Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 510
Filtrar
Más filtros

Banco de datos
Tipo del documento
Intervalo de año de publicación
2.
Tissue Antigens ; 80(2): 119-25, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22568758

RESUMEN

Minor histocompatibility (H) antigens are targets of graft-vs-host disease and graft-vs-tumor responses after human leukocyte antigen matched allogeneic hematopoietic stem cell transplantation. Recently, we reported a strategy for genetic mapping of linkage disequilibrium blocks that encoded novel minor H antigens using the large dataset from the International HapMap Project combined with conventional immunologic assays to assess recognition of HapMap B-lymphoid cell line by minor H antigen-specific T cells. In this study, we have constructed and provide an online interactive program and demonstrate its utility for searching for single-nucleotide polymorphisms (SNPs) responsible for minor H antigen generation. The website is available as 'HapMap SNP Scanner', and can incorporate T-cell recognition and other data with genotyping datasets from CEU, JPT, CHB, and YRI to provide a list of candidate SNPs that correlate with observed phenotypes. This method should substantially facilitate discovery of novel SNPs responsible for minor H antigens and be applicable for assaying of other specific cell phenotypes (e.g. drug sensitivity) to identify individuals who may benefit from SNP-based customized therapies.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas , Prueba de Histocompatibilidad/métodos , Antígenos de Histocompatibilidad Menor/inmunología , Polimorfismo de Nucleótido Simple , Programas Informáticos , Linfocitos B/inmunología , Línea Celular , Mapeo Cromosómico , Minería de Datos , Genotipo , Proyecto Mapa de Haplotipos , Humanos , Internet , Desequilibrio de Ligamiento , Antígenos de Histocompatibilidad Menor/genética , Fenotipo , Linfocitos T/inmunología , Trasplante Homólogo
3.
Domest Anim Endocrinol ; 74: 106522, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32841888

RESUMEN

Heat stress disrupts reproductive function in cattle. In summer, high ambient temperature and humidity elevate core body temperature, which is considered to be detrimental to reproductive abilities in cattle. Neurokinin B (NKB) is a factor that generates pulsatile GnRH and subsequent LH secretion in mammals. Recent studies have reported that NKB-neurokinin 3 receptor (NK3R) signaling is associated with heat-defense responses in rodents. The present study aimed to clarify the role of NKB-NK3R signaling in thermoregulation in cattle. We examined the effects of an NK3R-selective agonist, senktide, on vaginal temperature as an indicator of core body temperature in winter and summer. In both seasons, continuous infusion of senktide for 4 h immediately decreased vaginal temperature, and the mean temperature change in the senktide-treated group was significantly lower than that of both vehicle- and GnRH-treated groups. Administration of GnRH induced LH elevation, but there was no significant difference in vaginal temperature change between GnRH- and vehicle-treated groups. Moreover, we investigated the effects of senktide on ovarian temperature. Senktide treatment seemed to suppress the increase in ovarian temperature from 2 h after the beginning of administration, although the difference between groups was not statistically significant. Taken together, these results suggest that senktide infusion caused a decline in the vaginal temperature of cattle, in both winter and summer seasons, and this effect was not due to the gonadotropin-releasing action of senktide. These findings provide new therapeutic options for senktide to support both heat-defense responses and GnRH/LH pulse generation.


Asunto(s)
Temperatura Corporal/efectos de los fármacos , Bovinos/fisiología , Respuesta al Choque Térmico/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Receptores de Neuroquinina-3/agonistas , Sustancia P/análogos & derivados , Animales , Regulación de la Temperatura Corporal/efectos de los fármacos , Regulación de la Temperatura Corporal/fisiología , Femenino , Hormona Liberadora de Gonadotropina/fisiología , Hormona Luteinizante/fisiología , Neuroquinina B/fisiología , Ovario/fisiología , Fragmentos de Péptidos/uso terapéutico , Receptores de Neuroquinina-3/fisiología , Transducción de Señal/fisiología , Sustancia P/farmacología , Sustancia P/uso terapéutico , Vagina/fisiología
4.
J Exp Med ; 186(10): 1677-87, 1997 Nov 17.
Artículo en Inglés | MEDLINE | ID: mdl-9362528

RESUMEN

In this report, we establish a regulatory role of natural killer (NK) cells in experimental autoimmune encephalomyelitis (EAE), a prototype T helper cell type 1 (Th1)-mediated disease. Active sensitization of C57BL/6 (B6) mice with the myelin oligodendrocyte glycoprotein (MOG)35-55 peptide induces a mild form of monophasic EAE. When mice were deprived of NK cells by antibody treatment before immunization, they developed a more serious form of EAE associated with relapse. Aggravation of EAE by NK cell deletion was also seen in beta 2-microglobulin-/- (beta 2m-/-) mice, indicating that NK cells can play a regulatory role in a manner independent of CD8+ T cells or NK1.1+ T cells (NK-T cells). The disease enhancement was associated with augmentation of T cell proliferation and production of Th1 cytokines in response to MOG35-55. EAE passively induced by the MOG35-55-specific T cell line was also enhanced by NK cell deletion in B6, beta 2m-/-, and recombination activation gene 2 (RAG-2)-/- mice, indicating that the regulation by NK cells can be independent of T, B, or NK-T cells. We further showed that NK cells inhibit T cell proliferation triggered by antigen or cytokine stimulation. Taken together, we conclude that NK cells are an important regulator for EAE in both induction and effector phases.


Asunto(s)
Encefalomielitis Autoinmune Experimental/inmunología , Células Asesinas Naturales/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/biosíntesis , Antígenos/inmunología , Antígenos Ly , Antígenos de Superficie , Citocinas/biosíntesis , Encefalomielitis Autoinmune Experimental/etiología , Encefalomielitis Autoinmune Experimental/genética , Femenino , Interferón gamma/deficiencia , Interferón gamma/genética , Lectinas Tipo C , Activación de Linfocitos/inmunología , Depleción Linfocítica , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Datos de Secuencia Molecular , Subfamilia B de Receptores Similares a Lectina de Células NK , Proteínas/inmunología , Células TH1/metabolismo , Microglobulina beta-2/deficiencia , Microglobulina beta-2/genética
5.
J Exp Med ; 162(3): 1025-43, 1985 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-3897446

RESUMEN

Both connective tissue mast cells and mast cells grown in vitro are derived from multipotential hematopoietic stem cells, but these two mast cell populations exhibit many differences in morphology, biochemistry, and function. We investigated whether the phenotype of cultured mast cells or their progeny was altered when the cells were transferred into different locations in vivo. Cultured mast cells were immature by ultrastructure, and stained with alcian blue but with neither safranin or berberine sulfate, a fluorescent dye that binds to the heparin of connective tissue mast cell granules. By contrast, mast cells recovered from the peritoneal cavity of congenitally mast cell-deficient (WB X C57BL/6)F1-W/Wv (WBB6F1-W/Wv) mice 10 wk after intraperitoneal injection of cultured WBB6F1-+/+ or C57BL/6-bgJ/bgJ mast cells stained with both safranin and berberine sulfate. Staining with berberine sulfate was prevented by treatment of the cells with heparinase but not chondroitinase ABC, suggesting that the adoptively transferred mast cell population had acquired the ability to synthesize and store heparin. Furthermore, the recovered mast cells were indistinguishable by ultrastructure from the normal mature peritoneal mast cells of WBB6F1-+/+ mice, and contained substantially more histamine than mast cells studied directly from culture. Intravenous injection of cultured mast cells resulted in the development of safranin-and berberine sulfate-positive mast cells in the peritoneal cavity, spleen, skin, and glandular stomach muscularis propria. Mast cells also developed on the glandular stomach mucosa, but these cells stained with alcian blue rather than safranin, and did not stain with berberine sulfate. This result suggests that cultured mast cells can give rise to mast cells of either the connective tissue type or mucosal phenotype, depending on anatomical location. Furthermore, transplantation of cultured mast cells into WBB6F1-W/Wv mice had no measurable effect on the anemia of the recipient mice, suggesting a possible strategy for repairing the mast cell deficiency of WBB6F1-W/Wv mice without affecting other bone marrow-derived populations such as erythrocytes. Intravenous injection of representative connective tissue type mast cells (30-50% pure peritoneal mast cells derived from WBB6F1-+/+ mice) gave results similar to those obtained with cultured mast cells: mast cells developing in the peritoneal cavity, skin, spleen, and glandular stomach muscularis propria of WBB6F1-W/Wv recipients stained with safranin and berberine sulfate, whereas mast cells developing in the mucosa of the glandular stomach stained only with alcian blue.(ABSTRACT TRUNCATED AT 400 WORDS)


Asunto(s)
Trasplante de Médula Ósea , Mastocitos/ultraestructura , Animales , Diferenciación Celular , Células Cultivadas , Tejido Conectivo/patología , Inmunización Pasiva , Síndromes de Inmunodeficiencia/genética , Síndromes de Inmunodeficiencia/inmunología , Síndromes de Inmunodeficiencia/veterinaria , Inyecciones/métodos , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Mastocitos/clasificación , Mastocitos/trasplante , Ratones , Ratones Mutantes/inmunología , Membrana Mucosa/patología , Enfermedades de los Roedores/genética , Enfermedades de los Roedores/inmunología , Piel
6.
Domest Anim Endocrinol ; 68: 83-91, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-30908995

