RESUMEN
Heat-inducibility of two reporter constructs expressing lacZ gene under the control of mouse and Xenopus hsp70 promoters was tested in zebrafish (Danio rerio) embryos using a transient expression system. Cells expressing beta-galactosidase were stained blue by histochemical staining and their average number per embryo was used as an indicator of the expression level of the reporter gene. Both constructs were heat-inducible in the embryonic tissues and showed similar heat dependence (increasing expression levels from 35-36 degrees C up to 39 degrees C with an apparent decrease at 40 degrees C), resembling that of the zebrafish hsp70 genes. However, their induction kinetics were different, which might be due to differences in their 5' UTRs. Spatial expression patterns of the two hsp/lacZ constructs and an endogenous hsp70 gene were mostly similar on the RNA level. These results indicate that our approach is applicable for in vivo analysis of the heat-shock response and that exogenous heat-shock promoters may be useful for inducible expression of transgenes in fish.
Asunto(s)
Regulación de la Expresión Génica , Genes Reporteros , Proteínas HSP70 de Choque Térmico/genética , Regiones Promotoras Genéticas , beta-Galactosidasa/genética , Animales , Animales Modificados Genéticamente , Embrión no Mamífero , Escherichia coli/genética , Vectores Genéticos , Calor , Ratones , Secuencias Reguladoras de Ácidos Nucleicos , Xenopus , Pez Cebra/embriologíaRESUMEN
We searched for sex-specific DNA sequences in the male and female genomes of African catfish, Clarias gariepinus (Burchell, 1822) by comparative random amplified polymorphic DNA (RAPD) assays performed on pooled DNA samples. Two sex-linked RAPD markers were identified from the male DNA pool and confirmed on individual samples, showing good agreement with phenotypic sex. Both markers were isolated, cloned and characterized. The first marker (CgaY1) was nearly 2.6 kb long, while the length of second one (CgaY2) was 458 bp. Southern blot analysis with a CgaY1 probe showed strong hybridizing fragments only in males and not in females under stringent conditions, indicating the presence of multiple copies of CgaY1 in the male genome. When tested by zoo blot on the genomes of two closely related species from the Clariidae family, CgaY1 hybridized to the DNA of Heterobranchus longifilis and generated a faint male-specific band at low stringency. CgaY2 produced similar hybridization pattern in both sexes of C. gariepinus, C. macrocephalus and H. longifilis. Specific primers were designed to the sequences and the markers were amplified in multiplex PCR reactions together with a control band common to all individuals. This allowed for rapid, molecular sexing of the species on the basis of a simple three band (male) versus one band (female) pattern. According to our knowledge these are the first sex-specific DNA markers isolated from a siluroid fish species.
Asunto(s)
Bagres/genética , Marcadores Genéticos/genética , Cromosoma Y/genética , Animales , Clonación Molecular , Femenino , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Técnica del ADN Polimorfo Amplificado Aleatorio , Procesos de Determinación del SexoRESUMEN
A study of the volumes of the right and left breasts of 598 subjects was undertaken using biostereometric analysis. This measurement uses close-range stereophotogrammetry to characterize the shape of the breast, and is noncontact, noninvasive, accurate, and rapid with respect to the subject involvement time. Using chi-square tests, volumes and volumetric differences between breast pairs were compared with handedness, perception of breast size by each subject, age, and menstrual status. No significant relationship was found between the handedness, age, or menstrual status of the subject and the breast volume. Several groups of subjects were accurate in their perception of breast size difference. Analysis did confirm the generally accepted clinical impression of left-breast volume dominance. These results are shown to be consistent with those of a previous study using 248 women.
Asunto(s)
Antropometría/métodos , Mama/anatomía & histología , Fotogrametría , Adulto , Anciano , Imagen Corporal , Femenino , Lateralidad Funcional , Humanos , Menstruación , Persona de Mediana EdadRESUMEN
A study of volumes of the right and left breasts of 248 subjects was undertaken using biostereometric analysis. This measurement technique uses close-range stereophotogrammetry to characterize the shape of the breast and is noncontact, noninvasive, accurate, and rapid with respect to the subject involvement time. Volumes and volumetric differences between breast pairs were compared, using chi-square tests, with handedness, perception of breast size by each subject, age, and menstrual status. No significant relationship was found between the handedness of the subject and the larger breast volume. Several groups of subjects based on age and menstrual status were accurate in their perception of breast size difference. Analysis did not confirm the generally accepted clinical impression of left breast volume dominance. Although a size difference in breast pairs was documented, neither breast predominated.
Asunto(s)
Mama/anatomía & histología , Fotogrametría/métodos , Adulto , Anciano , Femenino , Humanos , Persona de Mediana Edad , Fotogrametría/instrumentaciónRESUMEN
A measurement technique involving the use of close-range stereophotogrammetry has been developed for the study of breast volume and volume distribution (shape) comparison. This biostereometric method has been shown to be able to measure volume to within 0.63% of that determined by water displacement. This study describes the rapid and non-invasive data acquisition methodology as well as the mathematical algorithms used to determine breast volume. Additionally, the paper discusses the application of this process to documenting the presence or absence of total volume and volume distribution differences existing in normal breasts and those containing benign and malignant tumors.