RESUMO
Unpreventable allograft rejection is one of the main problems in pancreatic islet transplantation (PIT). Therefore, it is imperative to develop a more effective immunosuppressive strategy. The blockade of transcription factors has been a central part of T cell-depleting immunosuppressive therapies, as typified by the use of calcineurin inhibitors. The inhibition of activator protein-1 (AP-1) offers a novel strategy for immunosuppression in PIT, although to date, no reports on the effects of AP-1 inhibition are available. In this study, we investigated the immunosuppressive effects of T-5224, a c-Fos/AP-1-selective inhibitor, on murine T cells activated by αCD3+αCD28 mAbs. T-5224 inhibited proliferation, CD25 up-regulation, and the production of IL-2 and interferon-γ. In addition, T-5224 blocked the nuclear translocation of c-Fos/AP-1 in activated murine T cells. In BALB/c (H-2(d) )-to-C57BL/6J (H-2(b) ) mouse PIT, the 2-week administration of T-5224 prolonged survival of 600 islet allografts in a dose-dependent manner. When combined with a 2-week low-dose tacrolimus, the T-5224 treatment markedly prolonged allograft survival to over 300 days, while the efficacy was indeterminate when transplanted islet allograft mass was reduced to 300. We conclude that the c-Fos/AP-1 inhibition by T-5224 is a potentially attractive strategy for allogeneic PIT.
Assuntos
Benzofenonas/uso terapêutico , Rejeição de Enxerto/prevenção & controle , Imunossupressores/uso terapêutico , Transplante das Ilhotas Pancreáticas/imunologia , Isoxazóis/uso terapêutico , Animais , Benzofenonas/farmacologia , Rejeição de Enxerto/imunologia , Imunossupressores/farmacologia , Isoxazóis/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-fos/antagonistas & inibidores , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Linfócitos T/metabolismo , Fator de Transcrição AP-1/antagonistas & inibidores , Transplante HomólogoRESUMO
A strategy for inhibiting CD40 has been considered as an alternative approach for immunosuppression because of undesirable effects of anti-CD154 monoclonal antibodies (mAbs). Previously, we demonstrated that ASKP1240, which is a fully human anti-CD40 mAb, significantly prolonged kidney and liver allograft survival in cynomolgus monkeys without causing thromboembolic complications. Herein, we evaluated the effect of ASKP1240 on pancreatic islet transplantation (PITx) in cynomolgus monkeys. Diabetes was induced by total pancreatectomy, and islet allografts were transplanted into the liver. Following PITx (8201-12 438 IEQ/kg), blood glucose levels normalized promptly in all animals. Control islet allografts were rejected within 9 days (n = 3), whereas ASKP1240 (10 mg/kg) given on postoperative days 0, 4, 7, 11 and 14 (induction treatment, n = 5) significantly prolonged graft survival time (GST) to >15, >23, 210, 250 and >608 days, respectively. When ASKP1240 (5 mg/kg) was administered weekly thereafter up to post-PITx 6 months (maintenance treatment, n = 4), GST was markedly prolonged to >96, >115, 523 and >607 days. During the ASKP1240 treatment period, both anti-donor cellular responses and development of anti-donor antibodies were abolished, and no serious adverse events were noted. ASKP1240 appears to be a promising candidate for immunosuppression in clinical PITx.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Ligante de CD40/imunologia , Diabetes Mellitus Tipo 1/terapia , Sobrevivência de Enxerto , Transplante das Ilhotas Pancreáticas , Ilhotas Pancreáticas/imunologia , Animais , Anticorpos Monoclonais/farmacocinética , Ligante de CD40/antagonistas & inibidores , Ligante de CD40/metabolismo , Diabetes Mellitus Tipo 1/etiologia , Diabetes Mellitus Tipo 1/metabolismo , Humanos , Tolerância Imunológica , Ilhotas Pancreáticas/metabolismo , Macaca fascicularis , Masculino , Pancreatectomia/efeitos adversos , Distribuição Tecidual , Transplante HomólogoRESUMO
BACKGROUND: Dehydroxymethylepoxyquinomicin (DHMEQ) is a newly developed compound that inhibits nuclear factor κB activation and is reported to ameliorate animal models of various inflammatory diseases without significant adverse effects. Because nuclear factor κB is a transcription factor that plays a critical role in the pathophysiology of asthma, DHMEQ may be of therapeutic benefit in asthma. OBJECTIVE: The purpose of this study was to evaluate the effects of DHMEQ on airway inflammation and remodelling in murine models of asthma. METHODS: The BALB/c mice were sensitized and then challenged acutely or chronically with ovalbumin and administered DHMEQ intraperitoneally before each challenge. Inflammation of airways, lung histopathology and airway hyper responsiveness to methacholine challenge were evaluated. In addition, the effect of DHMEQ on production of cytokines and eotaxin-1 by murine splenocytes, human peripheral blood mononuclear cells and bronchial epithelial cells was investigated. RESULTS: Airway hyper responsiveness was ameliorated in both acutely and chronically challenged models by treatment with DHMEQ. DHMEQ significantly reduced eosinophilic airway inflammation and levels of Th2 cytokines in bronchoalveolar lavage fluid in the acute model. It also inhibited parameters of airway remodelling including mucus production, peribronchial fibrosis and the expression of α-smooth muscle actin. Moreover, the production of Th2 cytokines from murine splenocytes and human peripheral blood mononuclear cells and the production of eotaxin-1 by bronchial epithelial cells were inhibited by DHMEQ. CONCLUSIONS AND CLINICAL RELEVANCE: These results indicate that DHMEQ inhibits allergic airway inflammation and airway remodelling in murine models of asthma. DHMEQ may have therapeutic potential in the treatment of asthma.
Assuntos
Remodelação das Vias Aéreas/efeitos dos fármacos , Asma/tratamento farmacológico , Benzamidas/farmacologia , Cicloexanonas/farmacologia , Inflamação/tratamento farmacológico , NF-kappa B/antagonistas & inibidores , Animais , Asma/metabolismo , Asma/patologia , Benzamidas/administração & dosagem , Líquido da Lavagem Broncoalveolar/imunologia , Células Cultivadas , Cicloexanonas/administração & dosagem , Citocinas/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Eosinófilos/imunologia , Eosinófilos/patologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Humanos , Inflamação/metabolismo , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Hipersensibilidade Respiratória/tratamento farmacológico , Hipersensibilidade Respiratória/metabolismoRESUMO
BACKGROUND: Mesothelin is expressed in various types of malignant tumour, and we recently reported that expression of mesothelin was related to an unfavourable patient outcome in pancreatic ductal adenocarcinoma. In this study, we examined the clinicopathological significance of the mesothelin expression in gastric cancer, especially in terms of its association with the staining pattern. METHODS: Tissue specimens from 110 gastric cancer patients were immunohistochemically examined. The staining proportion and intensity of mesothelin expression in tumour cells were analysed, and the localisation of mesothelin was classified into luminal membrane and/or cytoplasmic expression. RESULTS: Mesothelin was positive in 49 cases, and the incidence of mesothelin expression was correlated with lymph-node metastasis. Furthermore, luminal membrane staining of mesothelin was identified in 16 cases, and the incidence of luminal membrane expression was also correlated with pT factor, pStage, lymphatic permeation, blood vessel permeation, recurrence, and poor patient outcome. Multivariate analysis showed that luminal membrane expression of mesothelin was an independent predictor of overall patient survival. CONCLUSION: We described that the luminal membrane expression of mesothelin was a reliable prognostic factor in gastric cancer, suggesting the functional significance of membrane-localised mesothelin in the aggressive behaviour of gastric cancer cells.
Assuntos
Proteínas Ligadas por GPI/metabolismo , Neoplasias Gástricas/metabolismo , Idoso , Biomarcadores Tumorais/metabolismo , Células Epiteliais/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Linfonodos/metabolismo , Metástase Linfática , Masculino , Mesotelina , Pessoa de Meia-Idade , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Análise de SobrevidaRESUMO
Blockade of the CD40-CD154 costimulatory signal is an attractive strategy for immunosuppression and tolerance induction in organ transplantation. Treatment with anti-CD154 monoclonal antibodies (mAbs) results in potent immunosuppression in nonhuman primates (NHPs). Despite plans for future clinical use, further development of these treatments was halted by complications. As an alternative approach, we have been focusing on the inhibition of the counter receptor, CD40 and have shown that a novel human anti-CD40 mAb, ASKP1240, markedly prolongs renal allograft survival in NHPs, although allografts eventually underwent chronic allograft nephropathy. On the basis of our previous findings that a CD40-CD154 costimulation blockade induces tolerance to hepatic, but not cardiac, allografts in rodents, we tested here our hypothesis that a blockade of CD40 by ASKP1240 allows acceptance of hepatic allografts in NHPs. A 2-week ASKP1240 induction treatment prolonged liver allograft survival in NHPs; however, the graft function deteriorated due to chronic rejection. In contrast, a 6-month ASKP1240 maintenance monotherapy efficiently suppressed both cellular and humoral alloimmune responses and prevented rejection on the hepatic allograft. No serious side effects, including thromboembolic complications, were noted in the ASKP1240-treated monkeys. We conclude that CD40 blockade by ASKP1240 would be a desirable immunosuppressant for clinical liver transplantation.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos CD40/imunologia , Transplante de Fígado , Animais , Anticorpos Monoclonais Humanizados , Humanos , Memória Imunológica , Teste de Cultura Mista de Linfócitos , Macaca fascicularis , Masculino , Transplante HomólogoRESUMO
An indirect inguinal hernia containing the entire uterus, ovaries, and Fallopian tubes is extremely rare in pediatrics. The present report describes the very rare case of a 1-month-old girl with an irreducible indirect inguinal hernia containing the entire uterus, ovaries, and Fallopian tubes, and the successful surgical treatment of simple herniorraphy. We review the literature on this type of relationship between indirect inguinal hernia and hernial visceras of the uterus, ovaries, and Fallopian tubes and discuss the clinical features of this complication. Furthermore, the possible cause of indirect inguinal hernia containing the uterus, ovaries, and Fallopian tubes was explored.
Assuntos
Tubas Uterinas/diagnóstico por imagem , Hérnia Inguinal/diagnóstico por imagem , Ovário/diagnóstico por imagem , Útero/diagnóstico por imagem , Feminino , Hérnia Inguinal/complicações , Hérnia Inguinal/cirurgia , Herniorrafia , Humanos , Lactente , UltrassonografiaRESUMO
Blockade of CD40-CD154 signaling pathway is an attractive strategy to induce potent immunosuppression and tolerance in organ transplantation. Due to its strong immunosuppressive effect shown in nonhuman primate experiments, anti-CD154 monoclonal antibodies (mAbs) have been tried in clinical settings, but it was interrupted by unexpected thromboembolic complications. Thus, inhibition of the counter molecule, CD40, has remained an alternative approach. In the previous preliminary study, we have shown that 4D11, a novel fully human anti-CD40 mAb, has a fairly potent immunosuppressive effect on kidney allograft in nonhuman primates. In this study, we aimed to confirm the efficacy and untoward events of the 2-week induction and 180-day maintenance 4D11 treatments. In both, 4D11 significantly suppressed T-cell-mediated alloimmune responses and prolonged allograft survival. Addition of weekly 4D11 administration after the induction treatment further enhanced graft survival. Complete inhibition of both donor-specific Ab and anti-4D11 Ab productions was obtained only with higher-dose maintenance therapy. No serious side effect including thromboembolic complications was noted except for a transient reduction of hematocrit in one animal, and decrease of peripheral B-cell counts in all. These results indicate that the 4D11 appears to be a promising candidate for immunosuppression in clinical organ transplantation.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antígenos CD40/imunologia , Rejeição de Enxerto/prevenção & controle , Transplante de Rim/imunologia , Animais , Anticorpos Monoclonais/sangue , Anticorpos Monoclonais/imunologia , Antígenos CD40/antagonistas & inibidores , Ligante de CD40/antagonistas & inibidores , Ligante de CD40/imunologia , Rejeição de Enxerto/imunologia , Humanos , Terapia de Imunossupressão , Imunossupressores/uso terapêutico , Macaca fascicularis , Masculino , Modelos Animais , Transdução de Sinais/imunologiaRESUMO
IGF2, a maternally imprinted foetal growth factor gene, is implicated in many childhood tumours including hepatoblastoma (HB); however, the genetic and epigenetic alterations have not comprehensively been studied. We analysed the methylation status of the H19 differentially methylated region (DMR), loss of heterozygosity (LOH) and allelic expression of IGF2 in 54 HB tumours, and found that 12 tumours (22%) with LOH, 9 (17%) with loss of imprinting (LOI) and 33 (61%) with retention of imprinting (ROI). Biallelic and monoallelic IGF2 expressions correlated with hypermethylation and normal methylation of H19 DMR, respectively, in two tumours with LOI and seven tumours with ROI. Quantitative RT-PCR analysis showed minimal expression of H19 mRNA and substantial expression of IGF2 mRNA in tumours with LOH or LOI, and substantial expression of both H19 and IGF2 mRNAs in tumours with ROI. Increased IGF2 expression with predominant embryonic P3 transcript was found in the majority of HBs with ROI and foetal livers. In contrast to the earlier reports, our findings suggest that the disruption of the enhancer competition model reported in Wilms' tumour may also occur in HB. Both frequencies of LOH and LOI seem to be lower in HB than in Wilms' tumour, reflecting the different tissue origins.
Assuntos
Metilação de DNA , Impressão Genômica , Hepatoblastoma/genética , Fator de Crescimento Insulin-Like II/genética , Neoplasias Hepáticas/genética , Adolescente , Criança , Pré-Escolar , Proteínas de Ligação a DNA/genética , Humanos , Lactente , Perda de Heterozigosidade , Regiões Promotoras Genéticas , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , beta Catenina/genéticaRESUMO
A 20-year-old female developed a relapse of B-precursor acute lymphoblastic leukemia (ALL) as a mass in her left breast after 6 years of maintained continuous complete remission. No leukemic lesions were identified in other sites such as the bone marrow or cerebrospinal fluid. The relapsed leukemic cells in the breast revealed the same immunophenotypes (CD10(+), CD19(+), CD20(+), HLA-DR(+), CD34(+)) as those of the onset ALL cells in the bone marrow. A literature survey found 10 other cases of ALL relapse in the breast without bone marrow involvement, mostly consisting of adolescent girls. Including the present report, a total of 11 cases were analyzed; the onset ages of ALL were a median of 16.5 (range 5-50) years old and the ages of relapse in the breast a median of 20 (range 12-51) years old. Data suggest that, although rare, the breast could become one of the extramedullary relapse sites of ALL developed in adolescent girls.
Assuntos
Neoplasias da Mama/secundário , Recidiva Local de Neoplasia , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , Adulto , Feminino , HumanosRESUMO
OBJECTIVE: We have established immortalized human hepatocytes by transduction of HPV16 E6/E7 and hTERT (HHE6E7T-1). The cells retained the characteristics of differentiated hepatocytes, but the functional characteristics such as albumin secretion, ureogenesis, and glyconeogenesis decreased gradually as the passages progressed beyond 200 population doublings (PDs). In this report, we transplanted minimally differentiated HHE6E7T-1 cells into the spleens of acute liver failure severe combined immunodeficiency (SCID) mice to examine the potential of the cells to redifferentiate in vivo. MATERIALS AND METHODS: Acute liver failure was induced in SCID mice by intraperitoneal injection of 400 mg/kg acetaminophen. Two hours later, HHE6E7T-1 cells at 200 PDs were transplanted: group 1 (n = 10) 50 microL phosphate-buffered saline (PBS); group 2 (n = 9), lysate of 1 x 10(6) HHE6E7T-1 cells at 200 PDs resuspended in 50 microL PBS; and group 3 (n = 8), 1 x 10(6) HHE6E7T-1 cells. Survival rates at 7 days after transplantation were compared. Blood glucose levels, plasma ammonia levels, and spleen histology were examined at 24 hours after transplantation. RESULTS: Survivals in each group were: 30% for group 1, 33% for group 2, and 100% for group 3. The survival of group 3 was significantly higher than groups 1 or 2 (P < .01). Plasma ammonia levels in group 3 (200 +/- 34 microg/dL) were significantly lower than those in group 1 (325 +/- 92 microg/dL; P < .05). Blood glucose levels in group 3 (110 +/- 20 mg/dL) were significantly higher than those in group 1 (83 +/- 14 mg/dL; P < .05). Upon histologic examination of spleen, the clusters of HHE6E7T-1 cells were clearly identified. CONCLUSIONS: The immortalized human hepatocytes, HHE6E7T-1 at 200 PDs, improved the survival of acute liver failure mice through possible redifferentiation in vivo.
Assuntos
Acetaminofen/toxicidade , Transplante de Células/métodos , Hepatócitos/transplante , Falência Hepática/terapia , Baço , Doença Aguda , Animais , Transplante de Células/mortalidade , Sobrevivência de Enxerto , Humanos , Falência Hepática/induzido quimicamente , Masculino , Camundongos , Camundongos SCID , Análise de Sobrevida , Transplante Heterólogo , Resultado do TratamentoRESUMO
Human neuroblastoma remains enigmatic because it often shows spontaneous regression and aggressive growth. The prognosis of advanced stage of sporadic neuroblastomas is still poor. Here, we investigated whether genomic and molecular signatures could categorize new therapeutic risk groups in primary neuroblastomas. We conducted microarray-based comparative genomic hybridization (array-CGH) with a DNA chip carrying 2464 BAC clones to examine genomic aberrations of 236 neuroblastomas and used in-house cDNA microarrays for gene-expression profiling. Array-CGH demonstrated three major genomic groups of chromosomal aberrations: silent (GGS), partial gains and/or losses (GGP) and whole gains and/or losses (GGW), which well corresponded with the patterns of chromosome 17 abnormalities. They were further classified into subgroups with different outcomes. In 112 sporadic neuroblastomas, MYCN amplification was frequent in GGS (22%) and GGP (53%) and caused serious outcomes in patients. Sporadic tumors with a single copy of MYCN showed the 5-year cumulative survival rates of 89% in GGS, 53% in GGP and 85% in GGW. Molecular signatures also segregated patients into the favorable and unfavorable prognosis groups (P=0.001). Both univariate and multivariate analyses revealed that genomic and molecular signatures were mutually independent, powerful prognostic indicators. Thus, combined genomic and molecular signatures may categorize novel risk groups and confer new clues for allowing tailored or even individualized medicine to patients with neuroblastoma.
Assuntos
Perfilação da Expressão Gênica , Neuroblastoma/diagnóstico , Neuroblastoma/genética , Análise de Sequência com Séries de Oligonucleotídeos , Criança , Pré-Escolar , Aberrações Cromossômicas , Cromossomos Humanos Par 17 , Análise por Conglomerados , Amplificação de Genes , Humanos , Lactente , Recém-Nascido , Proteína Proto-Oncogênica N-Myc , Neuroblastoma/classificação , Neuroblastoma/mortalidade , Proteínas Nucleares/genética , Proteínas Oncogênicas/genética , Prognóstico , Risco , Análise de SobrevidaRESUMO
AIM: There is little reported evidence on the effect of preoperative chemical bowel preparation by the administration of oral antibiotics in patients with biliary atresia (BA). This study was designed to examine and compare the alterations of microbial flora in feces cultures before and after the administration of oral antibiotics, and to examine fluid cultures from the tip of the jejunal limb at bile duct reconstruction by Kasai hepatoportoenterostomy (HPJ), and to evaluate the effects of preoperative oral antibiotics on postoperative cholangitis. MATERIAL AND METHODS: Between January 2003 and July 2005, 6 infants with BA underwent surgical correction, and were treated preoperatively with polymyxin B (80 000 U/kg/day in 3 divided doses for 2 days) and metronidazole (10 mg/kg/day in 2 divided doses for 2 days). Quantitative feces cultures, both before administration of oral antibiotics and at Kasai HPJ, were obtained in patients with BA. Furthermore, fluid cultures from the tip of the jejunal limb at bile duct reconstruction by Kasai HPJ were obtained to examine alterations of bacteria and fungi. RESULTS: Bacterial colonization of the tip of the jejunal limb did not occur at the Kasai HPJ. The most commonly encountered organisms were ENTEROCOCCUS and intestinal bacterial flora. Fungus was not detected in the feces before or after the administration of oral antibiotics. CANDIDA was detected in the tip of the jejunal limb after Kasai HPJ in only one patient. CONCLUSIONS: The authors propose that colonic-type flora are generally found in the feces before and after the administration of oral antibiotics, and no bacteria are detected in the bilioenteric conduits.
Assuntos
Anti-Infecciosos/uso terapêutico , Atresia Biliar/cirurgia , Colo/microbiologia , Enterococcus/isolamento & purificação , Jejunostomia/métodos , Jejuno/microbiologia , Portoenterostomia Hepática/métodos , Colangite/microbiologia , Colangite/prevenção & controle , Contagem de Colônia Microbiana , Quimioterapia Combinada , Fezes/microbiologia , Feminino , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/prevenção & controle , Humanos , Lactente , Recém-Nascido , Masculino , Metronidazol/uso terapêutico , Polimixina B/uso terapêutico , Complicações Pós-Operatórias/microbiologia , Complicações Pós-Operatórias/prevenção & controle , Cuidados Pré-Operatórios/métodos , Estudos Retrospectivos , Fatores de TempoRESUMO
Thymomas are rare mediastinal tumors in pediatrics and cystic thymoma, is characterized by a predominantly cystic formation, is extremely rare. This report describes a 13-year-old girl with cystic thymoma, who was treated by video-assisted thoracoscopic surgery. The microscopic findings were characteristic of cystic thymoma. The literature on cystic thymoma is reviewed here and the clinical characteristics of cystic thymoma are discussed.
Assuntos
Timoma/diagnóstico , Neoplasias do Timo/diagnóstico , Adolescente , Feminino , Humanos , Cirurgia Torácica Vídeoassistida , Timoma/diagnóstico por imagem , Timoma/patologia , Timoma/cirurgia , Neoplasias do Timo/diagnóstico por imagem , Neoplasias do Timo/patologia , Neoplasias do Timo/cirurgia , Tomografia Computadorizada por Raios XRESUMO
Because it is difficult to distinguish preoperatively between a prenatally diagnosed choledochal cyst (CC) and type-1 cystic biliary atresia (BA) by ultrasound scanning or magnetic resonance imaging, some mode of discriminating between the 2 entities is required. The aim of this study was to investigate the immunohistological differences between prenatally diagnosed CC and type-1 cystic BA, using liver biopsy specimens immunostained for CD56. Five children with prenatally diagnosed CC and two children with prenatally diagnosed type-1 cystic BA were identified by fetal ultrasonography between 1985 and 2004. The control group included two children who were operated on at an earlier period due to postnatally diagnosed BA. Liver wedge biopsy in the right lobe was performed at the time of the radical operation. Histological findings of the CD56-stained liver biopsy specimens were classified into 4 categories each, with particular focus on staining distribution and intensity. The staining distribution was classified according to the scale 0 = no staining; 1 = some staining of bile ducts/ductules but staining in less than one-third of portal tracts; 2 = staining in one-third to less than two-thirds of portal tracts; and 3 = staining in more than two-thirds of portal tracts. Staining intensity was scored as follows: 0 = no staining, 1 = weak staining, 2 = moderate staining, and 3 = strong staining. The staining intensity and distribution in the CC group was zero in all 5 cases. The type-1 cystic BA group consisted of patients with scale 1 or 3 staining distribution and score 1 or 2 staining intensity. In the control group, staining distribution was 1 or 3, and staining intensity was 1 or 3. These results indicate that CD56-positive biliary duct cells are present in prenatally diagnosed type-1 cystic BA. The authors suggest that exploratory laparotomy might be avoided and, instead, immunohistological examination using liver biopsy specimens may be a reliable test for the differential diagnosis of CC and type-1 cystic BA in prenatally diagnosed neonates.
Assuntos
Atresia Biliar/diagnóstico , Antígeno CD56/metabolismo , Cisto do Colédoco/diagnóstico , Doenças Fetais/diagnóstico , Fígado/metabolismo , Fígado/patologia , Biópsia , Pré-Escolar , Colangiopancreatografia por Ressonância Magnética , Diagnóstico Diferencial , Feminino , Humanos , Imuno-Histoquímica , Lactente , Gravidez , Estudos Retrospectivos , Ultrassonografia Pré-NatalRESUMO
We have previously defined a homozygously deleted region at chromosome 1p36.2-p36.3 in human neuroblastoma cell lines, NB-1 and NB-C201, and identified six genes including DFF45/ICAD within this region. In this study, we found that NB-C201 cells are much more resistant to various genotoxic stresses such as cisplatin (CDDP) than CHP134 and SH-SY5Y cells that do not have the homozygous deletion. To examine a role(s) of DFF45 in the regulation of apoptosis in response to CDDP, we have established stably DFF45-expressing NB-C201 cell clones (DFF45-1 and DFF45-3) and a control cell clone (NB-C201-C) using a retrovirus-mediated gene transfer. In contrast to NB-C201-C cells, DFF45-3 cells displayed apoptotic nuclear fragmentation in response to CDDP. Although CDDP-induced proteolytic cleavage of procaspase-3 and DFF45 in DFF45-3 cells, we could not detect a typical apoptotic DNA fragmentation. Additionally, deletion analysis revealed that C-terminal region of DFF45 is required for inducing nuclear fragmentation. Unexpectedly, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays demonstrated that DFF45 has undetectable effect on CDDP sensitivity of NB-C201 cells. Taken together, our present results suggest that DFF45/DFF40 system may be sufficient for CDDP-induced nuclear fragmentation but not DNA cleavage.
Assuntos
Proteínas Reguladoras de Apoptose/genética , Cisplatino/farmacologia , Fragmentação do DNA/efeitos dos fármacos , Deleção de Genes , Antineoplásicos/farmacologia , Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Northern Blotting , Western Blotting , Linhagem Celular Tumoral , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/genética , Sobrevivência Celular/efeitos dos fármacos , Clivagem do DNA/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neuroblastoma/genética , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Fatores de Tempo , TransfecçãoRESUMO
Nuclear factor kappa B (NF-kappaB), which exists as heterodimeric complexes composed of p50 and p65, has been shown to play an important role in cell survival processes. In the present study, we found for the first time that NF-kappaB has an ability to induce the ubiquitin-dependent proteasomal degradation of proapoptotic p73alpha. The activation of NF-kappaB in tumor necrosis factor alpha (TNF-alpha)-stimulated H1299 cells resulted in a significant reduction in the amounts of the endogenous p73alpha. Consistent with these results, TNF-alpha-mediated downregulation of p73alpha was observed in wild-type (WT) mouse embryonic fibroblasts (MEFs) but not in p65-deficient MEFs. Ectopic expression of NF-kappaB decreased a half-life of p73alpha by increasing its ubiquitination levels, and thereby inhibiting the transcriptional activity as well as proapoptotic function of p73alpha, whereas NF-kappaB had undetectable effects on p53. Immunoprecipitation experiments demonstrated that, under our experimental conditions, NF-kappaB does not bind to p73alpha in mammalian cultured cells. In contrast to WT p65, the COOH-terminal deletion mutant of p65 (p65DeltaC) failed to reduce the expression levels of p73alpha, suggesting that NF-kappaB-mediated proteolytic degradation of p73alpha requires the transcriptional activity of NF-kappaB. Taken together, our present results imply that NF-kappaB-mediated degradation of proapoptotic p73 is a novel inhibitory mechanism of p73 that regulates cell survival and death.
Assuntos
Proteínas de Ligação a DNA/metabolismo , NF-kappa B/metabolismo , Proteínas Nucleares/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Ubiquitina/metabolismo , Animais , Células COS , Linhagem Celular Tumoral , Chlorocebus aethiops , Regulação para Baixo , Embrião de Mamíferos/citologia , Fibroblastos/metabolismo , Deleção de Genes , Humanos , Camundongos , Subunidade p50 de NF-kappa B/genética , Subunidade p50 de NF-kappa B/metabolismo , Processamento de Proteína Pós-Traducional , Sinaptotagmina I/genética , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Proteína Tumoral p73RESUMO
PURPOSE: The aim of this study was to determine an appropriate management plan for childhood and adolescent FNH, in particular to establish an algorithm for preoperative diagnosis and treatment. PATIENTS AND METHODS: Between 1985 and 2003, 4 children with FNH were diagnosed. Of these 4 patients, 3 (Group A) underwent tumor resection, and 1 (Group B) was treated by conservative management. Clinical data, pathological findings and follow-up were evaluated retrospectively. RESULTS: The 3 patients in Group A were symptomatic, while the 1 patient in Group B was asymptomatic. In 3 of 4 patients, a homogeneous tumor with a central stellate area was noted on abdominal ultrasonography, CT scan and MR imaging. In case 2, SPIO-enhanced MR imaging was useful for differentiating FNH from hepatocellular carcinoma. Though percutaneous needle biopsy was performed in case 3, a pathologically definitive diagnosis was impossible. An open biopsy was performed in case 4 and FNH was diagnosed. In case 4 treated by conservative management, the tumor size did not change during the 7 years after the diagnosis of FNH. CONCLUSION: FNH is usually treated conservatively because of the good evolutionary outcome of the lesion. Surgery is indicated in cases of complications, compressed adjacent organs, lesion progression, or for symptomatic patients. We advocate the use of less invasive SPIO-enhanced MR imaging instead of open biopsy when the diagnosis of focal liver lesions is not clear after contrast-enhanced CT scan and non-enhanced MR imaging.
Assuntos
Hiperplasia Nodular Focal do Fígado/terapia , Adolescente , Biópsia por Agulha , Criança , Hiperplasia Nodular Focal do Fígado/diagnóstico , Hiperplasia Nodular Focal do Fígado/patologia , Hiperplasia Nodular Focal do Fígado/cirurgia , Humanos , Imageamento por Ressonância Magnética , Estudos Retrospectivos , Tomografia Computadorizada por Raios XRESUMO
Recessive dystrophic epidermolysis bullosa (RDEB) is an inherited disorder of squamous epithelium that results in dystrophic scarring of the skin after minor trauma. RDEB is classified into two subtypes: Hallopeau-Siemens (HS) and non-Hallopeau-Siemens (nHS). Although severe scarring of the skin is the most common and obvious complication of the disease, esophageal scarring with formation of long strictures may also develop. Treatment options for esophageal stenosis in patients with RDEB include steroids, hyperalimentation, esophageal dilation and replacement. This report describes a child who was dilated immediately after diagnosis of severe esophageal stenosis subsequent to nHS-RDEB and managed successfully. Endoscopic esophageal balloon dilation under fluoroscopic control was very useful for detecting the region of stenosis and bougienage. The literature on such injuries is reviewed here, and the problems associated with the treatment of children with esophageal stenosis associated with RDEB are discussed.
Assuntos
Cateterismo/métodos , Epidermólise Bolhosa Distrófica/complicações , Estenose Esofágica/terapia , Criança , Transtornos de Deglutição/etiologia , Transtornos de Deglutição/terapia , Estenose Esofágica/etiologia , Estenose Esofágica/patologia , Esofagoscopia , Humanos , MasculinoRESUMO
AIM: This analysis was performed to evaluate clinical factors such as local tumor control, control of distant metastasis, survival, and complications in children and adolescents with thyroid carcinoma. MATERIALS AND METHODS: From 1960 to 2003, 21 children and adolescents (16 girls and 5 boys, aged 4 to 15 years) were treated for thyroid carcinoma in our departments. Clinical data, operative findings, pathological findings, operative procedures, intra- and postoperative complications and follow-up were evaluated. Follow-up of patients ranged from 3 to 42 years (median, 20 years). RESULTS: Papillary carcinoma was found in 20 patients (95%) and follicular carcinoma in one patient. Cervical lymph node metastases were found in 12 out of 21 patients (57.1%) when thyroid carcinoma was diagnosed, and 5 patients (23.8%) had pulmonary metastasis. In patients with pulmonary metastasis, total thyroidectomy and bilateral radical lymph node resection was performed in 3 patients, lobectomy of the thyroid and radical lymph node resection in one patient, and partial thyroidectomy in one patient. All three patients who underwent total thyroidectomy are still alive. Internal irradiation by 131I and thyroid hormone replacement therapy was administered to 2 of 3 patients with pulmonary metastasis. CONCLUSION: Despite metastasis in the lymph nodes and even the lungs, the prognosis for patients with papillary carcinoma is good. Total thyroidectomy and positive 131I therapy are recommended for childhood and adolescent thyroid carcinoma with pulmonary metastasis.
Assuntos
Carcinoma Papilar/terapia , Neoplasias da Glândula Tireoide/terapia , Adenocarcinoma Folicular/patologia , Adenocarcinoma Folicular/terapia , Adolescente , Carcinoma Papilar/patologia , Carcinoma Papilar/secundário , Criança , Pré-Escolar , Feminino , Humanos , Neoplasias Pulmonares/secundário , Metástase Linfática , Masculino , Recidiva Local de Neoplasia , Radioterapia Adjuvante , Estudos Retrospectivos , Análise de Sobrevida , Neoplasias da Glândula Tireoide/patologia , TireoidectomiaRESUMO
Because it is difficult to distinguish preoperatively between prenatally diagnosed choledochal cyst (CC) and type I cystic biliary atresia (BA) by ultrasound scanning or magnetic resonance imaging (MRI), some modality of discrimination for the 2 entities is required. The aim of this study was to investigate the histological differences between prenatally diagnosed CC and type I cystic BA using liver biopsy specimens. Four children with prenatally diagnosed CC and two children with prenatally diagnosed type I cystic BA were identified by fetal ultrasonography between 1985 and 2003. The control group included two children who were operated on at an earlier period due to postnatally diagnosed BA. Liver wedge biopsy in the right lobe was performed at the time of the radical operation. Histological findings of the H & E-stained liver biopsy specimens were classified into 4 grades (Grade 0, no abnormality; Grade 1, ductular proliferation without bridging fibrosis; Grade 2, ductular proliferation with bridging fibrosis; and Grade 3, liver cirrhosis). The CC group consisted of one case of Grade 0 and three cases of Grade 1. The type I cystic BA group consisted of one case each of Grade 2 and Grade 3. In the control group, both cases were Grade 2. The authors suggest that exploratory laparotomy might be avoided and, instead, histologic examination using liver biopsy specimens can be a reliable test for the differential diagnosis of CC and type I cystic BA in prenatally diagnosed neonates.