RESUMEN

Pulsatile gonadotropin-releasing hormone (GnRH) secretion is indispensable for reproduction in mammals. Kisspeptin neurons in the hypothalamic arcuate nucleus (ARC), referred to as KNDy neurons because of the coexpression of neurokinin B and dynorphin A, are considered as components of the GnRH pulse generator that produces rhythmic GnRH secretion. The present study aimed to investigate if peripheral administration of PF-4455242, a κ-opioid receptor (KOR, a dynorphin A receptor) antagonist, facilitates pulsatile luteinizing hormone (LH) secretion and GnRH pulse generator activity in estrogen-treated ovariectomized Shiba goats to determine the possibility of using KOR antagonists to artificially control ovarian activities. PF-4455242 was intravenously infused for 4 h (1 or 10 µmol/kg body weight/4 h) or as a single subcutaneous injection (1 or 10 µmol/kg body weight). In a separate experiment, the same KOR antagonist (10 µmol/kg body weight/4 h) was intravenously infused during the recording of multiple unit activity (MUA) in the ARC that reflects the activity of the GnRH pulse generator to test the effects of KOR antagonist administration on GnRH pulse generator activity. Intravenous infusion and single subcutaneous injection of the KOR antagonist significantly increased the frequency of LH pulses compared with controls. Intravenous infusion of KOR antagonist also significantly increased the frequency of episodic bursts in the MUA. The present study demonstrates that peripherally administered KOR antagonist stimulates pulsatile LH secretion by acting on the GnRH pulse generator, and peripheral administration of PF-4455242 can be used to facilitate pulsatile LH secretion, which in turn facilitates ovarian activities in farm animals.


Asunto(s)
Compuestos de Bifenilo/farmacología , Estrógenos/administración & dosificación , Cabras/fisiología , Hormona Liberadora de Gonadotropina/metabolismo , Receptores Opioides kappa/antagonistas & inhibidores , Sulfonamidas/farmacología , Animales , Compuestos de Bifenilo/administración & dosificación , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Inyecciones Intravenosas , Inyecciones Subcutáneas , Ovariectomía/veterinaria , Sulfonamidas/administración & dosificación
7.
Curr Top Microbiol Immunol ; 314: 251-67, 2007.
Artículo en Inglés | MEDLINE | ID: mdl-17593664

RESUMEN

CDld-restricted invariant natural killer T (NKT) cells emerge as unique lymphocyte subsets implicated in the regulation of autoimmunity. Abnormalities in the numbers and functions of NKT cells have been observed in patients with diverse autoimmune diseases as well as in animal models of autoimmune diseases. NKT cells recognize glycolipid antigens presented by the nonpolymorphic MHC class I-like protein CD1d and participate in various kinds of immunoregulation due to a potent ability to produce a variety of cytokines. In this review, we examine the potential roles of NKT cells in the regulation and pathogenesis of autoimmune disease and the recent advances in glycolipid therapy for autoimmune disease models.


Asunto(s)
Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/fisiopatología , Células Asesinas Naturales/inmunología , Animales , Artritis Experimental/inmunología , Artritis Experimental/fisiopatología , Autoinmunidad , Colitis/inmunología , Colitis/fisiopatología , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 1/fisiopatología , Modelos Animales de Enfermedad , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/fisiopatología , Humanos , Lupus Eritematoso Sistémico/inmunología , Lupus Eritematoso Sistémico/fisiopatología , Ratones , Ratones Endogámicos BALB C
8.
Leukemia ; 32(12): 2729-2730, 2018 12.
Artículo en Inglés | MEDLINE | ID: mdl-30232464

RESUMEN

Owing to the insufficient specificity of the anti-myeloproliferative leukemia protein (MPL) antibody in the original version of this Article, Figure 6 and parts of Figures 2a, 4e, and 5a do not represent the correct information. The corrected version of Figure 6 is in this correction and those of Figures 2a, 4e, and 5a are shown in the supplemental information.

9.
J Clin Invest ; 74(4): 1229-37, 1984 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-6480826

RESUMEN

We analyzed the heterogeneity of apo E in very low density lipoprotein from 58 hyperlipidemic subjects with or without atherosclerosis, 69 patients with ischemic heart disease, and 100 apparently healthy individuals. Apo E gene frequencies in the group of healthy individuals were comparable with those in German and American populations. The distribution of six common apo E phenotypes in the groups of hyperlipidemia and ischemic heart disease was similar to that in the healthy group. In addition to the three major isoforms of apolipoprotein E (apo E-4, E-3, and E-2) and the new one (apo E-5) which was recently found in this laboratory, we have discovered an additional series of components, which showed themselves as at least three bands on an isoelectric focusing gel in the region of E-VII through E-V, in four patients with hyperlipidemia and atherosclerosis. The new series of apo E components, named apo E-Suita, was identical with the ordinary apo E in its interaction with heparin-Sepharose gel and with anti-apo E antibody. The most basic component of apo E-Suita (E-VII) was the unsialylated form and other components (E-VI and E-V), the sialylated forms. Family studies revealed that apo E-Suita was determined by inheritance of a new apo E allele located at the same locus as the hitherto known apo E components. Apo E-5 and apo E-Suita isoproteins had isoelectric points more basic than apo E-3, the parent type, by two and four units of charge, respectively. While the apo E-Suita isoprotein had the same molecular weight as ordinary major apo E isoproteins, the molecular weight of the apo E-5 isoprotein was approximately 1,500-2,000 lower than the other apo E isoproteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The incidence of abnormal apo E components, including apo E-5 and apo E-Suita, was high among patients with hyperlipidemia and ischemic heart disease (7:127), while we could not find such components among 100 healthy individuals. Moreover, five of seven patients with the abnormal apo E had overt atherosclerotic disease. The findings suggest that these kinds of apolipoprotein mutation are closely related to the development of atherosclerosis.


Asunto(s)
Apolipoproteínas E/genética , Arteriosclerosis/genética , Hiperlipoproteinemia Tipo III/genética , Mutación , Reacciones Antígeno-Anticuerpo , Apolipoproteínas E/inmunología , Apolipoproteínas E/aislamiento & purificación , Cisteamina/farmacología , Femenino , Frecuencia de los Genes , Humanos , Lipoproteínas VLDL/aislamiento & purificación , Masculino , Neuraminidasa/farmacología , Fenotipo , Pruebas de Precipitina
10.
J Clin Invest ; 107(5): R23-9, 2001 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-11238569

RESUMEN

Multiple sclerosis (MS) is an autoimmune disease characterized by clinical relapse and remission. Because of the potential role of natural killer (NK) cells in the regulation of autoimmunity, we have examined cytokine profile and surface phenotype of NK cells in the peripheral blood of MS. Here we demonstrate that NK cells in the remission of MS are characterized by a remarkable elevation of IL-5 mRNA and a decreased expression of IL-12Rbeta2 mRNA, as well as a higher expression of CD95. Moreover, the NK cells from MS in remission produced much larger amounts of IL-5 than did those from controls after stimulation with phorbol myristate acetate (PMA) and ionomycin. These features are reminiscent of those of NK type 2 (NK2) cells that can be induced in a condition favoring functional deviation of T cells toward Th2. Remarkably, the NK cells lose the NK2-like property when relapse of MS occurs, but regain it after recovery. We also found that NK2 cells induced in vitro inhibit induction of Th1 cells, suggesting that the NK2-like cells in vivo may also prohibit autoimmune effector T cells. Taken together, it is possible that NK cells play an active role in maintaining the remission of MS.


Asunto(s)
Autoinmunidad , Células Asesinas Naturales/inmunología , Esclerosis Múltiple/inmunología , Adulto , Citocinas/genética , Citocinas/metabolismo , Femenino , Humanos , Interleucina-5/genética , Interleucina-5/metabolismo , Interleucina-5/farmacología , Células Asesinas Naturales/clasificación , Activación de Linfocitos/efectos de los fármacos , Masculino , Esclerosis Múltiple/tratamiento farmacológico , Fenotipo , Receptores de Citocinas/genética , Receptores de Citocinas/metabolismo , Recurrencia , Inducción de Remisión , Células TH1/efectos de los fármacos , Células TH1/inmunología , Transcripción Genética , Receptor fas/metabolismo
11.
J Clin Invest ; 105(7): 977-84, 2000 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-10841661

RESUMEN

In multiple sclerosis (MS) patients who carry the Class II major histocompatibility (MHC) type HLA-DR2, T cells specific for amino acids 95-116 in the proteolipid protein (PLP) are activated and clonally expanded. However, it remains unclear whether these autoreactive T cells play a pathogenic role or, rather, protect against the central nervous system (CNS) damage. We have addressed this issue, using mice transgenic for the human MHC class II region carrying the HLA-DR2 (DRB1* 1502) haplotype. After stimulating cultured lymph node cells repeatedly with PLP95-116, we generated 2 HLA-DR2-restricted, PLP95-116-specific T-cell lines (TCLs) from the transgenic mice immunized with this portion of PLP. The TCLs were CD4+ and produced T-helper 1 (Th1) cytokines in response to the peptide. These TCLs were adoptively transferred into RAG-2/2 mice expressing HLA-DR2 (DRG1* 1502) molecules. Mice receiving 1 of the TCLs developed a neurological disorder manifested ataxic movement without apparent paresis on day 3, 4, or 5 after cell transfer. Histological examination revealed inflammatory foci primarily restricted to the cerebrum and cerebellum, in association with scattered demyelinating lesions in the deep cerebral cortex. These results support a pathogenic role for PLP95-116-specific T cells in HLA-DR2+ MS patients, and shed light on the possible correlation between autoimmune target epitope and disease phenotype in human CNS autoimmune diseases.


