RESUMO
Treatment of a p-azobenzoate (PAB) derivative of a copolymer of D-glutamic acid and D-lysine (D-GL) induced a profound state of unresponsiveness to PAB-reactive helper T lymphocytes generated in PAB-mouse gamma globulin (MGG)-primed mice. This unresponsiveness in T lymphocytes was specific for PAB-reactive cells, since the bacterial alpha-amylase-, keyhole limpet hemocyanin-, or ovalbumin-primed helper T lymphocytes were not suppressed by PAB-D-GL treatment. Taking advantage of the relative ease with which PAB-D-GL can induce specific unresponsiveness to helper T lymphocytes in an animal previously primed with PAB-MGG, it was possible to approach certain questions concerning the mechanisms of tolerance-induction and the fate of tolerant helper T lymphocytes in the PAB-D-GL model by utilizing a classical adoptive cell transfer systemmelimination of the possibility of carry-over of the tolerogen with cells or of the generation of suppressor cells as the result of PAB-D-GL treatment as an explanation of the suppression of helper T-cell activity strongly inplicates the existence of a central intracellular mechanism of specific tolerance on the helper T-cell level. The possibility that suppression of the activity of PAB-reactive helper T lymphocytes by PAB-D-GL reflects simple blocking of surface receptor molecules on T lymphocytes was ruled out as it was found that the helper activity of PAB-reactive cells was minimally suppressed even when PAB-D-GL was directly exposed in vitro to helper T lymphocytesmmoreover, the most conclusive evidence on te the tolerant state induced by in vivo exposure of primed T cells to PAB-D-GL. It appears, therefore, that specific tolerance induced by PAB-D-GL' TO PAB-reactive helper T lymphocytes is an example of irreversible inhibition of T-cell reactivity to antigen, reflecting yet to be determined events at the intra- and subcellular levels.
Assuntos
Proteínas de Transporte/imunologia , Haptenos , Tolerância Imunológica , Linfócitos T/imunologia , Amilases/imunologia , Animais , Formação de Anticorpos , Células Produtoras de Anticorpos/imunologia , Compostos Azo/imunologia , Benzoatos/imunologia , Sítios de Ligação de Anticorpos , Membrana Celular/imunologia , Proteínas do Sistema Complemento/metabolismo , Adjuvante de Freund , Glutamatos , Hemocianinas/imunologia , Soros Imunes , Esquemas de Imunização , Imunização Passiva/métodos , Lisina/imunologia , Camundongos , Peptídeos/imunologia , Tripsina/farmacologia , gama-GlobulinasRESUMO
OBJECTIVE: Psychotic disorders are a significant risk factor for suicide, especially among young people. Psychotic-like experiences (PLEs) in the general population may share an etiological background with psychotic disorders. Therefore, the present study examined the association between PLEs and risk of suicide in a community sample of adolescents. METHOD: Psychotic-like experiences, suicidal feelings, and self-harm behaviors were studied using a self-report questionnaire administered to 5073 Japanese adolescents. Depression and anxiety were evaluated using the 12-item General Health Questionnaire (GHQ). RESULTS: The presence of PLEs was significantly associated with suicidal feelings (OR = 3.1, 95% CI = 2.2-4.5) and deliberate self-harm behaviors (OR = 3.1, 95% CI = 2.0-4.8) after controlling for the effects of age, gender, GHQ-12 score, victimization, and substance use. Suicidal feelings and behaviors were more prevalent in subjects with a greater number of PLEs. CONCLUSION: Psychotic-like experiences may increase the risk of suicidal problems among adolescents.
Assuntos
Ansiedade/psicologia , Depressão/psicologia , Transtornos Psicóticos/psicologia , Comportamento Autodestrutivo/psicologia , Suicídio/psicologia , Adolescente , Ansiedade/epidemiologia , Criança , Vítimas de Crime/psicologia , Depressão/epidemiologia , Feminino , Humanos , Japão , Masculino , Transtornos Psicóticos/epidemiologia , Fatores de Risco , Comportamento Autodestrutivo/epidemiologia , Transtornos Relacionados ao Uso de Substâncias/psicologiaRESUMO
Splenic tumors are occasionally found in clinical practice but the diagnosis is often difficult if only serologic and imaging tests are used. Therefore, pathologic sampling is required in such cases. Endoscopic ultrasonography (EUS) provides a good image of the spleen through the gastric wall, and a transgastric EUS-guided fine needle aspiration (EUS-FNA) biopsy may be easier than the percutaneous approach. Furthermore, a large-gauge needle may raise the capability of EUS-FNA for the histopathologic diagnosis. The aim of this study was to evaluate the yield of EUS-FNA using a large-gauge needle for a splenic tumor. Five patients with splenic tumor were subjected to EUS-FNA with a 19-gauge needle to obtain histopathologic materials. A pathologic sample was obtained in all cases, and the diagnoses were lymphoma (n = 2), sarcoidosis (n = 2), and inflammatory pseudotumor (n = 1). EUS-FNA using a 19-gauge needle is safe and useful for the diagnosis of splenic tumors.
Assuntos
Biópsia por Agulha Fina/métodos , Endossonografia , Granuloma de Células Plasmáticas/patologia , Linfoma/patologia , Sarcoidose/patologia , Esplenopatias/patologia , Idoso , Feminino , Seguimentos , Granuloma de Células Plasmáticas/diagnóstico por imagem , Humanos , Linfoma/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Agulhas , Projetos Piloto , Valor Preditivo dos Testes , Estudos Prospectivos , Sarcoidose/diagnóstico por imagem , Esplenopatias/diagnóstico por imagemRESUMO
OBJECTIVES: Liver transplantation remains the only curative therapy for decompensated liver cirrhosis. However, it has several limitations, and not all patients can receive liver transplants. Therefore, liver regenerative therapy without liver transplantation is considered necessary. In this study, we attempted minimally invasive liver regenerative therapy by peripheral vein infusion of bone marrow-derived mesenchymal stem cells (BMSCs) cultured from a small amount of autologous bone marrow fluid and evaluated the effects of BMSCs on hepatocarcinogenesis in a mouse model. METHODS: C57BL/6 male mice were injected intraperitoneally with N-nitrosodiethylamine once at 2 weeks of age, followed by carbon tetrachloride twice a week from 6 weeks of age onwards, to create a mouse model of highly oncogenic liver cirrhosis. From 10 weeks of age, mouse isogenic green fluorescent protein-positive BMSCs (1.0 × 106/body weight) were infused once every 2 weeks, for a total of 5 times, and the effects of frequent BMSC infusion on hepatocarcinogenesis were evaluated. RESULTS: In the histologic evaluation, no significant differences were observed between the controls and BMSC-administered mice in terms of incidence rate, number, or average size of foci and tumors. However, significant suppression of fibrosis and liver injury was confirmed in the group that received BMSC infusions. DISCUSSION: Considering that BMSC infusion did not promote carcinogenesis, even in the state of highly oncogenic liver cirrhosis, autologous BMSC infusion might be a safe and effective therapy for human decompensated liver cirrhosis.
Assuntos
Transplante de Medula Óssea/métodos , Cirrose Hepática/cirurgia , Neoplasias Hepáticas Experimentais/prevenção & controle , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/citologia , Animais , Carcinogênese , Células Cultivadas , Cirrose Hepática/complicações , Cirrose Hepática/patologia , Neoplasias Hepáticas Experimentais/etiologia , Neoplasias Hepáticas Experimentais/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
OBJECTIVE: To determine whether standardised low-resolution brain electromagnetic tomography modified for a quantifiable method (sLORETA-qm) can be used for quantitative analysis in magnetoencephalography (MEG). METHODS: Somatosensory evoked fields (SEFs) were obtained from 10 hemispheres of five healthy volunteers stimulated on the median nerve at 0.75, 1.0, 1.25, 1.5, 1.75 and 2.0 x threshold of thenar muscle twitch (TMT). N20 m intensity changes were analysed quantitatively using sLORETA-qm. Then, SEFs were measured with stimulation on the median nerve at 1.5 x TMT from 47 hemispheres in 24 subjects. sLORETA-qm intensity and the equivalent current dipole (ECD) moment of N20 m were calculated, and relationships between the values were evaluated. RESULTS: sLORETA-qm intensity increased linearly with stimulus intensity between 0.75 and 1.5 x TMT, and tended to reach a plateau or decrease at higher stimulus intensities. The distribution of sLORETA-qm intensity after natural logarithmic transformation was normal and a close correlation was found between the ECD moment and sLORETA-qm intensity (r(s)=0.91, p<0.001). CONCLUSIONS: The results of this study focusing on N20 m suggested that sLORETA-qm is reliable for quantitative analysis of MEG as well as ECD models. SIGNIFICANCE: sLORETA-qm appears promising for quantitative analyses of MEG for which ECD models are inappropriate.
Assuntos
Mapeamento Encefálico , Encéfalo/fisiologia , Potenciais Somatossensoriais Evocados/fisiologia , Magnetoencefalografia/métodos , Magnetoencefalografia/normas , Adulto , Relação Dose-Resposta à Radiação , Estimulação Elétrica/métodos , Lateralidade Funcional , Humanos , Modelos Lineares , Masculino , Nervo Mediano/fisiologia , Nervo Mediano/efeitos da radiaçãoRESUMO
We report a dural arteriovenous fistula (AVF) that developed at a site on the midline dorsal surface of the dura mater that had been damaged by repeated lumbar punctures. A 61-year-old male patient had undergone repeated lumbar punctures and discectomy for severe lumbago 40 years before the present admission. After surgery, the lumbago symptoms resolved. However, 30 years after the operation, he started to experience dysaesthesia, motor weakness in both legs, and urinary disturbance. Physical examination revealed bilateral leg weakness, diminished deep tendon reflexes in the patellar and Achilles tendons bilaterally, and decreased superficial sensation below L1. Magnetic resonance imaging revealed swelling with intramedullary high intensity and multiple flow voids around the conus and spinal cord on T(2)-weighted images, and adhesive arachnoiditis. Spinal angiography revealed an AVF between the left lateral sacral artery and the S1 radicular vein at the site of the previous operation. Surgery was conducted to carry out excision of the dural AVF at the shunting point, the arterialized intradural vein, and lysis of the arachnoiditis. This case of dural AVF may have been caused by repeated lumbar punctures.
Assuntos
Fístula Arteriovenosa/etiologia , Fístula Arteriovenosa/patologia , Dura-Máter/patologia , Complicações Pós-Operatórias/patologia , Fístula Arteriovenosa/cirurgia , Angiografia Cerebral , Humanos , Laminectomia/efeitos adversos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Compressão da Medula Espinal/etiologia , Punção Espinal/efeitos adversosRESUMO
Correct sequences are prerequisite for quality control of therapeutic oligonucleotides. However, there is no definitive method available for determining sequences of highly modified therapeutic RNAs, and thereby, most of the oligonucleotides have been used clinically without direct sequence determination. In this study, we developed a novel sequencing method called 'hydrophobic tag sequencing'. Highly modified oligonucleotides are sequenced by partially digesting oligonucleotides conjugated with a 5'-hydrophobic tag, followed by liquid chromatography-mass spectrometry analysis. 5'-Hydrophobic tag-printed fragments (5'-tag degradates) can be separated in order of their molecular masses from tag-free oligonucleotides by reversed-phase liquid chromatography. As models for the sequencing, the anti-VEGF aptamer (Macugen) and the highly modified 38-mer RNA sequences were analyzed under blind conditions. Most nucleotides were identified from the molecular weight of hydrophobic 5'-tag degradates calculated from monoisotopic mass in simple full mass data. When monoisotopic mass could not be assigned, the nucleotide was estimated using the molecular weight of the most abundant mass. The sequences of Macugen and 38-mer RNA perfectly matched the theoretical sequences. The hydrophobic tag sequencing worked well to obtain simple full mass data, resulting in accurate and clear sequencing. The present study provides for the first time a de novo sequencing technology for highly modified RNAs and contributes to quality control of therapeutic oligonucleotides. Copyright © 2016 John Wiley & Sons, Ltd.
Assuntos
Oligonucleotídeos/análise , Aptâmeros de Nucleotídeos/análise , Sequência de Bases , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Interações Hidrofóbicas e Hidrofílicas , Peso Molecular , Espectrometria de Massas por Ionização por Electrospray , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidoresRESUMO
BACKGROUND: Human T-cell leukemia virus type 1 (HTLV-1) can infect a number of cells of different lineages in vitro, yet the immunophenotypes of most adult T-cell leukemia/lymphomas (ATLs) are restricted to CD4+. The apparent discrepancy between these findings is still largely unknown. PURPOSE: We report on a unique case of ATL in which the leukemia cells were positive for both T-cell and myeloid cell antigens. To characterize these cells, we isolated cell lines from this patient with ATL. METHODS: The fresh leukemia cells were cultured without the addition of interleukin-2. Cell cloning was carried out by limiting dilution. RESULTS: A cell line (MU) and its clonal sublines were established. MU cells showed the same chromosomal abnormalities and T-cell receptor beta-chain gene rearrangement pattern as those of fresh leukemia cells. MU cells were exclusively positive for a myeloid cell marker (CD13) but not for T-cell markers, despite the presence of T-cell receptor gene rearrangement. CONCLUSION: The established ATL cell line showed both T-cell and myeloid cell characteristics, which seems to be the first evidence for the close association of ATL cells with both lymphoid and myeloid features. The cell line may provide a new insight for the targets of HTLV-1 infection and transformation in vivo.
Assuntos
Antígenos de Diferenciação Mielomonocítica/análise , Leucemia-Linfoma de Células T do Adulto/patologia , Adulto , Antígenos CD/análise , Antígenos de Neoplasias/análise , Southern Blotting , Linfócitos T CD4-Positivos/patologia , DNA de Neoplasias/genética , Rearranjo Gênico , Humanos , Cariotipagem , Leucemia-Linfoma de Células T do Adulto/genética , Leucemia-Linfoma de Células T do Adulto/imunologia , Receptores de Lipopolissacarídeos , Masculino , Microscopia Eletrônica , Receptores de Antígenos de Linfócitos T/genética , Células Tumorais CultivadasRESUMO
A rare case of hemangioblastoma located in the region of hippocampus is reported. A 27-year-old female presented with a single episode of generalized convulsion. The vascular and cherry red color hemangioblastoma was resected by a temporo-zygomatic approach. There has been no recurrence of tumor at a follow-up of 11 years.
Assuntos
Hemangioblastoma/diagnóstico , Hipocampo , Adulto , Feminino , Lateralidade Funcional , Hemangioblastoma/diagnóstico por imagem , Hemangioblastoma/cirurgia , Humanos , Radiografia , Convulsões/etiologiaRESUMO
The antigen recognized by the monoclonal antibody MM2-9B6 is specific for melanomas originating in C57BL/6 mice. It is expressed by three melanomas of independent origin and not by normal or fetal tissues, by a wide variety of other nonmelanoma tumors in C57BL/6 mice, or even by melanomas syngeneic to other strains of mice. We have demonstrated that the expression of the relevant antigen is dependent on the replication and budding of a B-tropic ecotropic murine retrovirus. The relationship between the expression of this virus and carcinogenic progression may yield important insights into the cascade of events leading to neoplastic transformation.
Assuntos
Antígenos de Neoplasias/análise , Antígenos Virais/análise , Melanoma Experimental/imunologia , Retroviridae/imunologia , Animais , Anticorpos Monoclonais/imunologia , Transformação Celular Neoplásica , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Retroviridae/isolamento & purificaçãoRESUMO
Lymphoid cell subpopulations infiltrating into autografts of methylcholanthrene-induced sarcomas in rats immunized with autologous tumor cells were identified in terms of immunohistochemical and cytofluorographic techniques using various monoclonal antibodies raised against different classes of rat lymphohemopoietic cells. These antibodies included in this study directed to rat T-cell antigens corresponding to mouse Lyt-1 (RLyt-1) and Lyt-2,3 antigens (RLyt-2) and to W3/25 antigen expressed on a particular subset of rat T-cells with helper function, as well as to rat granulocyte-macrophage-specific antigen (RGM-1). Histological studies demonstrated that the autografts of highly antigenic tumors introduced to the primary hosts were completely rejected following massive immigration of lymphoid cells into the tumor sites, which was not observed in progressively growing, minimally antigenic tumors. These lymphoid cells found within regressing highly antigenic tumor autografts were identified mostly to be T-cells bearing RLyt-1 (approximately 70%), and more than two-thirds of these T-cells expressed RLyt-2 antigen. In contrast to T-cells, macrophages and B-cells, each of which could be recognized by the presence of either RGM-1 antigen or immunoglobulin on their cell surfaces, appeared to have a minimal role in the rejection of autochthonous tumors, as reflected by their less frequent appearance within the tumor tissues during the rejection process.
Assuntos
Rejeição de Enxerto , Linfócitos/imunologia , Neoplasias Experimentais/imunologia , Animais , Citometria de Fluxo , Transplante de Neoplasias , Neoplasias Experimentais/patologia , Ratos , Ratos Endogâmicos , Transplante Autólogo , Transplante IsogênicoRESUMO
Primary screening of a cDNA expression library of Taenia taeniaeformis oncospheres in lambda gt11 bacteriophage was carried out using rabbit anti-T, taeniaeformis oncosphere serum affinity-purified from oncosphere pellets. From approximately 1.6 x 10(5) plaques, 21 single clones that were positive with the affinity-purified antibodies were isolated. Sibling analysis revealed that 17 clones out of the 21 could be assigned to five different antigen families. Only family 1 was strongly recognized by a serum prepared in a rabbit against a partially purified host-protective oncosphere antigen fraction. The fragments of lambda DNA were inserted into a pGEX plasmid vector that encodes glutathione S-transferase (GST) of Schistosoma japonicum. Clones designated TtO-18, -49.53 (family 1), 46 (family 2), 15 (family 3), 40 (family 4) and 66 (family 5) were established as subclones in pGEX-1 plasmid vectors which produced GST fusion proteins. All GST fusion proteins were soluble and recognized by anti-GST and anti-TtO sera. Three vaccination experiments with these fusion proteins using specific-pathogen-free Wistar rats revealed that all three fusion proteins of family 1 were exclusively effective against T. taeniaeformis oncosphere challenge with approximately 95% and 91% reductions in cystic metacestode and total metacestode recoveries, respectively. Rats vaccinated with fusion proteins of family 1 produced antibodies which reacted with a 21-kDa oncosphere antigen component which appeared to be a major oncosphere stage-specific antigen.
Assuntos
Anticorpos Anti-Helmínticos/biossíntese , Antígenos de Helmintos/imunologia , Taenia/imunologia , Teníase/imunologia , Vacinas Sintéticas/imunologia , Animais , Anticorpos Anti-Helmínticos/imunologia , Clonagem Molecular , Glutationa Transferase/imunologia , Proteínas de Helminto/imunologia , Masculino , Ratos , Ratos Endogâmicos , Proteínas Recombinantes de Fusão/imunologia , Organismos Livres de Patógenos Específicos , VacinaçãoRESUMO
Human melanoma HMV-1 cells formed efficiently multicellular spheroids in media containing low concentrations (0.001-0.01%) of agarose (LCAM). In addition, the spheroids were prevented from deformation without apparent delay of the growth rate. On the other hand, spheroid formation was inhibited markedly by LCAM in the case of murine melanoma B16 cells. Also, LCAM could not prevent the B16 spheroids from deformation. These results indicate that LCAM acts differently depending on the cell types used for spheroid formation.
Assuntos
Meios de Cultura , Sefarose , Células Tumorais Cultivadas/citologia , Animais , Ciclo Celular/efeitos dos fármacos , Humanos , Camundongos , Sefarose/farmacologia , Células Tumorais Cultivadas/ultraestruturaRESUMO
Two hundred ninety-one normal postmenopausal women without hormone replacement therapy were studied for their blood levels of CA125. The levels ranged from 0.1 to 31.8 U/ml (mean+/-SEM 5.5+/-0.3 U/ml). Twenty patients with endometrial adenocarcinoma who had preoperative CA125 levels below 50 U/ml were also studied. The levels of CA125 in the latter group ranged from 2.1 to 43.0 U/ml (17.2+/-2.3 U/ml) and the difference of the mean value of the two groups was statistically significant (P < 0.001). The CA125 levels in normal postmenopausal women have a weak negative correlation with their age (CA125 = -0.025 x age + 6.9, P < 0.001), whereas in cancer patients, a positive correlation was seen (CA125 = 0.6 x age + 11.5, P < 0.05). Patients with endometrial cancer showed a decrease in their CA125 levels after a hysterectomy and salpingo-oophorectomy (6.5+/-1.2 U/ml, P < 0.0001). These results confirmed that the normal range of the CA125 of menopausal women is much lower than that of cycling women and each laboratory should establish its own normal range for the population. Also, it was suggested that CA125 levels which apparently fall within the normal range but are high, however, for the respective age, may indicate that an extensive search for an endometrial malignancy is necessary.
Assuntos
Adenocarcinoma/sangue , Biomarcadores Tumorais/sangue , Antígeno Ca-125/sangue , Neoplasias do Endométrio/sangue , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Pós-Menopausa , Período Pós-OperatórioRESUMO
The aim of this study is to determine the diagnostic value of cerebrospinal fluid (CSF) soluble CD27 (sCD27) as a tumor marker for primary central nervous system lymphoma (PCNSL). We examined sCD27 levels in CSF obtained from various types of brain tumor patients. Forty-two patients were studied (including 12 PCNSL patients) who had not received any therapy for their tumors. In all PCNSL cases, CSF sCD27 levels were more than 15 U/ml (median 84.5 U/ml, range 17-484 U/ml) and in other brain tumor cases, CSF sCD27 levels were all less than 15 U/ml. Our data suggest that CSF sCD27 levels are useful to distinguish PCNSL from other brain tumors.
Assuntos
Neoplasias do Sistema Nervoso Central/líquido cefalorraquidiano , Linfoma de Células B/líquido cefalorraquidiano , Linfoma Difuso de Grandes Células B/líquido cefalorraquidiano , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/líquido cefalorraquidiano , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias do Sistema Nervoso Central/patologia , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imuno-Histoquímica , Lactente , Linfoma de Células B/patologia , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/sangueRESUMO
When mice, previously given oral inoculation with viable oncospheres of the heterologous cestode species (Hymenolepis diminuta, H. microstoma, Taenia taeniaeformis) and the homologous one (H. nana), were challenged with oncospheres of H. nana 4 days after the primary inoculation, they showed strong and complete resistance to H. nana challenge, respectively. However, the resistance was not evoked in mice given either infective eggs of Toxocara canis or non-viable oncospheres of all cestode species examined. Congenitally athymic nude mice given viable oncospheres did not show any resistance to H. nana either. Eosinophil infiltration around cysticercoids of H. nana in the intestinal villi appeared to be more prominent in mice previously given viable oncospheres of H. diminuta than in mice given non-viable oncospheres or PBS only. Some of the eosinophils in the villus harboring cysticercoid(s) of H. nana invaded the epithelia in the former, whereas all eosinophils remained in the lamina propria in the latter. There was almost no eosinophil infiltration in nude mice. Microscopic observations revealed that oncospheres of H. diminuta, which require beetles as the intermediate host like H. microstoma, could invade the mouse intestinal tissue. Therefore, it is strongly suggested that the strong cross resistance to H. nana in mice, induced by oncospheres of all heterologous cestode species, is thymus-dependent and due to oncospheral invasion into the intestinal tissue of mice.
Assuntos
Himenolepíase/imunologia , Hymenolepis/imunologia , Animais , Eosinófilos/imunologia , Imunidade Ativa , Imunocompetência , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Taenia/imunologiaRESUMO
Messenger RNA differential display is conducted to search for genes that are expressed in a region-specific pattern in the rodent brain. Eleven novel gene fragments are isolated. One of these genes which we call pancortin, based on its predominant mRNA expression in the cerebral cortex of the adult, is studied. These pancortin cDNA clones are grouped into four different types of cDNA, designated as pancortin-1 to -4. All pancortin cDNAs share a common sequence in the middle of their structure, having two alternative sequences at both 5'- and 3'-ends, respectively. Deduced amino acid sequence shows that all pancortins have sequences of hydrophobic amino acids at N-terminus and no obvious membrane spanning regions. In situ hybridization histochemistry using oligonucleotide probes specific for 5'- and 3'-end variable parts has revealed that these four pancortin mRNAs are expressed differentially in the adult rodent brain. Robust expression of pancortin-1 and -2 mRNA is observed in the cerebral cortex (including the hippocampus and the olfactory bulb). However, little of pancortin-3 and -4 mRNA is observed there. In the cortex, some neurons are stained by an antibody raised against Pancortin. Immuno-electron microscopic study has revealed that Pancortin-like immunoreactive products are localized mainly in the endoplasmic reticulum and not in the Golgi apparatus indicating that Pancortins are the endoplasmic reticulum-anchored proteins. Our results suggest that each Pancortin is differentially regulated and may perform different functions in the brain.
Assuntos
Encéfalo/metabolismo , Proteínas da Matriz Extracelular/química , Glicoproteínas/biossíntese , Glicoproteínas/química , Proteínas do Tecido Nervoso/biossíntese , Transcrição Gênica , Sequência de Aminoácidos , Animais , Sequência de Bases , Encéfalo/anatomia & histologia , Clonagem Molecular , Feminino , Hibridização In Situ , Masculino , Camundongos , Camundongos Endogâmicos ICR , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/química , Sondas de Oligonucleotídeos , Especificidade de Órgãos , Sondas RNA , RNA Mensageiro/biossíntese , Ratos , Ratos Wistar , Proteínas Recombinantes/biossínteseRESUMO
BACKGROUND: Non-polio enterovirus infections are recognized in children during summer-fall seasons and they sometimes cause large outbreaks. We experienced a nosocomial infection in the neonatal nursery and echovirus type 7 was isolated from samples of four patients. OBJECTIVES: We diagnosed the horizontal infection of four neonates by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) and the nucleotide sequence. STUDY DESIGN: Total RNA was extracted from clinical isolates, serum samples and cerebrospinal fluid (CSF). We amplified enterovirus genome in the 5'-noncoding region by nested PCR and determined the nucleotide sequences. RESULTS: Enterovirus genome was detected in all isolates, in the acute-phase sera in all four patients and in the CSF in one patient by the first PCR. By using nested PCR, the genome was detected from convalescent-phase sera in two patients. All enterovirus genome obtained from the nursery outbreak showed the same sequences with 100% homology. CONCLUSION: We demonstrated the clinical advantages of RT-nested PCR from serum samples and the analysis of nucleotide sequencing gave the supportive evidence of identification of transmission pathway.
Assuntos
Infecção Hospitalar , Surtos de Doenças , Infecções por Echovirus/virologia , Enterovirus Humano B/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Regiões 5' não Traduzidas/genética , Sequência de Bases , Líquido Cefalorraquidiano/virologia , DNA Complementar , Infecções por Echovirus/epidemiologia , Enterovirus Humano B/classificação , Enterovirus Humano B/genética , Humanos , Recém-Nascido , Epidemiologia Molecular , Dados de Sequência Molecular , Berçários Hospitalares , Faringe/virologia , RNA Viral/sangue , Reto/virologia , Sensibilidade e Especificidade , Análise de Sequência de DNARESUMO
To show the clinical importance of cells containing FXIIIa in pulmonary fibrosis induced by bleomycin, the distributions of FXIIIa and collagenous components were investigated immunohistochemically in both normal lung tissues and those affected by bleomycin. In the normal tissues FXIIIa-containing cells were sparse, but they were numerous in the pulmonary fibrotic tissues, especially in the subpleural area and around the blood vessels of alveolar septa, where slight to moderate fibrosis was seen, and in the intra-alveolar fibrinous exudate. In the collagenous scar-like areas, however, these cells were fewer in number and their FXIIIa expression was depleted. These findings suggest that cells containing FXIIIa have an important role in the development of pulmonary fibrosis induced by bleomycin.
Assuntos
Bleomicina/efeitos adversos , Pulmão/enzimologia , Fibrose Pulmonar/enzimologia , Transglutaminases/análise , Adulto , Idoso , Feminino , Humanos , Técnicas Imunoenzimáticas , Masculino , Pessoa de Meia-Idade , Fibrose Pulmonar/induzido quimicamenteRESUMO
Wistar rats inoculated intraperitoneally with 10 viable metacestodes of Taenia crassiceps without adjuvant once on day 0 showed strong resistance to challenge with 200 eggs of T. taeniaeformis on day 30. When rats were killed one month after challenge, there were 80.4% and 46.1% reductions in the number of cystic and total metacestodes of T. taeniaeformis in the liver, respectively. When five rats were killed 16 months after challenge, they showed almost complete immunity against the challenge, with 99.4% and 91.1% reductions in the number of cystic and total metacestodes, respectively. There were only a few degenerated, pin-point metacestodes of T. taeniaeformis in the liver of all five rats; one harbored one cystic metacestode as well. However, there were no such reductions in rats injected initially with cyst fluid antigens of T. crassiceps with Freund's complete adjuvant. An additional experiment was carried out using 500 eggs of T. taeniaeformis in order to confirm the vaccine effect against higher egg dose. There were 96.6%, 87.9%, 83.9%, and 79.3% reductions in the number of cystic metacestodes in rats initially inoculated with 10 viable, 10 formalized, and 10 frozen metacestodes, and injected with sodium deoxycholate-solubilized metacestode antigens, respectively. It is strongly suggested that rats singly dosed with 10 viable or non-viable, intact metacestodes of T. crassiceps without adjuvant became highly resistant to challenge infection with eggs of T. taeniaeformis, which resulted in almost no cystic metacestode establishment.