RESUMO
BACKGROUND: Renal abscesses are relatively uncommon in children, and usually due to Gram-negative rods or Staphylococcus aureus, whereas abscesses caused by Salmonella are very rare. CASE PRESENTATION: We present the case of a previously healthy 10-year-old boy who had a renal abscess due to Salmonella bareilly. He responded well to treatment with antibiotics, and computed tomography (CT)-guided drainage of the abscess. His blood, urine and abscess aspirate cultures were sterile, but a broad-range 16S rDNA polymerase chain reaction (PCR) assay of the aspirate followed by analysis of four Salmonella genes (fliC, fliD, sopE2, and spaO) identified S. bareilly as the causative agent. CONCLUSION: To the best of our knowledge, this is the first report of renal abscess caused by S. bareilly.
Assuntos
Abscesso Abdominal , Nefropatias , Abscesso/diagnóstico , Abscesso/tratamento farmacológico , Criança , Drenagem , Humanos , Nefropatias/diagnóstico , Masculino , Salmonella/genéticaRESUMO
In Japan, monovalent and pentavalent rotavirus (RV) vaccines were approved in 2011 and 2012, respectively. To monitor changes in the RV genotypes before and after vaccine introduction, we performed a prospective observational study among children (ï¼ 5 years) with gastroenteritis who tested RV-positive on antigen rapid tests. Stool samples were collected from 3 different sites in Japan: Tsu City, Mie Prefecture; Kurashiki City, Okayama Prefecture; and Isumi City, Chiba Prefecture. RV genotypes were determined using reverse transcription-polymerase chain reaction. In Tsu City, G3P[8] was dominant (61.0-77.1%) before vaccine introduction, but decreased after introduction. Meanwhile, in an inverse proportion to the decrease in G3P[8], G1P[8] increased until the 2013/14 season, when a sudden predominance of G2P[4] (100%) occurred. A similar trend was observed in Kurashiki City in terms of the extent of reduction in G3P[8] and the emergence of G2P[4]. In Isumi City, G1P[8] was dominant (70.3%) before vaccine introduction, and G9P[8] became predominant (83.3%) in the 2013/14 season. To determine whether the genotype changes are attributable to vaccines or natural epidemiological changes, ongoing continuous monitoring of the RV genotypes is required.
Assuntos
Genótipo , Infecções por Rotavirus/prevenção & controle , Infecções por Rotavirus/virologia , Vacinas contra Rotavirus/administração & dosagem , Vacinas contra Rotavirus/imunologia , Rotavirus/classificação , Rotavirus/genética , Pré-Escolar , Fezes/virologia , Feminino , Técnicas de Genotipagem , Humanos , Lactente , Recém-Nascido , Japão/epidemiologia , Masculino , Epidemiologia Molecular , Estudos Prospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologiaRESUMO
BACKGROUND: Rotavirus vaccines were introduced in Japan in November 2011. We evaluated the subsequent reduction of the health-care burden of rotavirus gastroenteritis. METHODS: We conducted active surveillance for rotavirus gastroenteritis among children under 5 years old before and after the vaccine introduction. We surveyed hospitalization rates for rotavirus gastroenteritis in children in Tsu City, Mie Prefecture, Japan, from 2007 to 2015 and surveyed the number of outpatient visits at a Tsu City clinic from 2010 to 2015. Stool samples were obtained for rotavirus testing and genotype investigation. We assessed rotavirus vaccine coverage for infants living in Tsu City. RESULTS: In the pre-vaccine years (2007-2011), hospitalization rates for rotavirus gastroenteritis in children under 5 years old were 5.5, 4.3, 3.1 and 3.9 cases per 1000 person-years, respectively. In the post-vaccine years (2011-2015), the rates were 3.0, 3.5, 0.8 and 0.6 cases per 1000 person-years, respectively. The hospitalization rate decreased significantly in the 2013-2014 and 2014-2015 seasons compared to the average of the seasons before vaccine introduction (P < 0.0001). In one pre-vaccine year (2010-2011), the number of outpatient visits due to the rotavirus infection was 66. In the post-vaccine years (2011-2015), the numbers for each season was 23, 23, 7 and 5, respectively. The most dominant rotavirus genotype shifted from G3P[8] to G1P[8] and to G2P[4]. The coverage of one dose of rotavirus vaccine in Tsu City was 56.5% in 2014. CONCLUSION: After the vaccine introduction, the hospitalization rates and outpatient visits for rotavirus gastroenteritis greatly decreased.