Asunto(s)
Epítopos de Linfocito T/inmunología , Antígenos HLA-DR/inmunología , Antígeno HLA-DR2/inmunología , Esclerosis Múltiple/inmunología , Proteína Proteolipídica de la Mielina/inmunología , Secuencia de Aminoácidos , Animales , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/inmunología , Encefalomielitis Autoinmune Experimental/inmunología , Femenino , Expresión Génica , Antígenos HLA-DR/biosíntesis , Antígenos HLA-DR/genética , Antígeno HLA-DR2/biosíntesis , Cadenas HLA-DRB1 , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Datos de Secuencia Molecular , Proteínas Nucleares , Fragmentos de Péptidos/inmunología , Linfocitos T Citotóxicos/inmunología
12.
Leukemia ; 31(12): 2709-2716, 2017 12.
Artículo en Inglés | MEDLINE | ID: mdl-28386106

RESUMEN

Myelofibrosis (MF) may be caused by various pathogenic mechanisms such as elevation in circulating cytokine levels, cellular interactions and genetic mutations. However, the underlying mechanism of MF still remains unknown. Recent studies have revealed that fibrocytes, the spindle-shaped fibroblast-like hematopoietic cells, and the thrombopoietin (TPO)/myeloproliferative leukemia protein (MPL; TPO receptor) signaling pathway play a certain role in the development of MF. In the present study, we aimed to investigate the relationship between fibrocytes and MPL activation. We showed that TPO or a TPO receptor agonist directly induces fibrocyte differentiation using murine fibrocyte cell lines and a murine MF model. Conversely, elimination of macrophages expressing MPL by clodronate liposomes reversed the MF phenotype of the murine model, suggesting that fibrocyte differentiation induced by MPL activation contributes to the progression of MF. Furthermore, we revealed that SLAMF7high MPLhigh monocytes in human peripheral blood mononuclear cells were possible fibrocyte precursors and that these cells increased in number in MF patients not treated with ruxolitinib. Our findings confirmed a link between fibrocytes and the TPO/MPL signaling pathway, which could result in a greater understanding of the pathogenesis of MF and lead to the development of novel therapeutic interventions.


Asunto(s)
Mielofibrosis Primaria/etiología , Mielofibrosis Primaria/metabolismo , Receptores de Trombopoyetina/metabolismo , Animales , Médula Ósea/metabolismo , Médula Ósea/patología , Diferenciación Celular , Línea Celular , Ácido Clodrónico/farmacología , Fibroblastos/citología , Fibroblastos/metabolismo , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/metabolismo , Humanos , Inmunohistoquímica , Janus Quinasa 2/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Monocitos/citología , Monocitos/metabolismo , Fenotipo , Mielofibrosis Primaria/patología , Factores de Transcripción STAT/metabolismo , Transducción de Señal , Trombopoyetina/metabolismo
13.
J Phys Condens Matter ; 18(26): 6109-16, 2006 Jul 05.
Artículo en Inglés | MEDLINE | ID: mdl-21690824

RESUMEN

We have examined the structure of a U(2)IrSi(3) compound exhibiting ferromagnetic cluster glass behaviour by means of electron diffraction observation and high-resolution electron microscopy. The structure of U(2)IrSi(3) has been proposed as a new one of the U(2)RuSi(3)-type with a short-range ordered double stacking sequence of the U(2)RuSi(3)-type structure along the c-axis, and long-range ordered atomic arrangements in the a-b plane. The calculated patterns reproduce the characteristic features of observed electron patterns well. The Fourier-filtered high-resolution image clearly exhibits a micro-domain structure, which is considered to relate directly to the origin of the observed cluster glass behaviour in U(2)IrSi(3).

14.
Cancer Res ; 61(3): 1029-37, 2001 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-11221829

RESUMEN

5-Fluorouracil (5-FU) is one of the most widely used anticancer agents for advanced colorectal carcinoma, but its response rate is only 15%. The "pharmacokinetic modulating chemotherapy" (PMC) regimen that we have advocated has proved to be highly effective in treating colorectal carcinoma. PMC consists of a continuous i.v. infusion of 5-FU over 24 h for 1day a week at 600 mg/m2/day, and an oral dose of uracil-tegafur (UFT), a 5-FU derivative, at 400 mg/day for 5-7 days per week, repeated every week for more than 6 months. Assays of 5-FU in 23 patients receiving this treatment showed serum concentrations ranging from 88 to 1,323 ng/ml. We then analyzed the effects of clinically relevant concentrations of 5-FU found in colorectal cancer patients treated with the PMC regimen on the growth of three human colorectal adenocarcinoma cell lines, SW480 and COLO320DM (mutant p53) and HCT116 (wild-type p53). Exposure of these three cell lines to 5-FU resulted in growth inhibition in a dose-dependent manner. Exposure to 100 ng/ml of 5-FU in SW480 and COLO320DM caused G1 arrest after 24 h and G2 arrest after 72-144 h, and only a minority of the cell population showed apoptotic features, which indicated that most of the cells were killed through mitotic catastrophe, nonapoptotic cell death. On the contrary, exposure to 1000 ng/ml of 5-FU in SW480 and COLO320DM resulted in G1-S-phase arrest and the induction of apoptosis throughout the experimental period. Nuclear cyclin B1 expression was markedly induced with exposure to 100 ng/ml of 5-FU in SW480 and COLO320DM; and expression of 14-3-3sigma protein, a cell cycle inhibitor in the GG phase, was induced in SW480. ICT116 responded to lower concentrations of 5-FU more rapidly: G2 arrest was seen after 24-72 h of exposure to 10 ng/ml of 5-FU, and G,1rrest was seen after 12-24 h of exposure to 100 ng/ml. These results show that 5-FU acts via two different pathways, depending on dose: (a) G,1S-phase cell cycle arrest and apoptosis at 1,000 ng/ml in SW480 and COLO320DM, and 100 ng/ml in HCT116; and (b) G2-M-phase cell cycle arrest and mitotic catastrophe at 100 ng/ll in SW480 and COLO320DM, and 10 ng/ml in HCT116. These results suggest that the efficacy of our PMC regimen is based on targeting at least two different phases of the cell cycle. In our clinical trial, we showed efficacy independent of p53 status, ascertained by cell kinetic analysis in vitro, which may lead to a novel concept of schedule-oriented biochemical modulation of this drug.


Asunto(s)
Adenocarcinoma/patología , Antimetabolitos Antineoplásicos/farmacología , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Neoplasias Colorrectales/patología , Fluorouracilo/farmacología , Adenocarcinoma/sangre , Adenocarcinoma/tratamiento farmacológico , Administración Oral , Antimetabolitos Antineoplásicos/administración & dosificación , Antimetabolitos Antineoplásicos/farmacocinética , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/farmacocinética , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Proteínas de Ciclo Celular/biosíntesis , División Celular/efectos de los fármacos , Neoplasias Colorrectales/sangre , Neoplasias Colorrectales/tratamiento farmacológico , ADN de Neoplasias/análisis , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Fluorouracilo/administración & dosificación , Fluorouracilo/sangre , Fluorouracilo/farmacocinética , Humanos , Infusiones Intravenosas , Tegafur/administración & dosificación , Células Tumorales Cultivadas , Uracilo/administración & dosificación
15.
Transl Psychiatry ; 6: e754, 2016 Mar 08.
Artículo en Inglés | MEDLINE | ID: mdl-26954981

RESUMEN

Despite novel antidepressant development, 10-30% of patients with major depressive disorder (MDD) have antidepressant treatment-resistant depression (TRD). Although new therapies are needed, lack of knowledge regarding the neural mechanisms underlying TRD hinders development of new therapeutic options. We aimed to identify brain regions in which spontaneous neural activity is not only altered in TRD but also associated with early treatment resistance in MDD. Sixteen patients with TRD, 16 patients with early-phase non-TRD and 26 healthy control (HC) subjects underwent resting-state functional magnetic resonance imaging. To identify brain region differences in spontaneous neural activity between patients with and without TRD, we assessed fractional amplitude of low-frequency fluctuations (fALFF). We also calculated correlations between the percent change in the Hamilton Rating Scale for Depression (HRSD17) scores and fALFF values in brain regions with differing activity for patients with and without TRD. Patients with TRD had increased right-thalamic fALFF values compared with patients without TRD. The percent change in HRSD17 scores negatively correlated with fALFF values in patients with non-TRD. In addition, patients with TRD showed increased fALFF values in the right inferior frontal gyrus (IFG), inferior parietal lobule (IPL) and vermis, compared with patients with non-TRD and HC subjects. Our results show that spontaneous activity in the right thalamus correlates with antidepressant treatment response. We also demonstrate that spontaneous activity in the right IFG, IPL and vermis may be specifically implicated in the neural pathophysiology of TRD.


Asunto(s)
Trastorno Depresivo Mayor/diagnóstico por imagen , Trastorno Depresivo Resistente al Tratamiento/diagnóstico por imagen , Lóbulo Parietal/diagnóstico por imagen , Corteza Prefrontal/diagnóstico por imagen , Tálamo/diagnóstico por imagen , Adulto , Antidepresivos/uso terapéutico , Encéfalo/diagnóstico por imagen , Encéfalo/fisiopatología , Estudios de Casos y Controles , Trastorno Depresivo Mayor/tratamiento farmacológico , Trastorno Depresivo Mayor/fisiopatología , Trastorno Depresivo Resistente al Tratamiento/tratamiento farmacológico , Trastorno Depresivo Resistente al Tratamiento/fisiopatología , Femenino , Neuroimagen Funcional , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Lóbulo Parietal/fisiopatología , Corteza Prefrontal/fisiopatología , Tálamo/fisiopatología
16.
Biochim Biophys Acta ; 1008(3): 287-92, 1989 Aug 14.
Artículo en Inglés | MEDLINE | ID: mdl-2547436

RESUMEN

An endonuclease endogenous to rat-liver nuclei has been purified by a series of chromatographic procedures and finally by isoelectric focusing (IEF) electrophoresis. The nuclease fraction prepared by the IEF electrophoresis (IEF fraction) showed a pI value of 5.7 and migrated as a single band to a molecular weight position of 46,000 on an SDS-polyacrylamide gel. The activity for single-stranded DNA was enhanced by 10 mM MgCl2 and/or by 5-15 mM MgCl2 in the presence of 2 mM CaCl2 (an optimum pH, 7.0), but was lowered by CaCl2 alone and inhibited strongly by ZnCl2 or MnCl2. The activity for duplex DNA was rather low, although an optimum condition was 10 mM MgCl2. In fact, even under this condition, the activity was about 40% lower than that for single-stranded DNA. Moreover, the IEF fraction formed single-strand nicks much more rapidly than double-strand cuts in pBR322 DNA, and preferentially produced deoxyadenosine 5'-monophosphate termini in the DNA. In addition, RNAase activity was also detected in this fraction.


Asunto(s)
Endonucleasas/aislamiento & purificación , Hígado/enzimología , Animales , Núcleo Celular/enzimología , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Endonucleasas/metabolismo , Focalización Isoeléctrica , Hígado/citología , Ratas , Espectrofotometría Ultravioleta , Especificidad por Sustrato
17.
Biochim Biophys Acta ; 1127(2): 116-23, 1992 Jul 29.
Artículo en Inglés | MEDLINE | ID: mdl-1643096

RESUMEN

Two sandwich-type enzyme immunoassays have been developed to measure apolipoproteins A-I and E in rabbit serum. Specific goat antibodies were purified by affinity chromatography and used both for coating and for preparing antibody-peroxydase conjugates. The sensitivity of these assays is sufficient to allow studies of apo A-I and E distribution in lipoproteins fractionated by gel filtration from 50 microliters of serum. In WHHL rabbits, apo A-I is 5-fold lower (5.2 +/- 2.5 mg/dl) and apo E is 8-fold higher (9.9 +/- 3.5 mg/dl) than in normolipidemic rabbits (29 +/- 4.3 mg/dl and 1.3 +/- 0.5 mg/dl, respectively). In hyperlipidemic rabbits, fed 2 months on a 0.5% cholesterol diet, the apo A-I level was similar (32 +/- 12 mg/dl) to that of normolipidemic rabbits, but the apo E level is 12-fold higher (15.1 +/- 5.5 mg/dl). In addition, HDL particles were enriched with cholesterol and apo E. The bulk of apo E and cholesterol is located in large beta-VLDL in diet-induced hyperlipidemia, whereas they are mainly located in smaller size beta-VLDL in WHHL rabbits. In normolipidemic rabbits apo E occurs mainly in HDL, and cholesterol is distributed in the main three lipoprotein fractions VLDL, LDL and HDL. Interestingly, HDL of WHHL rabbit are deficient in apo A-I. These results are compatible with profound perturbations of lipoprotein composition and metabolism in atherogenic hyperlipidemia.


Asunto(s)
Apolipoproteína A-I/análisis , Apolipoproteínas E/análisis , Ensayo de Inmunoadsorción Enzimática/métodos , Hiperlipidemias/sangre , Animales , Especificidad de Anticuerpos , Dieta , Hiperlipidemias/etiología , Lipoproteínas/química , Conejos , Sensibilidad y Especificidad
18.
Biochim Biophys Acta ; 875(2): 211-9, 1986 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-3942763

RESUMEN

The effect of apolipoproteins C-II and C-III on the lipoprotein lipase-catalyzed hydrolysis of apolipoprotein C-II-deficient triacylglycerol-rich lipoproteins and particles of trioleoylglycerol stabilized with a phosphatidylcholine monolayer was investigated. For both triacylglycerol-rich lipoproteins and artificial lipid particles, maximal lipoprotein lipase activity occurred at a constant apolipoprotein C-II/phospholipid mol ratio of 2.0 X 10(-4) and was independent of particle size, indicating that the amount of apolipoprotein C-II bound to the surface of the substrate is important for enzyme activation. The effect of apolipoprotein C-II on lipoprotein lipase activity with apolipoprotein C-II-deficient lipoproteins as substrate was to decrease the apparent Michaelis constant (Kmapp) from 7.1 to 1.0 mM with minor changes on the apparent maximal velocity (Vmax) (22.2 mmol free fatty acid released/h per mg enzyme). In contrast, apolipoprotein C-II increased the apparent Vmax from 2.4 to 20.0 mmol free fatty acid/h per mg enzyme for the lipoprotein lipase-catalyzed hydrolysis of trioleoylglycerol/phospholipid particles with little change in Kmapp (1.0 mM). Addition of apolipoprotein C-II-deficient triacylglycerol-rich lipoproteins or high-density lipoproteins to trioleoylglycerol/phospholipid particles in the presence of apolipoprotein C-II inhibited lipoprotein lipase activity. Lipoprotein lipase activity was also inhibited by the addition of a large excess of lipid-free apolipoprotein C-III to the artificial particles. The decrease in lipoprotein lipase activity correlated with the amount of bound apolipoprotein C-II. We suggest that the reported discrepancies on the effect of apolipoproteins C-II and C-III on lipoprotein lipase catalysis is related to differences in substrates and to the amount of added apolipoproteins.


Asunto(s)
Apolipoproteínas C/metabolismo , Lipoproteína Lipasa/metabolismo , Lipoproteínas/metabolismo , Fosfatidilcolinas/metabolismo , Trioleína/metabolismo , Apolipoproteína C-II , Humanos , Hidrólisis , Cinética
19.
Circulation ; 102(19 Suppl 3): III269-74, 2000 Nov 07.
Artículo en Inglés | MEDLINE | ID: mdl-11082400

RESUMEN

BACKGROUND: The molecular mechanism of neointimal hyperplasia after vein graft surgery remains elusive. Vacuolar H(+)-ATPase (V-ATPase) is involved in intracellular trafficking and may play a crucial role in neointimal cell growth. METHODS AND RESULTS: Cultured human saphenous vein segments developed neointimal formation within 10 days. Neointimal cells were positive for vimentin and alpha-smooth muscle actin but negative for desmin, which is indicative of myofibroblasts. Those myofibroblasts were found to have originated from periadventitial fibroblasts, which upregulated the expression of 16-kDa proteolipid of V-ATPase before proliferation and phenotypic modulation. Neointimal myofibroblast growth and survival were highly sensitive to inhibition of V-ATPase by bafilomycin A(1) (BA(1)), because the incorporation of [(3)H]thymidine into the myofibroblasts was significantly inhibited by nanomolar concentrations of BA(1) and apoptotic cell death was induced by a similar concentration range of BA(1). In contrast, endothelial cells and differentiated smooth muscle cells were resistant to apoptosis by BA(1). CONCLUSIONS: These results suggest that V-ATPase plays a crucial role in growth and phenotypic modulation of myofibroblasts that contributes to neointimal formation in cultured human saphenous vein.


Asunto(s)
Fibroblastos/enzimología , Macrólidos , Músculo Liso Vascular/enzimología , ATPasas de Translocación de Protón/metabolismo , Vena Safena/enzimología , Túnica Íntima/metabolismo , ATPasas de Translocación de Protón Vacuolares , Adulto , Anciano , Anciano de 80 o más Años , Antibacterianos/farmacología , Antígenos de Diferenciación/biosíntesis , Apoptosis/efectos de los fármacos , Bromodesoxiuridina , División Celular/efectos de los fármacos , Movimiento Celular , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Humanos , Técnicas In Vitro , Masculino , Persona de Mediana Edad , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Fenotipo , Subunidades de Proteína , Proteolípidos/biosíntesis , Vena Safena/citología , Timidina/metabolismo , Túnica Íntima/citología
20.
Inflamm Bowel Dis ; 11(12): 1038-43, 2005 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-16306765

RESUMEN

BACKGROUND: Interleukin-18 (IL-18) is a pleiotropic cytokine that induces the production of interferon (IFN)-gamma and also to regulate Th2 cytokines. Recently, association studies between IL-18 gene promoter polymorphisms and several Th1- or Th2-mediated inflammatory diseases were reported. In inflammatory bowel diseases (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), recent evidence suggests that IL-18 is involved in the pathogenesis. METHODS: Using DNA direct sequencing, we investigated IL-18 gene promoter polymorphisms at -607C/A and -137G/C. Allele, genotype, and haplotype frequencies were determined in 210 Japanese patients with UC, 205 patients with CD, and 212 controls. RESULTS: In UC, the -137C allele frequency was significantly higher in the proctitis-type patients than in controls (Pc = 0.0068). The -137 genotype frequency was also significantly different in the proctitis-type patients than in controls (Pc = 0.032). No other allele and genotype frequencies were significantly associated with UC after Bonferroni correction. Furthermore, the frequency of haplotype 2 (-607A, -137C), which had a lower promoter activity and IFN-gamma mRNA level than the other haplotypes as previously reported, was significantly higher in the proctitis-type patients than in controls (Pc = 0.01). In CD, we could not find any significant differences. CONCLUSIONS: IL-18 gene promoter polymorphisms may not be associated with disease susceptibility but related to the extent of disease in UC.


Asunto(s)
Colitis Ulcerosa/genética , Enfermedad de Crohn/genética , Predisposición Genética a la Enfermedad/genética , Interleucina-18/genética , Polimorfismo Genético/genética , Regiones Promotoras Genéticas/genética , Adolescente , Adulto , Anciano , Pueblo Asiatico , Femenino , Haplotipos , Humanos , Japón , Masculino , Persona de Mediana Edad
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA