Ce travail a été soutenu par le CDMRP, subventions USUHS G188LE, G188MG et G188QC (HL), et le Centre USUHS pour l&#39;étude du stress post-traumatique.

<ol>
	<li>Adrien, J., Dugovic, C., Martin, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep-wakefulness+patterns+in+the+helpless+rat.">Sleep-wakefulness patterns in the helpless rat.</a> Physiol. Behav. 49, 257-262 (1991).</li><li>Bai, X., Wu, J., Zhang, Q., Alesci, S., Manoli, I., Blackman, M. R., Chrousos, G. P., Goldstein, A. L., Rennert, O. M., Su, Y. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Third-generation+human+mitochondria-focused+cDNA+microarray+and+its+bioinformatic+tools+for+analysis+of+gene+expression.">Third-generation human mitochondria-focused cDNA microarray and its bioinformatic tools for analysis of gene expression.</a> Biotechniques. 42, 365-375 (2007).</li><li>Blaszczyk, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Startle+response+to+short+acoustic+stimuli+in+rats.">Startle response to short acoustic stimuli in rats.</a> Acta Neurobiol. Exp. (Wars.). 63, 25-30 (2003).</li><li>Braga, M. F., roniadou-Anderjaska, V., Manion, S. T., Hough, C. J., Li, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress+impairs+alpha(1A)+adrenoceptor-mediated+noradrenergic+facilitation+of+GABAergic+transmission+in+the+basolateral+amygdala.">Stress impairs alpha(1A) adrenoceptor-mediated noradrenergic facilitation of GABAergic transmission in the basolateral amygdala.</a> Neuropsychopharmacology. 29, 45-58 (2004).</li><li>Bremner, J. D., Vythilingam, M., Vermetten, E., Anderson, G., Newcomer, J. W., Charney, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+glucocorticoids+on+declarative+memory+function+in+major+depression.">Effects of glucocorticoids on declarative memory function in major depression.</a> Biol. Psychiatry. 55, 811-815 (2004).</li><li>Garrick, T., Morrow, N., Eth, S., Marciano, D., Shalev, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychophysiologic+parameters+of+traumatic+stress+disorder+in+rats.">Psychophysiologic parameters of traumatic stress disorder in rats.</a> Ann. N.Y. Acad. Sci. 821, 533-537 (1997).</li><li>Garrick, T., Morrow, N., Shalev, A. Y., Eth, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress-induced+enhancement+of+auditory+startle:+an+animal+model+of+posttraumatic+stress+disorder.">Stress-induced enhancement of auditory startle: an animal model of posttraumatic stress disorder.</a> Psychiatry. 64, 346-354 (2001).</li><li>Hsiao, Y. H., Su, Y. A., Tsai, H. D., Mason, J. T., Chou, M. C., Man, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+invasiveness+and+aggressiveness+in+breast+epithelia+with+cytoplasmic+p63+expression.">Increased invasiveness and aggressiveness in breast epithelia with cytoplasmic p63 expression.</a> Int. J. Biol. Sci. 6, 428-442 (2010).</li><li>Jiang, X., Xing, G., Yang, C., Verma, A., Zhang, L., Li, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress+Impairs+5-HT(2A)+Receptor-Mediated+Serotonergic+Facilitation+of+GABA+Release+in+Juvenile+Rat+Basolateral+Amygdala.">Stress Impairs 5-HT(2A) Receptor-Mediated Serotonergic Facilitation of GABA Release in Juvenile Rat Basolateral Amygdala.</a> Neuropsychopharmacology. , (2008).</li><li>Jiang, X., Zhang, Z. J., Zhang, S., Gamble, E. H., Jia, M., Ursano, R. J., Li, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=5-HT2A+receptor+antagonism+by+MDL+11,939+during+inescapable+stress+prevents+subsequent+exaggeration+of+acoustic+startle+response+and+reduced+body+weight+in+rats.">5-HT2A receptor antagonism by MDL 11,939 during inescapable stress prevents subsequent exaggeration of acoustic startle response and reduced body weight in rats.</a> J. Psychopharmacol. , (2009).</li><li>Maher, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+deprivation:+are+you+a+victim.">Sleep deprivation: are you a victim.</a> AAOHN. J. 54, 548 (2006).</li><li>Maier, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exposure+to+the+stressor+environment+prevents+the+temporal+dissipation+of+behavioral+depression/learned+helplessness.">Exposure to the stressor environment prevents the temporal dissipation of behavioral depression/learned helplessness.</a> Biol. Psychiatry. 49, 763-773 (2001).</li><li>North, C. S., Suris, A. M., Davis, M., Smith, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+validation+of+the+diagnosis+of+posttraumatic+stress+disorder.">Toward validation of the diagnosis of posttraumatic stress disorder.</a> Am. J. Psychiatry. 166, 34-41 (2009).</li><li>Orr, S. P., Metzger, L. J., Pitman, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychophysiology+of+post-traumatic+stress+disorder.">Psychophysiology of post-traumatic stress disorder.</a> Psychiatr. Clin. North Am. 25, 271-293 (2002).</li><li>Orr, S. P., Roth, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychophysiological+assessment:+clinical+applications+for+PTSD.">Psychophysiological assessment: clinical applications for PTSD.</a> J. Affect. Disord. 61, 225-240 (2000).</li><li>Pitman, R. K., Orr, S. P., Shalev, A. Y., Metzger, L. J., Mellman, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychophysiological+alterations+in+post-traumatic+stress+disorder.">Psychophysiological alterations in post-traumatic stress disorder.</a> Semin. Clin. Neuropsychiatry. 4, 234-241 (1999).</li><li>Servatius, R. J., Ottenweller, J. E., Natelson, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+startle+sensitization+distinguishes+rats+exposed+to+one+or+three+stress+sessions:+Further+evidence+toward+an+animal+model+of+PTSD.">Delayed startle sensitization distinguishes rats exposed to one or three stress sessions: Further evidence toward an animal model of PTSD.</a> Biological Psychiatry. 38, 539-546 (1995).</li><li>Su, Y. A., Zhang, Q., Su, D. M., Tang, M. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+Mitochondrion-Neuron+Focused+Microarray+(rMNChip)+and+Bioinformatics+Tools+for+Rapid+Identification+of+Differential+Pathways+in+Brain+Tissues.">Rat Mitochondrion-Neuron Focused Microarray (rMNChip) and Bioinformatics Tools for Rapid Identification of Differential Pathways in Brain Tissues.</a> Int. J. Biol. Sci. 7, 308-322 (2011).</li><li>Su, D. M., Zhang, Q., Wang, X., He, P., Zhu, Y. J., Zhao, J., Rennert, O. M., Su, Y. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+types+of+human+malignant+melanoma+cell+lines+revealed+by+expression+patterns+of+mitochondrial+and+survival-apoptosis+genes:+implications+for+malignant+melanoma+therapy.+Mol.">Two types of human malignant melanoma cell lines revealed by expression patterns of mitochondrial and survival-apoptosis genes: implications for malignant melanoma therapy. Mol.</a> Cancer Ther. , (2009).</li><li>Su, Y. A., Li, H., He, P., Webster, M. J., Zhang, Q., Zhang, L., Wang, X., Su, D. M., Ursano, R. J., Rennert, O. M. <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+stress-responsive+genes,+canonical+pathways,+molecular+networks+and+drug+targets+in+brain+tissues+of+human+and+animals+for+systems-biology+study+of+post-traumatic+stress+disorder.">Identification of stress-responsive genes, canonical pathways, molecular networks and drug targets in brain tissues of human and animals for systems-biology study of post-traumatic stress disorder.</a> , (2009).</li><li>Su, Y. A., Wu, J., Zhang, L., Zhang, Q., Su, D. M., He, P., Wang, B. D., Li, H., Webster, M. J., Rennert, O. M., Ursano, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dysregulated+mitochondrial+genes+and+networks+with+drug+targets+in+postmortem+brain+of+patients+with+posttraumatic+stress+disorder+(PTSD)+revealed+by+human+mitochondria-focused+cDNA+microarrays.">Dysregulated mitochondrial genes and networks with drug targets in postmortem brain of patients with posttraumatic stress disorder (PTSD) revealed by human mitochondria-focused cDNA microarrays.</a> Int. J Biol. Sci. 4, 223-235 (2008).</li><li>Su, Y. A., Yang, J., Tao, L., Nguyen, H., He, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Undetectable+and+Decreased+Expression+of+KIAA1949+(Phostensin)+Encoded+on+Chromosome+6p21.33+in+Human+Breast+Cancers+Revealed+by+Transcriptome+Analysis.">Undetectable and Decreased Expression of KIAA1949 (Phostensin) Encoded on Chromosome 6p21.33 in Human Breast Cancers Revealed by Transcriptome Analysis.</a> J. Cancer. 1, 38-50 (2010).</li><li>Yehuda, R., Yang, R. K., Buchsbaum, M. S., Golier, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alterations+in+cortisol+negative+feedback+inhibition+as+examined+using+the+ACTH+response+to+cortisol+administration+in+PTSD.">Alterations in cortisol negative feedback inhibition as examined using the ACTH response to cortisol administration in PTSD.</a> Psychoneuroendocrinology. 31, 447-451 (2006).</li><li>Zhang, Q., Wu, J., Nguyen, A., Wang, B. D., He, P., Laurent, G. S., Rennert, O. M., Su, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanism+underlying+differential+apoptosis+between+human+melanoma+cell+lines+UACC903+and+UACC903(+6)+revealed+by+mitochondria-focused+cDNA+microarrays.">Molecular mechanism underlying differential apoptosis between human melanoma cell lines UACC903 and UACC903(+6) revealed by mitochondria-focused cDNA microarrays.</a> Apoptosis. 13, 993-1004 (2008).</li></ol>

