RESUMEN
Plaque assays of human respiratory syncytial virus (HRSV) are time-consuming, requiring 4 to 7 days for plaque formation and several hours for dye staining. Here, we describe a simple method by which RSV plaques can be visualized and counted with the naked eye only 2 days after infection of HEp-2 cells. In this assay, the infected cells are stained with monoclonal antibodies and the plaques are developed using diaminobenzidine (DAB). We tested the accuracy of this new plaque assay by comparing the results obtained on days 1, 2, 3, 4, 5, and 6 post-infection. The whole procedure is significantly simpler than the traditional method, with an immunostaining process of around 1.5h. Our method is rapid, accurate, and simple; thus, it has the potential to significantly contribute to studies related to RSV disease.
Asunto(s)
Inmunohistoquímica/métodos , Virus Sincitial Respiratorio Humano/aislamiento & purificación , Ensayo de Placa Viral/métodos , Células Hep G2 , Humanos , Virus Sincitial Respiratorio Humano/inmunología , Factores de TiempoAsunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Cromosomas Humanos Par 12 , Cromosomas Humanos Par 3 , Fusión Génica , Reordenamiento Génico , Leucemia Mieloide Aguda/genética , Proteína del Locus del Complejo MDS1 y EV11/genética , Proteínas Proto-Oncogénicas c-ets/genética , Proteínas Represoras/genética , Translocación Genética , Deleción Cromosómica , Cromosomas Humanos Par 7/genética , Predisposición Genética a la Enfermedad , Humanos , Cariotipificación , Leucemia Mieloide Aguda/inducido químicamente , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa Multiplex , Fenotipo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteína ETS de Variante de Translocación 6RESUMEN
BACKGROUND: The putamen is frequently involved in cases of secondary choreoballism (CB). To date, no study has investigated clinical features of secondary CB such as vascular CB (vCB) and glycemic CB (gCB) in view of putaminal involvement. OBJECTIVES: Cases of CB with putaminal lesions from our hospital were identified in hospital records. Historical cases were obtained from the MEDLINE database. Cases of gCB are defined as those with CB, diabetes mellitus and high signal intensities (HSIs) in the putamen on T1 weighted imaging (T1WI). Cases of vCB are identified among those with CB and stroke involving the putamen. RESULTS: A total of 284 cases (in-hospital cases, 11 gCB and 3 vCB; historical cases, 225 gCB and 45 vCB) were included after excluding 23 glycemic cases without HSIs on T1WI and 53 cases with non-glycemic etiologies. Persistence of CB was longer than one month in 84 cases (gCB, 36.9%, and vCB, 63.0%). Extra-putaminal lesions occurred more frequently in vCB (71.1%) than gCB (50.7%). Age, cerebrovascular etiology and extra-putaminal lesions were found to be significant predictors for persistence of CB one month after onset. Female gender and extra-putaminal lesions were significant predictors for persistence of CB one year after onset. CONCLUSIONS: gCB was the primary common cause of secondary CB involving the putamen. Older age, female gender, vascular etiologies and extensive lesions (putaminal and extra-putaminal) were significant predictors of CB persistence.
Asunto(s)
Hiperglucemia/diagnóstico , Atrofia de Múltiples Sistemas/diagnóstico , Atrofia de Múltiples Sistemas/patología , Putamen/patología , Accidente Cerebrovascular/metabolismo , Factores de Edad , Anciano , Anciano de 80 o más Años , Glucemia/metabolismo , Diagnóstico Diferencial , Femenino , Humanos , Imagen por Resonancia Magnética/métodos , Masculino , Persona de Mediana Edad , Putamen/metabolismo , Factores Sexuales , Accidente Cerebrovascular/diagnósticoRESUMEN
A painless self-immunization method with effective and broad cross-protection is urgently needed to prevent infections against newly emerging influenza viruses. In this study, we investigated the cross-protection efficacy of trivalent influenza vaccine containing inactivated A/PR/8/34 (H1N1), A/Hong Kong/68 (H3N2) and B/Lee/40 after skin vaccination using microneedle patches coated with this vaccine. Microneedle vaccination of mice in the skin provided 100% protection against lethal challenges with heterologous pandemic strain influenza A/California/04/09, heterogeneous A/Philippines/2/82 and B/Victoria/287 viruses 8 months after boost immunization. Cross-reactive serum IgG antibody responses against heterologous influenza viruses A/California/04/09, A/Philippines/2/82 and B/Victoria/287 were induced at high levels. Hemagglutination inhibition titers were also maintained at high levels against these heterogeneous viruses. Microneedle vaccination induced substantial levels of cross-reactive IgG antibody responses in the lung and cellular immune responses, as well as cross-reactive antibody-secreting plasma cells in the spleen. Viral loads in the lung were significantly (p < 0.05) reduced. All mice survived after viral challenges. These results indicate that skin vaccination with trivalent vaccine using a microneedle array could provide protection against seasonal epidemic or new pandemic strain of influenza viruses.
Asunto(s)
Inmunoglobulina G/sangre , Subtipo H1N1 del Virus de la Influenza A/inmunología , Subtipo H3N2 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/administración & dosificación , Infecciones por Orthomyxoviridae/prevención & control , Animales , Femenino , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Subtipo H3N2 del Virus de la Influenza A/efectos de los fármacos , Inyecciones Intradérmicas , Ratones Endogámicos BALB C , Agujas , Infecciones por Orthomyxoviridae/inmunologíaRESUMEN
The radiologic findings of a single nodule from Pneumocystis jirovecii pneumonia (PJP) have been rarely reported. We described a case of granulomatous PJP manifesting as a solitary pulmonary nodule with a halo sign in a 69-year-old woman with diffuse large B cell lymphoma during chemotherapy. The radiologic appearance of the patient suggested an infectious lesion such as angioinvasive pulmonary aspergillosis or lymphoma involvement of the lung; however, clinical manifestations were not compatible with the diseases. The nodule was confirmed as granulomatous PJP by video-assisted thoracoscopic surgery biopsy.
Asunto(s)
Neumonía por Pneumocystis/diagnóstico por imagen , Neumonía por Pneumocystis/diagnóstico , Nódulo Pulmonar Solitario/microbiología , Anciano , Anticuerpos Monoclonales de Origen Murino/efectos adversos , Anticuerpos Monoclonales de Origen Murino/uso terapéutico , Antineoplásicos/efectos adversos , Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Biopsia/métodos , Ciclofosfamida/efectos adversos , Ciclofosfamida/uso terapéutico , Doxorrubicina/efectos adversos , Doxorrubicina/uso terapéutico , Femenino , Humanos , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Linfoma de Células B Grandes Difuso/microbiología , Pneumocystis carinii/patogenicidad , Tomografía de Emisión de Positrones , Prednisona/efectos adversos , Prednisona/uso terapéutico , Rituximab , Cirugía Torácica Asistida por Video , Tomografía Computarizada por Rayos X , Vincristina/efectos adversos , Vincristina/uso terapéuticoAsunto(s)
Hipergammaglobulinemia/patología , Inmunoglobulina G/análisis , Gastropatías/patología , Estómago/patología , Biopsia , Endoscopía del Sistema Digestivo , Fibrosis/patología , Mucosa Gástrica/inmunología , Humanos , Hipergammaglobulinemia/complicaciones , Masculino , Persona de Mediana Edad , Células Plasmáticas/inmunología , Gastropatías/etiologíaRESUMEN
BACKGROUND AND PURPOSE: Unstable carotid atherosclerotic plaques are characterized by cap rupture, leading to thromboembolism and stroke. Matrix metalloproteinases (MMPs) have been implicated in the progression of atherosclerosis and plaque rupture. The aim of this study was to assess the relationship between the expressions of MMP-2 and MMP-9 and carotid plaque instability. METHODS: Eighty atherosclerotic plaques were collected from 74 patients undergoing carotid endarterectomy. Clinical information was obtained from each patient, and plaque morphology was examined at the macroscopic and microscopic levels. The immunohistochemical expressions of MMPs were graded using semiquantitative scales. RESULTS: Macroscopic ulceration (84.6% versus 63.4%, p=0.042) and microscopic cap rupture (79.5% versus 51.2%, p=0.010) were more common in symptomatic than in asymptomatic patients. Immunoreactivities of MMP-2 and MMP-9 were increased in 40 and 36 atheromatous plaques, respectively. Macroscopic ulceration was strongly correlated with the expressions of MMP-2 (p<0.001) and MMP-9 (p=0.001). There were significant correlations between increased MMP-2 expression and cap rupture (p=0.002), intraplaque hemorrhage (p=0.039), and a thin fibrous cap (p=0.002), and between increased MMP-9 expression and cap rupture (p=0.010) and a large lipid core (p=0.013). CONCLUSIONS: Plaque rupture was significantly associated with the development of vascular events in carotid atherosclerotic disease. MMP-2 and MMP-9 are strongly correlated with plaque instability.
RESUMEN
OBJECTIVES: Alpha1-antitrypsin (AAT) is the main inhibitor of human neutrophil elastase, and plays a role in counteracting the tissue damage caused by elastase in local inflammatory conditions. The study evaluated the involvement of AAT in nasal allergic inflammation. METHODS: Forty subjects with mono-sensitization to Dermatophagoides pteronyssinus (Dpt) were enrolled. Twenty allergic rhinitis patients frequently complained of nasal symptoms such as rhinorrhea, stuffiness, sneezing, and showed positive responses to the nasal provocation test (NPT) with Dpt (Group I). The other 20 asymptomatic patients showed sensitization to Dpt but negative NPT (Group II). The levels of AAT, eosinophil cationic protein (ECP), and Dpt-specific IgA antibodies were measured in the nasal lavage fluids (NLFs), collected at baseline, 10 minutes, 30 minutes, 3 hours, and 6 hours after the NPT. Nasal mucosa AAT expression was evaluated with immunohistochemical staining from Group I and Group II. RESULTS: At baseline, only the Dpt-specific IgA level was significantly increased in the NLFs of Group I compared with Group II, while ECP and AAT levels were not significantly different between two groups. After Dpt provocation, AAT, ECP, and Dpt-specific IgA levels were significantly increased in the NLFs of Group I during the early and late responses. The protein expression level of AAT was mostly found in the infiltrating inflammatory cells of the nasal mucosa, which was significantly increased in Group I compared to Group II. CONCLUSION: The increment of AAT showed a close relationship with the activation of eosinophils induced by allergen-specific IgA in the NLFs of patients with allergic rhinitis after allergen stimulation. These findings implicate AAT in allergen-induced nasal inflammation.
RESUMEN
Oleamide (cis-9-octadecenamide) is an endogenous sleep-inducing fatty acid amide that accumulates in the cerebrospinal fluid of the sleep-deprived animals. Microglia are the major immune cells involved in neuroinflammation causing brain damage during infection, ischemia, and neurodegenerative disease. In this study, we examined the effects of oleamide on LPS-induced production of proinflammatory mediators and the mechanisms involved in BV2 microglia. Oleamide inhibited LPS-induced production of NO and prostaglandin E2 as well as expression of iNOS and COX-2. We showed that oleamide blocked LPS-induced NF-kappaB activation and phosphorylation of inhibitor kappaB kinase (IKK). We also showed that oleamide inhibited LPS-induced phosphorylation of Akt, p38 MAPK, and ERK, activation of PI 3-kinase, and accumulation of reactive oxygen species (ROS). Finally, we showed that a specific antagonist of the CB2 receptor, AM630, blocked the inhibitory effects of oleamide on LPS-induced production of proinflammatory mediators and activation of NF-kappaB. Taken together, our results suggest that oleamide shows an anti-inflammatory effect through inhibition of NF-kappaB activation in LPS-stimulated BV2 microglia.
Asunto(s)
Ciclooxigenasa 2/biosíntesis , Microglía/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Ácidos Oléicos/metabolismo , Animales , Línea Celular , Separación Celular , Activación Enzimática/fisiología , Citometría de Flujo , Expresión Génica , Immunoblotting , Inmunoprecipitación , Inflamación/metabolismo , Mediadores de Inflamación/metabolismo , Lipopolisacáridos/farmacología , Ratones , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: The purpose of this study is to show the importance of aneurysmal appearance of medium-sized bronchi observed at CT in the diagnosis of tuberculosis (TB). CONCLUSION: Aneurysmal appearance of medium-sized bronchi is a CT feature of tuberculosis. In TB-endemic areas, the aneurysmal appearance of medium-sized bronchi at CT may suggest the diagnosis of TB.
