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1.
Cell ; 153(1): 112-25, 2013 Mar 28.
Artículo en Inglés | MEDLINE | ID: mdl-23477864

RESUMEN

Influenza A viruses are a major cause of mortality. Given the potential for future lethal pandemics, effective drugs are needed for the treatment of severe influenza such as that caused by H5N1 viruses. Using mediator lipidomics and bioactive lipid screen, we report that the omega-3 polyunsaturated fatty acid (PUFA)-derived lipid mediator protectin D1 (PD1) markedly attenuated influenza virus replication via RNA export machinery. Production of PD1 was suppressed during severe influenza and PD1 levels inversely correlated with the pathogenicity of H5N1 viruses. Suppression of PD1 was genetically mapped to 12/15-lipoxygenase activity. Importantly, PD1 treatment improved the survival and pathology of severe influenza in mice, even under conditions where known antiviral drugs fail to protect from death. These results identify the endogenous lipid mediator PD1 as an innate suppressor of influenza virus replication that protects against lethal influenza virus infection.


Asunto(s)
Transporte Activo de Núcleo Celular , Ácidos Docosahexaenoicos/inmunología , Subtipo H1N1 del Virus de la Influenza A/fisiología , Subtipo H5N1 del Virus de la Influenza A/fisiología , Infecciones por Orthomyxoviridae/inmunología , Replicación Viral , Transporte Activo de Núcleo Celular/efectos de los fármacos , Animales , Línea Celular , Ácidos Docosahexaenoicos/análisis , Ácidos Docosahexaenoicos/farmacología , Humanos , Ratones , Infecciones por Orthomyxoviridae/tratamiento farmacológico , Infecciones por Orthomyxoviridae/virología , Replicación Viral/efectos de los fármacos
2.
Nat Immunol ; 14(6): 554-63, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23624557

RESUMEN

Microenvironment-based alterations in phenotypes of mast cells influence the susceptibility to anaphylaxis, yet the mechanisms underlying proper maturation of mast cells toward an anaphylaxis-sensitive phenotype are incompletely understood. Here we report that PLA2G3, a mammalian homolog of anaphylactic bee venom phospholipase A2, regulates this process. PLA2G3 secreted from mast cells is coupled with fibroblastic lipocalin-type PGD2 synthase (L-PGDS) to provide PGD2, which facilitates mast-cell maturation via PGD2 receptor DP1. Mice lacking PLA2G3, L-PGDS or DP1, mast cell-deficient mice reconstituted with PLA2G3-null or DP1-null mast cells, or mast cells cultured with L-PGDS-ablated fibroblasts exhibited impaired maturation and anaphylaxis of mast cells. Thus, we describe a lipid-driven PLA2G3-L-PGDS-DP1 loop that drives mast cell maturation.


Asunto(s)
Fosfolipasas A2 Grupo III/inmunología , Mastocitos/inmunología , Comunicación Paracrina/inmunología , Prostaglandina D2/inmunología , Receptores de Prostaglandina/inmunología , Animales , Western Blotting , Células de la Médula Ósea/inmunología , Células de la Médula Ósea/metabolismo , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Células Cultivadas , Fibroblastos/citología , Fibroblastos/inmunología , Fibroblastos/metabolismo , Perfilación de la Expresión Génica , Fosfolipasas A2 Grupo III/genética , Fosfolipasas A2 Grupo III/metabolismo , Humanos , Oxidorreductasas Intramoleculares/genética , Oxidorreductasas Intramoleculares/inmunología , Oxidorreductasas Intramoleculares/metabolismo , Lipocalinas/genética , Lipocalinas/inmunología , Lipocalinas/metabolismo , Mastocitos/metabolismo , Mastocitos/ultraestructura , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Electrónica de Transmisión , Análisis de Secuencia por Matrices de Oligonucleótidos , Comunicación Paracrina/genética , Prostaglandina D2/metabolismo , Receptores de Prostaglandina/genética , Receptores de Prostaglandina/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
3.
Cell ; 141(6): 1068-79, 2010 Jun 11.
Artículo en Inglés | MEDLINE | ID: mdl-20537373

RESUMEN

Elucidation of molecular mechanisms that regulate synapse formation is required for the understanding of neural wiring, higher brain functions, and mental disorders. Despite the wealth of in vitro information, fundamental questions about how glutamatergic synapses are formed in the mammalian brain remain unanswered. Glutamate receptor (GluR) delta2 is essential for cerebellar synapse formation in vivo. Here, we show that the N-terminal domain (NTD) of GluRdelta2 interacts with presynaptic neurexins (NRXNs) through cerebellin 1 precursor protein (Cbln1). The synaptogenic activity of GluRdelta2 is abolished in cerebellar primary cultures from Cbln1 knockout mice and is restored by recombinant Cbln1. Knockdown of NRXNs in cerebellar granule cells also hinders the synaptogenic activity of GluRdelta2. Both the NTD of GluRdelta2 and the extracellular domain of NRXN1beta suppressed the synaptogenic activity of Cbln1 in cerebellar primary cultures and in vivo. These results suggest that GluRdelta2 mediates cerebellar synapse formation by interacting with presynaptic NRXNs through Cbln1.