Assuntos
Hospitalização/estatística & dados numéricos , Infecções por Rotavirus/tratamento farmacológico , Vacinas contra Rotavirus/uso terapêutico , Pré-Escolar , Efeitos Psicossociais da Doença , Surtos de Doenças/prevenção & controle , Surtos de Doenças/estatística & dados numéricos , Feminino , Gastroenterite/tratamento farmacológico , Gastroenterite/epidemiologia , Humanos , Lactente , Japão/epidemiologia , Masculino , Vigilância da População , Rotavirus/efeitos dos fármacos , Rotavirus/genética , Rotavirus/patogenicidade , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/genéticaRESUMO
Although CD4(+)CD25(+) Treg (Treg) cells are known to modulate NK cell functions, the modulation mechanism of these cells in cord blood has not been fully clarified. The purpose of this study was to clarify the mechanism whereby cord blood Treg cells modulate cord NK cells. By performing various cultures of purified NK cells with or without autologous Treg cells, diminished inhibitory effects of cord Treg cells towards cord NK cell functions, including activation, cytokine production, and cytotoxicity, were observed. We also observed lower secretion of sTGF-beta1 and lower expression of mTGF-beta1 by cord Treg cells than by adult Treg cells. These data revealed the capability of adult Treg cells to suppress rhIL-2-stimulated NK cell function by TGF-beta1, both membrane-bound and soluble types. The reduced inhibitory capabilities of cord Treg cells compared with adult Treg cells is thought to be due to insufficient expression of TGF-beta1.
Assuntos
Sangue Fetal/citologia , Células Matadoras Naturais/imunologia , Linfócitos T Reguladores/imunologia , Fator de Crescimento Transformador beta1/biossíntese , Adulto , Antígenos CD/biossíntese , Antígenos de Diferenciação de Linfócitos T/biossíntese , Antígenos CD4/imunologia , Antígeno CD56/imunologia , Células Cultivadas , Técnicas de Cocultura , Citocinas/biossíntese , Humanos , Recém-Nascido , Interleucina-2/imunologia , Subunidade alfa de Receptor de Interleucina-2/imunologia , Lectinas Tipo C/biossíntese , Ativação Linfocitária/imunologia , Adulto JovemRESUMO
Infants born with intrauterine growth restriction are at increased risk for adverse cardiovascular outcomes in neonatal and later life. Although circadian rhythm is a prognostic marker of cardiovascular health, the concern over the circadian rhythm of these infants is rarely observed. To determine the influence of intrauterine growth retardation on the pattern of circadian rhythm, heart rate (HR) circadian rhythmicity was analyzed in 39 small for gestational age (SGA; birth weight and height below <-2.0 standard deviation score [SDS]) and 117 appropriate for gestational age (AGA; >-1.5 to <1.5 SDS) infants within 72 hours of birth using spectral analysis and cosinor analysis. Amplitude, midline estimating statistic of rhythm, and acrophase calculated from circadian rhythm were analyzed with clinical variables. A significant HR circadian rhythm was observed in 23.1% of the SGA and 24.8% of the AGA group without significant differences; however, SGA infants exhibited remarkable smaller amplitudes compared with AGA in all gestational age (GA) groups (p < 0.001). Amplitudes in AGA infants were positively correlated with the GA or body composition relevant variables (p < 0.001, respectively), but not SGA infants. The blunted HR circadian rhythmicity in SGA infants showed in this study might indicate the vulnerability to pathophysiological condition and could potentially refer to cardiovascular disease in later life.
Assuntos
Ritmo Circadiano/fisiologia , Retardo do Crescimento Fetal/fisiopatologia , Frequência Cardíaca/fisiologia , Recém-Nascido Pequeno para a Idade Gestacional/fisiologia , Adulto , Feminino , Sofrimento Fetal/epidemiologia , Humanos , Hipertensão Induzida pela Gravidez/epidemiologia , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Masculino , Pessoa de Meia-Idade , Gravidez , Análise EspectralRESUMO
CK2 is a highly conserved protein kinase involved in several cellular events. CK2 is expressed in platelets but its role in platelet activation remains poorly understood. In the present study, we tested the hypothesis that CK2 plays a role in platelet activation, particularly in the PAR1-dependent signal transduction pathway. The effect of CK2 and PI 3-kinase inhibitors on aggregation of platelets, activation of GPIIb/IIIa, activation and translocation of CK2 was examined. Platelets were incubated with the cell permeable CK2 inhibitors, DRB, DMAT and TBB and stimulated with the PAR1-AP (SFLLRNP). CK2 inhibitors showed the specific inhibitory pattern of platelet aggregation, characterized by a primary phase of aggregation followed by progressive disaggregation. CK2 inhibitors suppressed the activation of GPIIb/IIIa. PAR1-AP induced two-fold increase in CK2 activity and stimulated the translocation of CK2 from Triton X-100-soluble to -insoluble fraction. Preincubation of platelets with the PI 3-kinase inhibitor, wortmannin or LY294002, impaired PAR1-AP-induced aggregation of platelets. PAR1-AP-induced increase in CK2 activity and translocation of CK2 were inhibited by these treatments. Taken together, the present study demonstrated, for the first time, that PI 3-kinase-CK2 pathway plays an important role in the mechanism of PAR1-dependent platelet aggregation.
