Standardization of molecular monitoring for chronic myeloid leukemia in Latin America using locally produced secondary cellular calibrators

Residual disease in chronic myeloid leukemia (CML) patients undergoing therapy with tyrosine kinase inhibitors (TKIs) is measured by assessing the quantity of transcripts of the BCR-ABL1 fusion gene in peripheral white blood cells. This analysis is based on reverse-transcription quantitative PCR (RT–qPCR) technology; however, the wide array of methods used worldwide has led to large variation in quantitative BCR-ABL1 measurements, which hamper inter-laboratory comparative studiesFil: Ruiz, María Sol. Fundación Cáncer. Centro de Investigaciones Oncológicas; ArgentinaFil: Medina, M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Tapia, I.. Fundación Cáncer. Centro de Investigaciones Oncológicas; ArgentinaFil: Mordoh, Jose. Fundación Cáncer. Centro de Investigaciones Oncológicas; ArgentinaFil: Cross, N. C. P.. Universidad de Southampton Uk; Reino UnidoFil: Larripa, Irene Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Bianchini, Michele. Fundación Cáncer. Centro de Investigaciones Oncológicas; Argentin

Comparative study of gene expression by cDNA microarray in human colorectal cancer tissues and normal mucosa

The causative molecular pathways underlying the pathogenesis of colorectal cancer (CRC) need to be better characterized. The purpose of our study was to better understand the genetic mechanism of oncogenesis for human colorectal cancer and to identify new potential tumor markers of use in clinical practice. We used cDNA microarrays to compare gene expression profiles of colorectal biopsies from 25 CRC patients and 13 normal mucosa from adjacent non-cancerous tissues. Findings were validated by real-time PCR; in addition, western blotting and immunochemistry analysis were carried out as further confirmation of differential expression at a protein level. Comparing cancerous tissues with normal colonic mucosa we identified 584 known genes differentially expressed to a significant degree (p<0.001). Many of the transcripts that were more abundant in tumors than in non-neoplastic tissues appear to reflect important events for colon carcinogenesis. For example, a significant number of these genes serve as apoptotic inhibitors (e.g. BFAR, BIRC1, BIRC6). Furthermore, we observed the simultaneous up-regulation of HLA-E and the down-regulation of beta2-microglobulin; these genes strongly support a potential tumor escape strategy from immune surveillance in colon cancer tissues. Our study provides new gene candidates in the pathogenesis of human CRC disease. From our results we hypothesize that CRC cells escape immune surveillance through a specific gene expression alteration; moreover, over-expression of several survival genes seems to confer a more anti-apoptotic phenotype. These genes are involved in pathways not previously implicated in CRC pathogenesis and they may provide new targets for therapy.Fil: Bianchini, Michele. Fundación P/la Invest.y Prevención del Cancer. Centro de Investigaciones Oncologicas; ArgentinaFil: Levy, Estrella Mariel. Fundación P/la Invest.y Prevención del Cancer. Centro de Investigaciones Oncologicas; Argentina. Universidad de Buenos Aires; ArgentinaFil: Zucchini, Cinzia. Università di Bologna; ItaliaFil: Pinski, Victor. Universidad de Buenos Aires; ArgentinaFil: Macagno, Carlo. Universidad de Buenos Aires; ArgentinaFil: De Sanctis, Paola. Università di Bologna; ItaliaFil: Valvassori, Luisa. Università di Bologna; ItaliaFil: Carinci, Paolo. Università di Bologna; ItaliaFil: Mordoh, Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Fundación P/la Invest.y Prevención del Cancer. Centro de Investigaciones Oncologicas; Argentin

Volumetric Lissajous confocal microscopy with tunable spatiotemporal resolution

Dynamic biological systems present challenges to existing three-dimensional (3D) optical microscopes because of their continuous temporal and spatial changes. Most techniques are rigid in adapting the acquisition parameters over time, as in confocal microscopy, where a laser beam is sequentially scanned at a predefined spatial sampling rate and pixel dwell time. Such lack of tunability forces a user to provide scan parameters, which may not be optimal, based on the best assumption before an acquisition starts. Here, we developed volumetric Lissajous confocal microscopy to achieve unsurpassed 3D scanning speed with a tunable sampling rate. The system combines an acoustic liquid lens for continuous axial focus translation with a resonant scanning mirror. Accordingly, the excitation beam follows a dynamic Lissajous trajectory enabling sub-millisecond acquisitions of image series containing 3D information at a sub-Nyquist sampling rate. By temporal accumulation and/or advanced interpolation algorithms, the volumetric imaging rate is selectable using a post-processing step at the desired spatiotemporal resolution for events of interest. We demonstrate multicolor and calcium imaging over volumes of tens of cubic microns with 3D acquisition speeds of 30 Hz and frame rates up to 5 kHz

A Social Network-based Framework for Data Services Selection in Modern Web Application Design

Abstract. In recent years the design of enterprise Web applications is more and more based on the integration of resources delivered through data services from outside the organization boundaries. Searching and composing existing data services offer many advantages, namely, the availability of widespread solutions in the form of services and data shared over the Web, and reduced development costs. In this scenario, new methods for speeding up the design process are emerging and, in particular, developers&apos; social networks have been established, where developers follow other developers to learn from their choices in selecting suitable services. In this paper, we propose a framework to support data service selection for modern Web application design, by also considering the developers&apos; social network. The network of social relationships, properly weighted with the developers&apos; credibility, is used to compute developers&apos; rank. This rank qualifies developers&apos; experience in selecting data services

Sorting Nexin 1 Down-Regulation Promotes Colon Tumorigenesis

PURPOSE: Colon cancer is one of the most common human malignancies, yet studies have only begun to identify the multiple mechanisms that underlie the development of this tumor. In this study, we have identified a novel mechanism, dysregulation of endocytic sorting, which promotes colon cancer development. EXPERIMENTAL DESIGN: Immunohistochemical and microarray analyses were done on human colon cancer tissue specimens to determine the levels of one endocytic protein, sorting nexin 1 (SNX1). SW480 cells, a human colon cancer cell line that retains a relatively high level of SNX1 expression, were used to assess the effects of down-regulating this protein by small hairpin RNA. Activation of signal transduction cascades was evaluated in these cells using Western blotting, and multiple functional assays were done. RESULTS: We determined by immunohistochemistry that the level of SNX1 was significantly down-regulated in 75% of human colon cancers. In corroborative studies using microarray analysis, SNX1 message was significantly decreased (log(2) ratio less than -1) for 8 of 19 colon carcinomas. Cell lines with reduced SNX1 levels showed increased proliferation, decreased apoptosis, and decreased susceptibility to anoikis. They also showed increased activation of epidermal growth factor receptor and extracellular signal-regulated kinase 1/2 in response to epidermal growth factor. This increased activation was abolished by inhibition of endocytosis. CONCLUSIONS: These data suggest that loss of SNX1 may play a significant role in the development and aggressiveness of human colon cancer, at least partially through the mechanism of increased signaling from endosomes. Further, these findings suggest that dysregulation of endocytic proteins may represent a new paradigm in the process of carcinogenesis.Fil: Nguyen, Lananh N.. University of Washington; Estados UnidosFil: Holdren, Matthew S.. University of Washington; Estados UnidosFil: Nguyen, Anthony P.. Baylor College of Medicine; Estados UnidosFil: Furuya, Momoko H.. University of Washington; Estados UnidosFil: Bianchini, Michele. Fundación Cáncer. Centro de Investigaciones Oncológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Levy, Estrella Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación Cáncer. Centro de Investigaciones Oncológicas; ArgentinaFil: Mordoh, Jose. Fundación Cáncer. Centro de Investigaciones Oncológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Liu, Annie. University of Washington; Estados UnidosFil: Guncay, Gabriela D.. University of Washington; Estados UnidosFil: Campbell, Jean S.. University of Washington; Estados UnidosFil: Parks, W. Tony. University of Washington; Estados Unido

GPR activities in Italy: a review

Ground-penetrating radar has been increasingly played an important role over the last 15 years in Italy due to its high reliability in assisting the assessment of the built environment for civil engineering purposes, and in being used for geophysical investigations within many other fields of application. In line with this, original works involving fundamental aspects of this technique and implementing its use more practically in a number of interesting projects have been developed over years, both under a research and an enterprise point of view. This paper will endeavour to review the current status of ground-penetrating radar activities in Italy. Efforts have been devoted to single out the most interesting national research projects, both recent and ongoing, involving ground-penetrating radar in Italy, such as the ARCHEO project in the 90s, funded by the Italian Ministry for Universities, wherein a stepped frequency ultra-wide band radar suited for archaeological surveys was manufactured. In this framework, it is worth citing another important and more recent project, European Community funded, namely, ORFEUS, which started in the late 2006 with the overall aim of providing the capability to locate buried infrastructure accurately and reliably by means of a bore-head ground-penetrating radar for horizontal directional drilling. A review on the main use of this non-destructive technique in management activities of national resources and infrastructures has been also performed, ranging from the applications made by Anas S.p.A., i.e. the main management authority for the Italian road and motorway network, up to private enterprises specialized in both services providing and ground-penetrating radar manufacturing such as, to cite a few, Sineco S.p.A. and IDS Ingegneria dei Sistemi S.p.A., respectively. Current national guidelines, rules or protocols to be followed during radar surveys have been also reviewed. Unlike well-established international standards such as the ASTM D 4748-98 and the ASTM D 6432-99 dealing with, respectively, thicknesses evaluation of bound layers in road pavements, and equipment, field procedures and data-interpretation for the electromagnetic evaluation of subsurface materials, it has to be noted that the Italian body of laws and rules tackles the ground-penetrating radar applications under an indirect and partial approach. Despite of such situation, national guidelines concerning utilities-detection activities as well as other theoretical and practical guidelines established by the major Italian private enterprises on this field can be also considered highly relevant. Moreover, a further focus on the activities and main devices of the major Italian ground-penetrating radar manufacturers have been thoroughly described. Under a research and innovation perspective, the most important test sites, such as the site of the University of Salento to reconstruct archaeological and urban subsurface scenarios have been listed along with the main advances reached in integrating ground-penetrating radar with other non-destructive techniques, to inform and potentially improve the possibility of new developments and collaborations

Improved Diagnosis of the Transition to JAK2V617F Homozygosity: The Key Feature for Predicting the Evolution of Myeloproliferative Neoplasms

Most cases of BCR-ABL1-negative myeloproliferative neoplasms (MPNs), essential thrombocythemia, polycythemia vera and primary myelofibrosis are associated with JAK2V617F mutations. The outcomes of these cases are critically influenced by the transition from JAK2V617F heterozygosity to homozygosity. Therefore, a technique providing an unbiased assessment of the critical allele burden, 50% JAK2V617F, is highly desirable. In this study, we present an approach to assess the JAK2V617F burden from genomic DNA (gDNA) and complementary DNA (cDNA) using one-plus-one template references for allele-specific quantitative-real-time-PCR (qPCR). Plasmidic gDNA and cDNA constructs encompassing one PCR template for JAK2V617F spaced from one template for JAK2Wild Type were constructed by multiple fusion PCR amplifications. Repeated assessments of the 50% JAK2V617F burden within the dynamic range of serial dilutions of gDNA and cDNA constructs resulted in 52.5364.2% and 51.4664.21%, respectively. The mutation-positive cutoff was estimated to be 3.65% (mean +2 standard deviation) using 20 samples from a healthy population. This qPCR approach was compared with the qualitative ARMS-PCR technique and with two standard methods based on qPCR, and highly significant correlations were obtained in all cases. qPCR assays were performed on paired gDNA/cDNA samples from 20 MPN patients, and the JAK2V617F expression showed a significant correlation with the allele burden. Our data demonstrate that the qPCR method using one-plus-one template references provides an improved assessment of the clinically relevant transition of JAK2V617F from heterozygosity to homozygosity.Fil: Gonzalez, Mariana Selena. Consejo Nacional de Invest.cientif.y Tecnicas. Instituto de Medicina Experimental;Fil: de Brasi, Carlos Daniel. Consejo Nacional de Invest.cientif.y Tecnicas. Instituto de Medicina Experimental;Fil: Bianchini, Michele. DTO. DE GENETICA;Fil: Gargallo, Patricia Martha. INST. DE INVEST. HEMATOLOGICAS;Fil: Stanganelli, Carmen Graciela. Academia Nacional de Medicina de Buenos Aires. Instituto de Invest. Hematologicas "mariano R. Castex";Fil: Zalcberg, Ilana. Molecular Biology, Laboratory, Instituto Nacional do Caˆncer, Rio de Janeiro, Brazil;Fil: Larripa, Irene Beatriz. Consejo Nacional de Invest.cientif.y Tecnicas. Instituto de Medicina Experimental

Estimation of the Aerodynamic Force Induced by Vaneless Diffuser Rotating Stall in Centrifugal Compressor Stages

Abstract Rotating stall in centrifugal compressors not only adversely affects the performance before surge, but also can generate high subsynchronous vibrations, marking the minimum flow limit of a machine. Recent works presented an experimental approach to estimate the stall force induced by the unbalanced pressure field in a vaneless diffuser using dynamic pressure measurements. In this study, the results of a 3D-unsteady simulation of a radial stage model were used to estimate the stall force and to compare it with the approximation obtained with an "experimental-like" approach. Results showed that: a) the experimental approach, using an ensemble average approach for transposing data between time and space domains provides sufficiently accurate results; b) the momentum contribution, neglected in experiments, gives negligible contribution to the final intensity of the stall force