Fluorescent Labeling of Proteins and Its Application to SDS-PAGE and Western Blotting.

This chapter describes very simple fluorescent methods developed in our laboratory allowing the rapid monitoring of total protein patterns on both sodium dodecyl sulfate (SDS) polyacrylamide gels and western blots. The noncovalent dye Nile red (9-diethylamino-5H-benzo[α]phenoxazine-5-one) is used for the sensitive staining of proteins in SDS gels. This method is compatible with the electroblotting of protein bands and with the staining of the resulting blot with the covalent dye MDPF (2-methoxy-2,4-diphenyl-3(2H)-furanone).

A model for describing the eutrophication in a heavily regulated coastal lagoon. Application to the Albufera of Valencia (Spain).

A simplified two-dimensional eutrophication model was developed to simulate temporal and spatial variations of chlorophyll-a in heavily regulated coastal lagoons. This model considers the hydrodynamics of the whole study area, the regulated connexion of the lagoon with the sea, the variability of the input and output nutrient loads, the flux from the sediments to the water column, the phytoplankton growth and mortality kinetics, and the zooplankton grazing. The model was calibrated and validated by applying it to the Albufera of Valencia, a hypertrophic system whose connection to the sea is strongly regulated by a system of sluice-gates.

HER2 status determination using RNA-ISH--a rapid and simple technique showing high correlation with FISH and IHC in 141 cases of breast cancer.

Aims: the assessment of the human epidermic growth factor receptor 2 (HER2) is currently performed in most laboratories using two techniques: Fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC), and novel methodology is being investigated continuously in the assessment of HER2, such as SISH, CISH, DNA chips, ELISA or real time PCR to make assessment easier, faster or more accurate. RNA-ISH (RNA in Situ Hybridisation) is a new technique designed to detect mRNA expression levels, conducted by light microscope without the need for counting or grading systems in a total processing time of 4 hours. This study aims to determine if RNA-ISH is a viable and effective technique and a possible alternative to the currently used techniques by analysing and comparing genetic amplification (FISH) and protein levels (IHC) with mRNA over-expression (RNA-ISH) in 141 cases of breast cancer.

mRNA in situ hybridization (HistoSonda): a new diagnostic tool for HER2-status in breast cancer-a multicentric Spanish study.

Monoclonal therapies could represent baseline-personalized medicine for patients with neoplasia. One of the most successful examples is Trastuzumab, a humanized antibody against epidermal growth factor receptor 2. Human epidermal growth factor receptor 2 (HER2) is a trans-membrane tyrosine kinase coded by the gene HER2/neu and overexpressed in approximately 12% to 20% of infiltrating breast carcinomas.

[Classifying Mexican adolescents' high blood pressure, associated factors and importance].

Objective: Determining characteristics regarding risk and high blood pressure (HBP) frequency in a group of 12 to 16 year-old students from the city of León, Mexico, according to two types of classification: the 7th Joint National Committee (JNC-7) and 4th report of the working group for diagnosis, evaluation and treatment of HBP in children and adolescents.

Methods: This was an observational, descriptive cross-sectional study of 458 male students. HBP was determined according to international standards after parents and students' informed consent had been obtained and they answered a questionnaire for recording clinical-epidemiological data.

S6K1 deficiency protects against apoptosis in hepatocytes.

Unlabelled: The mammalian target of rapamycin (mTOR)/S6K1 signaling pathway controls cell growth and proliferation. To assess the importance of S6K1 in the balance between death and survival in the liver, we have generated immortalized hepatocyte cell lines from wild-type and S6K1-deficient (S6K1(-/-)) mice. In S6K1(-/-) hepatocytes, caspase-8 and the pro-apoptotic protein Bid were constitutively down-regulated as compared with wild-type.

Fluorescent labeling of proteins and its application to SDS-PAGE and western blotting.

This chapter describes very simple fluorescent methods developed in our laboratory allowing the rapid monitoring of total protein patterns on both sodium dodecyl sulfate (SDS) polyacrylamide gels and western blots. The noncovalent dye Nile red (9-diethylamino-5H-benzo[alpha]phenoxazine-5-one) is used for the sensitive staining of proteins in SDS gels. This method is compatible with the electroblotting of protein bands and with the staining of the resulting blot with the covalent dye MDPF (2-methoxy-2,4-diphenyl-3(2H)-furanone).

Levels of protein tyrosine phosphatase 1B determine susceptibility to apoptosis in serum-deprived hepatocytes.

Protein tyrosine phosphatase 1B (PTP1B) is a negative regulator of tyrosine kinase growth factor signaling. To assess the importance of PTP1B in the balance between death and survival in the liver, we have developed immortalized neonatal hepatocyte cell lines lacking (PTP1B(-/-)) or overexpressing (PTP1B(+/+PTP1B)) PTP1B. Early activation of caspase-3 occurred in PTP1B(+/+PTP1B) hepatocytes but was nearly abolished in PTP1B(-/-) cells.