Assuntos
Caseína Quinase II/sangue , Citoesqueleto/enzimologia , Fosfatidilinositol 3-Quinases/sangue , Agregação Plaquetária/fisiologia , Receptor PAR-1/sangue , Plaquetas/citologia , Plaquetas/efeitos dos fármacos , Plaquetas/enzimologia , Caseína Quinase II/antagonistas & inibidores , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Ativação Enzimática , Humanos , Imunoprecipitação , Inibidores de Fosfoinositídeo-3 Quinase , Agregação Plaquetária/efeitos dos fármacos , Contagem de Plaquetas , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/metabolismo , Inibidores de Proteínas Quinases/farmacologiaRESUMO
Disseminated squamous cell carcinoma (SCC) of the skin is exceedingly rare in children. SCC occurs after immunodeficiency from immunosuppression in organ transplant recipients or patients with HIV infection or leukaemia, but has not been reported in primary immunodeficiencies other than epidermodysplasia verruciformis. Interferon gamma receptor 2 (IFN gamma R2) deficiency is an exceedingly rare primary immunodeficiency, conferring almost selective predisposition to mycobacterial diseases. A disseminated, cutaneous SCC is described that occurred in a patient homozygous for a novel frameshift deletion at positions 949 and 950 (949delTG) in the IFNGR2 gene. The patient first presented at 1 year of age with disseminated Mycobacterium avium infection, with later infections of atypical mycobacteria (Mycobacterium fortuitum and Mycobacterium porcium). At 17 years of age, the patient developed multifocal SCC lesions on the face and both hands. Histopathological examination revealed well differentiated SCC. Despite local tumour excision, multiple lesions occurred and a large SCC on the right arm required amputation. The patient died at 20 years of age of disseminated SCC. Inherited disorders of IFN gamma mediated immunity may predispose patients to SCC.
Assuntos
Carcinoma de Células Escamosas/metabolismo , Receptores de Interferon/deficiência , Neoplasias Cutâneas/metabolismo , Adolescente , Sequência de Aminoácidos , Sequência de Bases , Carcinoma de Células Escamosas/patologia , Criança , Pré-Escolar , Análise Mutacional de DNA , Genótipo , Humanos , Lactente , Recém-Nascido , Masculino , Dados de Sequência Molecular , Receptores de Interferon/química , Receptores de Interferon/genética , Neoplasias Cutâneas/patologia , Coloração e RotulagemRESUMO
BACKGROUND: We carried out Sukusuku cohort, Mie (SCM), a long term cohort study of child development and investigated the feasibility and validity of this study. Then we focused on the characteristics of the enrolled families and verified the representativeness of the participants in SCM. METHODS: The characteristics of 185 families recruited from 3 hospitals were analyzed, and we verified the representativeness of these subjects. We also analyzed the factors that may influence the mental health of the mothers who are raising children. RESULTS: There were no significant differences between the subjects from the 3 hospitals in terms of the age distribution, academic background, occupation, and annual income of the participating families. At 42 months, the average developmental quotients for postural and motor, cognitive and adaptive, and speech and social development in the 140 infants were 98.6, 100.6, and 99.9, respectively. The overall developmental quotient for infants was 100.3 +/- 13.2; this score was within the standard range (55-132). The path-analysis model revealed that family function was an important factor influencing the mental health of mothers. CONCLUSIONS: The participant characteristics were thought to be generally representative, and we showed the validity and representativeness of the participants in this cohort study. The mental health analysis of mothers suggested that relieving mothers from child-rearing stress and maintaining family function were important for the maintenance and improvement of maternal mental health.
Assuntos
Relações Mãe-Filho , Mães/psicologia , Poder Familiar/psicologia , Adulto , Fatores Etários , Desenvolvimento Infantil , Estudos de Coortes , Família/psicologia , Feminino , Humanos , Lactente , Japão , Masculino , Saúde Mental , Pessoa de Meia-Idade , Projetos de Pesquisa , Fatores Socioeconômicos , Estresse Psicológico/etiologia , Adulto JovemRESUMO
BACKGROUND: We investigated the feasibility and validity of and systematized the methods used to enroll and retain participants requiring long-term interdisciplinary collaborations. We carried out this study in the Sukusuku cohort, Mie (SCM), as one of the regional research site of Japan Children's Study (JCS). METHODS: A total of 467 families who were screened between December 1, 2004 and December 31, 2005, in the Mie-chuo Medical Center and 2 other hospitals; these families were deemed eligible for the study. Of these, a total of 185 families (39.6%) participated in the 4-month observation. Of these families, 5 dropped out at month 9 of the observation; 9, at month 18; 17, at month 30; and 5, at month 42. The retention rates at 9, 18, 30, and 42 months of observation were 97.3%, 92.4%, 83.2%, and 80.5%, respectively. Reinstatement to a previous job was the most common reason for dropouts. RESULTS: We observed that informative consultation notes during observation were beneficial for the retention of participants, and these notes also helped in improving communication between the study subjects and the evaluators during subsequent visits. CONCLUSIONS: In this study, we did not perform the standard checks for child development alone but also investigated the motivating influence of research partnerships with participants. Further, these visits help maintain the motivation levels of the participants and encourage them to contribute for social causes. The results present integration models that can be applied in future relevant longitudinal cohort studies in Japan.
Assuntos
Estudos de Coortes , Pacientes Desistentes do Tratamento/estatística & dados numéricos , Seleção de Pacientes , Projetos de Pesquisa , Desenvolvimento Infantil , Família/psicologia , Estudos de Viabilidade , Humanos , Lactente , Japão , Motivação , Observação , Relações Pesquisador-SujeitoRESUMO
Protein C inhibitor (PCI) regulates the anticoagulant protein C pathway and also inhibits urinary plasminogen activator (uPA), a mediator of tumor cell invasion. In the present study, we evaluated the effect of human PCI and its inactive derivatives on tumor growth and metastasis of human breast cancer (MDA-231) cells, and on angiogenesis in vivo. The invasiveness of MDA-231 cells was inhibited by recombinant intact PCI, but not by reactive site-modified PCI (R354APCI) or by the N-terminal fragment of protease-cleaved PCI (NTPCI). The in vitro invasiveness of MDA-231 cells expressing intact PCI (MDA-PCI) was significantly decreased as compared to MDA-231 cells expressing R354APCI (MDA-R354APCI) or NTPCI (MDA-NTPCI). Further, in vivo growth and metastatic potential of MDA-PCI, MDA-R354APCI and MDA-NTPCI cells in severe combined immunodeficient (SCID) mice were significantly decreased as compared to MDA-Mock cells. Angiogenesis was also significantly decreased in Matrigel implant containing MDA-PCI, MDA-R354APCI or MDA-NTPCI cells as compared to that containing MDA-Mock cells. In vivo angiogenesis in rat cornea and in vitro tube formation were also inhibited by recombinant intact PCI, R354APCI and NTPCI. Furthermore, the anti-angiogenic activity of PCI was strong as cleaved antithrombin (AT), and slightly stronger than that of plasminogen activator inhibitor (PAI)-1 and pigment epithelium-derived factor (PEDF). Overall, this study showed that, in addition to a reactive site-dependent mechanism, PCI may also regulate tumor growth and metastasis independently of its protease inhibitory activity by inhibiting angiogenesis.
Assuntos
Neoplasias da Mama/patologia , Divisão Celular/fisiologia , Metástase Neoplásica , Neovascularização Patológica , Inibidor da Proteína C/fisiologia , Animais , Sequência de Bases , Western Blotting , Neoplasias da Mama/irrigação sanguínea , Linhagem Celular Tumoral , Primers do DNA , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Imuno-Histoquímica , Camundongos , Camundongos SCID , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Most patients who have undergone hematopoietic cell transplantation (HCT) lose specific immunity to measles. However, due to its immunosuppressive potential, it has been recommended that a live attenuated measles vaccination be administered two years following HCT. Measles virus (MV) glycoproteins including hemagglutinin (HA) are expressed on MV-infected dendritic cells (DCs), and they impair efficient antigen presentation between the DC and T cell. We produced a DC-based vaccine against MV by loading DCs with MV-infected autologous DCs. MV in the infected DCs was inactivated using ultraviolet-B. The DC-based vaccine neither expressed HA nor inhibited allogeneic T cell proliferation, while it induced the production of interferon-gamma (IFN-gamma) by autologous CD4 and CD8 naive T cells ex vivo. Importantly, the vaccine derived from patients who had undergone HCT also efficiently induced IFN-gamma producing cells. These findings indicate that our DC-based MV vaccine induces MV-specific immunity even in post-HCT patients without causing immunosuppression.
Assuntos
Células Dendríticas/citologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Transplante de Células-Tronco Hematopoéticas/métodos , Vacina contra Sarampo/química , Sarampo/prevenção & controle , Adolescente , Adulto , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Criança , Pré-Escolar , Glicoproteínas/química , Humanos , Terapia de Imunossupressão , Interferon gama/metabolismo , Pessoa de Meia-IdadeRESUMO
Programmed cell death, or apoptosis, is a physiological means of eliminating unwanted cells and maintaining immune homeostasis. One of the primary mechanisms is the Fas (CD95)/Fas ligand system. Its inactivation in normal cells and malignant cells may be involved in malignant trans-formation and refractory clinical course, respectively. We established a Fas resistant clone and evaluated the molecular basis for its mechanism of resistance. The Fas-sensitive leukemia cell line, MML-1, was established from a child with B-precursor acute lymphoblastic leukemia. A Fas resistant clone, MML-1R, was obtained by co-culture selection with anti-Fas antibody CH-11. Flow cytometry analysis showed both cell lines had equivalent expression of cell surface CD13, 15, 19, 22 and Fas receptor. Western blot analysis revealed equal expression of FADD (Fas-associated death domain protein), caspase-3 and -8. MML-1 was quite sensitive to both CH-11 and etoposide-induced apoptotis. By contrast, MML-1R had similar sensitivity to etoposide but no response to CH-11. Fas receptor mutation analysis showed a heterozygous death domain A --> G point mutation at 1009 bp, causing a switch from glutamine to glycine at amino acid 256. Immunoprecipitation assay showed decreased binding of Fas to FADD. We also found that etoposide bypassed Fas-FADD interaction in MML-1R by activating caspase-8 and caspase-3. These results indicate that Fas resistance can result from mutations of the gene encoding the Fas receptor which result in decreased FADD binding, thereby blocking formation of the death inducing signaling complex. Screening for similar Fas mutations in therapy resistant malignancies would lead to a better understanding of tumorigenesis and recurrence.
Assuntos
Anticorpos Monoclonais/farmacologia , Apoptose/efeitos dos fármacos , Mutação , Receptor fas/genética , Proteínas Adaptadoras de Transdução de Sinal/imunologia , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Sequência de Aminoácidos , Antígenos CD19/metabolismo , Antineoplásicos Fitogênicos/farmacologia , Sequência de Bases , Western Blotting , Caspase 3 , Caspase 8 , Caspases/metabolismo , Linhagem Celular Tumoral , Criança , Análise Mutacional de DNA , Relação Dose-Resposta a Droga , Resistencia a Medicamentos Antineoplásicos/genética , Ativação Enzimática/efeitos dos fármacos , Etoposídeo/farmacologia , Proteína de Domínio de Morte Associada a Fas , Citometria de Fluxo , Humanos , Antígenos CD15/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/imunologia , Receptor fas/imunologia , Receptor fas/metabolismoRESUMO
Infantile cytomegalovirus (CMV)-associated disease and juvenile myelomonocytic leukemia (JMML) frequently present with similar clinical features, and thus the differential diagnosis is often difficult. An early and definite diagnosis of these disorders is required because their therapeutic approaches are very different. The authors describe a 2-month-old Japanese girl with JMML and CMV infection. The CMV antigen was detected by immunologic staining of leukocytes using the peroxidase-labeled monoclonal antibody HRP-C7. To assess clonality, the X-chromosome inactivation pattern was evaluated using polymerase chain reaction analysis of the human androgen receptor gene with or without predigestion of chromosomal DNA with HhaI or HpaII. The patient showed evidence of monoclonal origin of mononuclear cells at diagnosis. Although CMV infection mimicking JMML has previously been reported in two patients, to the authors' knowledge this is the first report describing a firm and definitive diagnosis of JMML based on the study of X-chromosome inactivation patterns.
Assuntos
Infecções por Citomegalovirus/complicações , Leucemia Mielomonocítica Crônica/complicações , Antígenos Virais/análise , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Antivirais/uso terapêutico , Medula Óssea/patologia , Cromossomos Humanos X/genética , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/tratamento farmacológico , Evolução Fatal , Feminino , Ganciclovir/uso terapêutico , Humanos , Hibridização in Situ Fluorescente , Recém-Nascido , Leucemia Mielomonocítica Crônica/diagnóstico , Leucemia Mielomonocítica Crônica/tratamento farmacológico , Leucemia Mielomonocítica Crônica/patologia , Receptores Androgênicos/genéticaRESUMO
We examined the immunological abnormality in a patient with recurrent Mycobacterium avium infection. T cells from the patient showed decreased ability both to produce IFN-gamma and to proliferate in response to IL-12. Despite decreased expression of IL-12R beta1 and beta2 chains in the patient's PHA-activated T cells, there was no difference in IL-12-induced tyrosine and serine phosphorylation of STAT4 in PHA-activated T cells between the patient and healthy subjects, suggesting that IL-12R signals are transmitted to STAT4 in the patient's PHA-activated T cells. Using EMSA, confocal laser microscopy, and Western blotting, we demonstrated that the nuclear translocation of STAT4 in response to IL-12 is reduced in PHA-activated T cells from the patient when compared with those from healthy subjects. Leptomycin B was used to examine whether nuclear export of STAT4 is increased in the patient's T cells. However, leptomycin B treatment did not reverse impaired IL-12-induced nuclear accumulation of STAT4. Although the exact mechanism responsible for the impaired STAT4 nuclear translocation in this patient remains unclear, the absence of mutation in the IL-12Rbeta1, IL-12Rbeta2, STAT4, and STAT4-binding sequence of the IFN-gamma gene and preservation of STAT4 tyrosine and serine phosphorylation suggest the existence of a defective STAT4 nuclear translocation. This defect is likely responsible for the impaired STAT4 nuclear translocation in IL-12-stimulated T cells, leading to impairment of both IFN-gamma production and cell proliferation. To the best of our knowledge, this is the first report of a patient with atypical mycobacterial infection associated with impairment of STAT4 nuclear translocation.
Assuntos
Núcleo Celular/imunologia , Núcleo Celular/metabolismo , Proteínas de Ligação a DNA/metabolismo , Interleucina-12/fisiologia , Mycobacterium avium/imunologia , Transativadores/metabolismo , Tuberculose/imunologia , Transporte Ativo do Núcleo Celular/imunologia , Divisão Celular/imunologia , Proteínas de Ligação a DNA/antagonistas & inibidores , Regulação para Baixo , Humanos , Lactente , Interleucina-12/antagonistas & inibidores , Cinética , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , Transtornos Linfoproliferativos/genética , Transtornos Linfoproliferativos/imunologia , Transtornos Linfoproliferativos/patologia , Masculino , Microscopia Confocal , Fosforilação , Fito-Hemaglutininas/farmacologia , Receptores de Interleucina/biossíntese , Receptores de Interleucina/genética , Receptores de Interleucina-12 , Recidiva , Fator de Transcrição STAT4 , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/patologia , Transativadores/antagonistas & inibidores , Tuberculose/genética , Tuberculose/patologiaRESUMO
Protein C inhibitor (PCI), a member of the serine protease inhibitor family, is produced in various human tissues, including the liver, kidney and testis. In addition to inhibiting the anticoagulant protein C pathway, PCI also inhibits urinary plasminogen activator (uPA), which is a well-known mediator of tumor cell invasion. In the present study, to clarify the biologic significance of PCI in the kidney, we compared the expression of PCI between human renal cell carcinoma (RCC) tissue and nontumor kidney tissue. The PCI antigen level in RCC tissue was found to be significantly lower than in nontumor kidney tissue, and expression of PCI mRNA was detected in normal renal proximal tubular epithelial cells (RPTEC), but not in RCC or in an RCC cell line (Caki-1 cells). No differences were detected between the nucleotide sequence of the major cis-elements in the promoter region of the PCI gene from nontumor kidney and RCC tissues, RPTEC and Caki-1 cells, an RPTEC-derived RCC cell line. The in vitro invasiveness of Caki-1 cells transfected with a PCI expression vector was significantly decreased compared to mock-transfected Caki-1 cells, and it was blocked in the presence of anti-PCI antibody. Since PCI itself did not affect the proliferation rate of Caki-1 cells or cell expression of uPA in vitro, the effect of uPA, PCI, heat-inactivated PCI and plasminogen activator inhibitor (PAI)-1 on the invasive potential of cultured RCC cells was evaluated. The in vitro invasiveness of Caki-1 cells, which express uPA, was significantly enhanced by the addition of uPA, and it was inhibited by anti-uPA antibody, PCI and PAI-1, but not by heat-inactivated PCI. In addition, uPA activity was significantly decreased and uPA-PCI complex level was significantly increased in the culture medium of PCI expression vector-transfected Caki-1 cells as compared to mock-transfected Caki-1 cells. These findings strongly suggest that PCI regulates the invasive potential of RCC cells by inhibiting uPA secreted by these cells. The results of our study suggest that PCI might be a potential therapeutic agent for inhibiting renal tumor invasion.
Assuntos
Carcinoma de Células Renais/patologia , Neoplasias Renais/patologia , Inibidor da Proteína C/metabolismo , Inibidores de Serina Proteinase/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Renais/metabolismo , Células Cultivadas , Regulação para Baixo , Feminino , Temperatura Alta , Humanos , Rim/metabolismo , Rim/patologia , Neoplasias Renais/metabolismo , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Regiões Promotoras Genéticas/genética , Inibidor da Proteína C/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Inibidores de Serina Proteinase/genética , Transfecção , Ativador de Plasminogênio Tipo Uroquinase/genética , Ativador de Plasminogênio Tipo Uroquinase/metabolismoRESUMO
Neuroblastoma, originated from neural crest cells, is the most common extracranial solid tumor in childhood. In the present study, we evaluated in vitro the oncolytic effect of live-attenuated poliovirus on human neuroblastoma cell lines, and in vivo its therapeutic efficacy in human neuroblastoma-bearing athymic mice. Live-attenuated poliovirus killed 27 (93%) of 29 established neuroblastoma cell lines in vitro. It induced cleavage of eukaryotic translation initiation factor 4G, leading to cell death through a mechanism involving activation of caspase-9, caspase-3 and poly(ADP-ribose)polymerase. For the in vivo experiments, an animal model was established using athymic mice xenotransplanted with SJ-N-JF neuroblastoma cells on both flanks. Inoculation of live-attenuated poliovirus into one of the two tumors caused a dramatic and complete regression of both the inoculated and contralateral tumors. Live-attenuated poliovirus has potent oncolytic activity against human neuroblastomas in vitro and in vivo and it may be useful for the treatment of advanced and refractory neuroblastomas, however, further studies are necessary to evaluate the safety of method.
Assuntos
Neuroblastoma/tratamento farmacológico , Vacinas contra Poliovirus/uso terapêutico , Vacinas Atenuadas/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Caspase 3 , Caspase 9 , Caspases/metabolismo , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Fator de Iniciação Eucariótico 4G/metabolismo , Humanos , Camundongos , Camundongos Nus , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Vacinas contra Poliovirus/farmacologia , Poli(ADP-Ribose) Polimerases/metabolismo , Fatores de Tempo , Vacinas Atenuadas/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The natural anticoagulant-activated protein C may inhibit inflammation and fibrosis in the lung. Platelet-derived growth factor is involved in the pathogenesis of lung fibrosis. This study assessed the effect of activated protein C on platelet-derived growth factor expression in human cell lines and in an in vivo model of lung fibrosis. Activated protein C significantly inhibited the secretion and expression of platelet-derived growth factor in human lung cell lines, primary bronchial epithelial cells, and macrophages. In vitro studies also showed that the endothelial activated protein C receptor is expressed by lung epithelial cells and macrophages, and that this receptor and the proteolytic activity of activated protein are implicated in the inhibition of platelet-derived growth factor expression. In the in vivo model of lung fibrosis, intratracheal administration of activated protein C decreased the expression of platelet-derived growth factor and suppressed the development of lung fibrosis. Concomitant intratracheal administration of activated protein C and anti-endothelial activated protein C receptor or anti-platelet-derived growth factor suppressed the inhibitory activity of activated protein C in vivo. In brief, this study describes a novel biological function of activated protein C that may further explain its inhibitory activity on lung inflammation and fibrosis.
Assuntos
Fatores de Coagulação Sanguínea/farmacologia , Pulmão/efeitos dos fármacos , Fator de Crescimento Derivado de Plaquetas/efeitos dos fármacos , Fibrose Pulmonar/patologia , Análise de Variância , Animais , Sequência de Bases , Bleomicina , Fatores de Coagulação Sanguínea/genética , Northern Blotting , Células Cultivadas , DNA Complementar/análise , Modelos Animais de Doenças , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/fisiologia , Feminino , Regulação da Expressão Gênica , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Dados de Sequência Molecular , Fator de Crescimento Derivado de Plaquetas/fisiologia , Probabilidade , Fibrose Pulmonar/tratamento farmacológico , Distribuição Aleatória , Receptores de Superfície Celular/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e EspecificidadeRESUMO
BACKGROUND/AIMS: To elucidate the mechanism of liver dysfunction occurring in patients with sepsis, we evaluated the effect of bacterial lipopolysaccharide (LPS) on the expression of thrombomodulin (TM) in rat sinusoidal endothelial cells (SECs) and the therapeutic efficacy of exogenous recombinant TM. METHODS: We induced endotoxemia in rats by bolus intraperitoneal injection of LPS. TM antigen levels within tissues were assessed by immunohistochemistry. We measured TM in cultured SECs by enzyme immunoassay, functional analysis and real-time polymerase chain reaction (PCR). RESULTS: TM antigen and activity levels were significantly decreased in SECs isolated from LPS-treated rats after 3 and 6 h treatment, and recovered after 12 h treatment, correlating with immunohistochemical observations. In contrast, TM messenger RNA was decreased after 6 and 12 h treatment, and slightly recovered after 24 h treatment. TM expression in cultured SECs isolated from normal rats was also reduced after treatment with LPS and tumor necrosis factor (TNF)-alpha in vitro. The increased levels of serum fibrin degradation products (FDP), fibrin deposition within liver sinusoids, injury of SECs and liver dysfunction induced by LPS in our rat model was improved by recombinant TM treatment. CONCLUSIONS: Decreased TM expression in SECs of LPS-treated rats may result in intrasinusoidal microthrombus formation and subsequent liver dysfunction during sepsis.