Asunto(s)
Cerebelo/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Moléculas de Adhesión de Célula Nerviosa/metabolismo , Precursores de Proteínas/metabolismo , Receptores de Glutamato/metabolismo , Sinapsis , Animales , Proteínas de Unión al Calcio , Línea Celular , Células Cultivadas , Humanos , Ratones
4.
Cell ; 139(2): 352-65, 2009 Oct 16.
Artículo en Inglés | MEDLINE | ID: mdl-19837036

RESUMEN

Many eukaryotic proteins are attached to the cell surface via glycosylphosphatidylinositol (GPI) anchors. How GPI-anchored proteins (GPI-APs) are trafficked from the endoplasmic reticulum (ER) to the cell surface is poorly understood, but the GPI moiety has been postulated to function as a signal for sorting and transport. Here, we established mutant cells that were selectively defective in transport of GPI-APs from the ER to the Golgi. We identified a responsible gene, designated PGAP5 (post-GPI-attachment to proteins 5). PGAP5 belongs to a dimetal-containing phosphoesterase family and catalyzed the remodeling of the glycan moiety on GPI-APs. PGAP5 catalytic activity is a prerequisite for the efficient exit of GPI-APs from the ER. Our data demonstrate that GPI glycan acts as an ER-exit signal and suggest that glycan remodeling mediated by PGAP5 regulates GPI-AP transport in the early secretory pathway.


Asunto(s)
Retículo Endoplásmico/metabolismo , Aparato de Golgi/metabolismo , Hidrolasas Diéster Fosfóricas/metabolismo , Polisacáridos/metabolismo , Animales , Glicoproteínas/metabolismo , Glicosilfosfatidilinositoles/metabolismo , Humanos , Ratones , Hidrolasas Diéster Fosfóricas/genética
5.
Kyobu Geka ; 77(6): 446-449, 2024 Jun.
Artículo en Japonés | MEDLINE | ID: mdl-39009539

RESUMEN

Dextrocardia associated with situs inversus totalis is a rare congenital anomaly. We herein report a patient with this condition and ischemic coronay artery disease who underwent emergency surgical myocardial revascularization. A 76-year-old man was admitted to our hospital with a diagnosis of unstable angina pectoris. He had incessant ventricular fibrillation attack on the second day of hospitalization. Emergent coronary angiography revealed total obstruction of the anatomically left anterior descending and right coronary arteries, and severe stenosis in the anatomically left circumflex artery. Emergent off-pump coronary artery bypass using saphenous vein grafts was successfully performed. Intraoperatively, the main surgeon continuouly stood on the patient's right side except during anastosiso of a vein graft to the left circumflex artery. Postoperative course was almost uneventful aside from transient renal replacement and prolonged ventilation.


Asunto(s)
Puente de Arteria Coronaria Off-Pump , Situs Inversus , Humanos , Masculino , Anciano , Situs Inversus/complicaciones , Situs Inversus/cirugía , Situs Inversus/diagnóstico por imagen , Angiografía Coronaria , Dextrocardia/complicaciones , Dextrocardia/cirugía , Dextrocardia/diagnóstico por imagen
6.
Soft Matter ; 17(15): 4040-4046, 2021 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-33881047

RESUMEN

Flexibility, viscoelasticity and stress-strain relation in bending polymeric films are key factors in designing mechanically durable flexible electronic devices and soft robots. However, bending hysteresis, which appears as a precursor phenomenon of fracture and fatigue, remains unclear; no one quantitatively evaluated a bending curvature causing hysteresis. Herein, we report the bending hysteresis of polymeric films used as common substrates in flexible electronics by precisely monitoring bending curvatures. By real-time measuring curvatures of films upon bending and subsequent unbending, we have successfully determined the curvatures that cause the hysteresis. These curvatures also depend on a film thickness. Furthermore, we revealed that the occurrence of bending hysteresis is explained by bending strains that have a nonlinear relation with internal stresses. This enables us to predict strain limits that cause the bending hysteresis, based on a stress-strain curve of polymeric films.

7.
Palliat Support Care ; 19(3): 377-379, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-33947505

RESUMEN

OBJECTIVE: Cognitive dysfunction has a negative effect on cancer treatment; however, in a cancer setting, specific treatments can restore cognitive function. Such conditions are known as reversible dementia, with one of these being vitamin B12 (VB12) deficiency. However, there have been no reports of VB12 deficiency identified by preoperative evaluation in cancer patients. METHOD: We studied a patient who was referred to the Department of Psycho-oncology on suspicion of cognitive decline prior to lung cancer surgery. Preoperative evaluation revealed VB12 deficiency. RESULTS: The patient was an 82-year-old woman diagnosed with lung cancer. She also presented with cognitive decline and, therefore, was referred to the Department of Psycho-oncology for preoperative evaluation. The patient scored 19 points on a Mini-Mental State Examination (MMSE), which is indicative of cognitive decline. As the onset of symptoms occurred several months previously and they were subacute, the possibility of reversible dementia was considered. Extensive examination revealed VB12 deficiency, and VB12 replacement therapy normalized the MMSE score to 25 points before surgery. SIGNIFICANCE OF THE RESULTS: When cognitive decline is observed in cancer patients, it is necessary to actively evaluate the serum levels of some B vitamins, including VB12.


Asunto(s)
Disfunción Cognitiva , Demencia , Neoplasias Pulmonares , Deficiencia de Vitamina B 12 , Anciano de 80 o más Años , Disfunción Cognitiva/etiología , Demencia/complicaciones , Femenino , Humanos , Neoplasias Pulmonares/complicaciones , Vitamina B 12/uso terapéutico , Deficiencia de Vitamina B 12/complicaciones
8.
Kyobu Geka ; 74(12): 1032-1035, 2021 Nov.
Artículo en Japonés | MEDLINE | ID: mdl-34795149

RESUMEN

We described a hemodialysis patient with spontaneous renal rupture of acquired cystic kidney disease (ACKD) after off-pump coronary artery bypass grafting(OPCAB). A 53-year-old man was transfer to our hospital with a diagnosis of unstable angina pectoris. He became ventilated due to congestive heart failure 3 days and underwent OPCAB on schedule 6 days after hospitalization. He suddenly complainted of colicky frank pain 31 days after surgery. Computed tomography (CT) demonstrated bulky hematoma with swollen kidney. Treatment included blood transfusions, withdrawal of anticoagulation, administration of vitamin K, and bed rest. The patient's symptom disappeared. A month later CT showed that the hematoma had been absorbed.


Asunto(s)
Puente de Arteria Coronaria Off-Pump , Enfermedad de la Arteria Coronaria , Enfermedades Renales Quísticas , Puente de Arteria Coronaria , Enfermedad de la Arteria Coronaria/complicaciones , Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Enfermedad de la Arteria Coronaria/cirugía , Humanos , Riñón , Masculino , Persona de Mediana Edad , Resultado del Tratamiento
9.
Genes Cells ; 23(10): 880-892, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-30133879

RESUMEN

Yeasts have two classes of glycosylphosphatidylinositol (GPI)-anchored proteins; one is transferred to the cell wall, whereas the other is retained on the plasma membrane. The lipid moieties of the GPI in Saccharomyces cerevisiae consist of either phosphatidylinositol (PI) or inositolphosphorylceramide (IPC). Cwh43p is involved in the remodeling of lipid from PI to IPC. We found that the GPI lipid moiety of Cwp2p in wild-type cells is PI. To elucidate the physiological role of the lipid remodeling by Cwh43p, we investigated the distribution of Gas1p and Cwp2p by immunoblotting and found that Gas1p with the PI-form GPI lipid moiety in cwh43∆ mutant cells tends to be localized to the cell wall, suggesting that the IPC species in the GPI lipid moiety contributes to the retention of GPI-anchored proteins on the plasma membrane. We also found that CWH43 is genetically related to TED1, which encodes a protein involved in the removal of the ethanolamine phosphate from the second mannose residue in GPI glycan moieties. We propose possible models for the physiological function of Cwh43p and Ted1p in the transfer of GPI-anchored proteins from the plasma membrane to the cell wall.


Asunto(s)
Glicoesfingolípidos/metabolismo , Glicosilfosfatidilinositoles/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Membrana Celular/metabolismo , Pared Celular/metabolismo , Lípidos/fisiología , Glicoproteínas de Membrana/metabolismo , Proteínas de la Membrana/genética , Fosfatidilinositoles/metabolismo , Transporte de Proteínas , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética
10.
Kyobu Geka ; 72(13): 1081-1084, 2019 Dec.
Artículo en Japonés | MEDLINE | ID: mdl-31879384

RESUMEN

We presented a case with chronic occlusion of the left anterior descending artery(LAD) after mitral valve replacement (MVR). An 80-year-old man was transferred to our hospital with a diagnosis of acute myocardial infarction. Before coronary angiography, the patient suddenly fell in a state of cardiogenic shock. Echocardiography revealed acute mitral regurgitation due to anterior papillary muscle rupture. Although emergency MVR was performed, introduction of percutaneous cardiopulmonary support (PCPS) and intraaortic balloon pumping ( IABP) was needed to come off cardiopulmonary bypass. Postoperative coronary angiography revealed chronic occlusion of LAD. The left internal thoracic artery was anastomosed to LAD via anterior minithracotomy to avoid median re-sternotomy. The postoperative course was uneventful and the cardiac function improved.


Asunto(s)
Rotura Cardíaca , Insuficiencia de la Válvula Mitral , Anciano de 80 o más Años , Puente de Arteria Coronaria , Humanos , Masculino , Músculos Papilares , Toracotomía
11.
Kyobu Geka ; 72(1): 17-22, 2019 Jan.
Artículo en Japonés | MEDLINE | ID: mdl-30765624

RESUMEN

OBJECTIVES: Pulmonary lobectomy is the standard surgical procedure for resectable non-small cell lung cancer (NSCLC), while sublobar resection is an important surgical alternative for high-risk patients with comorbidities. We evaluated the treatment outcome and prognostic factors of sublobar resection in high-risk patients with NSCLC. METHODS: Eighty three high-risk patients who underwent compromised sublobar resection for clinical-N0 NSCLC with a solid appearance were retrospectively reviewed. A total of 47 wedge resections and 36 segmentectomies performed. RESULTS: Poor pulmonary function and synchronous or metachronous multiple lung cancer were found in 56.7% and 20.5% of patients respectively, all requiring sublobar resection. There were 21 instances of tumor recurrence and 24 deaths during a mean follow-up of 1,500 days. There was no local recurrence in the segmentectomy group. The 3-year recurrence free survival (RFS) and overall survival (OS) were 72.6% and 73.8% respectively. A multivariate analysis indicated that resection type and lymphatic invasion were independent prognostic factors for RFS. In the wedge resection group, a ratio of surgical margin to clinical tumor size greater than 1 (MT ratio≥1) was an independent prognostic factor for RFS( 87.1%,p=0.001). CONCLUSION: Segmentectomy leads to a favorable prognosis. MT ratio was independently associated with a longer RFS in the wedge resection group.


Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/cirugía , Neoplasias Pulmonares/cirugía , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Supervivencia sin Enfermedad , Humanos , Neoplasias Pulmonares/mortalidad , Análisis Multivariante , Recurrencia Local de Neoplasia/mortalidad , Estadificación de Neoplasias , Neumonectomía/mortalidad , Neumonectomía/estadística & datos numéricos , Pronóstico , Estudios Retrospectivos , Riesgo , Resultado del Tratamiento
12.
Kyobu Geka ; 71(13): 1068-1072, 2018 12.
Artículo en Japonés | MEDLINE | ID: mdl-30587743

RESUMEN

We describe a patient with stenosis of an open stent graft during total arch replacement of a rightsided aortic arch. A 67-year-old man was transferred to our hospital with a diagnosis of acute type A (DeBakey type II) aortic dissection. Computed tomography( CT) imaging revealed concomitant rightsided aortic arch (RAA) and Kommerell diverticulum at the descending thoracic aorta. Total arch replacement was performed using an open stent graft over the Kommerell diverticulum. While closing the sternum, blood pressure of the femoral artery dropped to a level much lower than that of radial artery. Transesophageal echocardiography revealed an apparently new intimal flap in the descending thoracic aorta. Therefore, an axilla-femoral artery bypass and a femoro-femoral artery crossover bypass were performed to improve malperfusion of the visceral organs and lower extremities. CT imaging immediately after these procedures revealed stenosis at the non-stented portion of the open stent graft, which was located at the steep-angled site of the RAA. The pressure gradient between the upper and lower extremities was around 100 mmHg on the day of surgery, but it disappeared on postoperative day 4. Improved graft stenosis was confirmed by CT before discharge.


Asunto(s)
Aneurisma de la Aorta Torácica/cirugía , Disección Aórtica/cirugía , Complicaciones Posoperatorias/cirugía , Stents , Enfermedad Aguda , Anciano , Disección Aórtica/diagnóstico por imagen , Aorta Torácica/diagnóstico por imagen , Aneurisma de la Aorta Torácica/diagnóstico por imagen , Arteria Axilar/cirugía , Presión Sanguínea , Implantación de Prótesis Vascular , Constricción Patológica/diagnóstico por imagen , Constricción Patológica/cirugía , Divertículo/diagnóstico por imagen , Divertículo/cirugía , Arteria Femoral/fisiopatología , Arteria Femoral/cirugía , Humanos , Masculino , Complicaciones Posoperatorias/diagnóstico por imagen , Arteria Radial/fisiopatología , Procedimientos Quirúrgicos Vasculares
13.
Pathol Int ; 67(2): 99-104, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-28093881

RESUMEN

Ciliated muconodular papillary tumor (CMPT) is a rare papillary tumor that arises in the peripheral lung fields and is associated with the proliferation of ciliate d and goblet cells and increased mucin production. We report a case of CMPT involving the rearrangement of the anaplastic lymphoma kinase (ALK) gene. The patient was an 84-year-old Japanese female who had exhibited a small nodular shadow on chest computed tomography during a regular checkup 10 years ago. She underwent a partial resection of segment S10 of the right lung. The cut surface of the surgical specimen revealed a well-circumscribed, jelly-like mass measuring 8 × 8 × 10 mm. Histologically, the tumor was composed of a mixture of ciliated, goblet, and basal cells arranged in a papillary pattern together with pools of mucin. A diagnosis of CMPT was made. The lung tumor cells were subjected to fluorescent in situ hybridization and highly sensitive immunohistochemical staining for the ALK protein, both of which produced positive results. CMPT usually follows a favorable course, but the exact nature of this tumor; i.e., whether it is benign or malignant, has not been established. This is the first reported case of an ALK-positive CMPT.


Asunto(s)
Carcinoma Papilar/patología , Neoplasias Pulmonares/patología , Proteínas Tirosina Quinasas Receptoras/genética , Anciano de 80 o más Años , Quinasa de Linfoma Anaplásico , Biomarcadores de Tumor , Carcinoma Papilar/genética , Femenino , Células Caliciformes/patología , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Neoplasias Pulmonares/genética , Mucinas , Neoplasias Primarias Secundarias/genética , Neoplasias Primarias Secundarias/patología , Neoplasias del Cuello Uterino/patología
14.
J Biol Chem ; 290(2): 1005-19, 2015 Jan 09.
Artículo en Inglés | MEDLINE | ID: mdl-25361768

RESUMEN

Cryptococcosis is an infectious disease caused by pathogenic fungi, such as Cryptococcus neoformans and Cryptococcus gattii. The ceramide structure (methyl-d18:2/h18:0) of C. neoformans glucosylceramide (GlcCer) is characteristic and strongly related to its pathogenicity. We recently identified endoglycoceramidase-related protein 1 (EGCrP1) as a glucocerebrosidase in C. neoformans and showed that it was involved in the quality control of GlcCer by eliminating immature GlcCer during the synthesis of GlcCer (Ishibashi, Y., Ikeda, K., Sakaguchi, K., Okino, N., Taguchi, R., and Ito, M. (2012) Quality control of fungus-specific glucosylceramide in Cryptococcus neoformans by endoglycoceramidase-related protein 1 (EGCrP1). J. Biol. Chem. 287, 368-381). We herein identified and characterized EGCrP2, a homologue of EGCrP1, as the enzyme responsible for sterylglucoside catabolism in C. neoformans. In contrast to EGCrP1, which is specific to GlcCer, EGCrP2 hydrolyzed various ß-glucosides, including GlcCer, cholesteryl-ß-glucoside, ergosteryl-ß-glucoside, sitosteryl-ß-glucoside, and para-nitrophenyl-ß-glucoside, but not α-glucosides or ß-galactosides, under acidic conditions. Disruption of the EGCrP2 gene (egcrp2) resulted in the accumulation of a glycolipid, the structure of which was determined following purification to ergosteryl-3ß-glucoside, a major sterylglucoside in fungi, by mass spectrometric and two-dimensional nuclear magnetic resonance analyses. This glycolipid accumulated in vacuoles and EGCrP2 was detected in vacuole-enriched fraction. These results indicated that EGCrP2 was involved in the catabolism of ergosteryl-ß-glucoside in the vacuoles of C. neoformans. Distinct growth arrest, a dysfunction in cell budding, and an abnormal vacuole morphology were detected in the egcrp2-disrupted mutants, suggesting that EGCrP2 may be a promising target for anti-cryptococcal drugs. EGCrP2, classified into glycohydrolase family 5, is the first steryl-ß-glucosidase identified as well as a missing link in sterylglucoside metabolism in fungi.


Asunto(s)
Celulasas/genética , Ceramidas/metabolismo , Criptococosis/enzimología , Cryptococcus neoformans/genética , Proteínas Fúngicas/genética , Glicósido Hidrolasas/metabolismo , Celulasas/metabolismo , Ceramidas/química , Criptococosis/patología , Cryptococcus neoformans/metabolismo , Cryptococcus neoformans/patogenicidad , Glucosilceramidas/metabolismo , Glucolípidos/metabolismo , Glicósido Hidrolasas/química , Glicósido Hidrolasas/genética , Humanos
15.
Glycoconj J ; 33(2): 169-80, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26883028

RESUMEN

In this study, we immunized Gb3/CD77 synthase gene (A4galt) knockout (KO) mice with glycosphingolipids (GSLs) extracted from 3 renal cell cancer (RCC) cell lines to raise monoclonal antibodies (mAbs) reactive with globo-series GSLs specifically expressed in RCCs. Although a number of mAbs reactive with globo-series GSLs were generated, they reacted with both RCC cell lines and normal kidney cells. When we analyzed recognized antigens by mAbs that were specifically reactive with RCC, but not with normal kidney cells at least on the cell surface, many of them turned out to be reactive with sulfoglycolipids. Eight out of 11 RCC-specific mAbs were reactive with SM2 alone, and the other 3 mAbs were more broadly reactive with sulfated glycolipids, i.e. SM3 and SM4 as well as SM2. In the immunohistochemistry, these anti-sulfoglycolipids mAbs showed RCC-specific reaction, with no or minimal reaction with adjacent normal tissues. Thus, immunization of A4galt KO mice with RCC-derived GSLs resulted in the generation of anti sulfated GSL mAbs, and these mAbs may be applicable for the therapeutics for RCC patients.


Asunto(s)
Anticuerpos Monoclonales de Origen Murino/inmunología , Anticuerpos Antineoplásicos/inmunología , Galactosiltransferasas/deficiencia , Inmunización , Neoplasias Renales , Animales , Línea Celular Tumoral , Glucolípidos/química , Glucolípidos/inmunología , Glucolípidos/farmacocinética , Humanos , Neoplasias Renales/química , Neoplasias Renales/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Ratones Desnudos
16.
Biol Pharm Bull ; 39(2): 278-84, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26830486

RESUMEN

Hepatotoxicity is a known side effect of nonsteroidal anti-inflammatory drugs (NSAIDs). In the present study, the effects of N-phenylanthranilic acid (NPA) scaffold NSAIDs on rat liver mitochondria were examined. Mefenamic acid (MEF, 200 µM) induced mitochondrial swelling, which was inorganic phosphate (Pi)-dependent and suppressed by cyclosporin A (CsA, 2.5 µM), similar to calcium-induced swelling. Mitochondrial swelling was also observed following the addition of 200 µM flufenamic acid (FLU), meclofenamic acid (MCL), and tolfenamic acid (TOL). Less swelling was observed with the addition of 200 µM diclofenac (DIC) or NPA. Diphenylamine (DPA)-induced swelling occurred in a Pi-independent manner and was not sensitive to CsA. The mechanism by which DPA interacted with the mitochondrial inner membrane differed from those of the other NPA scaffold NSAIDs. The addition of 50 µM MEF, MCL, TOL, and FLU had uncoupling effects in mitochondrial inner membrane. These NSAIDs dose-dependently obstructed electron transport in the respiratory chain. NSAIDs are known to have various dynamic structures, and the solvation free energies (dGWs: an index of stereo-hydrophobicity) of the conformers obtained were determined using a molecular orbital analysis. The relationship between the dynamic structures and swelling induced by NPA scaffold NSAIDs was also examined.


Asunto(s)
Antiinflamatorios no Esteroideos/síntesis química , Mitocondrias Hepáticas/efectos de los fármacos , ortoaminobenzoatos/química , Animales , Antiinflamatorios no Esteroideos/química , Transporte de Electrón/efectos de los fármacos , Masculino , Mitocondrias Hepáticas/metabolismo , Estructura Molecular , Nitrilos/farmacología , Consumo de Oxígeno/efectos de los fármacos , Ratas , Ratas Wistar
17.
Proc Natl Acad Sci U S A ; 110(12): 4714-9, 2013 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-23471986

RESUMEN

Although endogenous ligands for Toll-like receptor (TLR)4-myeloid differentiation factor 2 (MD2) have not been well-understood, we here report that a globo-series glycosphingolipid, globotetraosylceramide (Gb4), attenuates the toxicity of lipopolysaccharides (LPSs) by binding to TLR4-MD-2. Because α1,4-galactosyltransferase (A4galt)-deficient mice lacking globo-series glycosphingolipids showed higher sensitivity to LPS than wild-type mice, we examined mechanisms by which globo-series glycosphingolipids attenuate LPS toxicity. Cultured endothelial cells lacking A4galt showed higher expression of LPS-inducible genes upon LPS treatment. In turn, introduction of A4galt cDNA resulted in the neo expression of Gb4, leading to the reduced expression of LPS-inducible genes. Exogenous Gb4 induced similar effects. As a mechanism for the suppressive effects of Gb4 on LPS signals, specific binding of Gb4 to the LPS receptor TLR4-MD-2 was demonstrated by coprecipitation of Gb4 with recombinant MD-2 and by native PAGE. A docking model also supported these data. Taken together with colocalization of TLR4-MD-2 with Gb4 in lipid rafts after LPS stimulation, it was suggested that Gb4 competes with LPS for binding to TLR4-MD-2. Finally, administration of Gb4 significantly protected mice from LPS-elicited mortality. These results suggest that Gb4 is an endogenous ligand for TLR4-MD-2 and is capable of attenuating LPS toxicity, indicating the possibility for its therapeutic application in endotoxin shock.


Asunto(s)
Globósidos/inmunología , Antígeno 96 de los Linfocitos/inmunología , Complejos Multiproteicos/inmunología , Receptor Toll-Like 4/inmunología , Animales , Galactosiltransferasas/genética , Galactosiltransferasas/inmunología , Galactosiltransferasas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Regulación de la Expresión Génica/inmunología , Globósidos/genética , Globósidos/metabolismo , Lipopolisacáridos/toxicidad , Antígeno 96 de los Linfocitos/genética , Antígeno 96 de los Linfocitos/metabolismo , Microdominios de Membrana/genética , Microdominios de Membrana/inmunología , Microdominios de Membrana/metabolismo , Ratones , Ratones Mutantes , Complejos Multiproteicos/genética , Complejos Multiproteicos/metabolismo , Unión Proteica , Choque Séptico/inducido químicamente , Choque Séptico/genética , Choque Séptico/inmunología , Choque Séptico/metabolismo , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo
18.
Biochim Biophys Acta ; 1841(4): 610-9, 2014 Apr 04.
Artículo en Inglés | MEDLINE | ID: mdl-24418004

RESUMEN

Peroxisomes are subcellular organelles that function in multiple anabolic and catabolic processes, including ß-oxidation of very-long-chain fatty acids (VLCFA) and biosynthesis of ether phospholipids. Peroxisomal disorders caused by defects in peroxisome biogenesis or peroxisomal ß-oxidation manifest as severe neural disorders of the central nervous system. Abnormal peroxisomal metabolism is thought to be responsible for the clinical symptoms of these diseases, but their molecular pathogenesis remains to be elucidated. We performed lipidomic analysis to identify aberrant metabolites in fibroblasts from patients with Zellweger syndrome (ZS), acyl-CoA oxidase1 (AOx) deficiency, D-bifunctional protein (D-BP) and X-linked adrenoleukodystrophy (X-ALD), as well as in peroxisome-deficient Chinese hamster ovary cell mutants. In cells deficient in peroxisomal biogenesis, plasmenylethanolamine was remarkably reduced and phosphatidylethanolamine was increased. Marked accumulation of very-long-chain saturated fatty acid and monounsaturated fatty acids in phosphatidylcholine was observed in all mutant cells. Very-long-chain polyunsaturated fatty acid (VLC-PUFA) levels were significantly elevated, whilst phospholipids containing docosahexaenoic acid (DHA, C22:6n-3) were reduced in fibroblasts from patients with ZS, AOx deficiency, and D-BP deficiency, but not in fibroblasts from an X-ALD patient. Because patients with AOx deficiency suffer from more severe symptoms than those with X-ALD, accumulation of VLC-PUFA and/or reduction of DHA may be associated with the severity of peroxisomal diseases.


Asunto(s)
Acilcoenzima A/metabolismo , Ácidos Grasos Insaturados/metabolismo , Fosfatidilcolinas/metabolismo , Síndrome de Zellweger/metabolismo , Acilcoenzima A/deficiencia , Acilcoenzima A/genética , Adrenoleucodistrofia/genética , Adrenoleucodistrofia/metabolismo , Adrenoleucodistrofia/patología , Animales , Células Cultivadas , Cricetinae , Fibroblastos/metabolismo , Humanos , Oxidación-Reducción , Trastorno Peroxisomal/metabolismo , Trastorno Peroxisomal/patología , Peroxisomas/metabolismo , Síndrome de Zellweger/genética , Síndrome de Zellweger/patología
19.
Diabetologia ; 57(4): 819-31, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24374551

RESUMEN

AIMS/HYPOTHESIS: Obesity is associated with ageing and increased energy intake, while restriction of energy intake improves health and longevity in multiple organisms; the NAD(+)-dependent deacetylase sirtuin 1 (SIRT1) is implicated in this process. Pro-opiomelanocortin (POMC) and agouti-related peptide (AgRP) neurons in the arcuate nucleus (ARC) of the hypothalamus are critical for energy balance regulation, and the level of SIRT1 protein decreases with age in the ARC. In the current study we tested whether conditional Sirt1 overexpression in mouse POMC or AgRP neurons prevents age-associated weight gain and diet-induced obesity. METHODS: We targeted Sirt1 cDNA sequence into the Rosa26 locus and generated conditional Sirt1 knock-in mice. These mice were crossed with mice harbouring either Pomc-Cre or Agrp-Cre and the metabolic variables, food intake, energy expenditure and sympathetic activity in adipose tissue of the resultant mice were analysed. We also used a hypothalamic cell line to investigate the molecular mechanism by which Sirt1 overexpression modulates leptin signalling. RESULTS: Conditional Sirt1 overexpression in mouse POMC or AgRP neurons prevented age-associated weight gain; overexpression in POMC neurons stimulated energy expenditure via increased sympathetic activity in adipose tissue, whereas overexpression in AgRP neurons suppressed food intake. SIRT1 improved leptin sensitivity in hypothalamic neurons in vitro and in vivo by downregulating protein-tyrosine phosphatase 1B, T cell protein-tyrosine phosphatase and suppressor of cytokine signalling 3. However, these phenotypes were absent in mice consuming a high-fat, high-sucrose diet due to decreases in ARC SIRT1 protein and hypothalamic NAD(+) levels. CONCLUSIONS/INTERPRETATION: ARC SIRT1 is a negative regulator of energy balance, and decline in ARC SIRT1 function contributes to disruption of energy homeostasis by ageing and diet-induced obesity.


Asunto(s)
Hipotálamo/metabolismo , Leptina/farmacología , Sirtuina 1/metabolismo , Aumento de Peso/fisiología , Animales , Calorimetría Indirecta , Genotipo , Hipotálamo/efectos de los fármacos , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos C57BL , Reacción en Cadena de la Polimerasa , Sirtuina 1/genética , Aumento de Peso/genética
20.
J Biol Chem ; 288(51): 36741-9, 2013 Dec 20.
Artículo en Inglés | MEDLINE | ID: mdl-24220030

RESUMEN

Very long-chain fatty acids (VLCFAs, chain length >C20) exist in tissues throughout the body and are synthesized by repetition of the fatty acid (FA) elongation cycle composed of four successive enzymatic reactions. In mammals, the TER gene is the only gene encoding trans-2-enoyl-CoA reductase, which catalyzes the fourth reaction in the FA elongation cycle. The TER P182L mutation is the pathogenic mutation for nonsyndromic mental retardation. This mutation substitutes a leucine for a proline residue at amino acid 182 in the TER enzyme. Currently, the mechanism by which the TER P182L mutation causes nonsyndromic mental retardation is unknown. To understand the effect of this mutation on the TER enzyme and VLCFA synthesis, we have biochemically characterized the TER P182L mutant enzyme using yeast and mammalian cells transfected with the TER P182L mutant gene and analyzed the FA elongation cycle in the B-lymphoblastoid cell line with the homozygous TER P182L mutation (TER(P182L/P182L) B-lymphoblastoid cell line). We have found that TER P182L mutant enzyme exhibits reduced trans-2-enoyl-CoA reductase activity and protein stability, thereby impairing VLCFA synthesis and, in turn, altering the sphingolipid profile (i.e. decreased level of C24 sphingomyelin and C24 ceramide) in the TER(P182L/P182L) B-lymphoblastoid cell line. We have also found that in addition to the TER enzyme-catalyzed fourth reaction, the third reaction in the FA elongation cycle is affected by the TER P182L mutation. These findings provide new insight into the biochemical defects associated with this genetic mutation.


Asunto(s)
Ceramidas/metabolismo , Ácidos Grasos/biosíntesis , Discapacidad Intelectual Ligada al Cromosoma X/genética , Mutación Missense , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/metabolismo , Esfingomielinas/metabolismo , Estabilidad de Enzimas , Células HEK293 , Células HeLa , Homocigoto , Humanos , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo
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