RESUMEN
BACKGROUND: The products of cyp19, dax, foxl2, mis, sf1 and sox9 have each been associated with sex-determining processes among vertebrates. We provide evidence for expression of these regulators very early in salmonid development and in tissues outside of the hypothalamic-pituitary-adrenal/gonadal (HPAG) axis. Although the function of these factors in sexual differentiation have been defined, their roles in early development before sexual fate decisions and in tissues beyond the brain or gonad are essentially unknown. RESULTS: Bacterial artificial chromosomes containing salmon dax1 and dax2, foxl2b and mis were isolated and the regulatory regions that control their expression were characterized. Transposon integrations are implicated in the shaping of the dax and foxl2 loci. Splice variants for cyp19b1 and mis in both embryonic and adult tissues were detected and characterized. We found that cyp19b1 transcripts are generated that contain 5'-untranslated regions of different lengths due to cryptic splicing of the 3'-end of intron 1. We also demonstrate that salmon mis transcripts can encode prodomain products that present different C-termini and terminate before translation of the MIS hormone. Regulatory differences in the expression of two distinct aromatases cyp19a and cyp19b1 are exerted, despite transcription of their transactivators (ie; dax1, foxl2, sf1) occurring much earlier during embryonic development. CONCLUSIONS: We report the embryonic and extragonadal expression of dax, foxl2, mis and other differentiation factors that indicate that they have functions that are more general and not restricted to steroidogenesis and gonadogenesis. Spliced cyp19b1 and mis transcripts are generated that may provide regulatory controls for tissue- or development-specific activities. Selection of cyp19b1 transcripts may be regulated by DAX-1, FOXL2 and SF-1 complexes that bind motifs in intron 1, or by signals within exon 2 that recruit splicing factors, or both. The potential translation of proteins bearing only the N-terminal MIS prodomain may modulate the functions of other TGF ß family members in different tissues. The expression patterns of dax1 early in salmon embryogenesis implicate its role as a lineage determination factor. Other roles for these factors during embryogenesis and outside the HPAG axis are discussed.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Salmo salar/genética , Diferenciación Sexual/fisiología , Animales , Femenino , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica/genética , Regulación del Desarrollo de la Expresión Génica/fisiología , Hibridación Fluorescente in Situ , Masculino , Regiones Promotoras Genéticas/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Diferenciación Sexual/genéticaRESUMEN
Advancing Best Practices in Care Management and Professional Development.
Asunto(s)
Benchmarking/organización & administración , Enfermería en Salud Comunitaria , Agencias de Atención a Domicilio/organización & administración , Servicios de Atención de Salud a Domicilio/organización & administración , Rehabilitación , Competencia Clínica , Enfermería en Salud Comunitaria/organización & administración , Continuidad de la Atención al Paciente/organización & administración , Curriculum , Difusión de Innovaciones , Necesidades y Demandas de Servicios de Salud , Humanos , Liderazgo , Modelos Educacionales , Modelos de Enfermería , New York , Enfermeras Administradoras/organización & administración , Enfermeras Clínicas/organización & administración , Grupo de Atención al Paciente/organización & administración , Rol Profesional , Rehabilitación/educación , Rehabilitación/organización & administración , Desarrollo de Personal , Apoyo a la Formación ProfesionalRESUMEN
BACKGROUND: We have previously identified associations between major histocompatibility complex (MHC) class I and resistance towards bacterial and viral pathogens in Atlantic salmon. To evaluate if only MHC or also closely linked genes contributed to the observed resistance we ventured into sequencing of the duplicated MHC class I regions of Atlantic salmon. RESULTS: Nine BACs covering more than 500 kb of the two duplicated MHC class I regions of Atlantic salmon were sequenced and the gene organizations characterized. Both regions contained the proteasome components PSMB8, PSMB9, PSMB9-like and PSMB10 in addition to the transporter for antigen processing TAP2, as well as genes for KIFC1, ZBTB22, DAXX, TAPBP, BRD2, COL11A2, RXRB and SLC39A7. The IA region contained the recently reported MHC class I Sasa-ULA locus residing approximately 50 kb upstream of the major Sasa-UBA locus. The duplicated class IB region contained an MHC class I locus resembling the rainbow trout UCA locus, but although transcribed it was a pseudogene. No other MHC class I-like genes were detected in the two duplicated regions. Two allelic BACs spanning the UBA locus had 99.2% identity over 125 kb, while the IA region showed 82.5% identity over 136 kb to the IB region. The Atlantic salmon IB region had an insert of 220 kb in comparison to the IA region containing three chitin synthase genes. CONCLUSION: We have characterized the gene organization of more than 500 kb of the two duplicated MHC class I regions in Atlantic salmon. Although Atlantic salmon and rainbow trout are closely related, the gene organization of their IB region has undergone extensive gene rearrangements. The Atlantic salmon has only one class I UCA pseudogene in the IB region while trout contains the four MHC UCA, UDA, UEA and UFA class I loci. The large differences in gene content and most likely function of the salmon and trout class IB region clearly argues that sequencing of salmon will not necessarily provide information relevant for trout and vice versa.
Asunto(s)
Genes MHC Clase I , Salmo salar/genética , Secuencia de Aminoácidos , Animales , Presentación de Antígeno/genética , Evolución Molecular , Duplicación de Gen , Genoma , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Alineación de SecuenciaRESUMEN
A combination of gas chromatography (GC) and chemometrics was evaluated for its ability to differentiate between apple juice samples on the basis of apple variety and applied heat treatment. The heat treatment involved exposure of 15 mL juice samples for 30 s in a 900 W domestic microwave oven. The chromatographic results were subjected to two chemometric procedures: (1) partial least squares (PLS) regression and (2) linear discriminant analysis (LDA) applied to principal component (PC) scores. The percent correct classification of samples were obtained from PLS and LDA in terms of separation on the basis of apple variety and applied heat treatment. PLS gave the highest level of correct classification of the apple juice samples according to both variety and heat treatment, 92.5% correct classification in each case. When LDA was performed on the PC scores obtained from GC analysis, 87.5% and 80% of samples were correctly classified according to apple variety used and applied heat treatment, respectively.
Asunto(s)
Bebidas/clasificación , Cromatografía de Gases/métodos , Frutas/química , Malus/clasificación , Bebidas/análisis , Análisis Discriminante , Manipulación de Alimentos/métodos , Calor , Malus/químicaRESUMEN
The potential of combining solid-phase microextraction (SPME) with gas chromatography and chemometric data analysis to differentiate between pure strawberry samples (Fragraria ananassa) and strawberry samples adulterated with 10, 40, and 70% (v/v) apple purée was investigated. The method involved the extraction of aroma volatiles from the headspace of the purée samples using a SPME fiber followed by GC analysis with flame ionization detection. The principal component analysis (PCA) data matrix consisted of the relative percent peak areas of 37 compounds deemed to be significant in the differentiation of the samples on the basis of adulteration. The PCA results clearly showed that differentiation of the adulterated and unadulterated samples was possible, particularly at the higher levels of adulteration. Partial least-squares regression (PLSR) using a dummy set of Y variables (set to 0 for unadulterated and 1 for adulterated samples) resulted in clear discrimination between unadulterated purées and those containing 40 and 70% (v/v) apple. PLSR using a second set of Y variables, consisting of the actual level of adulteration, enabled quantification of apple purée with a standard error of prediction of 11.6%, implying a minimum detectable level of 25% (v/v) apple. GC-MS analysis enabled identification of the compounds with the greatest influence on sample differentiation. These compounds were identified as hexanoic acid, 2-hexenal, and alpha-farnesene, all of which are key aroma compounds in apples.
Asunto(s)
Cromatografía de Gases/métodos , Contaminación de Alimentos/análisis , Fragaria/química , Frutas/química , Malus/química , Manipulación de Alimentos , Odorantes/análisisRESUMEN
BACKGROUND: Molecular alterations such as DNA microsatellite instability (MSI/RER), single nucleotide polymorphism (SNP) and loss of heterozygosity (LOH) can occur throughout the genome and be associated with different types of cancer. In the present study, we aimed at detecting molecular alterations within the mismatch DNA repair genes in ovarian cancer (OC), using a sensitive, accurate and reliable protocol we have developed. MATERIALS AND METHODS: A combination of high-resolution GeneScan software analysis and automated DNA cycle sequencing was used. RESULTS: Negligible coding MSI was observed in selected sequences of mismatch DNA repair genes in our series of sixty-two ovarian tumors and matched blood DNAs. Unlike MSI, loss of one hMLH1 allele was scored in almost half (47%) of the informative cases. In addition, an SNP in hMSH3/intron 5 was found to be highly variable in OC patients. CONCLUSION: 1) Coding DNA instability is likely to be a very rare event in OC and, therefore, may not significantly contribute to the development of OC, and 2) the high frequency of LOH at hMLH1 observed in our ovarian tumors suggests that further investigation is needed to determine if such a trend exists in other mismatch DNA repair and/or critical genes.