Asunto(s)
Broncografía , Tomografía Computarizada por Rayos X , Tuberculosis Pulmonar/diagnóstico por imagen , Adulto , Anciano , Cicatriz/diagnóstico por imagen , Constricción Patológica/diagnóstico por imagen , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Granulocytic sarcoma (GS) is defined as a localized tumor mass composed of myeloid blasts and/or immature myeloid cells in an extramedullary site. Usually, GS occurs concomitantly with or after acute myelogenous leukemia (AML), myeloproliferative disorder, or myelodysplastic syndrome. In rare cases, it occurs as a "preleukemic" condition and may precede the onset of AML, which occurs within several months if the patient is not treated with AML-type systemic chemotherapy. Recently, we discovered one case of nonleukemic GS in the small intestine incidentally when intussusception was suspected. The patient visited the emergency department, in October 2006, with symptoms of small-bowel obstruction. Intussusception due to a small-intestinal mass was suspected after evaluation, and small-intestine segmental resection was performed. The patient had no previous history of leukemia, and immunohistochemical staining was used to diagnose GS. Bone-marrow biopsy performed subsequently revealed no lesions that could be suspected as leukemia. The patient received three cycles of chemotherapy, applied as for AML (cytosine arabinoside and anthracycline), and is currently, as of October 29, 2007, showing no other marked indisposition; he has been disease-free for 12 months.
Asunto(s)
Neoplasias Intestinales/diagnóstico , Intestino Delgado , Intususcepción/diagnóstico , Sarcoma Mieloide/diagnóstico , Humanos , Neoplasias Intestinales/cirugía , Masculino , Persona de Mediana Edad , Sarcoma Mieloide/cirugíaRESUMEN
AMP-activated protein kinase (AMPK) is a key regulator of energy homeostasis. Kainic acid (KA), a prototype excitotoxin is known to induce brain-derived neurotrophic factor (BDNF) in brain. In this study, we examined the role of AMPK in KA-induced BDNF expression in C6 glioma cells. We showed that KA and KA receptor agonist induced activation of AMPK and KA-induced AMPK activation was blocked by inhibition of Ca(2+)/calmodulin-dependent protein kinase kinase (CaMKK) beta. We then showed that inhibition of AMPK by compound C, a selective inhibitor of AMPK, or small interfering RNA of AMPKalpha1 blocked KA-induced BDNF mRNA and protein expression. Inhibition of AMPK blocked KA-induced phosphorylation of CaMKII and I kappaB kinase (IKK) in C6 cells. Finally, we showed that inhibition of AMPK reduced DNA binding and transcriptional activation of nuclear factor-kappaB (NF-kappaB) in KA-treated cells. These results suggest that AMPK mediates KA-induced BDNF expression by regulating NF-kappaB activation.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/metabolismo , Ácido Kaínico/farmacología , Complejos Multienzimáticos/metabolismo , FN-kappa B/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Quinasas Activadas por AMP , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Calcio/metabolismo , Quinasa de la Proteína Quinasa Dependiente de Calcio-Calmodulina/metabolismo , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Línea Celular Tumoral , Activación Enzimática , Glioma , Complejos Multienzimáticos/antagonistas & inhibidores , Fosforilación , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , ARN Mensajero/metabolismo , Ratas , Receptores de Ácido Kaínico/agonistas , Receptores de Ácido Kaínico/metabolismo , Activación TranscripcionalRESUMEN
AMP-activated protein kinase (AMPK) is a key regulator of energy homeostasis and its activation during T cell receptor stimulation has recently been reported. In this study, we examined the role of AMPK in interleukin (IL)-2 production in T cells. Inhibition of AMPK by compound C, a specific inhibitor of AMPK or small interfering RNA of AMPKalpha1 suppressed IL-2 production in Jurkat T cells and peripheral blood lymphocytes stimulated with PMA plus ionomycin (PMA/Io) or with monoclonal anti-CD3 plus anti-CD28. We then showed that AMPK inhibition reduced PMA/Io-induced IL-2 mRNA expression and IL-2 promoter activation. Moreover, inhibition of AMPK suppressed transcriptional activation of NF-AT and AP-1, but not NF-kappaB, in PMA/Io-activated Jurkat cells. Finally, we found that compound C inhibited PMA/Io-induced phosphorylation of p38, JNK, and GSK-3beta but not of ERK. These results suggest that AMPK mediates IL-2 production by regulating NF-AT and AP-1activation during T cell stimulation.
Asunto(s)
Interleucina-2/metabolismo , Complejos Multienzimáticos/fisiología , Factores de Transcripción NFATC/metabolismo , Proteínas Serina-Treonina Quinasas/fisiología , Linfocitos T/inmunología , Factor de Transcripción AP-1/metabolismo , Proteínas Quinasas Activadas por AMP , Regulación hacia Abajo , Humanos , Interleucina-2/genética , Células Jurkat , Complejos Multienzimáticos/antagonistas & inhibidores , Fosforilación/efectos de los fármacos , Regiones Promotoras Genéticas , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Serina-Treonina Quinasas/metabolismo , Linfocitos T/efectos de los fármacos , Linfocitos T/enzimología , Factor de Transcripción AP-1/agonistas , Activación TranscripcionalRESUMEN
Adenosine is an endogenous nucleoside that regulates many processes, including inflammatory responses, through activation of its receptors. Adenosine receptors have been reported to be expressed in microglia, which are major immune cells of brain, yet little is known about the role of adenosine receptors in microglial cytokine production. Thus, we investigated the effect of adenosine and adenosine A3 receptor ligands on LPS-induced tumor necrosis factor (TNF-alpha) production and its molecular mechanism in mouse BV2 microglial cells. Adenosine and Cl-IB-MECA, a specific adenosine A3 receptor agonist, suppressed LPS-induced TNF-alpha protein and mRNA levels. Moreover, MRS1523, a selective A3 receptor antagonist, blocked suppressive effects of both adenosine and Cl-IB-MECA on TNF-alpha. We further examined the effect of adenosine on signaling molecules, such as PI 3-kinase, Akt, p38, ERK1/2, and NF-kappaB, which are involved in the regulation of inflammatory responses. Adenosine inhibited LPS-induced phosphatidylinositol (PI) 3-kinase activation and Akt phosphorylation, whereas it had no effect on the phosphorylation of p38 and ERK1/2. We also found that adenosine as well as Cl-IB-MECA inhibited LPS-induced NF-kappaB DNA binding and luciferase reporter activity. Taken together, these results suggest that adenosine A3 receptor activation suppresses TNF-alpha production by inhibiting PI 3-kinase/Akt and NF-kappaB activation in LPS-treated BV2 microglial cells.
Asunto(s)
Adenosina/metabolismo , Microglía/metabolismo , FN-kappa B/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Receptor de Adenosina A3/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Adenosina/análogos & derivados , Adenosina/farmacología , Agonistas del Receptor de Adenosina A3 , Antagonistas del Receptor de Adenosina A3 , Animales , Línea Celular , Encefalitis/metabolismo , Encefalitis/fisiopatología , Activación Enzimática/efectos de los fármacos , Activación Enzimática/fisiología , Gliosis/metabolismo , Gliosis/fisiopatología , Mediadores de Inflamación , Lipopolisacáridos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/fisiología , Ratones , Microglía/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Piridinas/farmacología , ARN Mensajero/efectos de los fármacos , ARN Mensajero/metabolismo , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Accessory ovary is a rare gynecologic condition, and tumors arising in accessory ovaries are extremely rare. Accessory ovary may result from separation of migrating ovaries during embryogenesis and injuries such as inflammation and operation on normal ovary. Congenital malformations, most frequently malformations of the genitourinary organ, are seen in connection with the accessory ovary. We experienced the first case of two dermoid cysts developing in an accessory ovary located in the left infundibulopelvic ligament and another in the eutopic ovary at the same side concurrently. Here, we present this extremely rare case with a review of the literature.
Asunto(s)
Quiste Dermoide/diagnóstico , Quistes Ováricos/diagnóstico , Enfermedades del Ovario/diagnóstico , Ovario/patología , Adulto , Quiste Dermoide/patología , Femenino , Humanos , Quistes Ováricos/patología , Enfermedades del Ovario/congénito , Ovario/anomalías , Tomografía Computarizada por Rayos XRESUMEN
Cryptococcus albidus, a non-neoformans species of the genus Cryptococcus, is generally regarded as a rare cause of disease. There have been only 14 previously reported cases in which this organism has been isolated as a pathogen, none of which occurred in a renal transplant recipient. A 23-year-old renal transplant recipient taking medication consisting of cyclosporine and prednisolone was admitted with a 10-day history of dry cough, fever and progressive dyspnea. The next day, his respiratory status deteriorated dramatically, and he developed acute respiratory distress syndrome (ARDS) and fulminant septic shock. On the eighth hospital day, tender macules on both his shins coalesced to form erythematous patches. Cryptococcus albidus was isolated by skin biopsy and tissue culture. We report here the first case of disseminated cryptococcosis caused by C. albidus in a renal transplant recipient who had been successfully treated with fluconazole monotherapy.