Aucun conflit d&#39;intérêt déclaré.

Le diagnostic de SSPT est basé sur l&#39;auto signalé des symptômes psychiatriques (DSM-IV) par des sujets potentiels. Aucun biomarqueur bien définie est actuellement disponible pour accéder à l&#39;état physiopathologique du potentiel de patients atteints de SSPT. Le SSPT est un trouble provoqué par la vie en danger événements traumatiques et les symptômes psychiatriques principaux restent présents dans la vie de la victime pendant des mois et même des années après les événements initiaux. Les symptômes les plus importants et persistants ont révélé chez les patients atteints du SSPT sont hypervigilance, sursauts exagérés réponse retardée 14, 15, 16 et un compromis apparent de l&#39;axe HPA. Chez l&#39;homme ces symptômes persistent ou apparaissent avec un délai de trois mois, après l&#39;arrêt du stress traumatique 24. Notre modèle actuel pour les études de biomarqueurs de stress post-traumatique emploie retenue et chocs répétés queue pendant trois jours consécutifs (2-hr séances de 40 mA, tailshocks 2) - l&#39;incontournable choc arrière-modèle (ITS) chez les rats pesant 150 grammes. Ce modèle SA aété montré à imiter dans une large mesure la physiopathologie de stress post-traumatique 17, 7, 4, 10. Notre laboratoire et d&#39;autres laboratoires ont vérifié que le modèle SA de stress chez le rat induit des modifications comportementales et neurobiologiques qui sont similaires à ceux trouvés dans le SSPT sujets 17, 7, 10, 9. Plus précisément, ces rats stressés présentent (1) une réaction tardive et exagérées de sursaut apparaissant plusieurs jours après l&#39;arrêt de stress, ce qui compte tenu de l&#39;échelle de temps compressé de la vie du rat par rapport à un homme, correspond au retard de un à trois mois des symptômes chez les patients atteints de SSPT (DSM-IV-TR SSPT Criterian D / E 13), (2) améliorer corticostérone plasmatique (CORT) pour plusieurs (10) jours, en indiquant compromis de l&#39;axe hypothalamo-hypophysaire (HPA), et (3) le gain de poids du corps après un retard de stress l&#39;arrêt, ce qui correspond à un dysfonctionnement de la régulation du métabolisme du SSPT. Il n&#39;existe aucune preuve dans la littérature que l&#39;odeur de renard, l&#39;exposition des prédateurs, ou craignent potentialisée réaction de sursaut, d&#39;exposipeu ces phénotypes comportementaux persistants et neuroendocrinologic associés à l&#39;ESPT. Des rats exposés à une session de stress unique (1DS) ont montré transitoire, mais pas les anomalies persistantes affichées par les rats 3DS 17 La présente expérience a comparé la réaction de sursaut de la 3DS et les rats 1DS 4, 7 et 10 jours après l&#39;arrêt de stress. Conformément aux travaux antérieurs, a souligné rats exposés élevés les taux plasmatiques basales CORT le stress premier jour après 17 ans. Ces niveaux étaient sensibles à CORT le nombre d&#39;expositions aux facteurs de stress avec des niveaux plus élevés chez le rat CORT 3DS que dans 1DS rats. Quant aux réactions de sursaut, les rats 1DS présentent une réponse exagérée à la surprise de stress après 7 jours, alors que la sensibilisation sursaut ne devient apparente 10 jours après stress chez le rat 3DS. Ainsi, l&#39;apparition d&#39;une réponse exagérée de sursaut après la cessation de stress semble être liée au nombre d&#39;expositions de session stress. Le modèle 3DS stressé sembleêtre utile pour mieux comprendre l&#39;altération de l&#39;expression des biomarqueurs associés à des symptômes de stress post-traumatique et les principaux phénotypes comportementaux mesurables liés à la synchronisation après la cessation du stress. Résultats génomiques a présenté une preuve de principe pour l&#39;application des outils rMNChip et la bioinformatique pour identifier les voies différentielles, et des biomarqueurs génétiques et de protéines, ce qui facilitera grandement les systèmes biologiques, l&#39;étude et la compréhension des mécanismes moléculaires sous-jacents complexes et multifactorielles des troubles neurologiques, y compris le SSPT. Bien que notre paradigme ne pas plonger dans le plan cognitif et plus complexes aspects comportementaux de stress post-traumatique, nous constatons que les habitudes de sommeil altérés dans le modèle STI 1 correspondent à la difficulté à s&#39;endormir et à rester endormi et les cauchemars de patients atteints de SSPT 11 (DSM-IV-TR SSPT Critères D 13), et les carences en fuite / évitement apprentissage et l&#39;apprentissage d&#39;une tâche appétitive dans le modèle SA <sup&gt; 12 correspond à des concentrations faibles et des déficits de la mémoire de l&#39;ESPT 5 (DSM-IV-TR SSPT Critères C 13). Le modèle actuel est bien corrélée avec les symptômes caractéristiques clés du SSPT et constitue un bon modèle pour corréler biomarqueurs périphériques de stress post-traumatique, qui peut être recueillie auprès des patients, avec des centrales, des biomarqueurs mécanistes, qui ne peuvent pas 6.

biomarkers in an animal model for revealing neural, hematologic, and behavioral correlates of ptsd

Identification de biomarqueurs représentant l&#39;évolution de la physiopathologie du syndrome de stress post-traumatique (SSPT) est d&#39;une importance vitale, non seulement pour le diagnostic mais aussi pour objectif l&#39;évaluation de l&#39;efficacité thérapeutique et la résistance à un traumatisme. La recherche en cours vise à identifier des biomarqueurs moléculaires pour le SSPT, y compris les protéines de stress traumatiques induits, transcriptomes, les écarts génomiques et génétiques modulateurs, en utilisant des échantillons biologiques dans le sang des sujets, la salive, l&#39;urine et les tissus du cerveau post-mortem. Cependant, la corrélation de ces molécules biomarqueurs dans des échantillons post mortem ou périphériques à fonctions cérébrales altérées associées à des symptômes psychiatriques chez les SSPT n&#39;est toujours pas résolu. Ici, nous présentons un modèle animal de stress post-traumatique dans lequel les deux le sang périphérique et biomarqueurs cérébraux centraux, ainsi que phénotype comportemental, peut être recueillie et mesurée, ce qui permet la corrélation nécessaire des biomarqueurs centrales de stress post-traumatique, qui sont mécaniste et pathognomoniquemonique, mais ne peuvent pas être recueillies auprès des personnes, avec les biomarqueurs périphériques et les phénotypes comportementaux qui peuvent. Notre modèle animal de stress post-traumatique emploie retenue et chocs répétés queue pendant trois jours consécutifs - l&#39;incontournable choc arrière-modèle (ITS) chez les rats. Ce modèle SON imite la physiopathologie de stress post-traumatique 17, 7, 4, 10. Nous et d&#39;autres avons vérifié que le modèle ITS induit des altérations du comportement et neurobiologiques semblables à ceux trouvés dans le SSPT sujets 17, 7, 10, 9. Plus précisément, ces rats stressés présentent (1) une réaction tardive et exagérées de sursaut apparaissant plusieurs jours après l&#39;arrêt de stress, ce qui compte tenu de l&#39;échelle de temps compressé de la vie du rat par rapport à un homme, correspond au retard de un à trois mois des symptômes chez les patients atteints de SSPT (DSM-IV-TR SSPT Criterian D / E 13), (2) améliorer corticostérone plasmatique (CORT) pendant plusieurs jours, ce qui indique compromis de l&#39;axe hypothalamo-hypophysaire (HPA), et (3) un retard de bole gain de poids après l&#39;arrêt dy facteur de stress, ce qui indique un dysfonctionnement de la régulation métabolique. Les paradigmes expérimentaux utilisés pour ce modèle sont les suivants: (1) un paradigme impuissance acquise chez le rat dosé par mesure de la réponse de sursaut acoustique (ASR) et une cartographie de la masse corporelle, (2) microdissection du cerveau de rat dans des régions et des noyaux; ( 3) lié à une enzyme (ELISA) pour les niveaux sanguins de CORT, (4) une puce à l&#39;expression des gènes en plus liée à outils bioinformatiques 18. Cette biopuce, surnommé rMNChip, se concentre sur les mitochondries et les mitochondries liée à des gènes nucléaires chez le rat afin de répondre spécifiquement aux bioénergétique neuronales suppose être impliqués dans le SSPT.

Watch this Scientific Journal Video about Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD at JoVE.com

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Nous décrivons un modèle de rat de trouble de stress post-traumatique (SSPT) qui révèle les altérations persistantes dans la fonction neuroendocrine et le retard à long terme, l&#39;intervention crainte exagérée, caractéristiques des patients SSPT. Le modèle animal et les méthodes décrites ici sont utiles pour corréler des biomarqueurs dans les noyaux du cerveau, qui sont mécaniste, mais ne peut pas être mesurée chez les patients avec des biomarqueurs en globules blancs périphériques, ce qui peut.

Biomarqueurs dans un modèle animal pour Révéler Corrélats neuronaux, hématologiques et du comportement de stress post-traumatique

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

biomarkers-an-animal-model-for-revealing-neural-hematologic

Research

JoVE Journal

Medicine

15.9K vues.  Uniformed Services University of the Health Sciences, Bethesda, Maryland. Nous décrivons un modèle de rat de trouble de stress post-traumatique (SSPT) qui révèle les altérations persistantes dans la fonction neuroendocrine et le retard à long terme, l'intervention crainte exagérée, caractéristiques des patients SSPT. Le modèle animal et les méthodes décrites ici sont utiles pour corréler des biomarqueurs dans les noyaux du cerveau, qui sont mécaniste, mais ne peut pas être mesurée chez les patients avec des biomarqueurs en g...

Vidéo: Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

L&#39;utilisation de modèle animal de sepsis pour évaluer de nouvelles thérapies à base de plantes

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Recherche

Médecine

An Isolated Working Heart System for Large Animal Models

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart model, have been invaluable tools for studying cardiovascular function and disease1-15. Although the Langendorff heart preparation can be used for any mammalian heart, most studies involving this apparatus use small animal models (e.g., mouse, rat, and rabbit) due to the increased complexity of systems for larger mammals1,3,11. One major difficulty is ensuring a constant coronary perfusion pressure over a range of different heart sizes &#8211; a key component of any experiment utilizing this device1,11. By replacing the classic hydrostatic afterload column with a centrifugal pump, the Langendorff working heart apparatus described below allows for easy adjustment and tight regulation of perfusion pressures, meaning the same set-up can be used for various species or heart sizes. Furthermore, this configuration can also seamlessly switch between constant pressure or constant flow during reperfusion, depending on the user&#8217;s preferences. The open nature of this setup, despite making temperature regulation more difficult than other designs, allows for easy collection of effluent and ventricular pressure-volume data.

Most studies involving the Langendorff apparatus use small animal models due to the increased complexity of systems for larger mammals. We describe a Langendorff system for large animal models that allows for use across a range of species, including humans, and relatively easy data acquisition.

Un système de Coeur de travail isolé pour les grands modèles animaux

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart ...

Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and histopathologic features of the human disease. When GL261 cells are implanted intracranially in syngeneic C57BL/6 mice, the model has the added advantage of maintaining an intact immune microenvironment. Stable expression of luciferase in GL261 cells allows non-invasive cost effective bioluminescence monitoring of intracranial tumor growth. We have recently demonstrated that luciferase expression in GL261 cells does not affect the tumor growth properties, tumor cell immunomodulatory cytokine expression, infiltration of immune cells into the tumor, or overall survival of animals bearing the intracranial tumor. Therefore, it appears that the GL261 luciferase glioma model can be useful in the study of novel chemotherapeutic and immunotherapeutic modalities. Here we report the technique for generating stable luciferase expression in GL261 cells and how to study the in vitro and in vivo growth of the tumor cells by bioluminescence imaging.

GL261 glioma cells provide a useful immunocompetent animal model of glioblastoma. The goals of this protocol are to demonstrate proper techniques for monitoring intracranial tumor growth using in vivo bioluminescence imaging, and to verify the utility of luciferase-modified GL261 cells for studying tumor immunology and immunotherapeutic approaches for treating glioblastoma.

Bioluminescence imagerie d&#39;un modèle animal pour immunocompétentes glioblastome

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and ...

A Repetitive Concussive Head Injury Model in Mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

Une blessure à la tête modèle Répétitive commotionnant chez la souris

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle screws after spinal surgery, which is a serious concern. To address this problem, diverse types of pedicle screws have been examined to identify those with good fixation strength and osseointegration in spine bone. The porcine spine is a good alternative for the human spine in the evaluation of pedicle screws due to the anatomical size, mechanical characteristics, and cost. Although several studies have reported that pedicle screws are efficient in the porcine model, no study has described detailed protocols for the evaluation of a pedicle screw using the porcine model. Here, we describe a detailed method for evaluating transpedicular screws using an in vivo porcine lumbar spine model. The technical details for anesthesia, spine surgery, and harvest provided here will facilitate with the evaluation of the transpedicular screw fixation model.

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

Here, we introduce a method to evaluate transpedicular screws by using an in vivo porcine lumbar spine model.

Une méthode d&#39;anesthésie, de chirurgie et de récolte pour l&#39;évaluation des vis transpediculaires en utilisant un<em&gt; In vivo</em&gt; Modèle de la colonne lombaire porcine

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle ...

A Chronic Cardiac Ischemia Model in Swine Using an Ameroid Constrictor

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but regardless of the approach, an in vivo model is needed to test each treatment. The pig is one of the most used large animal models for cardiovascular disease. Its heart is very similar in anatomy and function to that of a human. The ameroid placement technique creates an ischemic area of the heart, which has many useful applications in studying myocardial infarction. This model has been used for surgical research, pharmaceutical studies, imaging techniques, and cell therapies.
There are several ways of inducing an ischemic area in the heart. Each has its advantages and disadvantages, but the placement of an ameroid constrictor remains the most widely used technique. The main advantages to using the ameroid are its prevalence in existing research, its availability in various sizes to accommodate the anatomy and size of the vessel to be constricted, the surgery is a relatively simple procedure, and the post-operative monitoring is minimal, since there are no external devices to maintain. This paper provides a detailed overview of the proper technique for the placement of the ameroid constrictor.

The purpose of this protocol is to demonstrate the placement of a delayed constricting device (an ameroid constrictor) around a coronary artery in a swine model. This device creates an ischemic area of the heart that is useful for studying new diagnostic imaging techniques and new methods of treatment.

Un modèle d’ischémie cardiaque chronique chez les porcs à l’aide d’un Constrictor Améroïde

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but ...

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent hemorrhages at a later stage. The exact effects of plasma coagulation on liver tissue are only poorly examined. In our porcine model, the coagulation effects can be examined close to the clinical application. A combined laser Doppler flowmeter and spectrophotometer documents microcirculation changes during coagulation at 8 mm tissue depth noninvasively, providing quantifiable information about hemostasis beyond the subjective clinical impression. The temperature at coagulation site is assessed with an infrared thermometer prior and post coagulation and with a thermographic camera during coagulation, a measurement of the gas beam temperature is not possible due to the upper threshold of the devices. The depth of coagulation is measured microscopically on hematoxylin/eosin stained sections after calibration with an object micrometer and gives an exact information about the power setting-coagulation depth-relation. The sealing effect is examined on the bile ducts as it is not possible for a plasma coagulator to seal larger blood vessels. Burst pressure experiments are carried out on explanted organs to rule out blood pressure related effects.

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Here we present a protocol to experimentally assess plasma coagulation in liver tissue in vivo. In a porcine model, microcirculation is examined by laser Doppler, coagulation depth is measured histologically, temperature at coagulation site by infrared thermometer and thermographic camera, and duct sealing effect is documented by burst pressure experiments.

Évaluation de la Coagulation du Plasma sur le tissu hépatique dans un grand modèle Animal In Vivo

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent ...

Intra-Operative Neural Monitoring of Thyroid Surgery in a Porcine Model

Intraoperative injury to the recurrent laryngeal nerve (RLN) can cause vocal cord paralysis, which interferes with speech and can potentially interfere with breathing. In recent years, intraoperative neural monitoring (IONM) has been widely adapted as an adjunct technique to localize the RLN, detect RLN injury, and predict vocal cord function during the operations. Many studies have also used animal models to investigate new applications of IONM technology and to develop reliable strategies for preventing intraoperative RLN injury. The aim of this article is to introduce a standard protocol for using a porcine model in IONM research. The article demonstrates the procedures for inducing general anesthesia, performing tracheal intubation, and experimental design to investigate the electrophysiological characteristics of RLN injuries. Applications of this protocol can improve overall efficacy in implementing the 3R principle (replacement, reduction and refinement) in porcine IONM studies.

This study aims to develop a standard protocol of intra-operative neural monitoring of thyroid surgery in a porcine model. Here, we present a protocol to demonstrate general anesthesia, to compare different types of electrodes, and to investigate the electrophysiological characteristics of the normal and injured recurrent laryngeal nerves.

Une surveillance neurale peropératoire de la chirurgie de la thyroïde dans un modèle porcin

Intraoperative injury to the recurrent laryngeal nerve (RLN) can cause vocal cord paralysis, which interferes with speech and can potentially interfere ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Mesure de la contractilité isométrique de l’artère mésentérique de souris à l’aide de fil myographie

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Manufacture of a Multi-Purpose Low-Cost Animal Bench-Model for Teaching Tracheostomy

Tracheostomy is one of the most frequent procedures, performed through various techniques in the intensive care unit and emergency situations. Despite this, there is a lack of training on this procedure that affects its outcome, which is also dependent on operator's dexterity. Here, we take the specific training and simulation into consideration. This article aims to describe every step of the manufacture of a new multi-purpose low-cost animal bench-model, with the support of video and images, and to obtain an opinion about the quality of this model by administering a questionnaire to professionals with experience in the procedures. 
Ten experts in the technique were enrolled. The model scored an average of 3.45/5 for its anatomical realism; 4.75/5 for its usefulness as a training tool for simulation courses and assessments. The time necessary to build the model was 15 minutes, and the cost amounted to 10&#8364;. The animal bench-model was considered a very useful simulator for tracheostomy training and assessments. Therefore, it could be used as a tool for medical courses and residencies.

This article illustrates every step of the manufacture of a new multi-purpose low-cost animal bench-model for subglottic airway access management. All the procedures are shown in the video. The model's realism and its suitability for training the given clinical maneuvers were assessed by independent senior otolaryngologists and anesthesiologists.

Fabrication d’un banc-modèle d’animaux à faible coût polyvalent pour l’enseignement de la trachéostomie

Tracheostomy is one of the most frequent procedures, performed through various techniques in the intensive care unit and emergency situations. Despite ...