Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 437
Filtrar
Más filtros

Bases de datos
Tipo del documento
Intervalo de año de publicación
1.
Lett Appl Microbiol ; 68(5): 415-422, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30636057

RESUMEN

Talaromyces species are typical fungi capable of producing the heat-resistant ascospores responsible for the spoilage of processed food products. Hydrophobins, which are unique to fungi, are small secreted proteins that form amphipathic layers on the outer surface of fungal cell walls. In this study, species-specific primer sets for detecting and identifying Talaromyces macrosporus and Talaromyces trachyspermus were designed based on hydrophobin gene sequences. A conventional polymerase chain reaction (PCR) assay using these primer sets produced species-specific amplicons for T. macrosporus and T. trachyspermus. The detection limit for each primer set was 100 pg template DNA. This assay also detected fungal DNA extracted from blueberries inoculated with T. macrosporus. Other heat-resistant fungi, including Byssochlamys, Neosartorya and Talaromyces species, which cause food spoilage, were not detected in PCR amplifications with these primer sets. Furthermore, a conventional PCR assay using a crude DNA extract as the template also yielded amplicons specific to T. macrosporus and T. trachyspermus. The simple and rapid PCR assay described herein is highly species-specific and can reliably detect T. macrosporus and T. trachyspermus, suggesting it may be relevant for the food and beverage industry. SIGNIFICANCE AND IMPACT OF THE STUDY: The heat-resistant ascospores of Talaromyces macrosporus and Talaromyces trachyspermus are responsible for food spoilage after pasteurization. Traditional methods for detecting fungal contamination based on morphological characteristics are time-consuming and exhibit low sensitivity and specificity. In this study, a conventional polymerase chain reaction (PCR) assay based on hydrophobin gene sequences was developed for the specific detection of T. macrosporus and T. trachyspermus. This detection method was simple, rapid and highly specific. These results suggest that the conventional PCR assay developed in this study may be useful for detecting T. macrosporus and T. trachyspermus in raw materials and processed food products.


Asunto(s)
ADN de Hongos/genética , Proteínas Fúngicas/genética , Reacción en Cadena de la Polimerasa/métodos , Talaromyces/genética , Arándanos Azules (Planta)/microbiología , Microbiología de Alimentos , Especificidad de la Especie , Esporas Fúngicas/química , Talaromyces/clasificación , Talaromyces/aislamiento & purificación
2.
Lett Appl Microbiol ; 66(1): 86-92, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-29108110

RESUMEN

Heat-resistant fungi occur sporadically and are a continuing problem for the food and beverage industry. The genus Talaromyces, as a typical fungus, is capable of producing the heat-resistant ascospores responsible for the spoilage of processed food products. Isocitrate lyase, a signature enzyme of the glyoxylate cycle, is required for the metabolism of non-fermentable carbon compounds, like acetate and ethanol. Here, species-specific primer sets for detection and identification of DNA derived from Talaromyces macrosporus and Talaromyces trachyspermus were designed based on the nucleotide sequences of their isocitrate lyase genes. Polymerase chain reaction (PCR) using a species-specific primer set amplified products specific to T. macrosporus and T. trachyspermus. Other fungal species, such as Byssochlamys fulva and Hamigera striata, which cause food spoilage, were not detected using the Talaromyces-specific primer sets. The detection limit for each species-specific primer set was determined as being 50 pg of template DNA, without using a nested PCR method. The specificity of each species-specific primer set was maintained in the presence of 1,000-fold amounts of genomic DNA from other fungi. The method also detected fungal DNA extracted from blueberry inoculated with T. macrosporus. This PCR method provides a quick, simple, powerful and reliable way to detect T. macrosporus and T. trachyspermus. SIGNIFICANCE AND IMPACT OF THE STUDY: Polymerase chain reaction (PCR)-based detection is rapid, convenient and sensitive compared with traditional methods of detecting heat-resistant fungi. In this study, a PCR-based method was developed for the detection and identification of amplification products from Talaromyces macrosporus and Talaromyces trachyspermus using primer sets that target the isocitrate lyase gene. This method could be used for the on-site detection of T. macrosporus and T. trachyspermus in the near future, and will be helpful in the safety control of raw materials and in food and beverage production.


Asunto(s)
Cartilla de ADN/genética , Reacción en Cadena de la Polimerasa/métodos , Talaromyces/genética , Talaromyces/aislamiento & purificación , Bebidas/microbiología , Arándanos Azules (Planta)/microbiología , ADN de Hongos/genética , Microbiología de Alimentos , Calor , Especificidad de la Especie , Esporas Fúngicas/química , Esporas Fúngicas/genética , Talaromyces/química , Talaromyces/clasificación
3.
Eur J Vasc Endovasc Surg ; 46(3): 321-8, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23880423

RESUMEN

OBJECTIVES: Many pancreaticoduodenal artery (PDA) aneurysms are associated with celiac artery (CA) stenosis. The pathogenesis of PDA aneurysm may be associated with hemodynamic changes due to CA stenosis/occlusion. The aim of this study was to assess the hemodynamic changes of celiaco-mesenteric anastomosis in patients with PDA aneurysms concomitant with CA occlusion using four-dimensional flow-sensitive magnetic resonance imaging (4D-Flow). METHODS: 4D-Flow was performed preoperatively on five patients. Seven age- and sex-matched individuals were used as controls. Hemodynamic parameters such as flow volume and maximum flow velocity in PDAs, gastroduodenal arteries, common hepatic arteries, and superior mesenteric arteries were compared between both groups. Wall shear stress (WSS) and oscillatory shear index (OSI) were mapped in both groups. RESULTS: In the patient group, 4D-Flow identified retrograde flow of both gastroduodenal arteries and common hepatic arteries. Heterogeneous distribution patterns of both WSS and OSI were identified across the entire PDA in the patient group. OSI mapping showed multiple regions with extremely high OSI values (OSI > 0.3) in all patients. All PDA aneurysms, which were surgically resected, were atherosclerotic. CONCLUSIONS: 4D-Flow identified hemodynamic changes in celiaco-mesenteric arteries in patients with PDA aneurysms with concomitant CA occlusion. These hemodynamic changes may be associated with PDA aneurysm formation.


Asunto(s)
Aneurisma/fisiopatología , Aneurisma/cirugía , Aterosclerosis/fisiopatología , Arteria Celíaca , Duodeno/irrigación sanguínea , Hemodinámica/fisiología , Arteria Hepática , Angiografía por Resonancia Magnética/métodos , Arteria Mesentérica Superior , Páncreas/irrigación sanguínea , Anastomosis Quirúrgica , Estudios de Casos y Controles , Medios de Contraste , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Masculino , Persona de Mediana Edad , Estadísticas no Paramétricas , Estrés Mecánico
4.
Kyobu Geka ; 65(8): 648-52, 2012 Jul.
Artículo en Japonés | MEDLINE | ID: mdl-22868422

RESUMEN

Severe atherosclerosis of the ascending aorta frequently causes difficulties during heart operations, hindering surgical maneuvers and potentially leading to systemic embolism. There have been several methods to solve these problems but the best way to treat patients requiring aortic valve replacement (AVR) has not been established yet. Surgical techniques for AVR in these patients include AVR under deep hypothermic circulatory arrest with or without endarterectomy of the ascending aorta or replacement of the ascending aorta. Endovascular clamping using a balloon is another approach but requires manipulation of the heavily calcified aorta that may result in a certain risk for stroke. Another option to avoid the ascending aorta and cross-clamping is the apicoaortic conduit. Recently introduced trans-catheter AVR (TAVR), especially trans-apical AVR, has been shown to be feasible in such patients. Larger studies and longer follow-up will be required to scientifically prove the superiority of trans-apical AVR over conventional surgical strategies in patients with porcelain aorta requiring AVR.


Asunto(s)
Válvula Aórtica/cirugía , Enfermedades de la Aorta/cirugía , Aterosclerosis/cirugía , Procedimientos Quirúrgicos Cardíacos/métodos , Prótesis Valvulares Cardíacas , Humanos
5.
Cytopathology ; 21(2): 103-10, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19863615

RESUMEN

OBJECTIVE: Extracellular matrix metalloproteinase inducer (EMMPRIN) and its induced matrix metalloproteinases (MMPs) play a crucial role in tumour progression, invasion and metastasis. EMMPRIN expression has been demonstrated in several tumours, but its expression profile in thyroid cancer remains unclear. METHODS: We evaluated the expression profile of EMMPRIN at various stages of differentiation of thyroid carcinoma, including 20 cases of well-differentiated papillary carcinoma (WDPC), 15 cases of papillary carcinoma with a poorly differentiated carcinoma component (PC/PDC) and four cases with an undifferentiated carcinoma (UDC) component, using paraffin-embedded sections for immunohistochemical stains. Also, we used 32 fine needle aspiration cytology and imprint smears from the same cases for immunocytochemical stains. The staining results were evaluated with a scoring system. RESULTS: Immunohistochemical staining showed that EMMPRIN expression was absent or weak in almost all WDPC specimens, whereas it was moderate or strong in PDC and UDC components. In tumours that showed a gradual morphological transformation from WDPC to PDC components, the expression of EMMPRIN was progressively stronger from the areas of WDPC to those of PDC. WDPC, PC/PDC and UDC had expression scores of 4.9, 45.0 and 245.7, respectively. Results of immunocytochemical staining showed almost the same staining profile as those of immunohistochemical staining. The cytological atypia of EMMPRIN-positive cells was greater than that of negative cells. CONCLUSION: These results indicated that EMMPRIN expression correlates significantly with the degree of dedifferentiation of thyroid carcinoma. This study demonstrates the feasibility of expression of EMMPRIN using fine needle aspiration samples. Therefore, immunocytochemical analysis of EMMPRIN may be a novel aid to evaluate the differentiation of thyroid carcinoma.


Asunto(s)
Adenocarcinoma Papilar/patología , Basigina/metabolismo , Neoplasias de la Tiroides/patología , Adenocarcinoma Papilar/metabolismo , Biomarcadores de Tumor/metabolismo , Biopsia con Aguja Fina , Desdiferenciación Celular , Humanos , Inmunohistoquímica , Neoplasias de la Tiroides/metabolismo
6.
J Cell Biol ; 133(4): 733-47, 1996 May.
Artículo en Inglés | MEDLINE | ID: mdl-8666660

RESUMEN

High level expression of the M2 ion channel protein of influenza virus inhibits the rate of intracellular transport of the influenza virus hemagglutinin (HA) and that of other integral membrane glycoproteins. HA coexpressed with M2 is properly folded, is not associated with GRP78-BiP, and trimerizes with the same kinetics as when HA is expressed alone. Analysis of the rate of transport of HA from the ER to the cis and medial golgi compartments and the TGN indicated that transport through the Golgi apparatus is delayed. Uncleaved HA0 was not expressed at the cell surface, and accumulation HA at the plasma membrane was reduced to 75-80% of control cells. The delay in intracellular transport of HA on coexpression of M2 was not observed in the presence of the M2-specific ion channel blocker, amantadine, indicating that the Golgi transport delay is due to the M2 protein ion channel activity equilibrating pH between the Golgi lumen and the cytoplasm, and not due to saturation of the intracellular transport machinery. The Na+/H+ ionophore, monensin, which also equilibrates pH between the Golgi lumen and the cytoplasm, caused a similar inhibition of intracellular transport as M2 protein expression did for HA and other integral membrane glycoproteins. EM data showed a dilation of Golgi cisternae in cells expressing the M2 ion channel protein. Taken together, the data suggest a similarity of effects of M2 ion channel activity and monensin on intracellular transport through the Golgi apparatus.


Asunto(s)
Aparato de Golgi/metabolismo , Hemaglutininas Virales/metabolismo , Virus de la Influenza A/metabolismo , Canales Iónicos/metabolismo , Proteínas de la Matriz Viral/metabolismo , Animales , Transporte Biológico/efectos de los fármacos , Bovinos , Línea Celular , Membrana Celular/metabolismo , Chlorocebus aethiops , Retículo Endoplásmico/metabolismo , Chaperón BiP del Retículo Endoplásmico , Aparato de Golgi/ultraestructura , Células HeLa , Glicoproteínas Hemaglutininas del Virus de la Influenza , Hemaglutininas Virales/genética , Hexosaminidasas , Humanos , Concentración de Iones de Hidrógeno , Canales Iónicos/genética , Cinética , Monensina/farmacología , Pliegue de Proteína , Precursores de Proteínas/metabolismo , Proteínas de la Matriz Viral/genética
7.
Nihon Hoshasen Gijutsu Gakkai Zasshi ; 64(7): 877-8, 2008 Jul 20.
Artículo en Japonés | MEDLINE | ID: mdl-18719306

RESUMEN

PURPOSE/AIM OF THE EXHIBIT: The purpose of this exhibit is: 1. To explain "resampling", an image data processing, performed by the digital radiographic system based on flat panel detector (FPD). 2. To show the influence of "resampling" on the basic imaging properties. 3. To present accurate measurement methods of the basic imaging properties of the FPD system. CONTENT ORGANIZATION: 1. The relationship between the matrix sizes of the output image and the image data acquired on FPD that automatically changes depending on a selected image size (FOV). 2. The explanation of the image data processing of "resampling". 3. The evaluation results of the basic imaging properties of the FPD system using two types of DICOM image to which "resampling" was performed: characteristic curves, presampled MTFs, noise power spectra, detective quantum efficiencies. CONCLUSION/SUMMARY: The major points of the exhibit are as follows: 1. The influence of "resampling" should not be disregarded in the evaluation of the basic imaging properties of the flat panel detector system. 2. It is necessary for the basic imaging properties to be measured by using DICOM image to which no "resampling" is performed.


Asunto(s)
Intensificación de Imagen Radiográfica/métodos , Chicago , Congresos como Asunto , Radiología , Sociedades Médicas
8.
J Clin Invest ; 82(1): 282-6, 1988 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-3292586

RESUMEN

The present study was designed to test whether there are diurnal changes in the firing rate of sympathetic nerves to brown adipose tissue and whether these diurnal rhythms influenced the response to insulin injected into the suprachiasmatic nucleus or ventromedial hypothalamic nucleus (VMH). Food intake was highest at the beginning of the dark period (1800-2200 hours) and lowest during the daylight hours (0600-1000 and 1200-1600 hours). The basal sympathetic firing rate was highest at noon (1000-1200 hours) when food intake was lowest. At midnight, when food intake was highest, sympathetic firing rate was lowest. Injection of insulin (77, 144, and 288 pmol) into the VMH produced a dose-dependent depression of sympathetic firing rate at each of the four measurement periods (0400-0600 hours, 1000-1200 hours, 1600-1800 hours, and 2200-2400 hours), but the magnitude of the effect was greater at noon than at night. In contrast, insulin injections into the suprachiasmatic nucleus decreased the sympathetic firing rate at noon but produced a significant increase in the sympathetic firing rate at night. These data show that a diurnal rhythm exists for the sympathetic firing rate. The decrease in firing rate in response to insulin when injected into the VMH is in the same direction but varies in magnitude throughout the day, whereas the responsiveness of the suprachiasmatic nucleus to injections of insulin shows a reversal of response in relation to day/night cycles. The highly significant inverse relationship between basal sympathetic firing rate and food intake suggests that sympathetic activity may be part of an important control system for energy balance.


Asunto(s)
Ritmo Circadiano , Conducta Alimentaria/fisiología , Insulina/administración & dosificación , Núcleo Supraquiasmático/fisiología , Sistema Nervioso Simpático/fisiología , Núcleo Hipotalámico Ventromedial/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Glucemia/metabolismo , Mapeo Encefálico , Relación Dosis-Respuesta a Droga , Femenino , Insulina/sangre , Ratas
9.
Kidney Int Suppl ; (102): S12-5, 2006 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-16810304

RESUMEN

Hyperplasia of the parathyroid gland (PTG) is associated not only with excessive secretion of parathyroid hormone (PTH) but also with changes in the parathyroid cell (PTC) characteristics (i.e. hyperproliferative activity, and low contents of vitamin D and calcium-sensing receptors). Control of PTG hyperplasia is most important in the management of secondary hyperparathyroidism, but the advanced stage of hyperplasia is considered irreversible. In the present study, dialysis patients with PTG hyperplasia underwent direct injection of calcitriol or maxacalcitol (OCT) into the PTG. Ultrasonography showed that this treatment had significantly reduced PTG volume and tissue analysis using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method and DNA electrophoresis indicated that cellular apoptosis had been induced. The mechanism of apoptosis was evaluated in detail in uremic rats fed a high-phosphate diet. OCT or its vehicle was directly injected into the rats' PTGs. In the PTGs treated by OCT, there was a significantly increased number of TUNEL-positive PTCs and DNA electrophoresis revealed the characteristic ladder pattern of DNA fragmentation, both findings indicative of apoptosis. There was also a significant upregulation of both vitamin D and Ca-sensing receptors in the PTCs and a clear shift of the Ca-PTH response curve to the left and downward. None of these findings was observed in the PTGs treated by vehicle. This novel treatment is successful in causing regression of PTG hyperplasia. Thus, it is expected to significantly reduce the PTH level and ameliorate the abnormal bone turnover and mineral metabolism.


Asunto(s)
Antineoplásicos/administración & dosificación , Calcitriol/análogos & derivados , Calcitriol/administración & dosificación , Fragmentación del ADN/efectos de los fármacos , Fallo Renal Crónico/patología , Glándulas Paratiroides/patología , Vitaminas/administración & dosificación , Animales , Femenino , Humanos , Hiperplasia/sangre , Hiperplasia/tratamiento farmacológico , Hiperplasia/etiología , Hiperplasia/patología , Fallo Renal Crónico/sangre , Fallo Renal Crónico/complicaciones , Fallo Renal Crónico/terapia , Masculino , Glándulas Paratiroides/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores Sensibles al Calcio/metabolismo , Vitamina D
10.
Cancer Res ; 52(22): 6153-7, 1992 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-1423258

RESUMEN

Mouse monoclonal antibodies against human (beta 1-4)galactosyl-transferase (GalT) purified from human ovarian tumor effusion fluids were prepared and characterized. GalT purified from normal human plasma showed a single diffused band in nondenaturing polyacrylamide gel electrophoresis, but GalT purified from human ovarian tumor effusion fluids showed several oligomeric bands and a monomeric band in nondenaturing polyacrylamide gel electrophoresis. These oligomeric bands were dissociated into monomer by urea treatment and polymerized by a 2-mercaptoethanol treatment. Nine monoclonal antibodies (MAb) were prepared by immunization of purified GalT from human ovarian tumor effusion fluids and classified into three groups. Type I MAbs (MAb8611, MAb8913, and MAb8919) reacted only to the GalT monomer. Type II MAbs (MAb4880, MAb8507, and MAb8628) reacted to both the GalT monomer and the GalT polymer. Type III MAbs (MAb7907, MAb8513, and MAb8677) reacted only to the GalT polymer. These MAbs except MAb7907 could recover GalT enzyme activity from effusion fluids by immunoprecipitation. A fraction passed through MAb8513 affinity chromatography still showed reactivity to MAb8919, demonstrating that an epitope of MAb8513 resides on a minor part of GalT. A sandwich immunoassay (MAb8513-MAb8628HRP) was developed, and serum samples from ovarian cancer patients and benign ovarian patients were tested. The levels of sandwich immunoassay of serum samples from cancer were elevated significantly compared to those from benign and did not necessarily correlate to total GalT enzyme activity in serum samples. These results suggested that MAb8513 (Type III) might recognize a unique GalT associated with tumor (GAT).


Asunto(s)
Anticuerpos Monoclonales/inmunología , Líquido Ascítico/enzimología , Biomarcadores de Tumor/inmunología , Galactosiltransferasas/inmunología , Isoenzimas/inmunología , Proteínas de Neoplasias/inmunología , Neoplasias Ováricas/enzimología , Animales , Anticuerpos Monoclonales/aislamiento & purificación , Especificidad de Anticuerpos , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/aislamiento & purificación , Electroforesis en Gel de Poliacrilamida , Femenino , Galactosiltransferasas/sangre , Galactosiltransferasas/aislamiento & purificación , Humanos , Inmunización , Isoenzimas/sangre , Isoenzimas/aislamiento & purificación , Ratones , Ratones Endogámicos BALB C , Peso Molecular , Proteínas de Neoplasias/sangre , Proteínas de Neoplasias/aislamiento & purificación , Neoplasias Ováricas/sangre
11.
Mech Dev ; 107(1-2): 25-38, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11520661

RESUMEN

Vertebrate endoderm development has recently become the focus of intense investigation. We have identified a novel sox gene, 226D7, which is important in zebrafish endoderm development. 226D7 was isolated by an in situ hybridization screening for genes expressed in the yolk syncytial layer (YSL) at the blastula stage. 226D7 is expressed mainly in the YSL at this stage and, during gastrulation, its expression is also detected in the forerunner cells and endodermal precursor cells. The expression of 226D7 is positively regulated by Nodal signaling. The knockdown of 226D7 using morpholino antisense oligonucleotides results in a lack of sox17-expressing endodermal precursor cells during gastrulation, and, consequently, lacks endodermal derivatives such as gut tissue. The effect is strictly restricted to the endodermal lineage, while the mesoderm is normally formed, a phenotype that is nearly identical to that of the casanova mutant (Dev. Biol. 215 (1999) 343). We further demonstrate that overexpression of 226D7 increases the number of sox17-expressing endodermal progenitor cells without upregulating the expression of the Nodal genes, cyclops and squint. Region-specific knockdown and overexpression of 226D7 by injection into the YSL suggest that 226D7 in the YSL is not involved in endoderm formation and 226D7 in the endoderm progenitor cells is important for endoderm development. Taken together, our data demonstrate that 226D7 is a downstream target of Nodal signal and a critical transcriptional regulator of early endoderm formation.


Asunto(s)
Proteínas de Unión al ADN , Desarrollo Embrionario , Endodermo/fisiología , Proteínas del Grupo de Alta Movilidad/genética , Factores de Transcripción/genética , Factor de Crecimiento Transformador beta/fisiología , Proteínas de Pez Cebra , Pez Cebra/embriología , Pez Cebra/genética , Secuencia de Aminoácidos , Animales , Blastocisto/metabolismo , Linaje de la Célula , Embrión no Mamífero/metabolismo , Endodermo/citología , Gástrula/metabolismo , Regulación del Desarrollo de la Expresión Génica , Proteínas del Grupo de Alta Movilidad/química , Proteínas del Grupo de Alta Movilidad/fisiología , Hibridación in Situ , Datos de Secuencia Molecular , Proteína Nodal , Oligonucleótidos Antisentido , Fenotipo , Proteínas/genética , Factores de Transcripción SOX , Factores de Transcripción SOXF , Alineación de Secuencia , Transducción de Señal , Células Madre/metabolismo , Factores de Transcripción/química , Factores de Transcripción/fisiología , Factor de Crecimiento Transformador beta/genética , Regulación hacia Arriba
12.
Mech Dev ; 104(1-2): 113-5, 2001 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11404086

RESUMEN

BTG/tob family proteins are thought to be a potential tumor suppressor due to their anti-proliferative activity. We cloned zebrafish btg-b, a member of the BTG1/2 subfamily, using in situ hybridization screening. The tissue-specific expression of btg-b is first observed in the organizer region at the early gastrula stage. Later in development, the forebrain, the hindbrain, the polster and the paraxial mesoderm transiently express btg-b. Recently, mouse Btg1 and Btg2 have been shown to be a cofactor for Hoxb9. Double in situ hybridization with zebrafish btg-b and hoxb9a indicates that the expression domains of these two genes overlap in the posterior paraxial mesoderm.


Asunto(s)
Proteínas Portadoras/biosíntesis , Proteínas Portadoras/genética , Ciclo Celular/genética , Proteínas de Pez Cebra , Pez Cebra/embriología , Secuencia de Aminoácidos , Animales , Northern Blotting , Encéfalo/metabolismo , Clonación Molecular , ADN Complementario/metabolismo , Embrión no Mamífero/metabolismo , Gástrula/metabolismo , Biblioteca de Genes , Hibridación in Situ , Mesodermo/metabolismo , Datos de Secuencia Molecular , Filogenia , Estructura Terciaria de Proteína , Mapeo de Híbrido por Radiación , Factores de Tiempo
13.
Mech Dev ; 121(7-8): 915-32, 2004 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-15210196

RESUMEN

The medaka is becoming an attractive model organism for the study of vertebrate early development and organogenesis and large-scale mutagenesis projects that are aimed at creating developmentally defective mutants are now being conducted by several groups in Japan. To strengthen the study of medaka developmental genetics, we have conducted a large-scale isolation of ESTs from medaka embryos and developed tools that facilitate mutant analysis. In this study, we have characterized a total of 132,082 sequences from both ends of cloned insert cDNAs from libraries generated at different stages of medaka embryo development. Clustering analysis with 3-prime sequences finally identified a total of 12,429 clusters. As a pilot analysis, 924 clusters were subjected to in situ hybridization to determine the spatial localization of their transcripts. Using EST sequence data generated in the present study, a 60-mer oligonucleotide microarray with 8,091 unigenes (Medaka Microarray 8K) was constructed and tested for its usefulness in expression profiling. Furthermore, we have developed a rapid and reliable mutant mapping system using a set of mapped EST markers (M-marker 2003) that covers the entire medaka genome. These resources will accelerate medaka mutant analyses and make an important contribution to the medaka genome project.


Asunto(s)
Etiquetas de Secuencia Expresada , Oryzias/embriología , Oryzias/genética , Animales , Mapeo Cromosómico , Biblioteca de Genes , Marcadores Genéticos , Hibridación in Situ , Familia de Multigenes , Mutación , Análisis de Secuencia por Matrices de Oligonucleótidos , Análisis de Secuencia de ADN
14.
FASEB J ; 15(14): 2757-9, 2001 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-11606484

RESUMEN

Hypoxic induction of the early growth response-1 (Egr-1) transcription factor initiates proinflammatory and procoagulant gene expression. Orthotopic/isogeneic rat lung transplantation triggers Egr-1 expression and nuclear DNA binding activity corresponding to Egr-1, which leads to increased expression of downstream target genes such as interleukin-1b, tissue factor, and plasminogen activator inhibitor-1. The devastating functional consequences of Egr-1 up-regulation in this setting are prevented by treating donor lungs with a phosphorothioate antisense oligodeoxyribonucleotide directed against the Egr-1 translation initiation site, which blocks expression of Egr-1 and its gene targets. Post-transplant graft leukostasis, inflammation, and thrombosis are consequently diminished, with marked improvement in graft function and recipient survival. Blocking expression of a proximal transcription factor, which activates deleterious inflammatory and coagulant effector mechanisms, is an effective molecular strategy to improve organ preservation.


Asunto(s)
Proteínas de Unión al ADN/fisiología , Proteínas Inmediatas-Precoces , Inflamación/fisiopatología , Trasplante de Pulmón , Trombosis/fisiopatología , Factores de Transcripción/fisiología , Animales , Northern Blotting , Western Blotting , ADN sin Sentido/farmacología , Proteínas de Unión al ADN/genética , Proteína 1 de la Respuesta de Crecimiento Precoz , Fibrina/efectos de los fármacos , Fibrina/metabolismo , Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Supervivencia de Injerto/efectos de los fármacos , Supervivencia de Injerto/fisiología , Interleucina-1/genética , Inhibidor 1 de Activador Plasminogénico/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Transducción de Señal , Tromboplastina/genética , Factores de Transcripción/genética
16.
Arch Intern Med ; 145(4): 731-3, 1985 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-3985735

RESUMEN

In the clinical course of a patient with progressive facial hemiatrophy associated with ipsilateral body atrophy (total hemiatrophy), signs and symptoms of localized scleroderma were noted. The patient subsequently was found to have Schönlein-Henoch purpura with renal involvement and, later, paroxysmal nocturnal hemoglobinuria. To our knowledge, such an association has not been reported before.


Asunto(s)
Hemiatrofia Facial/complicaciones , Glomerulonefritis/complicaciones , Hemoglobinuria Paroxística/complicaciones , Vasculitis por IgA/complicaciones , Esclerodermia Localizada/complicaciones , Adulto , Atrofia/complicaciones , Femenino , Glomerulonefritis/patología , Humanos
17.
Cardiovasc Res ; 47(1): 74-80, 2000 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-10869532

RESUMEN

OBJECTIVE: Heat shock protein 72 (HSP72) is involved in the myocardial self-preservation system under several conditions such as ischemia-reperfusion injury or late preconditioning. However, its mechanism is not fully understood. Ecto-5'-nucleotidase is a key enzyme for synthesizing adenosine and plays an important role in ischemic preconditioning. In this study, we tested the hypothesis that ecto-5'-nucleotidase plays a role in the cardioprotection of HSP72. METHODS: Rat hearts (H group, n=6) were transfected with HSP72 gene by an intracoronary infusion of hemagglutinating virus of Japan (HVJ)-liposome complex. Control hearts (C group, n=6) were transfected with the beta-galactosidase gene. Following 30 min of normothermic ischemia, grafts were reperfused using Langendorff apparatus. RESULTS: The activity of ecto-5'-nucleotidase was significantly higher in H group than C group both before and after ischemia-reperfusion (H vs. C; 0.51+/-0.05 vs. 0.29+/-0.06, and 1.41+/-0.15 vs. 0.85+/-0.11 nmol/mg protein/min, P<0.05). H group also showed significant better functional recoveries than C group (P<0.05), as well as less creatine phosphokinase leakage (4.4+/-2.8 vs. 14.2+/-3.4 mU/min, P<0.05) and higher adenosine release (247.5+/-35.1 vs. 54.3+/-1.7 pmol/min, P<0.05). Administration of alpha,beta-methylene adenosine diphosphate (AMP-CP), an inhibitor of ecto-5'-nucleotidase, significantly diminished the tolerance to ischemia-reperfusion injury in H group (P<0.05). CONCLUSION: These results demonstrated that ecto-5'-nucleotidase activated by an overexpression of HSP72 attenuated ischemia-reperfusion injury in the rat myocardium. They suggest that ecto-5'-nucleotidase plays a role in the cardioprotective effects of HSP72 in rat hearts.


Asunto(s)
Adenosina Trifosfatasas/fisiología , Proteínas de Choque Térmico/genética , Daño por Reperfusión Miocárdica/metabolismo , Adenosina Difosfato/análogos & derivados , Adenosina Difosfato/farmacología , Adenosina Trifosfatasas/antagonistas & inhibidores , Adenosina Trifosfatasas/metabolismo , Análisis de Varianza , Animales , Western Blotting , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Expresión Génica , Proteínas del Choque Térmico HSP72 , Proteínas de Choque Térmico/metabolismo , Masculino , Perfusión , Ratas , Ratas Sprague-Dawley , Transfección
18.
Neurosci Biobehav Rev ; 19(3): 469-78, 1995.
Artículo en Inglés | MEDLINE | ID: mdl-7566748

RESUMEN

D-glucose solution injected into the portal vein influences efferent tonic activities of the vagal nerve innervating the stomach. This suggested the existence of a neural connection between hepatic vagal branch afferents and gastric vagal efferents in the brain. Considering this observation together with findings indicating that electrical stimulation of the proximal cut end of the hepatic vagal branch changes acidity in the gastric perfusing fluid or pressure within the stomach, it has been presumed that hepatic afferent signals related to glucose may regulate the motor or secretory function of the stomach through a change in central nervous activity. Recently active interaction between the portal and medullary glucose signals in gastric function was discovered, and analysis of the characteristic features of the system is in progress.


Asunto(s)
Glucemia/fisiología , Motilidad Gastrointestinal/fisiología , Sistema Porta/fisiología , Estómago/fisiología , Animales , Humanos
19.
Endocrinology ; 121(6): 1960-5, 1987 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-3119312

RESUMEN

An amidated peptide derived from cleavage of the C-terminal of prepro-GRF [prepro-GRF-(78-107)NH2] and designated CTPG or anorectin was injected ip and into the third ventricle of rats, and food intake and body weight were measured. An acute injection of 0.2, 0.5, or 1.0 microgram CTPG (anorectin; 60, 150, or 300 pmol) into the third ventricle produced a significant dose-related reduction in food intake in the hungry rat during the 12-h dark period. Water intake was suppressed 30% by 1.0 microgram (300 pmol), but the lower doses had no effect. The highest dose (1 microgram or 300 pmol) produced a small rise in glucose concentration 5 min after injection into the third ventricle, which returned to the control value by 15 min. Anorectin increased the sympathetic efferent firing rate 3, 6, and 24 h after a single injection. Intraperitoneal injection of 2.0, 5.0, and 20 micrograms, however, had no significant effect on food or water intake. Chronic infusion into the third ventricle at a rate of 50 ng/h for 7 days produced a persistent reduction in food intake and a steady fall in body weight. The chronically infused rats had a significantly higher level of sympathetic activity, as measured by the firing rate of sympathetic nerves innervating the interscapular brown adipose tissue. These studies raise the possibility that anorectin released during processing of GRF might be involved in the modulation of feeding behavior.


Asunto(s)
Apetito/efectos de los fármacos , Hormona Liberadora de Hormona del Crecimiento/farmacología , Precursores de Proteínas/farmacología , Animales , Glucemia/metabolismo , Ventrículos Cerebrales/efectos de los fármacos , Ventrículos Cerebrales/fisiología , Esquema de Medicación , Hormona Liberadora de Hormona del Crecimiento/administración & dosificación , Inyecciones Intraventriculares , Insulina/sangre , Masculino , Precursores de Proteínas/administración & dosificación , Ratas , Ratas Endogámicas , Sistema Nervioso Simpático/efectos de los fármacos , Sistema Nervioso Simpático/fisiología , Factores de Tiempo
20.
J Cereb Blood Flow Metab ; 11(3): 472-8, 1991 May.
Artículo en Inglés | MEDLINE | ID: mdl-2016355

RESUMEN

In the focal infarction model of the rat middle cerebral artery (MCA), the thalamus of the occluded side becomes gradually atrophic, mainly because of retrograde degeneration. We determined whether basic fibroblast growth factor (bFGF) administered intracisternally could prevent this thalamic atrophy. We occluded the left MCA through a small cranial opening, and animals were then divided into two groups. One group received intracisternal injections of recombinant bFGF (1 microgram dissolved in 0.1 ml of saline with 2% rat serum) starting 1 day after occlusion and repeated once a week to a total dose of 4 micrograms by four injections. The other group received vehicle solution by the same schedule. The animals were perfused and fixed at 28 days after occlusion, and histological examination was made at the level of the caudoputamen and thalamus. In the bFGF-treated rats, the area of the posterior ventral thalamus of the occluded side was 93% of that of the contralateral side, i.e., significantly larger than in the normal saline-treated rats (75%, p less than 0.01). The infarction size was not statistically different in the two groups. Microscopic observation indicated that normal-saline-treated animals showed shrinkage and disappearance of thalamic neurons, whereas bFGF-treated groups showed preservation of thalamic neurons. Computerized analysis of the cell size substantiated this observation. To assess the effect of bFGF on astrocytes, bFGF or vehicle solution was injected into normal rats, and their histology was evaluated at 1, 2, and 4 weeks after injection. The bFGF-injected group showed a significant increase in glial fibrillary acidic protein-positive astrocytes in the brain tissue facing the ventriculocisternal system.(ABSTRACT TRUNCATED AT 250 WORDS)


Asunto(s)
Infarto Cerebral/patología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Tálamo/patología , Animales , Astrocitos/química , Astrocitos/patología , Atrofia , Recuento de Células , Femenino , Proteína Ácida Fibrilar de la Glía/análisis , Hipocampo/patología , Inmunohistoquímica , Ratas , Ratas Endogámicas , Proteínas Recombinantes
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA