We describe herein the combination of electrochemical immunosensors using single-wall carbon nanotube (SWNT) forest platforms with multi-label secondary antibody-nanotube bioconjugates for highly sensitive detection of a cancer biomarker in serum and tissue lysates. Greatly amplified sensitivity was attained by using bioconjugates featuring horseradish peroxidase (HRP) labels and secondary antibodies (Ab 2 ) linked to carbon nanotubes (CNT) at high HRP/Ab 2 ratio. This approach provided a detection limit of 4 pg mL −1 (100 amol mL −1 ), for prostate specific antigen (PSA) in 10 μL of undiluted calf serum, a mass detection limit of 40 fg. Accurate detection of PSA in human serum samples was demonstrated by comparison to standard ELISA assays. PSA was quantitatively measured in prostate tissue samples for which PSA could not be differentiated by the gold standard immunohistochemical staining method. These easily fabricated SWNT immunosensors show excellent promise for clinical screening of cancer biomarkers and point-of-care diagnostics.
The incidence and mortality rates of prostate cancer are significantly higher in African-American men when compared with European-American men. We tested the hypothesis that differences in tumor biology contribute to this survival health disparity. Using microarray technology, we obtained gene expression profiles of primary prostate tumors resected from 33 African-American and 36 European-American patients. These tumors were matched on clinical variables. We also evaluated 18 nontumor prostate tissues from seven AfricanAmerican and 11 European-American patients. The resulting datasets were analyzed for expression differences on the gene and pathway level comparing African-American with European-American patients. Our analysis revealed a significant number of genes, e.g., 162 transcripts at a false-discovery rate of V5% to be differently expressed between African-American and European-American patients. Using a disease association analysis, we identified a common relationship of these transcripts with autoimmunity and inflammation. These findings were corroborated on the pathway level with numerous differently expressed genes clustering in immune response, stress response, cytokine signaling, and chemotaxis pathways. Several known metastasis-promoting genes, including autocrine mobility factor receptor, chemokine (C-X-C motif) receptor 4, and matrix metalloproteinase 9, were more highly expressed in tumors from African-Americans than EuropeanAmericans. Furthermore, a two-gene tumor signature that accurately differentiated between African-American and European-American patients was identified. This finding was confirmed in a blinded analysis of a second sample set. In conclusion, the gene expression profiles of prostate tumors indicate prominent differences in tumor immunobiology between African-American and European-American men. The profiles portray the existence of a distinct tumor microenvironment in these two patient groups. [Cancer Res 2008;68(3):927-36]
A study of process-induced stress and deformation in thick-section thermosetting composite laminates is presented. A methodology is proposed for predict ing the evolution of residual stress development during the curing process. A one- dimensional cure simulation analysis is coupled to an incremental laminated plate theory model to study the relationships between complex gradients in temperature and degree of cure, and process-induced residual stress and deformation. Material models are proposed to describe the mechanical properties, thermal and chemical strains of the thermoset resin during cure. These material models are incorporated into a micromechanics model to pre dict the effective mechanical properties and process-induced strains of the composite dur ing cure. Thermal expansion and cure shrinkage contribute to changes in material specific volume and represent important sources of internal loading included in the analysis. Tem perature and degree of cure gradients that develop during the curing process represent fun damental mechanisms that contribute to stress development not considered in traditional residual stress analyses of laminated composites. Model predictions of cure dependent epoxy modulus and curvature in unsymmetric graphite/epoxy laminates are correlated with experimental data. The effects of processing history (autoclave temperature cure cy cle), laminate thickness, resin cure shrinkage and laminate stacking sequence on the evo lution of process-induced stress and deformation in thick-section glass/polyester and graphite/epoxy laminates during cure are studied. The magnitude of process-induced residual stress is sufficient to mitiate transverse cracks and delammations. The results clearly indicate that the mechanics and performance of thick-section thermoset laminates are strongly dependent on processing history.
Measurement of gene-expression profiles using microarray technology is becoming increasingly popular among the biomedical research community. Although there has been great progress in this field, investigators are still confronted with a difficult question after completing their experiments: how to validate the large data sets that are generated? This review summarizes current approaches to verifying global expression results, discusses the caveats that must be considered, and describes some methods that are being developed to address outstanding problems.
The reproducibility of conventional two-dimensional (2D) gel electrophoresis can be improved using differential in-gel electrophoresis (DIGE) Proteomics (1) includes the systematic cataloging of protein expression on a large scale, providing complementary information to that obtained from mRNA profiling by microarray (2, 3). Such studies could lead to the molecular characterization of cellular events associated with cancer progression, cellular signaling, and developmental stages (4 -7). Proteomics studies of clinical tumor samples have led to the identification of cancer-specific protein markers, which provide a basis for developing new methods for early diagnosis and early detection and clues to understand the molecular characterization of cancer progression (5, 8 -10).,A mainstay of conventional proteomics is high resolution 2D 1 gel electrophoresis (11, 12) followed by protein identification using mass spectrometry (13-15). The state of the art 2D gel system can be loaded with a few milligrams of protein and separates thousands of protein spots (5, 16). Although the technique has been widely used and successfully applied in a variety of biological systems, several technical limitations exist. Because of subtle changes in experimental conditions, the protein expression patterns on a single 2D gel usually cannot be fully duplicated, which makes it difficult to find the proteins changed between gels and to quantify the changes in protein expression. Although a comparison of protein expression profiles from regular 2D gel electrophoresis can be carried out with the assistance of various software programs, it typically requires some computerized justification of 2D gel images so that two images can be superimposed and compared. These difficulties limit the speed and accuracy of quantitation of protein spots in 2D gel electrophoresis.The differential in-gel electrophoresis (DIGE) technique recently introduced by Amersham Biosciences, Inc. is aimed at improving reproducibility. The concept of DIGE was originally developed by Minden and colleagues (17). To analyze the samples in DIGE, two pools of protein extracts are labeled covalently with fluorescent cyanine dyes, Cy3 and Cy5, re-
A critical review of three classic papers by B Hopkinson, RM Davies, and H Kolsky, and the state-of-the-art in Hopkinson bar experimental techniques is presented. The validity and applicability of the assumption made in the 1D Hopkinson bar theory are discussed. Fundamentals of the Hopkinson bar experimental procedure are outlined including bar calibration, specimen design, pulse shaping, and data analysis following the essential dispersion correction methodology. Additional data tables necessary for dispersion correction are provided to cover a wide range of Poisson's ratio. In addition to the elastic-plastic metals, methodologies for soft and hard materials are also discussed. This paper provides the necessary information for conducting a Hopkinson bar experiment and analyzing the experimental data. This review article cites 61 references.
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