Differential expression of Axl and Gas6 in renal cell carcinoma reflecting tumor advancement and survival

PURPOSE: Overexpression of the receptor tyrosine kinase Axl is implicated in several cancers. Therefore, we conducted this study to determine the expression of Axl and its ligand Gas6 in various renal cell carcinoma (RCC) types and in oncocytoma. EXPERIMENTAL DESIGN: Real-time quantitative reverse transcription-PCR was used to quantify tumor mRNA levels for Axl and Gas6 in a cohort (n = 221) of RCC patients. Serum levels of soluble sAxl and Gas6 proteins were measured using specific ELISA assays (n = 282). The presence of Axl protein in tumor tissue was evaluated by immunohistochemistry (n = 294). Results were correlated to tumor-associated variables, clinical biochemical tests, and patient survival. RESULTS: Tumor Axl mRNA levels correlated independently to survival when assessed against tumor stage and grade. In the study group, the median cancer-specific survival of all RCC patients during 307 months of follow-up was 55 months (confidence interval, +/-40.4). The 25% of patients with lowest tumor Axl mRNA levels had significantly better survival than the rest (P = 0.0005), with 70% of the patients still alive at the end of follow-up. In contrast, in patients with medium-high Axl mRNA, only 25% were alive at the end of follow-up. Tumor Gas6 mRNA levels correlated to survival, tumor-associated variables, and disease severity as did serum levels of soluble sAxl and Gas6 protein. However, no correlation between Axl protein in tumor tissue and survival was found. CONCLUSIONS: Axl and Gas6 expression in RCC are associated with tumor advancement and patient survival. In particular, low tumor Axl mRNA levels independently correlated with improved survival

Mapping cells and sub-cellular organelles on 2-D gels: ‘new tricks for an old horse’

AbstractNowadays, investigators in all fields are faced with the identification of unknown, up- or down-regulated, modified proteins that they are trying to identify. Two-dimensional (2-D) gel electrophoresis, with its ability to resolve several thousand proteins, is an extremely powerful technique. The current resolution and reproducibility of 2-D gel technology and the establishment of computer assisted 2-D gel protein databases have paved new ways for the identification of proteins

Quality assurance of the solar UV network in the Antarctic

Measuring ultraviolet radiation in the Antarctic region, where weather conditions are extremely challenging, is a demanding task. Proper quality control of the measurements and quality assurance of the data, which are the basis of all scientific use of data, has to be especially well planned and executed. In this paper we show the importance of proper quality assurance and describe the methods used to successfully operate the NILU-UV multichannel radiometers of the Antarctic network stations at Ushuaia, 54S, and Marambio, 64S. According to our experience, even though multichannel instruments are supposed to be rather stable as a function of time, severe drifts can occur in the sensitivity of the channels under these harsh conditions. During 2000–2003 the biggest drifts were 35%, both at Ushuaia and Marambio, with the sensitivity of the channels dropping at different rates. Without proper corrections in the data, this would have seriously affected the calculated UV dose rates. As part of the quality assurance of the network a traveling reference NILU-UV, which was found to be stable, was used to transfer the desired irradiance scale to the site NILU-UV data. Relative lamp tests were used to monitor the stability of the instruments. Each site NILU-UV was scaled channel by channel to the traveling reference by performing solar comparisons. The method of scaling each channel separately was found to be successful, even though the differences between the raw data of the site NILU-UV and the reference instruments were, before the data correction, as much as 40%. After the correction, the mean ratios of erythemally weighted UV dose rates measured during the solar comparisons in 2000–2003 between the reference NILU-UV and the site NILU-UV were 1.007 ± 0.011 and 1.012 ± 0.012 for Ushuaia and Marambio, respectively, when the solar zenith angle varied up to 80. These results make possible the scientific use of NILU-UV data measured simultaneously at quite different locations, e.g., the Antarctic and Arctic, and the method presented is also practicable for other multichannel radiometer networks.S, and Marambio, 64S. According to our experience, even though multichannel instruments are supposed to be rather stable as a function of time, severe drifts can occur in the sensitivity of the channels under these harsh conditions. During 2000–2003 the biggest drifts were 35%, both at Ushuaia and Marambio, with the sensitivity of the channels dropping at different rates. Without proper corrections in the data, this would have seriously affected the calculated UV dose rates. As part of the quality assurance of the network a traveling reference NILU-UV, which was found to be stable, was used to transfer the desired irradiance scale to the site NILU-UV data. Relative lamp tests were used to monitor the stability of the instruments. Each site NILU-UV was scaled channel by channel to the traveling reference by performing solar comparisons. The method of scaling each channel separately was found to be successful, even though the differences between the raw data of the site NILU-UV and the reference instruments were, before the data correction, as much as 40%. After the correction, the mean ratios of erythemally weighted UV dose rates measured during the solar comparisons in 2000–2003 between the reference NILU-UV and the site NILU-UV were 1.007 ± 0.011 and 1.012 ± 0.012 for Ushuaia and Marambio, respectively, when the solar zenith angle varied up to 80. These results make possible the scientific use of NILU-UV data measured simultaneously at quite different locations, e.g., the Antarctic and Arctic, and the method presented is also practicable for other multichannel radiometer networks.S. According to our experience, even though multichannel instruments are supposed to be rather stable as a function of time, severe drifts can occur in the sensitivity of the channels under these harsh conditions. During 2000–2003 the biggest drifts were 35%, both at Ushuaia and Marambio, with the sensitivity of the channels dropping at different rates. Without proper corrections in the data, this would have seriously affected the calculated UV dose rates. As part of the quality assurance of the network a traveling reference NILU-UV, which was found to be stable, was used to transfer the desired irradiance scale to the site NILU-UV data. Relative lamp tests were used to monitor the stability of the instruments. Each site NILU-UV was scaled channel by channel to the traveling reference by performing solar comparisons. The method of scaling each channel separately was found to be successful, even though the differences between the raw data of the site NILU-UV and the reference instruments were, before the data correction, as much as 40%. After the correction, the mean ratios of erythemally weighted UV dose rates measured during the solar comparisons in 2000–2003 between the reference NILU-UV and the site NILU-UV were 1.007 ± 0.011 and 1.012 ± 0.012 for Ushuaia and Marambio, respectively, when the solar zenith angle varied up to 80. These results make possible the scientific use of NILU-UV data measured simultaneously at quite different locations, e.g., the Antarctic and Arctic, and the method presented is also practicable for other multichannel radiometer networks.. These results make possible the scientific use of NILU-UV data measured simultaneously at quite different locations, e.g., the Antarctic and Arctic, and the method presented is also practicable for other multichannel radiometer networks.Fil: Lakkala, K.. Finnish Meteorological Institute; FinlandiaFil: Redondas, A.. Instituto Nacional de Meteorología; EspañaFil: Meinander, O.. Finnish Meteorological Institute; FinlandiaFil: Torres ,Carlos. Instituto Nacional de Meteorología; EspañaFil: Koskela, T.. Finnish Meteorological Institute; FinlandiaFil: Cuevas, Eduardo. Instituto Nacional de Meteorología; EspañaFil: Taalas, P.. Finnish Meteorological Institute; FinlandiaFil: Dahlback, A.. University of Oslo; NoruegaFil: Deferrari, Guillermo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; ArgentinaFil: Edvardsen, K.. Instituto Noruego de Investigación del Aire; NoruegaFil: Ochoa, H.. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentin

Plasma concentrations of Gas6 and sAxl correlate with disease activity in systemic lupus erythematosus

Objectives. SLE is a systemic autoimmune disease with an annual incidence of 3.8 per 100 000. Several pathogenic mechanisms are believed to be operating in SLE, including an impaired clearance of apoptotic cells, activation of the type I IFN pathway and generation of autoimmune leucocytes. Growth arrest-specific protein 6 (Gas6) and its receptor Axl are known to regulate inflammation and may be implicated in lupus pathogenesis. We have recently developed immunological methods to quantify the vitamin-K-dependent protein Gas6 and its soluble receptor sAxl in human plasma, which we have used to investigate the role of Gas6 and soluble Axl in SLE

Multiple var2csa-Type PfEMP1 Genes Located at Different Chromosomal Loci Occur in Many Plasmodium falciparum Isolates

BACKGROUND:The var2csa gene encodes a Plasmodium falciparum adhesion receptor which binds chondroitin sulfate A (CSA). This var gene is more conserved than other PfEMP1/var genes and is found in all P. falciparum isolates. In isolates 3D7, FCR3/It4 and HB3, var2csa is transcribed from a sub-telomeric position on the left arm of chromosome 12, but it is not known if this location is conserved in all parasites. Genome sequencing indicates that the var2csa gene is duplicated in HB3, but whether this is true in natural populations is uncertain. METHODOLOGY/PRINCIPAL FINDINGS:To assess global variation in the VAR2CSA protein, sequence variation in the DBL2X region of var2csa genes in 54 P.falciparum samples was analyzed. Chromosome mapping of var2csa loci was carried out and a quantitative PCR assay was developed to estimate the number of var2csa genes in P.falciparum isolates from the placenta of pregnant women and from the peripheral circulation of other malaria patients. Sequence analysis, gene mapping and copy number quantitation in P.falciparum isolates indicate that there are at least two loci and that both var2csa-like genes can be transcribed. All VAR2CSA DBL2X domains fall into one of two distinct phylogenetic groups possessing one or the other variant of a large (approximately 26 amino acid) dimorphic motif, but whether either motif variant is linked to a specific locus is not known. CONCLUSIONS/SIGNIFICANCE:Two or more related but distinct var2csa-type PfEMP1/var genes exist in many P. falciparum isolates. One gene is on chromosome 12 but additional var2csa-type genes are on different chromosomes in different isolates. Multiplicity of var2csa genes appears more common in infected placentae than in samples from non-pregnant donors indicating a possible advantage of this genotype in pregnancy associated malaria

Adaptation of an Evaluation System for e-Health Environments

Proceedings of: 14th International Conference, KES 2010, Cardiff, UK, September 8-10, 2010The increase in ageing of European population implies a high cost in economy and society in any European country and it can be reduced if we pay attention and develop home care systems. Evaluation of these systems is a critical and challenging issue but seldom tackled. It is important before evaluating a system to figure out what is the evaluation goal. In our case, such a goal is to evaluate enhanced user experience and beyond the evaluation goal it is also a central concern about what to evaluate. In this paper we propose a multi-agent home care system where we describe how agents coordinate their decisions to provide e-services to patients when at home after hospitalization. Finally we center our proposal on the adaptation of an evaluation system, previously developed, to support the challenges of an e-Health environment and also the multi-user evaluation. These evaluation methods (online/offline) will provide user's (patients, patient's relatives and healthcare professionals) feedback into the system.This work was supported in part by Projects CICYT TIN2008-06742-C02-02/ TSI, CICYT TEC2008-06732-C02-02/TEC, CAM CONTEXTS (S2009/ TIC-1485) and DPS2008-07029-C02-02.Publicad

MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR

Background Methylation of the O6-methylguanine-DNA methyltransferase (MGMT) gene promoter is a favorable prognostic factor in glioblastoma patients. However, reported methylation frequencies vary significantly partly due to lack of consensus in the choice of analytical method. Method We examined 35 low- and 99 high-grade gliomas using quantitative methylation specific PCR (qMSP) and pyrosequencing. Gene expression level of MGMT was analyzed by RT-PCR. Results When examined by qMSP, 26% of low-grade and 37% of high-grade gliomas were found to be methylated, whereas 97% of low-grade and 55% of high-grade gliomas were found methylated by pyrosequencing. The average MGMT gene expression level was significantly lower in the group of patients with a methylated promoter independent of method used for methylation detection. Primary glioblastoma patients with a methylated MGMT promoter (as evaluated by both methylation detection methods) had approximately 5 months longer median survival compared to patients with an unmethylated promoter (log-rank test; pyrosequencing P = .02, qMSP P = .06). One third of the analyzed samples had conflicting methylation results when comparing the data from the qMSP and pyrosequencing. The overall survival analysis shows that these patients have an intermediate prognosis between the groups with concordant MGMT promoter methylation results when comparing the two methods. Conclusion In our opinion, MGMT promoter methylation analysis gives sufficient prognostic information to merit its inclusion in the standard management of patients with high-grade gliomas, and in this study pyrosequencing came across as the better analytical method

Cleavage Requirements for Activation of Factor V by Factor Xa

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65728/1/j.1432-1033.1997.00012.x.pd

Identifying novel genetic determinants of hemostatic balance

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73734/1/j.1538-7836.2005.01461.x.pd