les avantages de l&#39;imagerie thermique dynamique. Il est à noter que la résolution spatiale de la caméra IR (nombre de pixels dans la matrice à plan focal IR) est critiquetique quand exigeants de petites lésions. Tant la résolution spatiale et la sensibilité à la température des caméras infrarouges début était limité, ce qui explique aussi les difficultés de détection précoce du mélanome dans le passé. Les principales différences entre notre approche et avant les tentatives d&#39;imagerie thermique - qui étaient un succès modéré - sont les séquences d&#39;étalonnage et les étapes de traitement d&#39;images qui nous permettent de mesurer avec précision les différences de température dans ce système, en plus du processus d&#39;imagerie dynamique qui repose sur un refroidissement actif.

<table border="1"><tbody><tr><td> Nom de l&#39;équipement </td><td> Entreprise </td><td> Numéro de catalogue </td></tr><tr><td> Merlin MWIR caméra </td><td> FLIR </td><td> - </td></tr><tr><td> Canon PowerShot G11 </td><td> Canon </td><td> - </td></tr><tr><td> DermLite Foto système </td><td> DermLite </td><td> - </td></tr><tr><td> Vortex </td><td> Exair </td><td> - </td></tr><tr><td> Réservoirs d&#39;air </td><td> Airgas </td><td> - </td></tr></tbody></table>

quantitative visualization and detection of skin cancer using dynamic thermal imaging

En 2010, environ 68.720 mélanomes seront diagnostiqués dans les seuls États-Unis, avec près de 8650 ayant entraîné la mort 1. À ce jour, le seul traitement efficace contre le mélanome reste l&#39;exérèse chirurgicale, par conséquent, la clé de la survie prolongée est la détection précoce 2,3. Compte tenu du grand nombre de patients diagnostiqués chaque année et les limitations d&#39;accès aux soins spécialisés rapidement, le développement de l&#39;objectif in vivo instruments de diagnostic pour faciliter le diagnostic est essentielle. De nouvelles techniques pour détecter les cancers de la peau, en particulier les outils non invasifs de diagnostic, sont à l&#39;étude dans de nombreux laboratoires. Avec les méthodes chirurgicales, des techniques telles que la photographie numérique, dermoscopie, les systèmes d&#39;imagerie multispectrale (MelaFind), systèmes à laser (microscopie confocale à balayage laser, l&#39;imagerie de perfusion par laser Doppler, la tomographie par cohérence optique), l&#39;échographie, l&#39;imagerie par résonance magnétique, sont à l&#39;essai . Chaque technique offre des avantages uniques d&#39;unnd inconvénients, dont un grand nombre constituent un compromis entre l&#39;efficacité et la précision plutôt que la facilité d&#39;utilisation et de coût. Les détails de ces techniques et les comparaisons sont disponibles dans la littérature 4. Infrarouge (IR) d&#39;imagerie a été montré pour être une méthode utile pour diagnostiquer les signes de certaines maladies par la mesure de la température cutanée locale. Il ya un grand nombre de preuves montrant que la maladie ou l&#39;écart par rapport au fonctionnement normal sont accompagnés par des changements de la température du corps, ce qui à nouveau affecter la température de la peau 5,6. Des données précises sur la température du corps humain et de la peau peuvent fournir une foule d&#39;informations sur les processus responsables de la production de chaleur et la thermorégulation, en particulier l&#39;écart des conditions normales, souvent causés par la maladie. Cependant, l&#39;imagerie IR n&#39;a pas été reconnue en médecine en raison de l&#39;utilisation prématurée de la technologie 7,8 il ya plusieurs décennies, quand temprécision de la mesure rature et la résolution spatiale étaient insuffisantes et des outils sophistiqués de traitement d&#39;image n&#39;étaient pas disponibles. Cette situation a changé de façon spectaculaire à la fin des années 1990 et 2000. Les progrès en instrumentation infrarouge, la mise en œuvre des algorithmes de traitement numérique d&#39;images et d&#39;imagerie dynamique IR, ce qui permet aux scientifiques d&#39;analyser non seulement l&#39;espace, mais aussi le comportement temporel thermique de la peau 9, percées permis sur le terrain. Dans notre recherche, nous explorons la possibilité de l&#39;imagerie infrarouge, combinée avec des études théoriques et expérimentales, comme une solution rentable, non invasive, en technique de mesure in vivo optique pour la détection des tumeurs, en mettant l&#39;accent sur ​​le dépistage et la détection précoce du mélanome 10-13 . Dans cette étude, nous montrons les données obtenues dans une étude patiente dans laquelle les patients porteurs d&#39;une lésion pigmentée avec une indication clinique pour la biopsie sont sélectionnés pour l&#39;imagerie. Nous avons comparé la différence dans les réponses thermiques entresaine et malignes des tissus et comparé nos données avec les résultats des biopsies. Nous avons conclu que l&#39;activité métabolique accrue de la lésion mélanome peut être détectée par l&#39;imagerie infrarouge dynamique.

Watch this Scientific Journal Video about Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging at JoVE.com

Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

Nous avons démontré que les lésions malignes pigmentées avec une activité métabolique accrue générer des quantités mesurables de la chaleur et de la mesure de la réponse thermique transitoire de la peau à une excitation de refroidissement permet l&#39;identification quantitative de mélanome et autres cancers cutanés (vs non-prolifération naevus) à un stade précoce stade de la maladie.

Visualisation quantitative et la détection de cancer de la peau Utilisation dynamique imagerie thermique

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment ...

quantitative-visualization-detection-skin-cancer-using-dynamic

Research

JoVE Journal

Medicine

16.6K vues.  The Johns Hopkins University. Nous avons démontré que les lésions malignes pigmentées avec une activité métabolique accrue générer des quantités mesurables de la chaleur et de la mesure de la réponse thermique transitoire de la peau à une excitation de refroidissement permet l'identification quantitative de mélanome et autres cancers cutanés (vs non-prolifération naevus) à un stade précoce stade de la maladie.

Vidéo: Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&amp;#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

Mesure précise et simple de la cytokine pro-inflammatoire IL-1β en utilisant un dosage de sang total Stimulation

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

Recherche

Médecine

Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex process that can involve many tissue types, span variable time periods, and often occur deep within organs, making it difficult to investigate and quantify. In addition, the efficacy of the metastatic process is influenced by multiple steps in the metastatic cascade making it difficult to evaluate the contribution of a single aspect of tumor cell behavior. As a consequence, metastasis assays are frequently performed in experimental animals to provide a necessarily realistic context in which to study metastasis. Unfortunately, these models are further complicated by their complex physiology. The chick embryo is a unique in vivo model that overcomes many limitations to studying metastasis, due to the accessibility of the chorioallantoic membrane (CAM), a well-vascularized extra-embryonic tissue located underneath the eggshell that is receptive to the xenografting of tumor cells (figure 1). Moreover, since the chick embryo is naturally immunodeficient, the CAM readily supports the engraftment of both normal and tumor tissues. Most importantly, the avian CAM successfully supports most cancer cell characteristics including growth, invasion, angiogenesis, and remodeling of the microenvironment. This makes the model exceptionally useful for the investigation of the pathways that lead to cancer metastasis and to predict the response of metastatic cancer to new potential therapeutics. The detection of disseminated cells by species-specific Alu PCR makes it possible to quantitatively assess metastasis in organs that are colonized by as few as 25 cells. Using the Human Epidermoid Carcinoma cell line (HEp3) we use this model to analyze spontaneous metastasis of cancer cells to distant organs, including the chick liver and lung. Furthermore, using the Alu-PCR protocol we demonstrate the sensitivity and reproducibility of the assay as a tool to analyze and quantitate intravasation, arrest, extravasation, and colonization as individual elements of metastasis.

Using quantitative PCR, we demonstrate how the well-established chick CAM model can be used to quantitatively analyze the metastasis of human tumor cells to distant organs.

Analyse quantitative des métastases du cancer en utilisant un modèle d&#39;embryon aviaire

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex ...

