RESUMO
AIM: To determine the usefulness of three-dimensional reversed fast imaging with steady-state precession diffusion-weighted imaging (3D-PSIF DWI) for the detection of middle ear cholesteatoma. MATERIALS AND METHODS: The study population consisted of 81 patients who underwent 3D-PSIF-DWI at 3 T. They included cholesteatoma in 73 cases, otitis media in five, and cholesterol granuloma in three. Two observers independently performed qualitative evaluations for the detection of cholesteatoma and measured apparent diffusion coefficient (ADC) values and ADC ratios of the lesions. Kappa (κ) statistics, the intraclass correlation coefficient (ICC), the independent t-test, and receiver operating characteristic (ROC) analysis were used for statistical analysis. Pair-wise comparison of the ROC curves was performed using the area under the ROC curve (AUC). RESULTS: Interobserver agreement and ICC for the qualitative and quantitative evaluations were excellent (κ=0.92 and ICC=0.90-0.92, respectively). The ADC value and the ADC ratio were significantly lower for cholesteatoma than non-cholesteatoma lesions (p<0.0001). In <5 mm cholesteatoma group, the diagnostic performance of the ADC value (AUC=0.97) and the ADC ratio (AUC=1) was significantly superior to qualitative 3D-PSIF-DWI (AUC=0.76; p=0.0001 and <0.0001, respectively). For ≥5 mm cholesteatoma group, there were no significant differences in diagnostic performance among the three parameters. CONCLUSION: 3D-PSIF-DWI sequence is useful for the detection of middle ear cholesteatomas, especially <5 mm lesions.
Assuntos
Colesteatoma da Orelha Média/diagnóstico , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Colesteatoma da Orelha Média/cirurgia , Protocolos Clínicos , Imagem de Difusão por Ressonância Magnética/métodos , Feminino , Humanos , Imageamento Tridimensional , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Recidiva , Estudos Retrospectivos , Adulto JovemRESUMO
Human noggin (NOG) is a responsible gene for multiple synostosis syndrome (SYNS1) and proximal symphalangism (SYM1), two conditions that are recently known to be within a wider range of clinical manifestations of stapes ankylosis with symphalangism. This study was performed to determine the range of phenotype caused by NOG mutations, using Japanese patients with various phenotypes including sporadic inherited SYM1, dominantly inherited SYM1, stapes ankylosis with broad thumb and toes (Teunissen and Cremer syndrome). In addition, 33 patients with typical otosclerosis (without symphalangism) were studied. Direct sequencing analysis disclosed three novel mutations of the NOG gene in three SYM1 families. None of the otosclerosis patients without symphalangism had NOG mutations, indicating that NOG mutations may be restrictively found within patients with various skeletal abnormalities. These results together with the literature review indicated that there are no clear genotype-phenotype correlations for NOG mutations. With regard to surgical outcome, most of the patients in these three families with NOG mutations showed remarkable air-bone gap recovery after stapes surgery. Molecular genetic testing is useful to differentiate syndromic stapes ankylosis from otosclerosis, and even mild skeletal anomalies can be a diagnostic indicator of NOG-associated disease.
Assuntos
Anquilose/genética , Proteínas de Transporte/genética , Predisposição Genética para Doença/genética , Hiperopia/genética , Artropatias/congênito , Fenótipo , Sindactilia/genética , Adulto , Idoso , Anquilose/patologia , Povo Asiático/genética , Ossos do Carpo/anormalidades , Primers do DNA/genética , Feminino , Articulações dos Dedos/anormalidades , Articulações dos Dedos/patologia , Deformidades Congênitas do Pé , Deformidades Congênitas da Mão , Perda Auditiva Condutiva/genética , Perda Auditiva Condutiva/patologia , Humanos , Hiperopia/patologia , Artropatias/genética , Artropatias/patologia , Masculino , Pessoa de Meia-Idade , Mutação/genética , Otosclerose/genética , Linhagem , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Estribo/anormalidades , Estribo/patologia , Sindactilia/patologia , Sinostose , Ossos do Tarso/anormalidades , Polegar/anormalidades , Polegar/patologia , Dedos do Pé/anormalidades , Dedos do Pé/patologiaRESUMO
BACKGROUND: Subjective tinnitus is a common symptom, and there is often an underlying otological cause. This study investigated the degree of tinnitus-related annoyance in patients with chronic otitis media and analysed whether associations with tinnitus severity exist. METHOD: The multinational collaborative Chronic Otitis Media Questionnaire-12 study collected prospective data on 478 adult patients suffering from chronic otitis media across 9 otology referral centres in 8 countries. Based on this dataset, we investigated tinnitus severity using participant responses to item 7 of a native version of the Chronic Otitis Media Questionnaire-12. RESULTS: With respect to tinnitus severity, 23.8 per cent, 17.4 per cent, 15.5 per cent, and 43.4 per cent of participants reported no, minor, moderate, and major inconvenience or greater, respectively. The absence of ear discharge, absence of cholesteatoma, and poorer disease-specific health-related quality-of-life were associated with increased tinnitus severity in patients with chronic otitis media, whereas age, hearing disability and geographical region showed no association. CONCLUSION: This analysis provided novel insight into potential risk factors for tinnitus in patients with chronic otitis media.
Assuntos
Otite Média , Zumbido , Humanos , Adulto , Zumbido/epidemiologia , Zumbido/etiologia , Estudos Prospectivos , Otite Média/complicações , Otite Média/epidemiologia , Inquéritos e Questionários , Doença Crônica , Fatores de RiscoRESUMO
Galanin receptor 1 (GALR1) maps to a common region of 18q loss in head and neck squamous cell carcinomas and is frequently inactivated by methylation. To investigate effects of GALR1 and its signaling pathways, we stably expressed hemaglutinin-tagged GALR1 in a human oral carcinoma cell line (UM-SCC-1-GALR1) that expresses no endogenous GALR1. In transfected cells, galanin induced activation of the extracellular-regulated protein kinase-1/2 (ERK1/2) and suppressed proliferation. Galanin stimulation mediated decreased expression of cyclin D1 and increased expression of the cyclin-dependent kinase inhibitors (CKI), p27(Kip1) and p57(Kip2). Pretreatment with the ERK1/2-specific inhibitor U0126 prevented these galanin-induced effects. Phosphatidylinositol 3-kinase (PI3K) pathway activation did not differ in UM-SCC-1-GALR1 and UM-SCC-1-mock cells after galanin treatment. Pertussis toxin and LY294002 inhibition demonstrated that galanin and GALR1 induce ERK1/2 activation via Galphai, not the PI3K pathway-linked to the Gbetagamma subunit. Galanin and GALR1 also inhibit colony formation and tumor growth in vivo. Our results implicate GALR1, a Gi protein-coupled receptor, as a tumor suppressor gene that inhibits cell proliferation via ERK1/2 activation.
Assuntos
Carcinoma de Células Escamosas/patologia , Proliferação de Células/efeitos dos fármacos , Proteínas Inibidoras de Quinase Dependente de Ciclina/metabolismo , Galanina/farmacologia , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Receptor Tipo 1 de Galanina/fisiologia , Carcinoma de Células Escamosas/enzimologia , Ensaio de Unidades Formadoras de Colônias , Ciclina D1/metabolismo , Ativação Enzimática/efeitos dos fármacos , Proteína Ligante Fas/metabolismo , Humanos , Immunoblotting , Neoplasias Bucais/enzimologia , Neoplasias Bucais/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Fosfatidilinositol 3-Quinases/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Transfecção , Células Tumorais CultivadasRESUMO
The c-kit proto-oncogene encodes a receptor tyrosine kinase. Binding of c-kit ligand, stem cell factor (SCF) to c-kit receptor (c-kitR) is known to activate c-kitR tyrosine kinase, thereby leading to autophosphorylation of c-kitR on tyrosine and to association of c-kitR with substrates such as phosphatidylinositol 3-kinase (PI3K). In a human mast cell leukemia cell line HMC-1, c-kitR was found to be constitutively phosphorylated on tyrosine, activated, and associated with PI3K without the addition of SCF. The expression of SCF mRNA transcript in HMC-1 cells was not detectable by means of PCR after reverse transcription (RT-PCR) analysis, suggesting that the constitutive activation of c-kitR was ligand independent. Sequencing of whole coding region of c-kit cDNA revealed that c-kit genes of HMC-1 cells were composed of a normal, wild-type allele and a mutant allele with two point mutations resulting in intracellular amino acid substitutions of Gly-560 for Val and Val-816 for Asp. Amino acid sequences in the regions of the two mutations are completely conserved in all of mouse, rat, and human c-kit. In order to determine the causal role of these mutations in the constitutive activation, murine c-kit mutants encoding Gly-559 and/or Val-814, corresponding to human Gly-560 and/or Val-816, were constructed by site-directed mutagenesis and expressed in a human embryonic kidney cell line, 293T cells. In the transfected cells, both c-kitR (Gly-559, Val-814) and c-kitR (Val-814) were abundantly phosphorylated on tyrosine and activated in immune complex kinase reaction in the absence of SCF, whereas tyrosine phosphorylation and activation of c-kitR (Gly-559) or wild-type c-kitR was modest or little, respectively. These results suggest that conversion of Asp-816 to Val in human c-kitR may be an activating mutation and responsible for the constitutive activation of c-kitR in HMC-1 cells.
Assuntos
Leucemia de Mastócitos/genética , Mutação Puntual , Proteínas Proto-Oncogênicas/genética , Proto-Oncogenes , Receptores Proteína Tirosina Quinases/genética , Receptores de Fator Estimulador de Colônias/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , Primers do DNA , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Interleucina-3/farmacologia , Camundongos , Dados de Sequência Molecular , Fosfotirosina , Reação em Cadeia da Polimerase , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-kit , Receptores Proteína Tirosina Quinases/biossíntese , Receptores Proteína Tirosina Quinases/metabolismo , Receptores de Fator Estimulador de Colônias/biossíntese , Receptores de Fator Estimulador de Colônias/metabolismo , Proteínas Recombinantes/farmacologia , Transfecção , Células Tumorais Cultivadas , Tirosina/análogos & derivados , Tirosina/análiseRESUMO
Recently, we reported that hepatic allografts are permanently accepted when transplanted after intraportal injection (IP) of donor spleen cells (SPCs). The mechanism of this effect was investigated using various protocols for antigen injection. ACI (RT1a) and Buffalo (RT1b) rats were used as donors and recipients, respectively. Experimental groups were divided into the following groups depending on treatment: IP, intravenous injection (IV), splenectomy (Spx), and intravenous injection after splenectomy (IV+Spx). Accumulation of donor antigen in the various organs was examined by injecting 51Cr-labeled SPCs. Cellular and humoral responses after SPC injection was assessed by delayed-type hypersensitivity response and complement-dependent cytotoxicity (CDC) assay. Heterotopic cardiac transplantation and orthotopic hepatic transplantation were performed 10 days after each treatment. The accumulation ratio in the liver was significantly higher in the IP and IV+Spx groups than in the IV group. Delayed-type hypersensitivity responses were lower in the IP and IV+Spx groups than in the IV or Spx groups. The CDC titer 7 days after inoculation was significantly lower in the IP and IV+Spx groups than in the IV group. In order to examine whether the treatment actively suppressed the immune response, the animals were rechallenged with SPCs given intravenously 10 days after the initial injection. Elevation of CDC titer was suppressed in the IP and IV+Spx groups, but not in the IV and Spx groups. Significant prolongation of cardiac allograft survival was not observed in the IV and Spx groups. Prolongation was observed in the IP and IV+Spx groups. The survival of hepatic allografts in the IV group was decreased. No significant prolongation of graft survival was observed in the Spx group. In contrast, all hepatic allotransplants in the IP and IV+Spx groups survived over 45 days. These findings suggest that the accumulation of donor SPCs in the liver may play an important role in inducing unresponsiveness after intraportal injection of donor SPCs.
Assuntos
Terapia de Imunossupressão/métodos , Isoantígenos/administração & dosagem , Transplante de Fígado/imunologia , Animais , Estudos de Avaliação como Assunto , Sobrevivência de Enxerto , Transplante de Coração/imunologia , Hipersensibilidade Tardia/prevenção & controle , Injeções Intravenosas , Isoanticorpos/biossíntese , Transplante de Fígado/patologia , Masculino , Veia Porta , Ratos , Ratos Endogâmicos ACI , Ratos Endogâmicos BUF , Baço/citologia , Baço/imunologia , Esplenectomia , Fatores de Tempo , Doadores de Tecidos , Transplante HomólogoRESUMO
This study provides evidence of antibodies playing an important role in hamster-to-rat cardiac xenograft rejection and discusses the use of 15-deoxyspergualin (DSG) to suppress this first-set rejection, as well as hyperacute rejection induced by sensitization. The effect of recipient splenectomy (Spx) as an adjuvant to DSG to control first set xenograft rejection was also examined. When hyperimmune serum was taken from control recipients at rejection time and injected i.v. into new recipients of cardiac xenografts, hyperacute graft rejection resulted. Survival depended on the amount of serum injected and ranged from 14.7 +/- 2.5 min with 3 ml of serum to 233.3 +/- 61.1 min with 0.5 ml. Experiments on first-set xenograft rejection revealed that a dose of 2.5 mg/kg/day DSG could prolong xenograft survival from 3.4 +/- 0.5 days in untreated controls to 9.5 +/- 2.6 days (P less than 0.01). A dose of 5 mg/kg/day DSG, though it increased graft survival to 16.4 +/- 5.9 days, proved to be toxic to the recipients. Spx alone prolonged xenograft survival to 5.2 +/- 0.4 days, and, when combined with 2.5 mg/kg/day DSG administration, prolonged graft survival to 22.1 +/- 5.5 days (P less than 0.01 vs. DSG alone). The appearance of cytotoxic antibodies was delayed, and titers decreased from 1:256 in untreated controls to 1:16-1:64 both in the group that underwent Spx only and in the group that received 2.5 mg/kg/day DSG. Combined treatment suppressed antibody response for more than two weeks. Experiments on hyperacute rejection induced by sensitization revealed that 1 ml of hamster whole blood transfused into prospective heart recipients 1 week before grafting resulted in graft loss in 18.2 +/- 6.1 min. Pretransplant transfusion and concomitant daily administration of 5 mg/kg/day DSG until one day after grafting not only prevented hyperacute rejection but prolonged graft survival to 7.0 +/- 0.7 days. This survival was significantly longer than with DSG alone (4.2 +/- 0.8 days, P less than 0.01). We concluded that the marked suppression of antibody formation by DSG plays a major role in preventing first-set xenograft rejection and hyperacute rejection induced by sensitization.
Assuntos
Formação de Anticorpos/efeitos dos fármacos , Rejeição de Enxerto/efeitos dos fármacos , Guanidinas/farmacologia , Transplante de Coração , Imunossupressores/farmacologia , Animais , Cricetinae , Feminino , Imunização , Masculino , Mesocricetus , Ratos , Ratos Endogâmicos , Esplenectomia , Transplante HeterólogoRESUMO
The effect of donor-specific blood transfusion (DST), in combination with pretransplant immunosuppression with deoxyspergualin (DSG), on hamster-to-Wistar rat cardiac xenograft survival was assessed. While DST given on day -6 sensitized the recipients, resulting in hyperacute xenograft rejection, the addition of 5 mg/kg/k/day DSG from the day of transfusion to the day of grafting not only prevented hyperacute rejection but resulted in prolongation of graft survival from 3.4 +/- 0.5 days in untreated controls to 7.0 +/- 0.7 days (P less than 0.01). In contrast, DSG alone as pretransplant immunosuppression had no beneficial effect and rejection occurred in 4.0 +/- 0.7 days. This effect appears to be at least donor species-specific, in the sense that ACI cardiac allograft survival was not prolonged when transplanted into xenotransfused and DSG-treated Wistar recipients. DST alone resulted in marked increase in antibody titers, showing the value of 1:512 or more on transplantation day. On the other hand, combined treatment suppressed the titers to 1:1-1:4 on that day. An adoptive cell transfer system was used to analyze the mechanisms underlying this effect. When sublethally irradiated secondary hosts were transferred with 5 x 10(7) lymph node cells (LNCs) harvested on day 0 from xenotransfused and DSG-treated rats, the test heart xenograft survived longer than the irradiated and nontransferred controls, suggesting the presence of suppressor cells. Further in vitro studies demonstrate that LNCs from DST+DSG-treated rats response less in a mixed lymphocyte reaction to hamster LNCs (41% on day 0 [P less than 0.01]), compared with the controls. In coculture experiments, the LNCs from treated recipients suppressed the response of unmodified Wistar LNCs to hamster LNCs by 76% on day 0 compared with the positive controls (P less than 0.01). On the other hand, the transfer of serum taken from treated rats on day 0 did not lead to prolongation of test heart xenografts in syngeneic naive hosts. These findings suggest that the mechanisms underlying the hyporesponsiveness induced by pretreatment with DST and DSG include the induction of suppressor cells, although a degree of clonal deletion can not be ruled out. The generation of serum suppressor factors seems to have no role in this phenomenon.
Assuntos
Transfusão de Sangue , Guanidinas/farmacologia , Transplante de Coração/imunologia , Linfócitos T Reguladores/imunologia , Transplante Heterólogo/imunologia , Animais , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Cricetinae , Transplante de Coração/patologia , Linfonodos/citologia , Linfonodos/imunologia , Teste de Cultura Mista de Linfócitos , Linfócitos/efeitos dos fármacos , Mesocricetus , Ratos , Ratos Endogâmicos , Linfócitos T Reguladores/efeitos dos fármacosRESUMO
Interleukin 6 has recently been noted to be present during the rejection response to grafted organs. In this study, we investigated biliary and serum interleukin 6 levels following liver transplantation in rats. IL-6 levels in bile and serum of naive rats were below 0.6 U/ml and 0.5 +/- 0.2 U/ml (mean +/- SD), respectively. Both biliary and serum IL-6 levels showed high values (greater than 10.0 U/ml and greater than 1.6 U/ml, respectively) on the day after transplantation, which seemed to reflect the inflammatory status caused by the surgical stress. Later samplings showed that the kinetics of serum IL-6 differed among the animals without any definite feature related to graft rejection. In contrast, biliary IL-6 levels correlated well with the severity of the rejection response as determined histologically. Biliary IL-6 levels started to rise at the onset of the rejection response (6.6 +/- 0.6 U/ml), increased further with its progression (19.3 +/- 7.8 U/ml), and then finally fell in the terminal stage (less than 2.0 U/ml). Elevation of biliary IL-6 was observed at an early stage when abnormalities could be detected histologically but not in liver function tests and bile flow. Therefore, biliary IL-6 levels may be of value for the early diagnosis of rejection following liver transplantation.
Assuntos
Bile/química , Interleucina-6/análise , Transplante de Fígado/imunologia , Animais , Rejeição de Enxerto/fisiologia , Sobrevivência de Enxerto , Interleucina-6/sangue , Masculino , Ratos , Ratos Endogâmicos ACI , Ratos Endogâmicos Lew , Transplante Homólogo/imunologia , Transplante Isogênico/imunologiaRESUMO
T-lymphocyte activation requires engagement of the T cell receptor with antigen-major histocompatibility complex, and simultaneous ligation of costimulatory pathways via the lymphocyte receptors CD2 and CD28/ CTLA4. Anti-CD2 monoclonal antibody (mAb) blocks the interaction of the antigen-presenting cell receptor CD48 with its ligand CD2, whereas CTLA4Ig binds with high affinity to the antigen-presenting cell ligands B7-1 and B7-2, blocking their interaction with CD28/CTLA4. We tested the immunosuppressive effects of simultaneously blocking both costimulatory pathways. Using donor C57BL/6J (H2b) hearts transplanted to CBA/J (H2k) recipients, anti-CD2 mAb plus CTLA4Ig administered at the time of transplantation prolonged cardiac allograft mean survival time to >120 days compared with untreated controls (12.2+/-0.5 days, P<0.01), anti-CD2 mAb alone (24.8+/-1.0 days, P<0.01), or CTLA4Ig alone (55.0+/-2.0 days, P<0.01). Retransplantation of these recipients with donor-specific and third-party grafts demonstrated that hyporesponsiveness and tolerance were achieved. In vitro stimulation of lymphocytes from tolerant recipients with donor-specific alloantigen resulted in normal cytotoxic T lymphocyte and mixed lymphocyte reaction responses, showing that clonal deletion or anergy did not occur, but that graft adaptation or suppression likely helped to maintain long-term graft survival. In vitro combinations of anti-CD2 mAb and CTLA4Ig suppressed the generation of allogeneic cytotoxic T lymphocytes (58%) and the mixed lymphocyte reaction (36%); CTLA4Ig was more effective in this regard and the two agents were not synergistic. Anti-CD2 mAb and CTLA4Ig suppressed mitogen-driven proliferation in differential fashions, suggesting that they affected independent signaling pathways. Anti-CD2 mAb and CTLA4Ig also inhibited interleukin (IL)-2, IL-4, and IL-2 receptor (CD25). These data indicate that anti-CD2 mAb plus CTLA4Ig induces hyporesponsiveness and tolerance. The mechanism is likely related to the initial disruption of independent pathways of T-lymphocyte activation leading to antigen-specific long-term graft survival.
Assuntos
Antígenos de Diferenciação/farmacologia , Antígenos CD2/efeitos dos fármacos , Antígenos CD28/efeitos dos fármacos , Dessensibilização Imunológica , Imunoconjugados , Sistemas do Segundo Mensageiro/efeitos dos fármacos , Linfócitos T/imunologia , Linfócitos T/ultraestrutura , Abatacepte , Animais , Anticorpos Monoclonais/farmacologia , Antígenos CD , Antígenos CD2/imunologia , Antígenos CD28/imunologia , Antígeno CTLA-4 , Citotoxicidade Imunológica , Epitopos , Feminino , Sobrevivência de Enxerto/efeitos dos fármacos , Sobrevivência de Enxerto/imunologia , Transplante de Coração/imunologia , Humanos , Isoantígenos/imunologia , Ativação Linfocitária/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos , Gravidez , Ratos , Linfócitos T/efeitos dos fármacos , Fatores de TempoRESUMO
Thymidine phosphorylase (dThdPase) is the rate-limiting enzyme that metabolizes 5'-deoxy-5-fluorouridine (5'-dFUrd, doxifluridine), an intermediate metabolite of capecitabine, to the active drug 5-fluorouracil (5-FUra), while dihydropyrimidine dehydrogenase (DPD) catabolizes 5-FUra to an inactive molecule. The susceptibility of tumors to fluoropyrimidines is reported to correlate with tumor levels of these enzymes. To obtain some insight into the tumor types susceptible to fluoropyrimidine therapy, we measured expression levels of these two enzymes in various types of human cancer tissues (241 tissue samples) by the ELISA methods. DPD exists in all the cancer types studied, such as bladder, breast, cervical, colorectal, esophageal, gastric, hepatic, pancreatic, prostate, and renal cancers. Among them, the cervical, hepatic, pancreatic, esophageal, and breast cancer tissues expressed high levels of DPD (median >70 U/mg protein), while high concentrations of the dThdPase were expressed in esophageal, cervical, breast, and pancreatic cancers and hepatoma (median >150 U/mg protein). The dThdPase/DPD ratio, which was reported to correlate with the susceptibility of human cancer xenografts to capecitabine, was high in esophageal, renal, breast, colorectal, and gastric cancers (median ratio of >1.5). In any of these three parameters, the inter-patient DPD variability for each cancer type was much larger than the DPD variability among cancer types; highest/lowest ratios for dThdPase, DPD, and dThdPase/DPD were 10-321, 7-513, and 2-293, respectively. These results indicate that measurements of the three parameters, DPD, dThdPase and dThdPase/DPD, would be useful criteria for selecting cancer patients suitable for fluoropyrimidine therapy rather than for selecting cancer types.
Assuntos
Neoplasias/enzimologia , Oxirredutases/metabolismo , Timidina Fosforilase/metabolismo , Animais , Antineoplásicos/uso terapêutico , Capecitabina , Desoxicitidina/análogos & derivados , Desoxicitidina/uso terapêutico , Di-Hidrouracila Desidrogenase (NADP) , Ensaio de Imunoadsorção Enzimática , Feminino , Fluoruracila/análogos & derivados , Humanos , Japão , Masculino , Neoplasias/tratamento farmacológico , Oxirredutases/análise , Timidina Fosforilase/análise , Transplante HeterólogoRESUMO
Expression of mRNA for pituitary adenylate cyclase-activating polypeptide (PACAP) was detected in the cochlea of rats using a reverse transcription-polymerase chain reaction and in situ hybridization. Examination of in situ hybridization demonstrated that cells in the spiral ganglion, and marginal cells in the stria vascularis expressed mRNA for PACAP. These findings suggest that PACAP may play an important role in regulating cochlear functions.
Assuntos
Cóclea/fisiologia , Neuropeptídeos/genética , Animais , Expressão Gênica/fisiologia , Hibridização In Situ , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , RNA Mensageiro/análise , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Antigen pretreatment through the oral or intraportal route has been reported to suppress antibody formation or delayed-type hypersensitivity (DTH) response to the same antigens. However, this effect on allografted organs has not been well studied. In this study we evaluated the efficacy for suppression of antibody formation and DTH response to the allogeneic antigens and tried to prolong liver allograft survival in rats. Male ACI (RT1a) rats were used as donors and male BUF (RT1b) rats as recipients. Intraportal or intravenous injection of spleen cells (SPCs) (5 x 10(7)) was performed through the mesenteric vein or the tail vein, respectively. The anti-ACI DTH response was tested by ear challenge of ACI SPCs. Cytotoxic antibody was assessed by complement-dependent cytotoxicity assay. ACI rat liver was transplanted orthotopically to BUF rat by the cuff technique 10 days after SPC injection. Cytotoxic antibody titer rose to X2(6) to X2(8) at 7 to 10 days after intravenous injection of SPCs. However, intraportal injection of the cells rarely caused a rise in antibody titer and even strongly suppressed subsequent antibody formation induced by intravenous injection when given 10 days before. The DTH response was also suppressed by intraportal injection of SPCs, with a mean value of 0.18 +/- 0.13 mm versus 0.67 +/- 0.19 mm for controls or 0.46 +/- 0.04 mm with intravenous injection. Liver-allografted rats died between 9 and 11 days, averaging 10.1 +/- 0.7 days in the control group. All seven transplants injected intraportally with donor SPCs survived more than 100 days, whereas six of eight rats injected intravenously with donor SPCs died of bleeding from the surface of the liver grafts within 12 hours after grafting, with signs similar to those of hyperacute rejection. Four of five rats injected intraportally with F344 (third-party) SPCs died of acute rejection in the same way the control rats died. The liver allograft-bearing rats had permanently accepted ACI skin grafts when tested 60 days after liver transplantation but rejected F344 skin grafts in the normal fashion. Intraportal injection of donor SPCs markedly suppressed the antibody formation, as well as the DTH response, and completely blocked liver allograft rejection. Moreover, the liver allograft-bearing rats proved to be fully tolerant of the donor antigen. This method might be a promising modality for inducing donor-specific tolerance in liver transplantation.
Assuntos
Sobrevivência de Enxerto , Terapia de Imunossupressão/métodos , Transplante de Fígado , Baço/citologia , Animais , Formação de Anticorpos , Antígenos/fisiologia , Hipersensibilidade Tardia/imunologia , Injeções Intravenosas , Fígado/citologia , Tecido Linfoide/transplante , Masculino , Veia Porta , Ratos , Ratos Endogâmicos BUF , Ratos Endogâmicos F344 , Ratos Endogâmicos , Baço/imunologia , Transplante HomólogoRESUMO
Intracellular free Ca2+ concentration ([Ca2+]i) in spiral ganglion cells (SGC) of guinea pig was measured using the fura-2 fluorescence method. Application of high-K+ external solution increased [Ca2+]i. [Ca2+]i elevation was suppressed by either depletion of external Ca2+ or application of the dihydropyridine Ca2+ channel antagonist, nicardipine. One of the putative hair cell afferent nerve neurotransmitters, L-glutamate (Glu), induced [Ca2+]i elevation in SGC. Pharmacological studies suggested that the Glu receptor may be a non-NMDA subtype. Glu-induced response was suppressed by either depletion of external Ca2+ or application of nicardipine, indicating that Glu-induced [Ca2+]i elevation is mainly carried out by the voltage-dependent Ca2+ channel in SGC.
Assuntos
Cálcio/análise , Gânglio Espiral da Cóclea/química , Animais , Cálcio/fisiologia , Canais de Cálcio/efeitos dos fármacos , Cóclea/fisiologia , Relação Dose-Resposta a Droga , Fura-2 , Ácido Glutâmico/farmacologia , Cobaias , Nicardipino/farmacologia , Gânglio Espiral da Cóclea/fisiologia , Fatores de TempoRESUMO
The time course of changes in perilymphatic glutamate release and their Ca2+-dependency were studied in the guinea pig cochlea during high K+-evoked depolarization. The glutamate concentration was analyzed continuously by an enzyme-linked fluorometric assay combined with microdialysis. Two peaks of glutamate increase were found in response to perfusion for 10 min. In the absence of Ca2+, the first peak was diminished, whereas the inhibition of the second peak was minimal.
Assuntos
Cóclea/efeitos dos fármacos , Ácido Glutâmico/metabolismo , Sistemas On-Line , Potássio/farmacologia , Animais , Cóclea/metabolismo , Fluorometria/métodos , Glutamato Desidrogenase , Cobaias , Potenciais da Membrana/efeitos dos fármacos , MicrodiáliseRESUMO
The time course of the changes in perilymphatic glutamate was determined during the application of kanamycin and ethacrynic acid, which are known to damage the hair cells in the inner ear. For the continuous recording of glutamate, the microdialysis technique combined with an enzyme-linked fluorometric assay was used. In guinea pigs receiving a loading dose of 800 mg/kg of kanamycin subcutaneously followed 3 h later by an i.v. injection of 40 mg/kg of ethacrynic acid, a marked glutamate release was clearly found about 2 h after the injection of ethacrynic acid. Injection of kanamycin or ethacrynic acid alone did not produce any change in the perilymphatic glutamate. The morphological changes induced by the administration of both drugs indicated that the collapsing hair cells might release glutamate into the perilymphatic space. The present findings provide additional evidence that glutamate acts as an aggravating factor in aminoglycoside-induced ototoxicity.
Assuntos
Ácido Glutâmico/fisiologia , Células Ciliadas Auditivas Internas/metabolismo , Células Ciliadas Auditivas Internas/fisiopatologia , Células Ciliadas Auditivas Externas/metabolismo , Células Ciliadas Auditivas Externas/fisiopatologia , Animais , Antibacterianos/toxicidade , Diuréticos/farmacologia , Ácido Etacrínico/farmacologia , Ácido Glutâmico/análise , Cobaias , Células Ciliadas Auditivas Internas/química , Células Ciliadas Auditivas Externas/química , Canamicina/toxicidade , Microdiálise/métodos , Perilinfa/química , Perilinfa/metabolismo , Estria Vascular/química , Estria Vascular/metabolismoRESUMO
We have tested the hypothesis that the cause of cochlear dysfunction associated with perilymphatic fistula (PLF) is closely related to endolymphatic hydrops (ELH). Using guinea pigs, we studied the tone-burst elicited compound action potential (CAP) and its modulation as caused by a 50 Hz biasing tone in experimental PLF. We compared these results with those of experimental ELH. Following perilymph aspiration through the perforated round window membrane, mild but significant elevations of CAP thresholds at tested frequencies were found. A reduction in the amplitude of cochlear microphonics (CM) for a 50 Hz sine wave appeared to correlate with these CAP threshold changes. However, there were no significant changes in the modulation effect of the 50 Hz biasing tone on the CAP elicited by an 8 kHz tone burst. This finding differed from that in ears with experimental ELH, in which significant reductions of both 50 Hz CM and the degree of CAP modulation were consistently observed. We concluded that it is unlikely that the underlying mechanisms of a modification to the low frequency response of the base of the cochlea following perilymph aspiration is linked to that of experimental ELH.
Assuntos
Cóclea/fisiopatologia , Endolinfa/fisiologia , Perilinfa/fisiologia , Estimulação Acústica , Potenciais de Ação , Animais , Modelos Animais de Doenças , Edema/fisiopatologia , Eletrofisiologia , Fístula/fisiopatologia , CobaiasRESUMO
Kanamycin (KM)-induced changes in expression of the gene for glutamate-aspartate transporter (GLAST) in the rat cochlea were analyzed by Northern blotting. With the administration of KM (600 mg/kg/day) once daily for 20 days, the expression of GLAST mRNA gradually increased and reached a peak on day 20. Although the expression of GLAST mRNA remained at a high level until 12 days after the completion of the KM treatment, it then fell to the normal level within 2 months. Such KM treatment resulted in loss of both inner and outer hair cells and a concomitant profound permanent threshold shift. The present findings suggest that during KM administration, high concentrations of extracellular glutamate released by collapsing hair cells induced GLAST mRNA expression. Increased GLAST mRNA might play an important role in the prevention of the secondary death of spiral ganglion neurons from glutamate neurotoxicity.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Antibacterianos/toxicidade , Cóclea/efeitos dos fármacos , Cóclea/metabolismo , Canamicina/toxicidade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sistema X-AG de Transporte de Aminoácidos , Animais , Ácido Aspártico/metabolismo , Sequência de Bases , Cóclea/lesões , Primers do DNA/genética , Potenciais Evocados Auditivos do Tronco Encefálico/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Ácido Glutâmico/metabolismo , Células Ciliadas Auditivas/efeitos dos fármacos , Células Ciliadas Auditivas/lesões , Células Ciliadas Auditivas/metabolismo , Masculino , Ratos , Ratos WistarRESUMO
We here report a 53-year-old woman who had undergone resection of a choledochal cyst and hepaticojejunostomy three years before. She was readmitted because of intermittent fever, and abdominal computed tomography revealed a 4-cm tumor in the head of the pancreas. We performed pancreatoduodenectomy, and examination of the resected specimen showed well-differentiated papillary adenocarcinoma. Only 5 cases of carcinoma occurring after the resection of a choledochal cyst have been reported, and to our knowledge, this is the second case of carcinoma of the head of the pancreas.
Assuntos
Adenocarcinoma Papilar/complicações , Cisto do Colédoco/complicações , Cisto do Colédoco/cirurgia , Neoplasias Pancreáticas/complicações , Complicações Pós-Operatórias/cirurgia , Adenocarcinoma Papilar/cirurgia , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Pancreáticas/cirurgia , PancreaticoduodenectomiaRESUMO
Sensorineural hearing loss (SNHL) has been documented in patients with otitis media. Despite a number of clinical and pathologic works dealing with this common problem, animal studies searching for possible relationships between the middle ear inflammation and cochlear function remain insufficient. Bacterial inoculation and ototoxins and inflammatory products in the middle ear cavity cause SNHL in rodents. Human serum albumin placed in the middle ear cavity in chinchillas also produces SNHL, owing to the effects of nonspecific inflammation in the middle ear cavity. Most of the middle ear inflammatory mediators enter the inner ear through the round window route, and alteration of the permeability of the round window membrane plays an important role in causing cochlear dysfunction. Although an immunologic response in the middle ear plays an important role in otitis media, the immunologic response in the inner ear as it relates to middle ear inflammatory mediators requires further study.