The recent development of tissue microarray technology has potentiated large-scale retrospective cohort studies using archival formalin-fixed, paraffin-embedded tissues. A major obstacle to broad ...acceptance of microarrays is that they reduce the amount of tissue analyzed from a whole tissue section to a disk, 0.6 mm in diameter, that may not be representative of the protein expression patterns of the entire tumor. In this study, we examine the number to disks required to adequately represent the expression of three common antigens in invasive breast carcinoma--estrogen receptor, progesterone receptor, and the Her2/neu oncogene--in 38 cases of invasive breast carcinoma. We compared the staining of 2 to 10 microarray disks and the whole tissue sections from which they were derived and determined that analysis of two disks is comparable to analysis of a whole tissue section in more than 95% of cases. To evaluate the potential for using archival tissue in such arrays, we created a breast cancer microarray of 8 to 11 cases from each decade beginning in 1932 to the present day and evaluated the antigenicity of these markers and others. This array demonstrates that many proteins retain their antigenicity for more than 60 years, thus validating their study on archival tissues. We conclude that the tissue microarray technique, with 2-fold redundancy, is a valuable and accurate method for analysis of protein expression in large archival cohorts.
The recent development of tissue microarrays-composed of hundreds of tissue sections from different tumors arrayed on a single glass slide-facilitates rapid evaluation of large-scale outcome studies. ...Realization of this potential depends on the ability to rapidly and precisely quantify the protein expression within each tissue spot. We have developed a set of algorithms that allow the rapid, automated, continuous and quantitative analysis of tissue microarrays, including the separation of tumor from stromal elements and the sub-cellular localization of signals. Validation studies using estrogen receptor in breast carcinoma show that automated analysis matches or exceeds the results of conventional pathologist-based scoring. Automated analysis and sub-cellular localization of beta-catenin in colon cancer identifies two novel, prognostically significant tumor subsets, not detected by traditional pathologist-based scoring. Development of automated analysis technology empowers tissue microarrays for use in discovery-type experiments (more typical of cDNA microarrays), with the added advantage of inclusion of long-term demographic and patient outcome information.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Summary Vascular endothelial growth factor has been shown to be up-regulated in breast cancers. Vascular endothelial growth factor receptors, VEGFR-1 and VEGFR-2, are the principal mediators of its ...effects. Together with VEGFR-1 and VEGFR-2, neuropilin-1 may act as a coreceptor for vascular endothelial growth factor. Although vascular endothelial growth factor exerts important effects on endothelial cells, VEGFRs are likely present on tumor cells as well. We used AQUA to analyze tumor-specific expression of vascular endothelial growth factor, VEGFR-1, VEGFR-2, and neuropilin-1 on a large cohort of breast cancer tissue microarray. Two-fold redundant arrays were constructed from 642 cases of primary breast adenocarcinomas. Automated image analysis with AQUA (Automated Quantitative Analysis) was then performed to determine a quantitative expression score. Scores from redundant arrays were normalized and averaged. Kaplan-Meier survival analysis showed that high levels of vascular endothelial growth factor, VEGFR-1, VEGFR-2, and neuropilin-1 were all significantly associated with survival (Miller Siegmeund corrected P = .0020, .0160, and .0320, respectively). In addition, vascular endothelial growth factor and neuropilin-1 retained a significant association with survival independent of other standard prognostic factors. Vascular endothelial growth factor, VEGFR-1 and -2, and neuropilin-1 are expressed to varying degrees in primary breast cancers and have prognostic significance. Further study of the functional significance of this finding is warranted as well as the prognostic value of these biomarkers in other tumor microenvironment-specific compartments (eg, vessels).
Nonambulatory dairy cattle pose a complex problem due to the challenges associated with prevention, appropriate treatment and management, and arriving at an accurate prognosis. There is a breadth of ...literature regarding this topic, of which there is currently no formal synthesis. The objective of this scoping review was to describe and characterize the literature investigating risk factors, sequela, preventions, treatments, and prognostic factors for nonambulatory conditions in dairy cattle, with the intent of qualitatively synthesizing knowledge of the topic and identifying gaps in the literature. A literature search was conducted in 6 databases and 2 conference proceeding archives, which returned 7,568 unique articles. Initial screening of abstracts resulted in 1,544 articles reviewed at the full-text stage, of which 379 were included for data extraction. Over 75% of the included literature was published after 1980, and the most common countries in which these studies took place were the United States (n = 72), Canada (18), Sweden (17), and Germany (17). Common eligibility criteria used for inclusion were geographic region (97) and parity (92). Of the 379 studies included in this review, 144 were randomized controlled trials and 235 were observational studies. The majority of the controlled trials assessed prevention of nonambulatory conditions (116), most commonly through supplementation of vitamin D (27) and calcium (25) or the provision of anionic salts (22). Of the 28 studies focusing on treatment of nonambulatory conditions, 26 focused on calcium administration. Becoming nonambulatory was evaluated as an outcome in 165 of the observational studies. Frequently measured risk factors for becoming nonambulatory included hematological variables, such as blood calcium (73), phosphorus (53) and magnesium (42), and other factors such as parity (35) and breed (22). Recovery from a nonambulatory condition was the outcome in 31 of the observational studies, with commonly measured prognostic indicators being calcium (9), phosphorus (9), and duration of recumbency (7). Nonambulatory disorders were measured as risk factors in 53 of the observational studies, with the most commonly assessed outcomes including disorders of the transition period (11), and death or euthanasia (11). The most common terms used to describe nonambulatory conditions were “milk fever” (199) and “parturient paresis” (147). These terms were only further defined with explicit symptomatic criteria in 193 of the 379 studies in this review. Recumbency was the most commonly used of these criteria (144), followed by inability to rise (55). Potential gaps in the literature concerning nonambulatory dairy cattle that were identified in the present review included investigation of prognostic indicators for recovery from nonambulatory conditions that are applicable on farm, treatment alternatives to calcium administration, and guidance regarding the appropriate usage of terms meant to categorize nonambulatory dairy cattle.
HSP90 chaperones molecules critical for cell survival and malignant progression, including mutated B-raf. HSP90-targeting agents are in clinical trials. No large studies have been conducted on ...expression of HSP90 in melanomas.
Tissue microarrays containing 414 nevi, 198 primary and 270 metastatic melanomas were assessed using our automated quantitative analysis (AQUA) method of in situ protein measurement; we use S-100 to define pixels as melanocytes (tumor mask) within the array spot, and measure HSP90 expression within the mask using Cy5-conjugated antibodies.
HSP90 expression was higher in melanomas than nevi (P<0.0001) and higher in metastatic than primary specimens (P<0.0001). No association was seen between high HSP90 expression and survival in the primary or metastatic patient subsets. In primary melanomas, high HSP90 expression was associated with higher Clark level (P=0.0167) and increased Breslow depth (P<0.0001).
HSP90 expression was significantly higher in tumors than nevi and was associated with disease progression, indicating that it might be a valuable drug target in melanoma, as well as a useful diagnostic marker. Prospective studies are needed to confirm the diagnostic role of HSP90, as well as the predictive role of HSP90 expression in patients treated with HSP90 inhibitors.
Growth factor receptor-bound protein-7 (Grb7) is an adapter-type signaling protein recruited to various tyrosine kinases, including HER2/neu. Grb7-specific inhibitors are in early development. As ...with other targeted therapies, response to therapy might be associated with target expression.
Tissue microarrays containing 638 primary breast cancer specimens with 15-year patient follow-up were employed to assess Grb7 expression using our Automated QUantitative Analysis method; cytokeratin defines pixels as breast cancer (tumor mask) within the histospot, and Grb7 expression within the mask is measured with Cy5-conjugated antibodies.
High Grb7 expression was strongly associated with decreased survival in the entire cohort and in the node-positive subset (P=0.0034 and P= 0.0019, respectively). On multivariable analysis, it remained an independent prognostic marker (P =0.01). High Grb7 was strongly associated with high HER2/neu, and coexpression of these molecules was associated with worse prognosis than HER2/neu overexpression alone.
High Grb7 defines a subset of breast cancer patients with decreased survival, indicating that Grb7 might be a valuable prognostic marker and drug target. Coexpression with HER2/neu indicates that cotargeting these molecules might be an effective approach for treating HER2/neu-positive breast cancers. Future studies using Grb7-targeting agents should include assessment of Grb7 levels.
It is often assumed that membership in a stigmatized group has negative consequences for the self-concept. However, this relationship is neither straightforward nor inevitable, and there is evidence ...suggesting that negative consequences may not necessarily occur (Psychol. Rev. 96(4) (1989) 608). This paper argues that the relationship has not been sufficiently theorized, and that a more detailed analysis is called for in order to understand the relationship between stigma and the self. The paper presents a critical examination of modified labeling theory (Am. Sociol. Rev. 52 (1987) 96), with examples from a study examining perceptions of stigma and their relationship to self-evaluation in women with chronic mental health problems. Open-ended interviews and qualitative analyses were used in preference to global measures of self-esteem. It was found that although the women were aware of society's unfavorable representations of mental illness, and the effects this had on their lives, they did not accept these representations as valid and therefore rejected them as applicable to the self. The participants did not deny their mental health problems, but their acceptance of labels was critical and pragmatic. Labels were rejected when they were perceived as carrying an unrealistic and negative stereotype, or when the women felt that their symptoms did not fit with the diagnostic criteria. The research illustrates the importance of considering people's subjective understandings of stigmatized conditions and societal reactions in order to understand the relation between stigma and the self.
beta-catenin has a role in cell adhesion and Wnt signaling. It is mutated or otherwise dysregulated in a variety of human cancers. In this study we assess beta-catenin alteration in 145 thyroid ...tumors samples from 127 patients. beta-catenin was localized using immunofluorescence and mutational analysis was performed by single-strand conformational polymorphism. Membrane beta-catenin expression was decreased in eight of 12 (66%) adenomas and in all 115 carcinomas (P: < 0.0001). Among carcinomas, reduced membrane beta-catenin was associated with progressive loss of tumor differentiation (P: < 0.0001). CTNNB1 exon 3 mutations and nuclear beta-catenin localization were restricted to poorly differentiated 7 of 28 (25%) and 6 of 28 cases (21.4%), respectively or undifferentiated carcinomas 19 of 29 (65.5%) and 14 of 29 (48.3%) cases, respectively. Poorly differentiated tumors always featured mutations involving Ser and Thr residues and were characterized by Thr to Ile amino acid substitutions (P: = 0.0283). The association between CTNNB1 exon 3 mutations and aberrant nuclear immunoreactivity (P: = 0.0020) is consistent with Wnt activation because of stabilizing beta-catenin mutations. Low membrane beta-catenin expression as well as its nuclear localization or CTNNB1 exon 3 mutations are significantly associated with poor prognosis, independent of conventional prognostic indicators for thyroid cancer but not of tumor differentiation. Analysis of beta-catenin dysregulation may be useful to objectively subtype thyroid neoplasms and more accurately predict outcomes.
Numerous studies have demonstrated that overexpression of Met, the hepatocyte growth factor(HGF) receptor, plays an important role in tumorigenesis. Met activation can either occur through ...ligand-independent or -dependent mechanisms, both of which are mediated by a series of proteases and modulators. We studied the protein expression of several components of the HGF/Met pathway on a cohort of 330 node-negative breast carcinomas using a tissue microarray annotated with 30-year, disease-specific patient follow-up data. We examined HGF, matriptase (an activator of HGF expressed on mammary epithelial cell surfaces), HAI-I (the cognate inhibitor of matriptase), and the Met receptor itself. Our studies demonstrate tight correlation between the expression of HGF, matriptase, and Met in breast carcinoma. High-level expression of Met, matriptase, and HAI-I were associated with poor patient outcome. Met and HAI-I showed independent prognostic value when compared with traditional breast markers in a multivariate analysis. Intriguingly, antibodies against the intracellular but not the extracellular domain of Met were prognostic, suggesting that overexpression of the cytoplasmic-tail of Met, perhaps through cleavage or truncating mutation, may play an important role in breast cancer progression.
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