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

In molecular imaging, positron emission tomography (PET) and optical imaging (OI) are two of the most important and thus most widely used modalities1-3. PET is characterized by its excellent sensitivity and quantification ability while OI is notable for non-radiation, relative low cost, short scanning time, high throughput, and wide availability to basic researchers. However, both modalities have their shortcomings as well. PET suffers from poor spatial resolution and high cost, while OI is mostly limited to preclinical applications because of its limited tissue penetration along with prominent scattering optical signals through the thickness of living tissues.
 Recently a bridge between PET and OI has emerged with the discovery of Cerenkov Luminescence Imaging (CLI)4-6. CLI is a new imaging modality that harnesses Cerenkov Radiation (CR) to image radionuclides with OI instruments. Russian Nobel laureate Alekseyevich Cerenkov and his colleagues originally discovered CR in 1934. It is a form of electromagnetic radiation emitted when a charged particle travels at a superluminal speed in a dielectric medium7,8. The charged particle, whether positron or electron, perturbs the electromagnetic field of the medium by displacing the electrons in its atoms. After passing of the disruption photons are emitted as the displaced electrons return to the ground state. For instance, one 18F decay was estimated to produce an average of 3 photons in water5. 
 Since its emergence, CLI has been investigated for its use in a variety of preclinical applications including in vivo tumor imaging, reporter gene imaging, radiotracer development, multimodality imaging, among others4,5,9,10,11. The most important reason why CLI has enjoyed much success so far is that this new technology takes advantage of the low cost and wide availability of OI to image radionuclides, which used to be imaged only by more expensive and less available nuclear imaging modalities such as PET.
 Here, we present the method of using CLI to monitor cancer drug therapy. Our group has recently investigated this new application and validated its feasibility by a proof-of-concept study12. We demonstrated that CLI and PET exhibited excellent correlations across different tumor xenografts and imaging probes. This is consistent with the overarching principle of CR that CLI essentially visualizes the same radionuclides as PET. We selected Bevacizumab (Avastin; Genentech/Roche) as our therapeutic agent because it is a well-known angiogenesis inhibitor13,14. Maturation of this technology in the near future can be envisioned to have a significant impact on preclinical drug development, screening, as well as therapy monitoring of patients receiving treatments.

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

Use of Cerenkov Luminescence Imaging (CLI) for monitoring preclinical cancer treatment is described here. This method takes advantage of Cerenkov Radiation (CR) and optical imaging (OI) to visualize radiolabeled probes and thus provides an alternative to PET in preclinical therapeutic monitoring and drug screening.

Luminescence Cerenkov Imaging (CLI) pour la surveillance de la thérapie du cancer

In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

Quantitative Multispectral Analysis Following Fluorescent Tissue Transplant for Visualization of Cell Origins, Types, and Interactions

With the desire to understand the contributions of multiple cellular elements to the development of a complex tissue; such as the numerous cell types that participate in regenerating tissue, tumor formation, or vasculogenesis, we devised a multi-colored cellular transplant model of tumor development in which cell populations originate from different fluorescently colored reporter gene mice and are transplanted, engrafted or injected in and around a developing tumor. These colored cells are then recruited and incorporated into the tumor stroma. In order to quantitatively assess bone marrow derived tumor stromal cells, we transplanted GFP expressing transgenic whole bone marrow into lethally irradiated RFP expressing mice as approved by IACUC. 0ovarian tumors that were orthotopically injected into the transplanted mice were excised 6-8 weeks post engraftment and analyzed for bone marrow marker of origin (GFP) as well as antibody markers to detect tumor associated stroma using multispectral imaging techniques. We then adapted a methodology we call MIMicc- Multispectral Interrogation of Multiplexed cellular compositions, using multispectral unmixing of fluoroprobes to quantitatively assess which labeled cell came from which starting populations (based on original reporter gene labels), and as our ability to unmix 4, 5, 6 or more spectra per slide increases, we&#39;ve added additional immunohistochemistry associated with cell lineages or differentiation to increase precision. Utilizing software to detect co-localized multiplexed-fluorescent signals, tumor stromal populations can be traced, enumerated and characterized based on marker staining.1

Complex tissue masses, from organs to tumors, are composed of various cellular elements. We elucidated the contribution of cellular phenotypes within a tissue utilizing multi-labeled fluorescent transgenic mice in combination with multiparameter immunofluorescent staining followed by spectral unmixing to decipher cell origin as well as cell characteristics based on protein expression.

Analyse quantitative multispectrale Après transplantation de tissu fluorescent pour la visualisation des origines cellulaires, types et Interactions

With the desire to understand the contributions of multiple cellular elements to the development of a complex tissue; such as the numerous cell types that ...

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

MicroRNAs (miRNAs) constitute a potent layer of gene regulation by guiding RISC to target sites located on mRNAs and, consequently, by modulating their translational repression. Changes in miRNA expression have been shown to be involved in the development of all major complex diseases. Furthermore, recent findings showed that miRNAs can be secreted to the extracellular environment and enter the bloodstream and other body fluids where they can circulate with high stability. The function of such circulating miRNAs remains largely elusive, but systematic high throughput approaches, such as miRNA profiling arrays, have lead to the identification of miRNA signatures in several pathological conditions, including neurodegenerative disorders and several types of cancers. In this context, the identification of miRNA expression profile in the cerebrospinal fluid, as reported in our recent study, makes miRNAs attractive candidates for biomarker analysis.
There are several tools available for profiling microRNAs, such as microarrays, quantitative real-time PCR (qPCR), and deep sequencing. Here, we describe a sensitive method to profile microRNAs in cerebrospinal fluids by quantitative real-time PCR. We used the Exiqon microRNA ready-to-use PCR human panels I and II V2.R, which allows detection of 742 unique human microRNAs. We performed the arrays in triplicate runs and we processed and analyzed data using the GenEx Professional 5 software.
Using this protocol, we have successfully profiled microRNAs in various types of cell lines and primary cells, CSF, plasma, and formalin-fixed paraffin-embedded tissues.

We describe a protocol of real time PCR to profile microRNAs in the cerebrospinal fluid (CSF). With the exception of RNA extraction protocols, the procedure can be extended to RNA extracted from other body fluids, cultured cells, or tissue specimens.

Liquide céphalo-rachidien profilage de microARN Utilisation quantitative PCR en temps réel

MicroRNAs (miRNAs) constitute a potent layer of gene regulation by guiding RISC to target sites located on mRNAs and, consequently, by modulating their ...

Measurement of Dynamic Scapular Kinematics Using an Acromion Marker Cluster to Minimize Skin Movement Artifact

The measurement of dynamic scapular kinematics is complex due to the sliding nature of the scapula beneath the skin surface. The aim of the study was to clearly describe the acromion marker cluster (AMC) method of determining scapular kinematics when using a passive marker motion capture system, with consideration for the sources of error which could affect the validity and reliability of measurements. The AMC method involves placing a cluster of markers over the posterior acromion, and through calibration of anatomical landmarks with respect to the marker cluster it is possible to obtain valid measurements of scapular kinematics. The reliability of the method was examined between two days in a group of 15 healthy individuals (aged 19-38 years, eight males) as they performed arm elevation, to 120&#176;, and lowering in the frontal, scapular and sagittal planes. Results showed that between-day reliability was good for upward scapular rotation (Coefficient of Multiple Correlation; CMC = 0.92) and posterior tilt (CMC = 0.70) but fair for internal rotation (CMC = 0.53) during the arm elevation phase. The waveform error was lower for upward rotation (2.7&#176; to 4.4&#176;) and posterior tilt (1.3&#176; to 2.8&#176;), compared to internal rotation (5.4&#176; to 7.3&#176;). The reliability during the lowering phase was comparable to results observed during the elevation phase. If the protocol outlined in this study is adhered to, the AMC provides a reliable measurement of upward rotation and posterior tilt during the elevation and lowering phases of arm movement.

This report presents details of how to adopt the acromion marker cluster method of obtaining scapular kinematics when using a passive marker motion capture device. As has been described in the literature, this method provides a robust, non-invasive, three-dimensional, dynamic and valid measurement of scapular kinematics, minimizing skin movement artifact.

Mesure de la dynamique Scapulaire cinématique utilisant un cluster Acromion Marker pour minimiser le mouvement de la peau Artefact

The measurement of dynamic scapular kinematics is complex due to the sliding nature of the scapula beneath the skin surface. The aim of the study was to ...

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.

The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.

Contraste dynamique amélioré imagerie par résonance magnétique d&#39;un modèle orthotopique cancer du pancréas de souris

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe ...

Diffuse Optical Spectroscopy for the Quantitative Assessment of Acute Ionizing Radiation Induced Skin Toxicity Using a Mouse Model

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and negatively impact patient quality of life and long term survival. Advances in the understanding of the biological pathways associated with normal tissue toxicities have allowed for the development of interventional drugs, however, current response studies are limited by a lack of quantitative metrics for assessing the severity of skin reactions. Here we present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe the instrumentation design of the DOS system as well as the inversion algorithm for extracting the optical parameters. Finally, to demonstrate clinical utility, we present representative data from a pre-clinical mouse model of radiation induced erythema and compare the results with a commonly employed visual scoring. The described DOS method offers an objective, high through-put evaluation of skin toxicity via functional response that is translatable to the clinical setting.

We present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe DOS instrumentation design, optical parameters extraction algorithms and the animal handling procedures required to yield representative data from a pre-clinical mouse model of radiation induced erythema.

Diffuse Spectroscopie optique pour l&#39;évaluation quantitative de la toxicité de la peau induit par les rayonnements ionisants aiguë Utilisation d&#39;un modèle de souris

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

Visualisation quantitative de l&#39;infiltration de leucocytes dans un modèle murin de myocardite fulminante par microscopie à feuilles lumineuses

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing continuous remodeling with the competing activity of osteoblasts, which produce the new bone matrix, and osteoclasts, whose function is to eliminate mineralized bone. [18F]-NaF is a positron emission tomography (PET) radiotracer that enables visualization of bone metabolism. [18F]-NaF is chemically absorbed into hydroxyapatite in the bone matrix by osteoblasts and can thus noninvasively detect osteoblastic activity, which is occult to conventional imaging techniques. Kinetic modeling of dynamic [18F]-NaF-PET data provides detailed quantitative measures of bone metabolism. Conventional semi-quantitative PET data, which utilizes standardized uptake values (SUVs) as a measure of radiotracer activity, is referred to as a static technique due to its snapshot of tracer uptake in time.&#160; Kinetic modeling, however, utilizes dynamic image data where tracer levels are continuously acquired providing tracer uptake temporal resolution. From the kinetic modeling of dynamic data, quantitative values like blood flow and metabolic rate (i.e., potentially informative metrics of tracer dynamics) can be extracted, all with respect to the measured activity in the image data. When combined with dual modality PET-MRI, region-specific kinetic data can be correlated with anatomically registered high-resolution structural and pathologic information afforded by MRI. The goal of this methodological manuscript is to outline detailed techniques for performing and analyzing dynamic [18F]-NaF-PET-MRI data. The lumbar facet joint is a common site of degenerative arthritis disease and a common cause for axial low back pain.&#160; Recent studies suggest [18F]-NaF-PET may serve as a useful biomarker of painful facetogenic disease.&#160; The human lumbar facet joint will, therefore, be used as a prototypical region of interest for dynamic [18F]-NaF-PET-MRI analysis in this manuscript.

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. We present detailed methodologies for performing and analyzing dynamic [18F]-NaF-PET-MRI data in a patient with facetogenic low back pain using the lumbar facet joints as a prototypical region of interest.

Analyse quantitative [18F]-Naf-PET-MRI pour l'évaluation du chiffre d'affaires dynamique d'os dans un patient présentant la douleur lombaire facetogenic

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing ...