RESUMO
Intestinal graft motility after small bowel transplantation (SBT) is poorly characterized. The aim of this study was to compare motor patterns with myenteric neuronal cell population as a parameter of graft viability at various degrees of acute cellular rejection (ACR). Three grades of ACR were achieved in orthotopic allografts. Syngeneic transplants and allografts with immunosuppression served as controls. Motor activities were recorded using strain gauge force transducers and analyzed visually. Quantifications of myenteric neurons in whole mounts of intestinal grafts were used to evaluate neuronal population. A typical migrating motor complex (MMC) was found in syngeneic and allogenic transplants with immunosuppression. A high prevalence of discrete clustered contractions (DCC) and nonpropagating contractions (NPC) without MMC was seen in moderately and severely rejected allografts. Neuronal cell loss in the allografts, which could be one of the causes of motor dysfunction, was noted in moderate rejection (19.3%) and progressed until severe rejection (60.1%). Monitoring motility patterns in SBT could be an effective tool for assessing intestinal rejection. Allograft dysmotility, such as absence of MMC and high prevalence of DCC or NPC, could be useful markers of progression of acute rejection and help guide treatment decisions.
Assuntos
Motilidade Gastrointestinal , Rejeição de Enxerto/diagnóstico , Intestino Delgado/fisiopatologia , Intestino Delgado/transplante , Neurônios/patologia , Animais , Intestino Delgado/inervação , Masculino , Ratos , Ratos Endogâmicos , Transplante HomólogoRESUMO
BACKGROUND: Regeneration of partial liver grafts is critical for successful living donor liver transplantation (LDLT), especially in adult recipients. The purpose of this study was to investigate the intraoperative hemodynamic changes in partial liver grafts and characterize their potential impact on post-transplant liver regeneration in LDLT. METHODS: We examined the portal venous flow (PVF) and hepatic arterial flow (HAF) to partial liver grafts by means of ultrasonic transit time flowmeter of donors immediately before graft retrieval and of the corresponding recipients after vascular reconstruction in 48 LDLT cases. We evaluated post-transplant liver regeneration according to the changes in graft liver volume between the time of transplantation and the 7th post-transplant day. RESULTS: There was a significant increase in PVF to the partial liver grafts in recipients (rPVF) compared with that in donors. In contrast, graft HAF in recipients significantly decreased compared with that in donors. The rPVF inversely correlated with graft weight (GW)-recipient body weight ratio (GRWR), whereas HAF volume showed no significant correlation. The rPVF/GW positively correlated with the rate of liver regeneration (GRR), which inversely correlated with GRWR. The rPVF/GW was significantly higher, and GRR tended to be larger in the small graft group than in the non-small graft group. CONCLUSIONS: Intraoperative portal hemodynamic changes in partial liver grafts strongly affect their post-transplant regeneration. In particular, in small liver grafts, an immediate and remarkable increase in graft PVF may contribute to rapid liver regeneration after LDLT if the increased PVF remains within a safe range.
Assuntos
Regeneração Hepática/fisiologia , Transplante de Fígado/métodos , Doadores Vivos , Veia Porta/fisiologia , Transplantes/irrigação sanguínea , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Hemodinâmica/fisiologia , Artéria Hepática/fisiologia , Humanos , Fígado/irrigação sanguínea , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Molecular genetic analyses have clarified that accumulation of genomic changes provides important steps in carcinogenesis and have identified a number of valuable genetic markers for certain cancers. To date, however, no prognostic markers have been identified for hepatocellular carcinoma (HCC). In this study, we used restriction landmark genomic scanning (RLGS), a new high-speed screening method for multiple genomic changes, to detect unknown genetic alterations in HCC. Thirty-one HCC samples and their normal counterparts were examined by RLGS. Eight spot changes were common in several cases, and all were seen only on the HCC profile. Five of these spots were detected in more than 12 of 31 cases (38.7%). Viral infection had no influence on changes in the RLGS spots. The disease-free survival rate for patients with > or =16 changed RLGS spots was significantly lower than that for patients with fewer changed RLGS spots (< or =15 spots) (P<0.001). In multivariate analysis, the number of changed spots was proven to retain an independent prognostic value (relative risk 1.095: P = 0.0031). These results suggest that the number of changed RLGS spots may be a useful biological marker for recurrence of HCC.
Assuntos
Carcinoma Hepatocelular/genética , Metilação de DNA , Análise Mutacional de DNA/métodos , DNA de Neoplasias/análise , Eletroforese em Gel Bidimensional , Amplificação de Genes , Neoplasias Hepáticas/genética , Metástase Neoplásica , Recidiva Local de Neoplasia , Técnica de Subtração , Adulto , Idoso , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/cirurgia , DNA de Neoplasias/genética , Densitometria , Intervalo Livre de Doença , Feminino , Genoma , Hepatectomia , Humanos , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/cirurgia , Masculino , Recidiva Local de Neoplasia/mortalidade , PrognósticoRESUMO
Dysregulation of apoptosis is one of the likely underlying mechanisms of neointimal thickening, a disorder in which proinflammatory cytokines may influence the function of vascular smooth muscle cells (VSMCs) and contribute to atherogenesis. One of these cytokines, tumor necrosis factor-alpha (TNF-alpha), induces 2 possibly conflicting pathways, 1 leading to the activation of nuclear factor-kappaB (NF-kappaB) and the other leading to caspase-mediated apoptosis. We investigated whether specific inhibition of NF-kappaB affects TNF-alpha-dependent apoptosis in human VSMCs. To inhibit NF-kappaB activation specifically, we constructed a recombinant adenovirus vector expressing a truncated form of the inhibitor protein IkappaBalpha (AdexIkappaBDeltaN) that lacks the phosphorylation sites essential for activation of NF-kappaB. The IkappaBDeltaN was overexpressed by adenoviral infection and was resistant to stimulus-dependent degradation. Electromobility gel shift and luciferase assays demonstrated that overexpression of IkappaBDeltaN inhibited NF-kappaB activation induced by TNF-alpha or interleukin-1beta (IL-1beta). In cells overexpressing IkappaBDeltaN, TNF-alpha dramatically induced apoptosis, whereas IL-1beta had no effect. The induction was suppressed by treatment with a selective inhibitor of the caspase-3 family, Z-DEVD-fmk, and the overexpression of IkappaBDeltaN induced TNF-alpha-mediated caspase-3 and caspase-2 activity. These results indicate that overexpression of IkappaBDeltaN induces TNF-alpha-dependent apoptosis by efficient and specific suppression of NF-kappaB and upregulation of caspase-3 and caspase-2 activity in human VSMCs. Our findings suggest that adenovirus-mediated IkappaBDeltaN gene transfer may be useful in the treatment of disorders associated with inflammatory conditions, such as the response to vascular injury and atherosclerosis.
Assuntos
Apoptose , Proteínas de Ligação a DNA/biossíntese , Proteínas I-kappa B , Músculo Liso Vascular/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Adenoviridae/genética , Aorta/metabolismo , Arteriosclerose/prevenção & controle , Western Blotting , Caspase 2 , Caspase 3 , Caspases/metabolismo , Células Cultivadas , Fragmentação do DNA , Proteínas de Ligação a DNA/antagonistas & inibidores , Proteínas de Ligação a DNA/genética , Eletroforese , Ativação Enzimática , Ensaio de Imunoadsorção Enzimática , Regulação da Expressão Gênica , Terapia Genética , Humanos , Interleucina-1/metabolismo , Interleucina-1/farmacologia , Luciferases/genética , Inibidor de NF-kappaB alfa , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Proteínas Repressoras/genética , Transfecção , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
BACKGROUND: We previously reported that intratracheal delivery of alloantigen induced regulatory cells in a mouse heart transplantation model. We investigated the roles of costimulatory pathways in the induction of regulatory cells by intratracheal delivery of alloantigen. METHODS: CBA (H-2k) mice were pretreated with intratracheal delivery of splenocytes (1 x 10(7)) from C57BL/10 (H-2b) mice and administration of monoclonal antibodies (mAb) specific for programmed death (PD)-1 and its ligands, programmed death-ligand (PD-L)1 and PD-L2, CD70, CD134 ligand (CD134L), CD153, CD137L, or receptor activator of nuclear factor-kappaB (NF-kappaB) (RANK). Seven days later, naive CBA mice underwent adoptive transfer of splenocytes (5 x 10(7)) from the pretreated CBA mice and transplantation of C57BL/10 heart. RESULTS: Adoptive transfer of splenocytes from CBA mice that had been pretreated with intratracheal delivery of C57BL/10 splenocytes significantly prolonged the survival of C57BL/10 allograft (median survival time [MST], 68 days) as compared with adoptive transfer from untreated CBA mice (MST, 12 days). Concomitant administration of control immunoglobulin (Ig)G, anti-PD-L2 mAb, or anti-CD137L along with intratracheal delivery did not significantly affect the prolongation (MST, 72, 68, and 65 days, respectively). In contrast, anti-PD-1, anti-PD-L1, anti-CD70, anti-CD134L, anti-CD153, or anti-RANK mAb abrogated the prolongation induced by adoptive transfer from the pretreated mice with intratracheal delivery (MST, 18, 17, 16, 14, 10, and 18 days, respectively). CONCLUSION: The PD-1/PD-L1, CD27/CD70, CD134/CD134L, CD30/CD153, and tumor necrosis factor (TNF)-related activation-induced cytokine (TRANCE)/RANK interactions are independently required for generation of regulatory cells by intratracheal delivery of alloantigen.
Assuntos
Sobrevivência de Enxerto/fisiologia , Isoantígenos/administração & dosagem , Transfusão de Linfócitos/métodos , Transferência Adotiva , Animais , Intubação Intratraqueal , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Modelos Animais , Transplante Homólogo/imunologiaRESUMO
The patient was a 45-year-old man with underlying alcoholic liver cirrhosis. Two years prior, he was repeatedly hospitalized for liver failure symptoms and requested a living-donor liver transplantation (LDLT) because of end-stage cirrhosis. A pretransplantation blood test revealed a high 1,3-beta-d-glucan (BDG) value of 102.0 pg/mL (reference value <20.0 pg/mL) and a high blood Aspergillus antigen (AsAg) value of 1.6 cutoff index (COI; reference value <0.5 COI). Contrast-enhanced thoracoabdominal-pelvic computed tomography (CT) and cranial magnetic resonance imaging revealed no fungal infection. However, latent fungal infection could not be ruled out, hence preoperative antifungal agent treatment was administered. BDG and AsAg levels showed a decreasing trend after treatment initiation. However, normalization did not occur; the BDG and AsAg levels were 25.8 pg/mL and 1.0 COI, respectively. Although the possibility of latent fungal infection was judged low, we prophylactically administered antifungal agents after LDLT. The BDG level consistently increased at 35-39 pg/mL until postoperative day 5 but subsequently normalized. The AsAg level was higher than the limit of detection at 5.0 COI on postoperative day 3 but normalized to 0.2 COI on postoperative day 5 and did not subsequently increase. The postoperative course was uneventful despite bacterial pneumonia and the patient was discharged on postoperative day 35. A histopathologic examination (Grocott methenamine silver staining) and a fungal polymerase chain reaction assay were performed for the resected liver, but the results of both were negative. At 9 postoperative months, the patient was making ambulatory follow-up visits. Currently, the BDG and AsAg values remain normal and clinical progress is favorable. We found no reports of LDLT for a recipient with a high preoperative BDG level and positive test result for AsAg. Thus, we report on such a case with a discussion of the literature on the causes of high preoperative BDG and AsAg values.
Assuntos
Antifúngicos/uso terapêutico , Antígenos de Fungos/sangue , Aspergilose/prevenção & controle , Aspergillus/imunologia , Transplante de Fígado , Complicações Pós-Operatórias/prevenção & controle , beta-Glucanas/sangue , Aspergilose/diagnóstico , Biomarcadores/sangue , Humanos , Doadores Vivos , Masculino , Pessoa de Meia-Idade , Cuidados Pós-Operatórios , Complicações Pós-Operatórias/diagnóstico , Cuidados Pré-Operatórios , ProteoglicanasRESUMO
The inflammatory cytokines interleukin (IL) 1 and tumor necrosis factor (TNF) may play an important role in hepatic ischemia-reperfusion (I/R) injury. To study the role of IL-1 in hepatic I-R injury, we investigated the effect of pretreatment with IL-1 receptor antagonist (IL-1ra) on the production of IL-1, TNF, histological findings in the liver, and the survival rate for 7 days. Rats were subjected to 90 min of partial liver warm ischemia by clamping the vessels of the left and middle lobes. In the IL-1ra-treated group, IL-1ra was given 5 min before liver ischemia was induced. IL-1alpha and TNF levels were determined in blood and liver at 0, 30, 90, and 180 min after reperfusion. In a second experiment to determine the effect of IL-1ra pretreatment on survival rate, after 90 min of partial liver ischemia, the right lateral and caudate lobes were excised, leaving only the ischemic lobes. In both groups, IL-1alpha was undetectable in blood, but increased in liver tissue. TNF increased in both blood and liver tissue as reperfusion time increased. Histological evidence of tissue injury was minimal in the IL-1ra-treated group. Furthermore, in the IL-1ra-treated group, the production of TNF decreased in both blood and liver tissue compared with the nontreated group. Survival rates in the IL-1ra-treated and nontreated group were 80% and 30%, respectively. The data demonstrated that the production of IL-1 and TNF increases in hepatic I-R injury and that pretreatment with IL-1ra protects the liver from ischemic insult, indicating an important role for IL-1 in I-R injury.
Assuntos
Interleucina-1/fisiologia , Isquemia/complicações , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/prevenção & controle , Sialoglicoproteínas/farmacologia , Fator de Necrose Tumoral alfa/biossíntese , Animais , Proteína Antagonista do Receptor de Interleucina 1 , Interleucina-1/análise , Fígado/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/farmacologiaRESUMO
BACKGROUND: Oxygen-derived free radicals (FRs) are critical mediators of ischemia/reperfusion injury. Inflammatory cytokines have been shown to play important roles in tissue injury. To examine the relationship between FRs and interleukin-1 (IL-1) in hepatic ischemia/reperfusion injury, we used interleukin-1 receptor antagonist (IL-1ra) to block endogenous IL-1 production in a rat model of hepatic ischemia/reperfusion. METHODS: Female SD rats were subjected to 30 min of hepatic ischemia followed by reperfusion. The animals were divided into two groups, control group and IL-1ra-treated group, according to the rinse solution. In both groups, FR production, histological changes, and interactions between leukocytes and endothelial cells were analyzed in the course of reperfusion. RESULTS: In the control group, production of FRs increased significantly after 60 min of reperfusion. After 60 and 180 min of reperfusion, histological examination showed atrophy and degeneration of hepatocytes. Hepatic microcirculation demonstrated a marked increase in the number of leukocytes adherent to endothelial cells and of injured cells after reperfusion. In the IL-1ra-treated group, IL-1ra pretreatment markedly reduced FR production after 60 min of reperfusion, the number of leukocytes adherent to endothelial cells, and tissue injury. CONCLUSION: These data clearly show an important role for IL-1 in the induction of FR production, leukocyte adhesion, and tissue injury after hepatic ischemia/reperfusion injury.
Assuntos
Interleucina-1/farmacologia , Fígado/irrigação sanguínea , Espécies Reativas de Oxigênio/fisiologia , Traumatismo por Reperfusão/fisiopatologia , Animais , Comunicação Celular , Endotélio/citologia , Feminino , Fluoroscopia , Leucócitos/citologia , Fígado/patologia , Medições Luminescentes , Luminol/farmacologia , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/patologia , Regulação para Cima/fisiologia , Veia Cava Inferior/fisiologiaRESUMO
BACKGROUND: Prostaglandin E1 (PGE1) has been reported to have a protective effect in experimental and clinical models of liver damage. The aim of this study was to elucidate the effects of the intraportal infusion of PGE1 on hepatic blood flow and graft viability after orthotopic liver transplantation in pigs. METHODS: First, the hepatic arterial flow (HAF), portal venous flow (PVF), and liver tissue blood flow (LTBF) were measured during the continuous intravenous or intraportal infusion of PGE1. Second, two groups of pigs underwent orthotopic liver transplantation: group A, untreated controls; and group B, animals that received intraportal PGE1 for 2 hr after vascular reconstruction of the allograft. Changes in HAF, PVF, LTBF, and hepatic function were measured. RESULTS: The intraportal infusion of PGE1 significantly increased HAF and had no effect on blood pressure, PVF, or LTBF. In group B, HAF and LTBF increased significantly with time. In group A, HAF remained unchanged and a decrease in LTBF was observed. Group B exhibited a higher arterial ketone body ratio and a greater bile flow compared with group A. A significant elevation in serum glutamic oxaloacetic transaminase concentration was observed in group A, but not in group B. CONCLUSIONS: This study demonstrates that the intraportal infusion of PGE1 improves hepatic allograft blood flow, predominantly through an effect on HAF, and may improve graft viability after orthotopic liver transplantation.
Assuntos
Alprostadil/administração & dosagem , Circulação Hepática/efeitos dos fármacos , Transplante de Fígado/imunologia , Transplante de Fígado/fisiologia , Animais , Feminino , Sobrevivência de Enxerto/efeitos dos fármacos , Infusões Intravenosas , Veia Porta , Fluxo Sanguíneo Regional/efeitos dos fármacos , Traumatismo por Reperfusão/prevenção & controle , SuínosRESUMO
Minimally invasive surgery has rapidly changed the performance of surgical practice in a wide range of surgical specialities in the last decade of the 20th century. The marked progress in endoscopic surgery has been conducted especially in general and digestive surgery. The Department of Surgery, School of Medicine, Keio University has contributed to the development and establishment of endoscopic surgery in every subspeciality of general and digestive surgery. Our achievements include the development of original methods for laparoscopic wedge gastrectomy and endoscopic thyroidectomy, establishment of surgical techniques in endoscopic surgery for esophageal, gastric, colorectal and hepatobiliary diseases, and the introduction of robotics and tele-communicative technologies to endoscopic surgery.
Assuntos
Laparoscopia , Doenças da Glândula Tireoide/cirurgia , Procedimentos Cirúrgicos do Sistema Digestório/efeitos adversos , Procedimentos Cirúrgicos do Sistema Digestório/métodos , Endoscopia/efeitos adversos , Endoscopia/métodos , Doenças do Esôfago/cirurgia , Humanos , Laparoscopia/efeitos adversos , Laparoscopia/métodos , Robótica , TelemedicinaRESUMO
BACKGROUND: Although high levels of interleukin-8 (IL-8) have been found in patients with sepsis and a monoclonal antibody (MoAb) against IL-8 has been successfully used in some animal models of inflammation, no specific therapeutic agent against IL-8 has been tested for the treatment of sepsis. We studied the effects of a MoAb against IL-8 in the treatment of endotoxic shock with a prospective randomized rabbit endotoxic shock model. METHODS: Twenty New Zealand white rabbits were anesthetized and divided into four groups: normal, anti-IL-8, control-Ab, and lipopolysaccharide (LPS). Anti-IL-8 and control-Ab groups received a MoAb (immunoglobulin G, 3 mg/kg) 5 minutes before the LPS injection. All groups, except the normal group, received a continuous 20-minute infusion of LPS (500 micrograms/kg). The normal group received NaCl (0.9%) rather than LPS. RESULTS: The 7-day survival rates were 100% for normal group, 80% for anti-IL-8 group, 40% for control-Ab group, and 0% for LPS group. Compared with the LPS group, anti-IL-8 rabbits had a smaller decrease in mean arterial blood pressure (p < 0.05) and increased urinary volume (p < 0.05). Anti-IL-8 rabbits had lower plasmatic levels of IL-1 beta, less free radical production (p < 0.05), and a higher survival rate (p < 0.01). CONCLUSIONS: IL-8 plays a significant role in endotoxic shock, and IL-8 blockage results in attenuation of the hypotensive and tachypneic effects of LPS, reduced free radical production, and an increased survival rate after lethal endotoxic shock.
Assuntos
Anticorpos Monoclonais/farmacologia , Hemodinâmica/fisiologia , Interleucina-8/imunologia , Choque Séptico/mortalidade , Choque Séptico/fisiopatologia , Animais , Anticorpos Monoclonais/uso terapêutico , Diurese , Feminino , Radicais Livres/metabolismo , Hematócrito , Hemodinâmica/imunologia , Interleucina-1/sangue , Interleucina-8/sangue , Contagem de Leucócitos , Lipopolissacarídeos , Neutrófilos/metabolismo , Coelhos , Espécies Reativas de Oxigênio/metabolismo , Choque Séptico/terapia , Taxa de Sobrevida , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Hepatic resection was performed in 125 patients. Ninety-three of the 125 patients had malignant neoplasms; primary liver carcinoma in 61, metastatic liver carcinoma in 15, carcinoma of the bifurcation of the hepatic ducts in 16, and carcinoma of the gallbladder in one. Performance of hepatic resection was complicated by the presence of liver cirrhosis and jaundice in 42 and 19 patients, respectively. Nine of the 125 patients died within 30 days of the operations, with an operative mortality of 7.2%. Eight of the nine deaths were due to liver failure in the cirrhotic patients who underwent resection of more than two segments of the liver. None of the jaundiced patients died postoperatively. The three-year actuarial survival rates of the patients with hepatocellular carcinoma, metastatic liver carcinoma, and carcinoma of the bifurcation of the hepatic ducts were 31%, 56%, and 21% respectively.
Assuntos
Carcinoma Hepatocelular/cirurgia , Hepatopatias/mortalidade , Neoplasias Hepáticas/cirurgia , Fígado/cirurgia , Adolescente , Adulto , Idoso , Feminino , Ducto Hepático Comum/cirurgia , Humanos , Japão , Icterícia/complicações , Icterícia/cirurgia , Cirrose Hepática/complicações , Cirrose Hepática/cirurgia , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , Complicações Pós-OperatóriasRESUMO
An aneurysm ruptured during superselective catheterization into the posterior cerebral artery for a left temporal arteriovenous malformation. The rupture may have been caused by stretching and displacement of the basilar and posterior cerebral arteries while the microcatheter with guide wire was advanced.
Assuntos
Aneurisma Roto/etiologia , Embolização Terapêutica/efeitos adversos , Aneurisma Intracraniano/complicações , Malformações Arteriovenosas Intracranianas/terapia , Adulto , Humanos , Malformações Arteriovenosas Intracranianas/complicações , MasculinoRESUMO
Cytoprotection by bifemelane hydrochloride was investigated immunohistochemically in the cerebral cortex of rats during ischemia. The middle cerebral artery was occluded for 30 min and then reperfused for 6 h. c-Fos-like immunoreactive neurons were found in layers II to VI of the cerebral cortex and were especially abundant in the parietal cortex and the piriform cortex on the side of the occlusion. In sham-operated control rats, a few c-Fos-like immunoreactive neurons were seen in the ipsilateral side of the cerebral cortex. In animals that had been injected with bifemelane hydrochloride (20 mg/kg, IP) 30 min before the onset of ischemia and 90 min after reperfusion, the number of c-Fos-like immunoreactive neurons was significantly reduced in the cerebral cortex. The results suggest that bifemelane hydrochloride can inhibit the ischemia-induced increase in c-Fos-like immunoreactivity in cerebral cortex neurons.
Assuntos
Compostos Benzidrílicos/farmacologia , Isquemia Encefálica/metabolismo , Córtex Cerebral/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Animais , Isquemia Encefálica/patologia , Isquemia Encefálica/fisiopatologia , Córtex Cerebral/patologia , Circulação Cerebrovascular , Imuno-Histoquímica , Masculino , Neurônios/metabolismo , Ratos , Ratos WistarRESUMO
Time-dependent changes in the tissue concentration of tyrosine hydroxylase (TH), adrenaline (A), noradrenaline (NA), and neuropeptide Y (NPY) in the early stages of cerebral ischemia were studied immunohistochemically in the amygdaloid complex of rats subjected to 1 h cerebral ischemia. Immunoreactivity to TH on the lesioned side reached a nadir at 12 h after cerebral ischemia, then gradually increased over 24 h to normal reactivity, but TH immunoreactivity between the ischemic side and the contralateral side was no different for up to 12 h after ischemia. The blood concentrations of NA and A were elevated to about twice the control concentration 12 h after ischemia, then gradually decreased back to normal. NPY immunoreactivity of both sides did not change for up to 6 h after ischemia, but NPY immunoreactivity on the lesioned side decreased over 12 h and maintained a plateau. These findings suggest that responses to cerebral ischemia between catecholamines and peptides are varied.
Assuntos
Ataque Isquêmico Transitório/metabolismo , Sistema Vasomotor/metabolismo , Animais , Epinefrina/sangue , Técnicas Imunoenzimáticas , Neuropeptídeo Y/sangue , Neuropeptídeo Y/metabolismo , Norepinefrina/sangue , Radioimunoensaio , Ratos , Ratos Wistar , Fatores de Tempo , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Time-dependent changes in c-fos-like immunoreactivity (c-fos-LI) were studied in the rat during focal cerebral ischemia and reperfusion after middle cerebral artery (MCA) occlusion. In the permanent ischemia model, the levels of c-fos-LI increased for the first 30 min of ischemia in neuronal nuclei in the lesioned hemisphere. They reached a maximum at 60 min. The level in the parietal cortex (PC) diminished considerably after 120 min, and in the cingulate cortex (CC) it gradually decreased to near the control value at 180 min. Regional cerebral blood flow (rCBF) in the PC fell to 32% and that in the CC fell to 64% of pre-ischemic values after MCA occlusion. Reperfusion induced strong expression of c-fos-LI in the PC and CC after 6 h of reperfusion that followed 30 min of ischemia. The c-fos-LI was effectively reduced by preadministration of the N-methyl-D-aspartate (NMDA) receptor antagonist, ketamine (100 mg/kg, IP). These findings suggest that the expression of c-fos after ischemia may be immediately activated through NMDA receptors and may spread to surrounding regions in a manner sensitive to reductions in rCBF. Reperfusion after ischemia also appears to cause activation of expression of c-fos and of intracellular signal transduction.
Assuntos
Isquemia Encefálica/metabolismo , Córtex Cerebral/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Traumatismo por Reperfusão/metabolismo , Animais , Isquemia Encefálica/patologia , Isquemia Encefálica/fisiopatologia , Córtex Cerebral/patologia , Circulação Cerebrovascular , Imuno-Histoquímica , Masculino , Proteínas Proto-Oncogênicas c-fos/antagonistas & inibidores , Ratos , Ratos Wistar , Traumatismo por Reperfusão/patologia , Traumatismo por Reperfusão/fisiopatologiaRESUMO
A new anchor system based on the ice nucleation protein (InaV) from Pseudomonas syringae INA5 was developed for cell surface display of functional organophosphorus hydrolase (OPH). The activity and stability of cells expressing the truncated InaV (INPNC)-OPH fusions were compared to cells with surface-expressed OPH using two other fusion anchors based on Lpp-OmpA and the truncated InaK protein. Whole cell activity was as much as 5-fold higher using the InaV anchor. Majority of the OPH activity was located on the cell surface as determined by protease accessibility and cell fractionation experiments. The surface localization of OPH was further verified by immunofluorescence microscopy. Constitutive expression of OPH on the surface using the InaV anchor resulted in no cell lysis or growth inhibition, in contrast to the Lpp-OmpA anchor. Suspended cultures also exhibited good stability, retaining almost 100% activity over a period of 3 weeks. Therefore, the InaV anchor system offers an attractive alternative to the currently available surface anchors, providing high-level expression and superior stability.
Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Esterases/metabolismo , Arildialquilfosfatase , Sequência de Bases , Membrana Celular/metabolismo , Primers do DNA , Esterases/genética , Microscopia de FluorescênciaRESUMO
The authors encountered the unusual case of a 57-year-old man with a right trochlear nerve neurinoma associated with a giant thrombosed dissecting aneurysm of the left vertebral artery. The right trochlear nerve neurinoma was subtotally removed via the right subtemporal transtentorial route. The trochlear nerve entered the tumor directly posterolaterally. The left vertebral artery was clipped just proximal to the thrombosed dissecting aneurysm via the left lateral suboccipital approach. The patient was discharged without neurologic deficits except for the right trochlear nerve palsy. To our knowledge, this seems to be the first reported case of such an occurrence.
Assuntos
Dissecção Aórtica/complicações , Neoplasias dos Nervos Cranianos/complicações , Neurilemoma/complicações , Trombose/complicações , Nervo Troclear , Artéria Vertebral , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
INTRODUCTION: A virally induced alloreactive memory seems to represent a potent barrier to tolerance induction but the combination of 15-deoxyspergualin (DSG), an inhibitor of NFkB translocation, with costimulation blockade (CB)-based chimerism as an induction regimen can overcome a preformed anti-donor memory response. In this study, we investigate the ability of DSG with CB to inhibit a naive alloimmune responses. METHODS: A BALB/c (H-2d) skin or heart was transplanted into a C57BL/6 (H-2b) recipient treated with anti-CD154 mAb (MR1; 500 mcg/d on days 0, 2, 4, 6) alone, DSG (5 mg/kg/d, days 0 to 7) alone, or both agents. Proliferation of alloreactive T cells after each treatment was also examined using a graft-versus-host disease (GvHD) model using the fluorescent dye CFSE. RESULTS: Treatment with DSG alone induced prolonged survival of the cardiac allografts (median survival time [MST]: 97.5 days). MR1 alone induced indefinite survival of cardiac allografts, although at 150 days after transplantation, the histology showed changes characteristic of chronic rejection, including interstitial fibrosis, infiltration of mononuclear cells, and intimal hyperplasia in coronary vessels. Combined treatment with DSG and MR1 induced donor-specific unresponsiveness in all recipients, graft histology showed only minimal infiltration. Treatment with DSG and MR1 also significantly prolonged the survival of skin allografts (MST: 31 days) compared with that of DSG or MR1 alone (MST: 17 and 14 days, respectively). In the GvHD model assessed with CFSE, the combined treatment was the more effective to suppress proliferation of alloreactive T cells while DSG alone inhibited proliferation more than MR1 alone. CONCLUSION: DSG potentiates anti-CD154 therapy to suppress the alloimmune response.
Assuntos
Guanidinas/farmacologia , Transplante de Coração/imunologia , Isoanticorpos/imunologia , Transplante de Pele/imunologia , Animais , Formação de Anticorpos/efeitos dos fármacos , Sobrevivência de Enxerto/imunologia , Memória Imunológica , Imunossupressores/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Transplante Homólogo/imunologiaRESUMO
Oxygen free radicals have been implicated in the genesis of traumatic brain injury and brain edema (BE). Recent studies have suggested that hydroxyl radical can initiate lipid peroxidation, thus producing lipid-free radicals that may become important sources of singlet oxygen. L-histidine, a singlet oxygen scavenger, potentially can be used to treat BE. In this study we investigated the effects of L-histidine and D-histidine on BE following cryogenic injury in rats. Male Wistar rats were anaesthetized with chloral hydrate. Vasogenic BE was produced by a cortical freezing lesion. Generation of singlet oxygen from photoactivation of rose bengal was studied by electron spin resonance (ESR). Animals were separated into four groups: sham rats (n = 5), saline-treated rats (n = 10), L-histidine treated rats (n = 6) and D-histidine treated rats (n = 7). Each agent (100 mg/kg) was administered intravenously at 30 minutes before lesion production. Animals were sacrificed at 24 hours after lesion production and the brain water content was determined by the dry-wet weight method. L-histidine had no effect on rectal and brain temperature. Election Spin Resonance studies demonstrated that L-histidine is a singlet oxygen scavenger. L-histidine but not D-histidine significantly attenuated BE following cryogenic injury (p < 0.05). In conclusion, L-histidine is useful in the treatment of traumatic BE.