Assuntos
Endotélio Vascular/metabolismo , Escherichia coli , Lipopolissacarídeos/farmacologia , Circulação Hepática , Trombomodulina/antagonistas & inibidores , Animais , Citocinas/sangue , Endotélio Vascular/citologia , Imuno-Histoquímica , Hepatopatias/etiologia , RNA/metabolismo , RNA Mensageiro/metabolismo , Ratos , Proteínas Recombinantes/farmacologia , Trombomodulina/genética , Trombomodulina/metabolismo , Trombose/etiologiaRESUMO
Enhanced expression of tissue factor (TF) is associated with the occurrence of coronary disease, strokes and arterial thrombosis. We demonstrated previously that adenosine inhibits TF expression in human umbilical vein endothelial cells (HUVECs) stimulated with inflammatory mediators. In the present study, we evaluated the mechanism of adenosine-induced inhibition of TF expression in HUVECs. The adenosine inhibitory activity on thrombin-induced TF expression in HUVECs was potentiated by the NO precursor, l-arginine, but it was significantly suppressed by the NO scavenger, 2(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide, and by endothelial NO synthase inhibitors, N(G)-monomethyl-l-arginine and N(G)-nitro-l-arginine methyl ester, in a dose-dependent manner. The concentrations of nitrites, cGMP and cAMP in the culture medium of HUVECs treated with a mixture of thrombin and adenosine were significantly higher compared with the culture medium of HUVECs treated with thrombin alone. Northern blotting showed that thrombin decreases and adenosine increases the eNOS mRNA expression in HUVECs. A cAMP-dependent protein kinase inhibitor suppressed NO-mediated TF inhibition in a dose-dependent manner. Overall, these results suggest that adenosine inhibits thrombin-induced TF expression in endothelial cells by a NO-mediated mechanism, and that increased intracellular formation of cAMP is implicated in this inhibitory activity of NO.
Assuntos
Adenosina/fisiologia , Endotélio Vascular/metabolismo , Óxido Nítrico/fisiologia , Trombina/fisiologia , Tromboplastina/metabolismo , Monofosfato de Adenosina/fisiologia , Arginina/fisiologia , Northern Blotting , Células Cultivadas , Relação Dose-Resposta a Droga , Endotélio Vascular/citologia , Inibidores Enzimáticos/farmacologia , Guanosina Monofosfato/fisiologia , Humanos , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase Tipo III , Inibidores de Fosfodiesterase/farmacologiaRESUMO
Pretreatment of IL-3 to Kasumi-1 human acute myeloid leukemia cells enhanced 1-B-D-arabinofuranosyl cytosine (ara-C) cytotoxicity 1.2. to 1.4-fold (median 1.3). To clarify the mechanism of interleukin-3 (IL-3) on ara-C cytotoxicity, we investigated the level of deoxycytidine kinase mRNA with the competitive polymerase chain reaction method and enzyme activities, the incorporation of [(3)H] ara-C into DNA and intracellular ara-cytidine triphosphate (CTP) levels with high-performance liquid chromatography and analyzed cell cycles. The level of deoxycytidine kinase mRNA showed a fourfold increase (88.3 plus minus 4.33 amol &mgr;g of total RNA) at 3 days after treatment with IL-3 compared to control (20.3 plus minus 4.33 amol &mgr;g). After IL-3 treatment, ara-C incorporation into the DNA was increased to 1.33 to 1.83-fold (median, 1.73-fold). The G0/G1 late-phase and S-phase percentages of cells were increased from 28.99 to 78.73% in the IL-3 treatment group as compared to control. These results indicate that IL-3 pretreatment increases the level of deoxycytidine kinase mRNA and ara-C incorporation into the DNA and also increases ratios of G0/G1 late-phase and S-phase subsequent to an enhancement of ara-C cytotoxicity against leukemia cells.