RESUMEN
Most pancreas and islet grafts are recovered from brain-dead (BD) donors. In this study we characterized the early inflammatory response induced by brain death in pancreata and islets from nonhuman primate donors and evaluated the effect of targeted anti-inflammatory intervention in the protection of pancreatic islets prior to transplantation. BD donors were monitored for 6 h and assigned to three experimental groups: group 1: BD-untreated donors (BD-UT) (n = 7), group 2: BD + donor pretreatment with IL-1ra (n = 6), and group 3: non-BD animals serving as controls (n = 7). We observed an IL-1ra-dependent reduction in the mobilization and activation of neutrophils from bone marrow and a significantly reduced accumulation of CD68(+) leukocytes in the pancreas and islets after brain death induction. Donor treatment with IL-1ra significantly decreased chemokine mRNA expression (MCP-1, IL-8, and MIP-1a) and attenuated the activation of circulating neutrophils and intraislet macrophages as demonstrated by a reduction in intracellular IL-1ß, IL-6, MCP-1, and MIP-1α expression. As a result, IL-1ra dramatically improved viability, mitochondrial membrane polarity, and islet engraftment in mice transplanted using a minimal islet mass. These results suggest that early immunomodulation targeting inflammation in the BD donor may represent an effective therapeutic strategy to improve islet quality and function prior to transplantation.
Asunto(s)
Islotes Pancreáticos/metabolismo , Receptores de Interleucina-1/antagonistas & inhibidores , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Muerte Encefálica , Diferenciación Celular , Movimiento Celular , Células Cultivadas , Quimiocinas/análisis , Quimiocinas/genética , Quimiocinas/metabolismo , Citocinas/análisis , Citocinas/genética , Citocinas/metabolismo , Diabetes Mellitus Experimental/terapia , Ensayo de Inmunoadsorción Enzimática , Hemodinámica , Inflamación/prevención & control , Islotes Pancreáticos/citología , Trasplante de Islotes Pancreáticos , Macaca mulatta , Macrófagos/citología , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos NOD , Ratones SCID , Neutrófilos/inmunología , Neutrófilos/metabolismo , Páncreas/patología , ARN Mensajero/metabolismo , Donantes de Tejidos , Trasplante HeterólogoRESUMEN
Treatment of anorectal foreign bodies is a frequent surgical and proctologic question. However, psychiatric implications of paraphilias are rarely discussed. Since the liberalisation of sexual behaviour in the 1970s, the reported number of severe complications due to paraphilias is increasing, and sexuality is performed until higher age. We describe the case of an 88 year old man who had undergone laparotomy for removal of foreign bodies. The initial diagnosis of a dementia with inappropriate sexual behaviour could not be confirmed as the paraphilia existed since his youth. Etiologic, diagnostic and therapeutic implications of the case are discussed.
Asunto(s)
Canal Anal/cirugía , Cuerpos Extraños/psicología , Cuerpos Extraños/cirugía , Trastornos Parafílicos/diagnóstico , Trastornos Parafílicos/psicología , Recto/cirugía , Conducta Autodestructiva/diagnóstico , Conducta Autodestructiva/psicología , Anciano de 80 o más Años , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/epidemiología , Enfermedad de Alzheimer/psicología , Comorbilidad , Cuerpos Extraños/epidemiología , Cuerpos Extraños/prevención & control , Humanos , Impotencia Vasculogénica/psicología , Tutores Legales/legislación & jurisprudencia , Tutores Legales/psicología , Masculino , Competencia Mental/legislación & jurisprudencia , Trastornos Parafílicos/epidemiología , Factores de Riesgo , Conducta Autodestructiva/prevención & control , Conducta Sexual , Negativa del Paciente al TratamientoRESUMEN
BACKGROUND: Donor brain death (BD) triggers a systemic inflammatory response that reduces organ quality and increases immunogenicity of the graft. We characterized the early innate immune response induced by BD in the liver and peripheral blood of hemodinamically stable non-human primates (NHP). METHODS: Rhesus macaques were assigned to either brain death or control group. BD was induced by inflation of a subdurally placed catheter and confirmed clinically and by cerebral angiography. Animals were monitored for 6 h after BD and managed to maintain hemodynamic stability. RESULTS: Cortisol, epinephrine, nor-epinephrine, and IL-6 levels were elevated immediately after BD induction. Neutrophils and monocytes significantly increased in circulation following BD induction, while dendritic cells were decreased at 6 h post-induction. Flow cytometry revealed increased expression of chemokine receptors CxCR1, CxCR2, CCR2, and CCR5 in peripheral blood leukocytes from NHP subjected to BD. Microarray analysis demonstrated a significant up-regulation of genes related to innate inflammatory responses, toll-like receptor signaling, stress pathways, and apoptosis/cell death in BD subjects. Conversely, pathways related to glucose, lipid, and protein metabolism were down-regulated. In addition, increased expression of SOCS3, S100A8/A9, ICAM-1, MHC class II, neutrophil accumulation, and oxidative stress markers (carboxy-methyl-lysine and hydroxynonenal) were detected by immunoblot and immunohistochemistry. CONCLUSIONS: Activation of the innate immune response after BD in association with a down-regulation of genes associated with cell metabolism pathways in the liver. These findings may provide a potential explanation for the reduced post-transplant function of organs from brain dead donors. In addition, this work suggests potential novel targets to improve donor management strategies.
Asunto(s)
Muerte Encefálica/inmunología , Hepatitis/inmunología , Inmunidad Innata/inmunología , Trasplante de Hígado , Hígado/inmunología , Donantes de Tejidos , Animales , Citocinas/sangre , Epinefrina/sangre , Perfilación de la Expresión Génica , Hepatitis/genética , Hidrocortisona/sangre , Inmunidad Innata/genética , Macrófagos del Hígado/inmunología , Linfocitos/inmunología , Macaca mulatta , Neutrófilos/inmunología , Norepinefrina/sangre , Receptores Toll-Like/metabolismoRESUMEN
Pro-inflammatory cytokines (PIC) impair islet viability and function by activating inflammatory pathways that induce both necrosis and apoptosis. The aim of this study was to utilize an in vitro rat islet model to evaluate the efficacy of a clinically approved IL-1 receptor antagonist (Anakinra) in blocking PIC induced islet impairment. Isolated rat islets were cultured for 48 h +/- PIC (IL-1beta, IFNgamma, and TNFalpha) and +/-IL-1ra then assayed for cellular integrity by flow cytometry, MAPK phosphorylation by proteome array, and gene expression by RT-PCR. Nitric oxide (NO) release into the culture media was measured by Griess reaction. Islet functional potency was tested by glucose stimulated insulin secretion (GSIS) and by transplantation into streptozotocin-induced diabetic NOD.scid mice. Rat islets cultured with PIC upregulated genes for NOS2a, COX2, IL6, IL1b, TNFa, and HMOX1. IL-1ra prevented the PIC induced upregulation of all of these genes except for TNFa. Inhibition of PIC induced iNOS by NG-monomethyl-L-arginine (NMMA) only blocked the increased expression of HMOX1. IL-1ra completely abrogated the effects of PIC with respect to NO production, necrosis, apoptosis, mitochondrial dysfunction, GSIS, and in vivo potency. IL-1ra was not effective at preventing the induction of necrosis or apoptosis by exogenous NO. These data demonstrate that Anakinra is an effective agent to inhibit the activation of IL-1beta dependent inflammatory pathways in cultured rat islets and support the extension of its application to human islets in vitro and potentially as a post transplant therapy.