RESUMEN
Trehalose 6,6'-dicorynomycolate (TDCM) was first characterized in 1963 as a cell surface glycolipid of Corynebacterium spp. by Ioneda and co-workers. TDCM shows potent anti-tumor activity due to its immunoadjuvant properties. Furthermore, the toxicity of TDCM in mice is much weaker than the related trehalose diester of mycolic acid; trehalose 6,6'-dimycolate (TDM, formerly known as cord factor). We have investigated the chemical modification of this class of compound to generate novel agents that display increased immunoadjuvant activity with minimal associated toxicity. During the course of this work we recently developed 6,6'-bis-O-(3-nonyldodecanoyl)-α,α'-trehalose (designated as vizantin). Our results show that vizantin exhibited a potent prophylactic effect on experimental lung metastasis of B16-F0 melanoma cells without a loss of body weight and death in mice. Furthermore, vizantin effectively stimulated human macrophages in an in vitro model, making it a promising candidate for a safe adjuvant in clinical applications. In order to elucidate the pharmacokinetics of vizantin, a probe molecule with similar activity was developed on the basis of a structure-activity relationship (SAR) study with vizantin. The distribution of the probe molecule after intravenous administration into a mouse was assessed by macro confocal microscopy, where it was found to accumulate in the lungs and liver.
Asunto(s)
Adyuvantes Inmunológicos/síntesis química , Glucolípidos/química , Trehalosa/análogos & derivados , Adyuvantes Inmunológicos/farmacocinética , Adyuvantes Inmunológicos/uso terapéutico , Animales , Línea Celular , Quimiocina CCL4/metabolismo , Corynebacterium/química , Glucolípidos/farmacocinética , Glucolípidos/uso terapéutico , Semivida , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/secundario , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Melanoma Experimental/patología , Ratones , Sondas Moleculares/química , Sondas Moleculares/metabolismo , Relación Estructura-Actividad , Trehalosa/química , Trehalosa/farmacocinética , Trehalosa/uso terapéutico , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Vizantin, 6,6'-bis-O-(3-nonyldodecanoyl)-α,α'-trehalose, was developed as a safe immunostimulator on the basis of a structure-activity relationship (SAR) study with trehalose 6,6'-dicorynomycolate (TDCM). It was possible to synthesize vizantin on a large scale more easily than in the case of TDCM, and the compound exhibited more potent prophylactic effect on experimental lung metastasis of B16-F0 melanoma cells. Because vizantin stimulated human macrophages, it is a promising candidate for clinical application.
Asunto(s)
Adyuvantes Inmunológicos/síntesis química , Antineoplásicos/síntesis química , Factores Cordón/síntesis química , Glucolípidos/síntesis química , Trehalosa/análogos & derivados , Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/farmacología , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Línea Celular Tumoral , Factores Cordón/química , Factores Cordón/farmacología , Glucolípidos/química , Glucolípidos/farmacología , Humanos , Técnicas In Vitro , Neoplasias Pulmonares/prevención & control , Neoplasias Pulmonares/secundario , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Melanoma Experimental/patología , Ratones , Ratones Endogámicos C57BL , Fagocitosis/efectos de los fármacos , Estereoisomerismo , Relación Estructura-Actividad , Trehalosa/síntesis química , Trehalosa/química , Trehalosa/farmacología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Oral mucosal melanoma (OMM) is a fatal sarcoma of unknown etiology. Histological morphology and genetic events are distinct from those of its cutaneous counterpart. Mutation and up-regulation of c-kit has been identified in OMM which may activate downstream molecules such as RAS and RAF. These molecules are involved in the mitogen-activated protein kinase (MAPK) pathway leading to tremendous cell proliferation and survival. NRAS and BRAF mutation and protein expression have been studied in other melanoma subtypes. The purpose of this study was to determine RAS protein expression and NRAS and BRAF mutation in 18 primary OMM cases using immunohistochemistry and mutation analysis. Results showed that RAS is intensely expressed in both in situ and invasive OMMs. However, NRAS mutation was only observed in 2/15 polymerase chain reaction (PCR) amplified cases both of which were silent mutations. On the other hand, BRAF missense mutations were observed only in 1/15 cases with PCR amplification. NRAS and BRAF mutations were independent from previously reported c-kit mutations. The classical V600E BRAF mutation was not found; instead a novel V600L was observed suggesting that the oncogenic event in OMM is different from that in skin melanoma. The low frequency of NRAS and BRAF mutations indicate that these genes are not common, but probable events in OMM pathogenesis, most likely independent of c-kit mutation.
Asunto(s)
Genes ras , Melanoma/genética , Mucosa Bucal/patología , Neoplasias de la Boca/genética , Mutación Missense , Mutación Puntual , Proteínas Proto-Oncogénicas B-raf/genética , Frecuencia de los Genes , Humanos , Inmunohistoquímica , Melanoma/patología , Neoplasias de la Boca/patología , Polimorfismo Conformacional Retorcido-Simple , Proteínas Proto-Oncogénicas B-raf/biosíntesis , Estudios Retrospectivos , Proteínas ras/biosíntesis , Proteínas ras/genéticaRESUMEN
Lymph node metastasis is considered a factor in determining the prognosis of squamous cell carcinoma (SCC). Both oral and cervical SCC tumor cells prefer lymph vessels as the route of metastasis. D2-40 is a specific marker of lymphatic endothelial cells. This study clarifies the distribution and characteristics of lymphatic vessels in oral and cervical SCCs. Immunohistochemistry was performed in 20 oral and 20 cervical SCCs (10 non-metastatic and 10 metastatic to lymph nodes) using D2-40, CD31, CD34, CD105 and double staining with D2-40 and keratin. Lymphatic vessel density (LVD) was also determined morphologically. Results showed that lymphatic vessels in both types of SCCs were distributed mainly at the superficial region beneath the epithelium. The LVD in each tumor was significantly higher compared to the corresponding normal mucosa. Moreover, the LVD in lymph node metastasis in each tumor was significantly higher compared to their non-metastatic counterparts. Cancer cell invasion was observed in the lymphatic vessels suggesting the existence of lymph node involvement during metastasis. The new lymphatic vessels that proliferated around the cancer nests in both SCCs have endothelial cell characteristics inferred to be associated with early lymphatic development and initial dissemination of cancer cells.
RESUMEN
Adipose tissue dysfunction has been associated with diabetogenic effects. The effects of repeated Cd exposure on adipocytes remain largely unknown. We administered Cd at doses of 0, 5, 10, and 20 micromol/kgbw sc for 2 weeks (3.5 times/week) to mice and assessed the possible alteration of epididymal white adipose tissue (WAT), including histological difference, adipocyte differentiation and functional capacity. Whereas hepatic weight did not differ between the control and Cd-exposed groups, WAT weight, as well as adipose cell mass, significantly decreased in a dose-dependent manner in Cd-treated mice. The Cd concentration in WAT significantly increased in Cd-treated groups after 2 weeks of exposure. Next, we examined the effects of Cd on adipocyte differentiation and hypertrophy. Cd exposure significantly decreased the paternally expressed gene 1/Mesoderm-specific transcript mRNA expression levels. Both peroxisome proliferator-activated receptor gamma2 and CCAAT/enhancer-binding protein alpha mRNA expression levels in WAT tended to decrease in the Cd-treated groups. Next, we determined the effects of Cd exposure on the mRNA expression levels of adipose-derived hormones, such as adiponectin and resistin. The adiponectin mRNA expression level in WAT decreased after both 6h and 2 weeks of exposure to a high dose of Cd, and the reduction in resistin mRNA expression levels was observed after 2 weeks of exposure. These results suggest that Cd exposure causes abnormal adipocyte differentiation, expansion, and function, which might lead to development of insulin resistance, hypertension, and cardiovascular disease.
Asunto(s)
Adipocitos/efectos de los fármacos , Adiponectina/metabolismo , Tejido Adiposo/metabolismo , Cadmio/toxicidad , Proteínas/metabolismo , Adipocitos/citología , Adipocitos/metabolismo , Adiponectina/genética , Tejido Adiposo/efectos de los fármacos , Animales , Ingestión de Alimentos/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Homeostasis/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Metalotioneína/metabolismo , Ratones , Ratones Endogámicos ICR , PPAR gamma/genética , PPAR gamma/metabolismo , Proteínas/genética , ARN Mensajero/metabolismo , Aumento de Peso/efectos de los fármacosRESUMEN
A 68-year-old man was admitted for bleeding tendency and generalized lymphadenopathy. A diagnosis of idiopathic thrombocytopenic purpura (ITP) associated with mantle cell lymphoma was made. The anti-CD20 monoclonal antibody rituximab 375 mg/m2 was given intravenously once weekly for four consecutive weeks. The patient's platelet counts increased gradually from 0.6 x 10(4)/microliter to 5.8 x 10(4)/microliter. At eighteen weeks after discontinuation of rituximab medication, his platelet count increased again to 10. 3 x 10(4)/microliter and this value has been sustained up to the time of writing. This suggests that rituximab is useful in the treatment of ITP associated with malignant lymphoma.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Linfoma de Células del Manto/complicaciones , Recuento de Plaquetas , Púrpura Trombocitopénica Idiopática/complicaciones , Púrpura Trombocitopénica Idiopática/tratamiento farmacológico , Anciano , Anticuerpos Monoclonales de Origen Murino , Humanos , Masculino , Púrpura Trombocitopénica Idiopática/sangre , RituximabRESUMEN
Previously, we cloned a protease gene of Aeromonas sobria, determined its nucleotide sequence and established a method of purifying its product. In this study, we examined the properties of the purified protease. The protease was temperature-labile and had an optimal pH of 7.5. Metallo-protease inhibitors and a cysteine protease inhibitor did not block the proteolytic activity of the enzyme. The treatment with reagents to modify sulfhydryl group did not reduce the activity. But, serine protease inhibitors did, showing that it was a serine protease. Subsequently, we examined the ability of the protease to enhance vascular permeability in dorsal skin. The protease showed activity and the reaction was inhibited by a simultaneously injected antihistaminic agent. Histopathological examination showed that mast cells appeared around the site where the protease was injected. These findings show that the vascular permeability-enhancing effect of the protease is due to histamine released at the site. Furthermore, we found that a soybean trypsin inhibitor (Kunitz) did not block the proteolytic action of the protease in vitro, but inhibited its vascular permeability-enhancing activity in skin. This suggests that a trypsin-like protease from skin mediates the activity of the protease to enhance its vascular permeability.